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Shade improves seedling quality of ornamental Cyclocarya species under plastic greenhouse cultivation

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  • Plastic greenhouse cultivation is a widespread and convenient way of cultivating high-quality seedlings, which are often damaged by higher temperatures during summer. Cyclocarya paliurus, a medical and ornamental species with low-quality seedlings, was investigated using three shading levels (treatment with no shade net (SH0), treatment with one layer of shade net (SH1), and treatment with two layers of shade net (SH2)). The growth and physiological responses of seedlings under plastic greenhouse cultivation were investigated from June to September. The results showed that the survival rate of seedlings reached 100%, and seedling growth and biomass were the best under SH2, with higher plant height and leaf area than that under other treatments. Water content of seedlings exhibit not difference between three shading levels, and leaves had the highest water content and total soluble sugar content. The chlorophyll content in the leaf increased, but malondialdehyde content decreased with increasing shading layers. Mineral content were in the following order: calcium > potassium > magnesium > sodium, and the translocation factor decreased with increasing shading layers. Antioxidant enzyme activities were in the following order: SOD > PPO > POD > CAT; their activities decreased with increasing shading layers, except for that of PPO. Various correlation existed between physiological response and seedling growth. Shade improved seedling quality through a series of physiological responses under plastic greenhouse conditions. This study provides a solid foundation for greenhouse cultivation of Cyclocarya species.
  • Bletilla Rchb. f. is one of the most economically valuable groups of orchids in the world. Due to its ornamental significance, the genus Bletilla occupies an important place in the worldwide horticultural market. Furthermore, in China, Japan, South Korea, and other Asian countries, it is highly valued for its medicinal use[1].

    There are eight species in the genus Bletilla, including Bletilla chartacea (King & Pantl.) Tang & F.T. Wang, Bletilla cotoensis Schltr., Bletilla foliosa (King & Pantl.) Tang & F.T. Wang, Bletilla formosana Schltr., Bletilla guizhouensis J. Huang & G.Z. Chen, Bletilla morrisonensis Schltr., Bletilla ochracea Schltr., and Bletilla striata Rchb.f.[2,3]. The distribution area spans from northern Myanmar in Asia to Japan via China[4]. Five species are native to China, namely, B. foliosa, B. formosana, B. guizhouensis, B. ochracea, and B. striata. In China, people have assigned various names to Bletilla based on its morphology and efficacy, such as baiji (白及/白芨), baigen (白根), baige (白给), baijier (白鸡儿), baijiwa (白鸡娃), diluosi (地螺丝), gangen (甘根), junkouyao (皲口药), lianjicao (连及草), and yangjiaoqi (羊角七)[5]. These diverse appellations highlight the importance of this genus in Chinese folk biological culture.

    The medicinal material known as 'baiji' in traditional Chinese medicine (TCM) is usually the dried tuber of B. striata, which is also the authentic product included in the Chinese Pharmacopoeia[6]. According to the Chinese Pharmacopoeia (2020), TCM baiji is sliced, dried, and crushed into a powder that can be used topically or internally, with a recommended dosage of 3–6 g at a time, offering astringent, hemostatic, detumescence, and myogenic effects. It is often used for conditions such as hemoptysis, hematemesis, traumatic bleeding, sores, and skin chaps[7]. Although only B. striata is the authentic product of TCM baiji, the other four Bletilla species native to China are also used as substitutes, and this practice is widespread[8].

    Modern research indicates that Bletilla contains a variety of chemical components, including benzol, dihydrophenanthrene, phenanthrene, and quinone derivatives. These components confer pharmacological effects on Bletilla, such as hemostasis, anti-tumor activity, and promotion of cell growth[9]. Due to its outstanding medicinal value, Bletilla can be found in nearly every corner of the traditional medicine market (Fig. 1). However, habitat destruction and uncontrolled mining have led to a significant reduction in the native populations of Bletilla, making its protection an urgent priority. Therefore, this paper provides a comprehensive review of relevant research up to August 2023, covering botanical characteristics, resource distribution, ethnobotanical uses, chemical components, pharmacological effects, clinical applications, and safety evaluations of Bletilla. The aim is to raise awareness and promote the protection and sustainable use of this genus.

    Figure 1.  Varieties of Bletilla at the traditional March Medicinal Market in Dali, Yunnan, China.

    The morphology of different Bletilla species is highly similar. The primary taxonomic feature distinguishing each species is the characteristics of the flower, particularly the lip of the flower, including its size, shape, and the number and shape of longitudinal ridges on the lip plate (Table 1, Fig. 2)[1014].

    Table 1.  The morphological differences among five species of Bletilla plants native to China.
    Morphological featureBletilla striataBletilla formosanaBletilla ochraceaBletilla foliosaBletilla guizhouensis
    Plant height (cm)18−6015−8025−5515−2045−60
    Rhizome shapeCompressedCompressedSomewhat compressedSubgloboseCompressed
    Rhizome diameter (cm)1−31−2About 21−1.53−4
    Stem characteristicsStoutEnclosed by sheathsStoutStout, shortThin
    Leaf shapeNarrowly oblongLinear-lanceolateOblong-lanceolateElliptic-lanceolateNarrowly lanceolate
    Leaf size (cm)8−29 × 1.5−46−40 × 0.5−4.58−35 × 1.5−2.85−12 × 0.8−325−45 × 1.2−4.5
    Flower colorPurplish red or pinkPale purple or pinkYellowPale purpleDeep purple
    Flower sizeLargeMediumMediumSmall to mediumLarge
    Inflorescence structureBranched or simpleBranched or simpleSimpleSimpleBranched
    Pedicel and ovary length (mm)10−248−12About 187−913−17
    Sepal shapeNarrowly oblongLanceolateLanceolateLinear-lanceolateOblong-elliptic
    Petal shapeSlightly larger than sepalsSlightly narrower than sepalsObliqueLanceolateOblong-elliptic
    Lip shapeObovate-ellipticBroadly ellipticNarrowly rhombic-obovateNarrowly oblongNarrowly oblong
    Lip colorWhite with purplish veinsWhitish to pale yellow with small dark purple spotsWhitish to pale yellow with small dark purple spotsWhite with purplish spots and purple edgeWhite with deep purple edge
    Number of lip Lamellae5 lamellae5 undulate lamellae5 longitudinal lamellae3 fimbriate lamellae7 longitudinal lamellae
    Column characteristicsSubterete, dilated towards apexSubterete, dilated towards apexSlender, dilated towards apexCylindric, dilated towards apexSuberect, with narrow wings
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    Figure 2.  (a)−(d) Bletilla striata (Thunb. ex Murray) Rchb. f. (e)−(h) Bletilla formosana (Hayata) Schltr. (i)−(l) Bletilla ochracea Schltr. (m), (n) Bletilla sinensis (Rolf) Schltr. (o), (p) Bletilla guizhouensis Jie Huang & G.Z. Chen (Photographed by Wang Meina, Zhu Xinxin, and He Songhua).

    The flowers of B. striata are large and purplish-red or pink, with narrowly oblong sepals and petals measuring 25−30 mm in length and 6−8 mm in width. They have acute apices, nearly as long as the sepals and petals. The lip is obovate or elliptic, predominantly white with purplish-red coloration and purple veins, measuring 23−28 mm in length, slightly shorter than the sepals and petals. The lip disc exhibits five longitudinal folds extending from the base to near the apex of the middle lobe, with waviness occurring only above the middle lobe[11]. In China, B. striata is found in regions such as Anhui, Fujian, Guangdong, Guangxi, Gansu, Guizhou, Hubei, Hunan, Jiangsu, Jiangxi, Shaanxi, Sichuan, and Zhejiang. It also occurs in the Korean Peninsula and Japan, thriving in evergreen broad-leaved forests, coniferous forests, roadside grassy areas, or rock crevices, at altitudes ranging from 100−3,200 m[12].

    B. ochracea's flowers are medium to large, featuring yellow or yellow-green exteriors on the sepals and petals, while the insides are yellow-white, occasionally nearly white. The sepals and petals are nearly equal in length, oblong, measuring 18−23 mm long and 5−7 mm wide, with obtuse or slightly pointed apices, often adorned with fine purple spots on the reverse side. The lip is elliptic, typically white or light yellow, measuring 15−20 mm in length and 8−12 mm in width, with three lobes above the middle. The lip disc is characterized by five longitudinally ridged pleats, with undulations primarily occurring above the middle lobe[13]. B. ochracea is native to southeastern Gansu, southern Shaanxi, Henan, Hubei, Hunan, Guangxi, Guizhou, Sichuan, and Yunnan, thriving in evergreen broad-leaved forests, coniferous forests, or beneath shrubs, in grassy areas or alongside ditches at altitudes ranging from 300−2,350 m[14].

    B. formosana's flowers come in shades of lavender or pink, occasionally white, and are relatively small. The sepals and petals are narrowly oblong, measuring 15−21 mm in length and 4−6.5 mm in width, and are nearly equal in size. The sepals have subacute apices, while the petal apices are slightly obtuse. The lip is elliptic, measuring 15−18 mm in length and 8−9 mm in width, with three lobes above the middle. The lip disc exhibits five longitudinal ridge-like pleats, which are wavy from the base to the top of the middle lobe[15]. B. formosana is indigenous to southern Shaanxi, southeastern Gansu, Jiangxi, Taiwan, Guangxi, Sichuan, Guizhou, central to northwest Yunnan, southeast Tibet (Chayu), and Japan. It is typically found in evergreen broad-leaved forests, coniferous forests, road verges, valley grasslands, grassy slopes, and rock crevices, at altitudes ranging from 600−3,100 m[16].

    The flowers of B. foliosa are small and lavender, with white sepals and petals featuring purple apices. The sepals are linear-lanceolate, measuring 11−13 mm in length and 3 mm in width, with subacute apices. The petals are lanceolate, also measuring 11−13 mm in length and 3 mm in width, with acute apices. The lip is white, oblong, adorned with fine spots, and features a purple apex. It measures 11−13 mm in length and 5−6 mm in width, tapering near the base and forming a scaphoid shape. The lip is anteriorly attenuated, unlobed, or abruptly narrowing with inconspicuous three lobes and exhibits fringe-like fine serrations along the edge. Three longitudinal ridge-like pleats are present on the upper lip disc[17]. B. foliosa typically grows on hillside forests, with its type specimen collected from Mengzi City, Honghe Hani and Yi Autonomous Prefecture, Yunnan Province, China[17].

    B. guizhouensis is a recently discovered species in Guizhou, China. In terms of shape, B. guizhouensis closely resembles B. striata, but it can be distinguished by its ovate-oblong buds, oblong dorsal sepals, obovate lips, and middle lobes of the lips, which are oval in shape. The disc of B. guizhouensis features seven distinct longitudinal lamellae, setting it apart from other known Bletilla species and establishing it as a distinct species[2]. Presently, B. guizhouensis has only been found in Guizhou, China, primarily thriving in evergreen broad-leaved forests at altitudes ranging from 900−1,200 m[3].

    Understanding the morphology, habitat, and distribution of Bletilla species is crucial for the conservation and propagation of these resources. To effectively implement plant conservation and breeding programs, a comprehensive understanding of the specific morphological characteristics, growth environments, and native habitats of these plants is essential, as without this knowledge, effective results cannot be achieved.

    The ethnobotanical uses of Bletilla worldwide primarily fall into two categories: ornamental and medicinal purposes. Bletilla orchids, renowned for their striking and distinct flowers, are commonly cultivated for ornamental purposes across many countries[18]. Valued for their aesthetic appeal, these orchids are frequently grown in gardens and utilized as potted plants. Among the various cultivars, B. striata stands out as the most favored choice for ornamental horticulture due to its ease of cultivation and adaptability to diverse climates[19,20].

    Contrastingly, in select Asian countries, Bletilla assumes a crucial role as a medicinal plant. For instance, influenced by TCM, the tuber of Bletilla also serves as a crude drug for hemostatic and anti-swelling purposes in Japan[21]. Likewise, traditional Korean medicine, deeply rooted in TCM principles, extensively documents the versatile use of Bletilla in addressing issues such as alimentary canal mucosal damage, ulcers, bleeding, bruises, and burns[22]. In Vietnam, Bletilla has been used as a medicinal herb for treating tumors and skin fissures, aligning with practices observed in the ethnic communities of southwest China[23].

    In China, Bletilla boasts a longstanding medicinal history, with numerous classical ancient Chinese medicine books containing detailed records of its medicinal applications[2432]. Even in contemporary society, many ethnic groups residing in mountainous areas in China continue to uphold the traditional medical practice of using Bletilla medicinally[31].

    In ancient Chinese medical literature, detailed records of Bletilla's morphology can be traced back to the late Han Dynasty, around 200 AD[24]. The Mingyi Bielu, a historical source, documented, 'Bletilla grows in the valley, with leaves resembling those of Veratrum nigrum L., and its root is white and interconnected. The ideal time for harvesting is September'. As awareness of the medicinal significance of Bletilla grew, successive dynastic-era Chinese medical texts consistently included descriptions of Bletilla's morphology (Table 2). In the Ming Dynasty, Li Shizhen compiled these earlier accounts of Bletilla's plant characteristics in his work, the Compendium of Materia Medica. He even provided an illustrative depiction of this plant genus (Fig. 3)[25].

    Table 2.  Morphological description of the plants belonging to Bletilla in the ancientChinese medicinal books.
    Dynasty (Year)TitleAuthorOriginal ChineseEnglish translation
    Late Han
    (184−220 AD)
    Mingyi Bielu/白给生山谷, 叶如藜芦,
    根白相连, 九月采
    Bletilla grows in the valley, with leaves like Veratrum nigrum L., root is white and connected. September is the time for harvesting.
    Wei-Jin period
    (220−420 AD)
    WuPu BencaoWu Pu白根, 茎叶如生姜, 藜芦,
    十月花, 直上, 紫赤色,
    根白连, 二月, 八月, 九月采
    Bletilla, stems and leaves like Zingiber officinale Roscoe and V. nigrum. It blooms in October and is purple and red, the inflorescence is vertical and upward. The roots are white and connected. It can be dug in February, August, and September.
    the Northern and Southern
    (420−589 AD)
    Bencao JizhuTao Hongjing近道处处有之, 叶似杜若,
    根形似菱米, 节间有毛
    It is everywhere near the road. The leaves are like Pollia japonica Thunb. The roots are like the fruit of Trapa natans L., and internode are many fibrous roots.
    Tang
    (618−907 AD)
    Su Jing, Zhangsun Wuji, etcTang materia medica生山谷, 如藜芦, 根白连, 九月采Born in the valley, with leaves like V. nigrum, root is white and connected. September is the time for harvesting.
    Song
    (960−1279 AD)
    Su SongCommentaries on the Illustrations白芨, 生石山上。春生苗,
    长一尺许, 似栟榈及藜芦,
    茎端生一台, 叶两指大, 青色,
    夏开花紫, 七月结实, 至熟黄黑色。
    至冬叶凋。根似菱米, 有三角白色, 角端生芽。二月, 七月采根
    Bletilla grow on the stone hill. It sprouts in spring and grows about a foot long. The seedlings are like Trachycarpus fortunei (Hook.) H. Wendl. and V. nigrum. The leaves are two finger-size. In summer, it blooms purple flowers and bears fruit in July. The ripe fruit is yellow-black. The leaves wither in winter. The root is like the fruit of T. natans, with three corners, white, and sprouting at the corners. The roots are dug in February and July.
    Ming
    (1368−1644 AD)
    Li ShizhenCompendium of Materia Medica一棵只抽一茎, 开花长寸许,
    红紫色, 中心如舌, 其根如菱米,
    有脐, 如凫茈之脐,
    又如扁扁螺旋纹, 性难干
    Only one stem per herb. The flower is more than one inch long, red and purple, and the center resembles a tongue. Its root is similar to the fruit of T. natans, possessing an umbilicus akin to that of Eleocharis dulcis (N. L. Burman) Trinius ex Henschel. It has spiral veins and is challenging to dry.
    −, Anonymous.
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    Figure 3.  Bletilla in Compendium of Materia Medica.

    Generally, ancient Chinese medical texts did not make clear distinctions between different Bletilla species. They collectively referred to plants with similar morphological traits as 'baige', 'baigen', 'baiji', 'gangen', 'lianjicao', or 'ruolan'. However, through textual analysis, it has been established that the descriptions of Bletilla in ancient texts before the Ming Dynasty largely align with Bletilla striata in terms of plant height, pseudo-bulb shape, leaf morphology, flower and fruit colors, and other characteristics. While the Bletilla portrayed in attached images may not precisely match B. striata in terms of morphology, considering the textual descriptions, it generally corresponds with B. striata. In writings from the Ming Dynasty and later periods, more specific descriptions of Bletilla emerged, encompassing details about its vascular arrangement, inflorescence, and flower structure, which consistently align with B. striata. Consequently, researchers have corroborated that the original plant of Bletilla described in ancient texts is Bletilla striata[24,33].

    According to the ancient Chinese medicinal books, Bletilla was used to treat a wide variety of conditions, including coughing, bruising, and bleeding, but their most mentioned use in ancient Chinese texts is for skin whitening and freckle removal[25]. Since ancient times, Bletilla species have been used consistently for skin care and whitening, and there are many well-known skincare products related to Bletilla. These Chinese formulae with Bletilla are similar to modern facial masks, face creams, facial cleanser, hand creams and other skin care products[26].

    For example, a prescription for 'facial fat (面脂)' in Medical Secrets from the Royal Library (752 AD) is made by boiling Bletilla with other traditional ingredients, and is applied to the face, resulting in skin whitening, freckle and wrinkle removal[27]. The 'Angelica dahurica cream (白芷膏)' in the General Medical Collection of Royal Benevolence (1111−1125 AD) is reputed to whiten facial skin through a seven-day treatment regiment, and contains Bletilla as the main botanical ingredient along with Angelica dahurica[28]. Jingyue Quanshu (1563-1640 AD) also contains a prescription called 'Yurong powder (玉容散)' for facial skin care. 'Yurong powder' is made of a fine powder of Bletilla, Nardostachys jatamansi (D. Don) DC., Anthoxanthum nitens (Weber) Y. Schouten & Veldkamp and other herbs[29]. Washing the face with Yurong powder in the morning and evening every day is said to make a person's face white without blemishes (Fig. 4)[29].

    Figure 4.  Yurong powder made of Bletilla and other traditional Chinese medicines in Jingyue Quanshu.

    In addition, in ancient Chinese medicine texts, Bletilla is also a well-known medicine for treating hematemesis, hemoptysis and bruises[23]. According to Shennong's Classic of Materia Medica (25−220), grinding the white fungus into fine powder and taking it after mixing with rice soup can be effective for treating lung damage and hematemesis[30]. Among the Prescriptions for Universal Relief (1406), 18 traditional Chinese medicines, such as Bletilla, are used to make 'snake with raw meat cream', which is said to be useful to treat carbuncles and incised wounds[31]. There is also a record of Bletilla powder treating lung heat and hematemesis in the Collected Statements on the Herbal Foundation (1624)[32].

    In ancient Chinese medicinal texts, most Bletilla are said to be useful for lung injury and hemoptysis, epistaxis, metal-inflicted wounds, carbuncles, burns, chapped hands and feet, whitening and especially for skin care. In the ancient medicinal texts, Bletilla is used alone or mixed with other traditional Chinese medicines. It is usually used in the form of a powder. The various medicinal effects of Bletilla described in these ancient texts suggest the great potential of this genus in clinical application, especially in the market of skin care products and cosmetics.

    As a skin care herb praised by ancient medical classics, 11 ethnic minorities in China, such as Bai, Dai, De'ang, Jingpo, Lisu, Miao, Mongolian, Mulao, Tu, Wa, and Yi still retain the traditional habit of using Bletilla for skin care in their daily life (Table 3). In addition to B. striata, B. formosana and B. ochracea are also used as substitutes. Although Chinese ethnic groups have different names for Bletilla spp., the skin care methods are basically the same. Dry Bletilla tubers are ground into a powder and applied to the skin[34], and this usage is also confirmed by the records in ancient medical texts[23, 24]. The various local names of Bletilla by different ethnic groups also indirectly suggests which ethnic groups play an important role in the traditional use. For example, Bai people called B. striata baijier (白鸡儿), goubaiyou (狗白尤), and yangjiaoqi (羊角七) (Table 3).

    Table 3.  The traditional medicinal knowledge of Bletilla in ethnic communities, China.
    Ethnic groupLatin nameLocal nameUsed partUse methodMedicinal effect
    AchangBletilla striata (Thunb. ex Murray) Rchb. F.BaijiTuberAfter the roots are dried, chew them orally or grind them into powder for external applicationTuberculosis, hemoptysis, bleeding from gastric ulcer, burns and scalds
    BaiBaijier, Goubaiyou, YangjiaoqiTuberTreatment of tuberculosis hemoptysis, bronchiectasis hemoptysis, gastric ulcer hemoptysis, hematochezia, skin cracking
    DaiYahejieTuberUsed for tuberculosis, tracheitis, traumatic injury, and detumescence
    De'angBageraoTuberTuberculosis, hemoptysis, bleeding from gastric ulcer, burns and scalds
    DongShaque, SanjueTuberTreat hematemesis and hemoptysis
    JingpoLahoiban, PusehzuotuberFor tuberculosis, bronchiectasis, hemoptysis, gastric ulcer, hematemesis, hematuria, hematochezia, traumatic bleeding, burns, impotence
    MengMoheeryichagan, NixingTuberFor tuberculosis hemoptysis, ulcer bleeding, traumatic bleeding, chapped hands, and feet
    MiaoBigou, Wujiu, SigouTuberUsed for hemoptysis of tuberculosis, bleeding of ulcer disease, traumatic bleeding, chapped hands, and feet
    MolaoDajiebaTuberTreat internal and external injuries caused by falls
    TibetanSanchabaijiTuberFresh chopped and soaked with honey; Powdered after sun-dried, then taken with honey and waterMainly used to treat cough, asthma, bronchitis, lung disease and a few gynecological diseases
    TuRuokeyeTuberAfter the roots are dried, chew them orally or grind them into powder for external applicationTreatment of tuberculosis, hemoptysis, bloody stool, chapped skin
    WaBaijiTuberAfter the roots are dried, chew them orally or grind them into powder for external applicationFor tuberculosis, hemoptysis, gastrointestinal bleeding, scald and burn
    YaoBiegeidaiTuberTreat gastric ulcer, pulmonary tuberculosis, cough, hemoptysis, and hematemesis
    YiDaibaij, Tanimobbaili, Niesunuoqi, AtuluoboTuberTreatment of tuberculosis, hemoptysis, golden wound bleeding, burns, chapped hands and feet
    ZhuangManggounuTuberTreat stomachache and hemoptysis
    BaiBletilla formosana (Hayata) Schltr.Baijier, YangjiaoqiTuberAfter the roots are dried, chew them orally or grind them into powder for external applicationIt is used for emesis, hemoptysis due to tuberculosis, and hemoptysis due to gastric ulcer. External application for treatment of incised wound
    MiaoLianwuTuberThe effect is the same as that of B. striata
    LisuHaibiqiuTuberIt can treat tuberculosis, hemoptysis, epistaxis, golden sore bleeding, carbuncle and swelling poison, scald by soup fire, chapped hands and feet
    YiNiesunuoqi, Yeruomaoranruo, Atuluobo, Ribumama, Atuxixi, Abaheiji, Binyue, ZiyouTuberIt is used for tuberculosis, hemoptysis, traumatic injury, treatment of frostbite, burn, scald, bed-wetting of children and other diseases
    BaiBletilla ochracea Schltr.Baijier, YangjiaoqiTuberAfter the roots are dried, chew them orally or grind them into powder for external applicationFor hematemesis, epistaxis, hemoptysis due to tuberculosis, hemoptysis due to gastric ulcer; External application of golden sore and carbuncle
    MengMoheeryichagan, NixingtuberThe effect is the same as that of B. striata
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    The formation of traditional medical knowledge among Chinese people is often directly related to the specific living environment and cultural background[34]. For example, the Bai, Dai, De'ang, Jingpo, Lisu Yi, Wa and other ethnic minorities live in mountainous areas. The cold weather in winter and year-round outdoor manual work makes it difficult to maintain their skin[35, 36]. In the face of this situation, the ethnic people who are concerned about their physical appearance have long ago chosen local Bletilla species for skin care, and have handed down this tradition for many generations[34]. This important traditional skin care tradition is worthy of further in-depth study.

    The six main classes of Bletilla chemical components, phenanthrene derivatives, phenolic acids, bibenzyls, flavonoids, triterpenoids, and steroids, have been described previously. Almost three hundred compounds have been isolated from Bletilla, including 116 phenanthrene derivatives, 58 phenolic acids, 70 bibenzyls, 8 flavonoids, 24 triterpenoids and steroid and 13 other compounds (Figs 514). Chemical structures of the isolates of Bletilla species most are phenanthrene derivatives, which have been demonstrated to possess various pharmacological activities.

    The prominent opioids oxycodone, hydrocodone, naloxone, and naltrexone are all phenanthrene derivatives[37]. Currently, phenanthrene derivatives (Fig. 5, 1 to 66) were isolated from B. formosana, B. ochracea, and B. striata. In 2022, 17 phenanthrene derivatives (117) were isolated from the ethyl acetate (EtOAc) extracts of B. striata tubers[38]. Then, other phenanthrene derivatives were isolated from Bletilla, such as dihydrophenanthrenes (1841), phenanthrenes (4266), biphenanthrenes (Fig. 6, 6789), dihydro/phenanthrenes with uniquestructures (90112) and phenanthraquinones (Fig. 7, 113116). Thus far, this genus has been documented to include these compounds, which have been shown to exhibit pharmacological actions[3945].

    Figure 5.  Phenanthrene derivatives from Bletilla species (1−66)[3841,4345,47,49,58,7072,7479].
    Figure 6.  Phenanthrene derivatives from Bletilla species (67−105)[41,43,49,5961,70,76,7986].
    Figure 7.  Phenanthrene derivatives from Bletilla species (106−116)[43,49,70,75,84,85,87].

    Phenolic acids are carboxylic acids created from the skeletons of either benzoic or cinnamic acids[4648]. Fifty-eight phenolic acids (Figs 810, 117 to 174) were isolated from B. formosana, B. ochracea, and B. striata.

    Figure 8.  Phenolic acids from Bletilla species (117−134)[1,5,36,4752,54,67,88,90].
    Figure 9.  Phenolic acids from Bletilla species (135−169)[39,45,4854,56,61,68,69,73,76,82,83,8994].
    Figure 10.  Phenolic acids from Bletilla species (170−174)[20,95].

    For example, compounds 121, 126, 139, 141, 148, 149, 154, 155 and 157 were isolated from the rhizomes of B. formosana[1,49,5052]. The structures of these compounds were determined, mostly from their NMR spectroscopy data. Additionally, protocatechuic (136) and vanillin (137) also have been isolated from B. striata[53]. Moreover, some bioactive components such as 2-hydroxysuccinic acid (164) and palmitic acid (165) have been discovered and identified from B. striata[20,5456].

    The bibenzyls were small-molecular substances with a wide range of sources, which were steroidal ethane derivatives and resembling the structural moiety of bioactive iso-quinoline alkaloids[57].

    For example, depending on their structural characteristics, 70 bibenzyl compounds (Fig. 11, 175 to 244) can be grouped into three groups, simple bibenzyls (175186, 233238), complex bibenzyls (189225) and chiral bibenzyls (226-232, 239-244)[5860].

    Figure 11.  Bibenzyls from Bletilla species (175-244)[1,4042,47,49,50,5860,70,73,76,9699].

    Flavonoids are among the most common plant pigments. Eight bibenzyls (Fig. 12, 245 to 252) have been isolated from B. formosana, B. ochracea, and B. striata. Apigenin (245) and 8-C-p-hydoxybenzylkaempferol (249) were isolated from the whole plant of B. formosana[45]. Bletillanol A (250), bletillanol B (251) and tupichinol A (252) were isolated from B. striata[61]. The names and chemical structures of the flavonoids reported from Bletilla are shown below (Fig. 12).

    Figure 12.  Flavonoids from Bletilla species (245–252)[45,61].

    Twenty-four triterpenoids and steroids (Fig. 13, 253 to 276) have been reported from Bletilla (Fig. 13), such as, tetracyclic triterpenes (253259) and pentacyclic triterpenes (189225) and chiral bibenzyls (260)[6264]. Steroids (261276) isolated from the Bletilla and have shown some bioactivity. For example, bletilnoside A (272) was isolated from Bletilla species and displayed anti-tumor activity[65,66].

    Figure 13.  Triterpenoids and steroid compounds from Bletilla species (253-276)[56,6265].

    Thirteen other compounds (Fig. 14, 277 to 289) were isolated from B. formosana, B. ochracea, and B. striata. These compounds included amino acids, indoles and anthraquinones[67,68]. For example, syringaresinol (285) and pinoresinol (286) have been described in the methanol extract of the tubers of B. striata[61].

    Figure 14.  Others compounds from Bletilla species (277−289)[50,54,61,62,67,68,94,97].

    Based on the information about the chemical constituents of Bletilla species, it appears that there is a substantial body of research on these compounds. However, there are some areas that may warrant further investigation and research. At first, it would be valuable to investigate potential synergistic effects and interactions between the different classes of compounds within Bletilla species, as some of the compounds may work together. Besides, it is worth considering the improvement of compound yield. Optimizing extraction methods and finding the most efficient and environmentally friendly techniques are vital for both research purposes and potential commercial applications. It is also important to take into account the variability in chemical composition among different Bletilla species and even within the same species from different cultivars.

    The rich and varied chemical components make the plants of Bletilla have a wide variety of pharmacological activities (Table 4). Many studies have shown that the plants of this genus have anti-inflammatory, antineoplastic, antiviral, antioxidant, hemostatic, antibacterial, and other biological activities, which help to support the traditional medicinal practice of Bletilla in folk medicine.

    Table 4.  Summary of the pharmacological activities of Bletilla species.
    Pharmacological activityTested substance/partTested system/organ/cellTested dose/dosing methodResultsRefs.
    Anti-inflammatoryEthanol extract of Bletilla striataRAW264.7 cells RAW264.7 cells were pre-treated with ethanol extract of B. striata for 1 h and then stimulated with LPS (200 ng/mL) for 12 h, 0.05% DMSO was applied as the parallel solvent control. The culture supernatant was collected for IL-6 and TNF-α detection.Ethanol extract of B striata significantly inhibited LPS-induced interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) expression at 2.5 µg/mL.[41]
    The ethyl acetate-soluble (EtOAc) extract of tubers of B. striataH2O2-induced PC12 cell injury modelPC12 cells were seeded in 96-well multiplates at a density of 1.5 × 105 cells/mL. After overnight incubation at 37 °C with 5% CO2, 10 μM test samples and H2O2 (final concentration of 450 μM) were added into the wells and incubated for another 12 h.It protected the cells with the cell viabilities of 57.86 ± 2.08%, 64.82 ± 2.84%, and
    64.11 ± 2.52%.
    [98]
    Ethanol extract of tubers of B. striataRAW264.7 cellsCells were treated with ethanol extracts (25 μM) dissolved in DMSO, in the presence of
    1 μg/mL lipopolysacchride
    (LPS) for 18 h
    The anti-inflammatory activity with IC50 of 2.86 ± 0.17 μM.[54]
    PE extract of the tubers of B. striataLPS-stimulated BV2 cellsCells treated with extract
    (0, 1, 10, 30, 100 μg/mL) and dihydropinosylvi (0, 1, 10, 30, 100 μM) in presence of LPS
    (1 μg/mL)
    The anti-inflammatory activity with IC50 values of 96.0 μM.[96]
    Ethanol extract of the roots of B. striataCox-1 and Cox-2Treated with the ethanol extracts at various concentrations
    (0, 1, 10, 100 μM)
    The compounds with sugar moieties displayed selective inhibition of Cox-2 (N90%).[38]
    B. striata polysaccharide (BSPb)Human mesangial cells (HMCs)HMCs were pre-treated with BSPb (5, 10, 20 μg/mL)BSPb efficiently mediated expression of NOX4 and TLR2, to attenuate generation of ROS and inflammatory cytokines.[12]
    Compounds extracted from the rhizomes of Bletilla ochraceaRAW264.7 cellsAfter 24 h preincubation,
    cells were treated with serial dilutions in the presence of
    1 μg/mL LPS for 18 h. Each compound was dissolved in DMSO and further diluted in medium to produce different concentrations. NO production in the supernatant was assessed by adding 100 μL of Griess regents.
    It showed the inhibitory effects with IC50 values in the range of 15.29–24.02 μM.[76]
    Compounds extracted from the rhizomes of B. ochraceaMurine monocytic RAW264.7 cellsAfter 24 h preinubation, RAW 264.7 cells were treated with compounds (25 μM) dissolved in DMSO, in the prenence of
    1 μg/mL LPS for 18 h. NO production in each well was assessed by adding 100 μL of Giress regent
    It showed the inhibitory effects with IC50 2.86 ± 0.17 μM.[86]
    Compounds extracted from the rhizomes of Bletilla formosanaElastase Release AssaysNeutrophils (6 × 105 cells/mL) were equilibrated in MeO-Suc-Ala-Ala-Pro-Val-p-nitroanilide (100 μM) at 37 °C for 2 min and then incubated with a test compound or an equal volume of vehicle (0.1% DMSO, negative control) for 5 min.Most of the isolated compounds were evaluated for their anti-inflammatory activities. The results showed that IC50 values for the inhibition of superoxide anion generation and elastase release ranged from 0.2 to 6.5 μM and 0.3 to 5.7 μM, respectively.[49]
    Anti-tumorTwo compounds from Bletilla striataA549 cellsCompounds were tested for their ability to induce ROS generation in A549 cells at concentrations of 20 two compounds for 24 h, the cells were harvested to evaluate the ROS production.The two compounds exhibited antiproliferative effects using the MTT test; these effects may be due to cell cycle arrest and inducing ROS generation.[87]
    Stilbenoids from B. striataBCRP-transduced K562 (K562/BCRP) cellsIt showed antimitotic activity and inhibited the polymerization of tubulin at IC50 10 μM.[78]
    Compounds extracted from the rhizomes of B. ochraceaThe human tumor cell lines HL-60 (acute leukemia), SMMC-7721 (hepatic cancer), A-549 (lung cancer), MCF-7 (breast cancer), and SW480 (colon cancer)100 μL of adherent cells were seeded into each well of 96-well cell culture plates. After 12 h of incubation at 37 °C, the test compound was added. After incubated for 48 h, cells were subjected to the MTS assay.All isolated metabolites except 7 were evaluated for cytotoxic activity against five human cancer cell lines (HL-60, SMMC7721, A-549, MCF-7 and SW480).[76]
    AntiviralThe tuber of B. striataMadin-Darby canine kidney model and embryonated eggs modelAs simultaneous treatment with 50% inhibition concentration (IC50) ranging from 14.6 ± 2.4 to 43.3 ± 5.3 μM.Phenanthrenes from B. striata had strong anti-influenza viral activity in both embryonated eggs and MDCK models.[107]
    The 95% ethanol
    Extract of B. striata
    BALB/C miceIt has significant anti-influenza
    virus effect in mice, which may be related to the increase of IL-2, INFα, INF-β and thus enhance the immune function of mice.
    [12]
    AntioxidantCompounds extracted from the rhizomes of B. formosanaDPPH radical-scavenging assaySolutions containing 160 μL of various concentrations of sample extract, 160 μL of various concentrations of BHA, 160 μL of various concentrations of ascorbic acid, and the control (160 μL of 75% methanol) were mixed separately with 40 μL of 0.8 mM DPPH dissolved in 75% methanol. Each mixture was shaken vigorously and left to stand for 30 min at room temperature in the dark.Tthe seedlings grown by tissue culture of B. formosan collected in Yilan County had the best antioxidant capacity. In addition, B. formosana collected in Taitung County had the best scavenging capacity in the tubers, leaves and roots.[93]
    Fibrous roots of B. striataDPPH model and superoxide anion systemThe ABTS+ solution was prepared by reacting 7 Mm ABTS with 2.45 mM potassium persulfate (final concentrations both dissolved in phosphate buffer, 0.2 M, pH 7.4) at room temperature for 12–16 h in the dark.It removed free radicals and inhibit tyrosinase activity.[33]
    B. striata extracts (BM60)The murine macrophage cells NR8383, male SD mice (180~200 g)NR8383 were pretreated with extracts (1, 10 and 100 g/mL) for 4 h and then 65 stimulated with 1 g/mL of LPS for 24 h. Acute lung injury was induced in mice by nonhexposure intratracheal instillation of LPS (3.0 mg/kg). Administration of the BM60 extract of 35, 70, and 140 mg/kg (L, M, H) was performed by oral gavages.The BM60 treatment reduced the production of NO in NR8383 macrophages. Treatments with BM60 at the doses of 35, 70, 140 mg/kg significantly reduced macrophages and
    neutrophils in the bronchoalveolar lavage fluid (BALF).
    [12]
    The crude
    polysaccharides obtained from B. striata
    DPPH free radical scavenging activityConcentration
    range of 2.5–5.0 mg/mL
    The IC50 of BSPs-H was 6.532 mg/mL.[35]
    HemostasisB. striata polysaccharide (BSP)Diabetes mellitus (DM) mouse models were induced by high fat-diet feeding combined with low-dose streptozocin injectionDM mouse models were induced by high fat-diet feeding combined with low-dose streptozocin injection. The BSP solutions were applied on the surface of each wound at a volume of 50 μl. RD mice were assigned as normal controls and received saline treatment (n = 6). All mice were treated with vehicle or BSP once daily from the day of wounding (d0) until 12 days later (d12).BSP administration accelerated diabetic wound healing, suppressed macrophage infiltration, promoted angiogenesis, suppressed NLRP3 inflammasome activation, decreased IL-1β secretion, and improved insulin sensitivity in wound tissues in DM mice.[112]
    B. striata Micron Particles (BSMPs)Tail amputation model and healthy male Sprague-Dawley (SD) rats
    (250 ± 20 g, 7 weeks of
    age)
    Rats were divided into six groups of five treated with cotton gauze and BSMPs (350–250, 250–180, 180–125, 125–75, and < 75 μm), respectively.Compared to other BSMPs of different size ranges, BSMPs of 350–250 μm are most efficient in hemostasis. As powder sizes decrease, the degree of aggregation between particles and hemostatic BSMP effects declines.[109]
    Rhizoma Bletillae polysaccharide (RBp)Adult male SD rats weighing 220 ± 20 gAfter incubation for 1 min at 37 °C, 300 μL of PRP was dealt with different concentrations of RBp (50, 100, 150, and 200 mg/L) under continuous stirring, and the vehicle was used as the blank control.RBp significantly enhanced the platelet aggregations at concentrations of 50−200 mg/L in a concentration-dependent manner.[113]
    AntibacterialBibenzyl derivatives from the tubers of Bletilla striataS. aureus ATCC 43300, Bacillus subtilis ATCC 6051, S. aureus ATCC 6538 and Escherichia coli ATCC 11775Using a microbroth dilution method, bacteria were seeded at
    1 × 106 cells per well (200 μL) in a
    96-well plate containing Mueller-Hinton broth with different concentrations (from 1 to 420 μg/mL, 300 μg/mL and so on;
    2-fold increments) of each test compound.
    It showed inhibitory activities with MIC of (3–28 μg/mL) against S. aureus ATCC6538[116]
    The crude extract of B. striataS. album, A. capillaris, C. cassiaThey were seeded at 1 × 106 cells per well (200 μL) in a 96-well plate containing Mueller−Hinton broth (meat extracts 0.2%, acid digest of casein 1.75%, starch 0.15%) with different concentrations (from 1 to 128 μg/mL; 2-fold increments) of each test compound.It showed S. album (0.10%), A. capillaris (0.10%), and C. cassia (0.10%) to have the strongest antibacterial properties.[118]
    The ethyl acetate-soluble (EtOAc) extract of tubers of B. striataS. aureus ATCC 43300, S. aureus ATCC 6538, and Bacillus subtilis ATCC 6051) and Escherichia coli ATCC 11775)Bacteria were seeded at 1 × 106 cells per well (200 μL) in a 96-well plate containing Mueller Hinton broth with different concentrations (from 1 to 420 μg/ml; 2-fold increments) of each test compound.The extract was effective against three Gram-positive bacteria with minimum inhibitory concentrations (MICs) of 52–105 μg/ml.[98]
    The phenanthrene fraction (EF60) from the ethanol extract of fibrous roots of Bletilla striata pseudobulbsS. aureus ATCC 25923, S. aureus ATCC 29213, S. aureus ATCC 43300, E. coli ATCC 35218, and P. aeruginosa ATCC 27853, Bacillus subtilis 168EF60 was active against all tested strains of Staphylococcus aureus, including clinical isolates and methicillin-resistant S. aureus (MRSA). The minimum inhibitory concentration (MIC) values of EF60 against these pathogens ranged from 8 to 64 μg/mL.EF60 could completely kill S. aureus ATCC 29213 at 2× the MIC within 3 h but could kill less than two logarithmic units of ATCC 43300, even at 4× the MIC within 24 h. The postantibiotic effects (PAE) of EF60 (4× MIC) against strains 29213 and 43300 were 2.0 and 0.38 h, respectively.[117]
    Anti-adhesiveBletilla striata extraction solutionPPA was induced by cecal wall abrasion, and Bletilla striata was injected to observe its efect on adhesion in ratsThe rats in the sham operation group was not treated; the other rats of the three experimental groups were intraperitoneally injected with 8 ml of phosphate-buffered saline (Control group), 15% Bletilla striata extraction solution (BS group), and 0.2% hyaluronic acid solution (HA group), respectively.Bletilla striata decreased the development of abdominal adhesion in abrasion-induced model of rats and reduced the expression of the important substance which increased in PPAs.[120]
    ImmunomodulatoryB. striata polysaccharide (BSPF2)Mouse spleen cellsTo observe the immune activity of BSPF2, mouse spleen cells were stimulated with BSPF2 at 10–100 g/mL for 72 h.Immunological assay results demonstrated that BSPF2 significantly induced the spleen cell proliferation in a dose-dependent manner.[121]
    Anti-pulmonary fibrosisB. striata polysaccharideClean grade male SD ratsSD rats were randomly divided into 5 groups, sham operation group (equal volume of normal saline), model group (equal volume of normal saline), tetrandrine positive control (24 mg/kg) group and white and Polysaccharide low
    (100 mg/kg) and high (400 mg/kg) dose groups.
    The Bletilla striata polysaccharide has remarkable regulation effect on anti-oxidation system and immune system, but cannot effectively prevent lung fibrosis.[127]
    Small molecule components of Bletilla striataClean grade male SD ratsSD rats were randomly divided into 5 groups, sham operation group (0.5 mL normal saline), model group (0.5 mL normal saline), and positive control group (tetrandrine 24 mg/kg) and low (20 mg/kg) and high (40 mg/kg) dosage groups of the small molecule pharmacological components of Bletilla, which were administered by gavage once a day for 2 consecutive months.The small molecule components of Bletilla striata can effectively prevent lung fibrosis though regulating the anti-oxidation system,immune system and cytokine level; SMCBS is one of the active components of Bletilla striata on silicosis therapy[124]
    —, not given.
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    Many phytochemicals have been well characterized to lessen swelling or inflammation[89]. A series of phenolic acid and polysaccharide compounds isolated from Bletilla demonstrated anti-inflammatory bioactivity against BV-2 microglial, RAW 264.7, and PC12 cells[96,100102]. For example, phochinenin K (106) exhibited growth inhibitory effects with an IC50 of 1.9 μM, and it is a possible candidate for development as neuroinflammation inhibitory agent[43]. Using the H2O2-induced PC12 cell injury model, (7S)-bletstrin E (242), (7R)-bletstrin F (243) and (7S)-bletstrin F (244) could clearly protect the cells with the cell viabilities of 57.86% ± 2.08%, 64.82% ± 2.84%, and 64.11% ± 2.52%, respectively[98]. With an IC50 of 2.86 ± 0.17 μM, 2,7-dihydroxy-4-methoxyphenanthrene (53) showed potential action against NO generation in RAW 264.7 macrophages[54]. The use of Bletilla in traditional skin care, it is said to function as an astringent, hemostatic and wound healing[33]. Modern medical pharmacology research has validated that this plant has antibacterial effects, which may may help to explain, in part, its traditional use in skin care[24].

    Though it's mentioned that some of these compounds from Bletilla have demonstrated anti-inflammatory action, more extensive studies are needed to fully understand their mechanisms of action, potential therapeutic applications, and safety profiles. Conducting in vivo studies and clinical trials can provide more concrete evidence of their effectiveness.

    There are important antineoplastic agents that have originated from plant natural products[103]. In recent years, several bibenzyl and flavonoid compounds have been discovered from Bletilla that have antineoplastic activity against A549 cells and other cells. For example, 7-hydroxy-2-methoxy-phenanthrene-3,4-dione (160) and 3′,7′,7-trihydroxy-2,2′,4′-trimethoxy-[1,8′-biphenanthrene]-3,4-dione (163) have shown strong antiproliferative effects and induced ROS production after 24 h in A549 cells[87]. The doxorubicin (Dox)/FA (folate)-BSP-SA (stearic acid) modified Bletilla striata polysaccharide micelles boosted the drug enrichment in tumors and improved the in vivo anticancer effects[104,105]. Micelles, nanoparticles, microspheres, and microneedles are examples of B. striata polysaccharide-based drug delivery systems that exhibit both drug delivery and anti-cancer functionality. These experiments confirmed that some of the compounds isolated from the Bletilla have potential activity for the treatment of cancer.

    However, most of the evidence presented in the previous studies is based on in vitro experiments or cell culture studies. It is highly necessary to use animal models to study the in vivo anti-tumor effects of Bletilla extracts or compounds. These studies can help evaluate the safety and effectiveness of treatments based on Bletilla. Additionally, through such methods, researchers can further investigate the mechanisms of Bletilla's anti-tumor activities, exploring how Bletilla compounds interact with cancer cells, immune responses, and signaling pathways involved in tumor growth and metastasis.

    Antiviral medications are essential for preventing the spread of illness, and are especially important nowadays with pandemics and drug-resistant viral strains[5, 6]. Therefore, it is vitally necessary to find novel, safe, and effective antiviral medications to treat or prevent viral infections[106]. B. striata plant contains compounds that have been recorded in ancient texts to cure cough, pneumonia, and skin rashes, and these may be related to potential antiviral constituents[23]. Some constituents of B. striata have antiviral activity, for example, phenanthrenes and diphenanthrenes from B. striata displayed potent anti-influenza viral in a Madin-Darby canine kidney model and embryonated eggs model, diphenanthrenes with parentally higher inhibitory activity than monophenanthrenes[107]. But more research is needed to further determine the antiviral activity of Bletilla, understand how Bletilla compounds interact with viral proteins or the host immune response, and conduct safety and toxicity studies, which are crucial for the development of related materials.

    Free radicals have the potential to exacerbate lipid peroxidation and harm cell membranes, which can lead to several prevalent human diseases, including cancer, cataracts, and coronary heart disease[108]. Research has shown that extracts from Bletilla possess strong antioxidant activity. However, this antioxidant activity can vary depending on the different growing environments of the plant. Additionally, the antioxidant capabilities of extracts from different parts of the Bletilla plant also vary[93]. Clinical studies have shown that traditional Chinese medicine formulas containing Bletilla can inhibit tyrosinase activity and possess antioxidant properties, thus resulting in skin-whitening effects[108]. Furthermore, some research reveals that the polysaccharides in the plant exhibit significant antioxidant activity, effectively scavenging free radicals and inhibiting tyrosinase activity[33]. This highlights the skin-whitening potential of the fibrous root of Bletilla striata, indicating promising prospects for the comprehensive utilization of the B. striata plant[33]. However, most studies on the pharmacological activities of Bletilla have focused solely on B. striata, neglecting other species within the genus. Different species may possess varying phytochemical compositions and antioxidant properties, which can lead to an incomplete understanding of the genus as a whole.

    Available hemostatic agents are expensive or raise safety concerns, and B. striata may serve as an inexpensive, natural, and promising alternative[109]. Polysaccharides of B. striata displayed hemostatic activity through inhibition of the NLRP3 inflammasome[110112]. The ADP receptor signaling pathways of P2Y1, P2Y12, and PKC receptors may be activated as part of the hemostasis[113]. Alkaloids from Bletilla have hemostatic activities through platelet deformation, aggregation, and secretion. In addition, polysaccharides of Bletilla striata have potential wound-healing medicinal value[110]. Currently, Bletilla plants have been used in various traditional systems, such as traditional Chinese medicine and Ayurveda, to control bleeding.

    Previous studies revealed that Bletilla displayed antibacterial effects[114]. For example, bletistrin F, showed inhibitory activities with MIC of (3–28 μg/mL) against S. aureus ATCC 6538[115,116]. Antimicrobial screening of Bletilla showed S. album (0.10%), A. capillaris (0.10%), and C. cassia (0.10%) to have the strongest antibacterial properties[117,118]. In addition, phenanthrenes are worthy of further investigation as a potential phytotherapeutic agent for treating infections caused by S. aureus and MRSA[119]. However, further in vivo studies on the antibacterial activity of Bletilla are lacking, which is needed for clinical application. For example, the specific mechanism of antibacterial activity of Bletilla still needs to be elucidated. While research on the antibacterial activity of Bletilla plants is promising, it faces several shortcomings and challenges that need to be addressed for a more comprehensive understanding of their potential therapeutic applications. Further studies with standardized methodologies, mechanistic insights, clinical trials, and consideration of ecological and safety concerns are essential to advance this field.

    There are other pharmacological activities of Bletilla, like anti-fibrosis activity, anti-adhesive activity, and immunomodulatory activity. For example, B. striata has been studied as a new and cheaper antiadhesive substance which decreased the development of abdominal adhesion abrasion-induced model in rats[120]. However, the natural resources of Bletilla are also getting scarcer. To preserve the sustainable development of Bletilla species, proper farming practices are required, along with the protection and economical use of these resources. The immunomodulatory activity of the Bletilla species was assessed using the 3H-thymidine incorporation method test, and BSP-2 increased the pinocytic capacity and NO generation, which improved the immunomodulatory function[121,122].

    B. striata extract was shown to have anti-pulmonary fibrosis effect[123]. B. striata polysaccharide can successfully prevent lung fibrosis through established by invasive intratracheal instillation method and evaluated by lung indexes[123,124]. Moreover, Bletilla species need further investigations to evaluate their long-term in vivo and in vitro activity before proceeding to the development of pharmaceutical formulation.

    While there is currently a deep understanding of the pharmacological activity of plants in the Bletilla genus, there are still many gaps that need to be addressed. To overcome these shortcomings, future research on the pharmacological activity of Bletilla species should emphasize comprehensive, well-designed studies with a focus on species-specific effects, mechanistic insights, and rigorous clinical trials. Additionally, collaboration among researchers, standardization of methods, and transparent reporting of results can help advance our understanding of the therapeutic potential of Bletilla plants. Researchers should also consider safety aspects and explore potential herb-drug interactions to ensure the responsible use of Bletilla-based therapies.

    There are several common clinical applications of Bletilla striata in TCM. The gum of B. striata has unique viscosity characteristics and can be used as thickener, lubricant, emulsifier and moisturizer in the petroleum, food, medicine, and cosmetics industries[125130]. B. striata is used as a coupling agent, plasma substitute, preparation adjuvant, food preservative and daily chemical raw material[131133]. In clinical practice, B. striata glue has also been proven to control the infections and is beneficial to the healing of burns and wounds[133135].

    In ethnic communities in Southwest China, the locals chew fresh Bletilla tubers directly or take them orally after soaking in honey to treat cough, pneumonia and other diseases[33, 34]. This traditional use is common in local communities in Southwest China, and suggests at the safety of Bletilla. However, current research shows it is still necessary to control the dosage when using Bletilla[136].

    Zebrafish embryos and larvae respond to most drugs in a manner similar to humans[137]. Militarine, the main active ingredient of Bletilla, was tested in a zebrafish embryo development assay at concentrations of 0.025 g/L and 0.05 g/L, and with the increased concentration, the heart rate of zebrafish embryos is slowed. Mortality and malformation rates of zebrafish embryos gradually increased with time and militarine concentration[138]. Although Bletilla species are safe at therapeutic dose ranges, further research on their safety is required[136]. More in-depth studies should be carried out on Bletilla to extract effective ingredients and make better preparations for clinical use[139].

    According to the traditional medicinal knowledge in ancient Chinese texts, Bletilla has been an important ingredient for skin care since ancient times. Many ethnic minority groups in China still retain the practice of using Bletilla for skin care, and the plant parts and preparation methods of use are consistent with the records in ancient texts. Almost 300 phytochemicals have been identified from Bletilla, and some of them possess important pharmacological activities, which support its traditional uses and suggest the important medicinal development potential of this genus. This review has demonstrated that Bletilla, as an important medicinal plant of Orchidaceae, still requires further research to fathom its medicinal potential.

    For instance, it is necessary to enhance the quality control procedures based on the chemical components and pharmacological activity of Bletilla. The chemical composition and pharmacological properties of Bletilla are critical areas of current research. According to previous studies, the main bioactive components of Bletilla can vary greatly according to its origin, harvest time, distribution, storage, and adulteration. However, variation in bioactivities caused by the differences in Bletilla constituentshave not been explored extensively yet. To develop clinical applications of Bletilla, it is crucial to further explore the mechanism of action between its chemical composition variation and its pharmacological actions.

    In addition, although the tuber has historically been the main medicinal part of Bletilla, research has shown that the chemical composition in other parts of Bletilla, such as stems, leaves, and flowers, also give these parts a variety of pharmacological activities. Further in-depth analysis of the chemical components and pharmacological activities of different parts of this genus is worthwhile, to explore the specific chemical basis of its pharmacological activities, develop related drugs, and promote clinical applications. For example, Bletilla polysaccharide has good hemostasis and astringent wound effects[110], so it may have the potential to be developed into a drug or related medical materials to stop bleeding and heal wounds.

    Finally, as a cautionary note, many unrestrained collections and the destruction of habitats have made the resources of wild Bletilla rarer. In addition to protecting the wild populations of Bletilla, appropriate breeding techniques should be adopted to meet the commercial needs of this economically important genus, thereby allowing its sustainable use in commerce.

    The authors confirm contribution to the paper as follows: study conception and design, funding acquirement: Long C; data analysis, draft manuscript preparation, literature review: Fan Y, Zhao J; manuscript revise and language editing: Wang M, Kennelly EJ, Long C. All authors reviewed the results and approved the final version of the manuscript.

    The raw data supporting the conclusion of this article will be made available by the authors, without undue reservation, to any qualified researcher. Requests to access these datasets should be directed to Yanxiao Fan (fanyanxiao0510@163.com).

    This research was funded by the Yunnan Provincial Science and Technology Talent and Platform Plan (202305AF150121), Assessment of Edible & Medicinal Plant Diversity and Associated Traditional Knowledge in Gaoligong Mountains (GBP-2022-01), the National Natural Science Foundation of China (32370407, 31761143001 & 31870316), China Scholarship Council (202206390021), and the Minzu University of China (2020MDJC03, 2022ZDPY10 & 2023GJAQ09).

  • The authors declare that they have no conflict of interest.

  • [1]

    Ali-Nezhad FM, Eskandari H. 2012. Effect of architectural design of greenhouse on solar radiation interception and crops growth conditions. International Journal of Agriculture and Crop Sciences 4:122−27

    Google Scholar

    [2]

    Aied KY, Wahab Z, Kamaruddin HR, Shaari AR. 2017. Growth response of eggplant (Solanummelongena L.) to shading and cultivation inside greenhouse in a tropical region. International Journal of Scientific and Engineering 8:89−101

    Google Scholar

    [3]

    Xu J, Li Y, Wang R, Liu W, Zhou P. 2015. Experimental performance of evaporative cooling pad systems in greenhouses in humid subtropical climates. Applied Energy 138:291−301

    doi: 10.1016/j.apenergy.2014.10.061

    CrossRef   Google Scholar

    [4]

    Rajametov SN, Yang EY, Jeong HB, Cho MC, Chae SY, et al. 2021. Heat treatment in two tomato cultivars: a study of the effect on physiological and growth recovery. Horticulturae 7:119

    doi: 10.3390/horticulturae7050119

    CrossRef   Google Scholar

    [5]

    Uzun S. 2006. The quantitative effects of temperature and light on the number of leaves preceding the first fruiting inflorescence on the stem of tomato (Lycopersicon esculentum, Mill.) and aubergine (Solanum melongena L. ). Scientia Horticulturae 109:142−46

    doi: 10.1016/j.scienta.2006.04.006

    CrossRef   Google Scholar

    [6]

    Rajasekar M, Arumugam T, Kumar SR. 2013. Influence of weather and growing environment on vegetable growth and yield. Journal of Horticulture and Forestry 5:160−67

    Google Scholar

    [7]

    Khawlhring N, Thanga LTJ, Lalnunmawia F. 2012. Plant performance of Anthurium andreanum as affected by shade conditions and different conventional nutrient sources. Journal of Horticulture and Forestry 4:22−26

    Google Scholar

    [8]

    Hatamian M, Salehi H. 2017. Physiological characteristics of two rose cultivars (Rosa hybrida L.) under different levels of dhading in greenhouse conditions. Journal of Ornamental Plants 7:147−55

    Google Scholar

    [9]

    Hasan MR, Chakrabarti R. 2009. Use of algae and aquatic macrophytes as feed in small-scale aquaculture. No. 531. Food and Agriculture Organization of the United Nations, Roma, Italy. 123 pp. https://www.fao.org/3/i1141e/i1141e.pdf

    [10]

    Rezai S, Nikbakht A, Etemadi N, Yousefi M, Majidi MM. 2015. Effect of different polyethylene covers and shade on morphological and physiological characteristics of dwarf Lisianthus cv. Matador. Journal of Science and Technology of Greenhouse Culture 6:135−44

    Google Scholar

    [11]

    Abduh MY, Ono JM, Khairani M, Manurung R. 2017. The influence of light intensity on the protein content of Azolla microphylla and pre-treatment with Saccharomyces cerevisiae to increase protein recovery. Journal of Applied Sciences Research 13:16−23

    Google Scholar

    [12]

    Yakhin OI, Lubyanov AA, Yakhin IA, Brown PH. 2016. Biostimulants in plant science: a global perspective. Frontiers in Plant Science 7:2049

    doi: 10.3389/fpls.2016.02049

    CrossRef   Google Scholar

    [13]

    Dong C, Wang G, Du M, Niu C, Bao Z, et al. 2020. Biostimulants promote plant vigor of tomato and strawberry after transplanting. Scientia Horticulturae 267:109355

    doi: 10.1016/j.scienta.2020.109355

    CrossRef   Google Scholar

    [14]

    Taiz L, Zieger E, 2002. Plant Physiology, 3 Edition. Sunderland: Sinauer Associates. 690 pp.

    [15]

    Estaji A, Souri MK, Omidbaigi R. 2011. Evaluation of different levels of nitrogen and flowerpruning on milk thistle (Silybum marianum L.) yield and fatty acids. Zeitschrift fur Arznei und Gewurzpflanzen 4:170−75

    Google Scholar

    [16]

    Zhou Y, Lam H, Zhang J. 2007. Inhibition of photosynthesis and energy dissipation induced by water and high light stresses in rice. Journal of Experimental Botany 58:1207−17

    doi: 10.1093/jxb/erl291

    CrossRef   Google Scholar

    [17]

    Fang S, Wang J, Wei Z, Zhu Z. 2006. Methods to break seed dormancy in Cyclocarya paliurus (Batal) Iljinskaja. Scientia Horticulturae 110:305−9

    doi: 10.1016/j.scienta.2006.06.031

    CrossRef   Google Scholar

    [18]

    Li J, Luo M, Luo Z, Guo A, Yang X, et al. 2019. Transcriptome profiling reveals the anti-diabetic molecular mechanism of Cyclocarya paliurus polysaccharides: anti-diabetic molecular mechanism of Cyclocarya paliurus polysaccharides. Journal of Functional Foods 55:1−8

    doi: 10.1016/j.jff.2018.12.039

    CrossRef   Google Scholar

    [19]

    Shang X, Tan J, Du Y, Liu X, Zhang Z. 2018. Environmentally-friendly extraction of flavonoids from Cyclocarya paliurus (Batal.) Iljinskaja leaves with deep eutectic solvents and evaluation of their antioxidant activities. Molecules 23:2110

    doi: 10.3390/molecules23092110

    CrossRef   Google Scholar

    [20]

    Xie J, Xie M, Nie S, Shen M, Wang Y, et al. 2010. Isolation, chemical composition and antioxidant activities of a water-soluble polysaccharide from Cyclocarya paliurus (Batal.) Iljinskaja. Food Chemistry 119:1626−32

    doi: 10.1016/j.foodchem.2009.09.055

    CrossRef   Google Scholar

    [21]

    Deng B, Shang X, Fang S, Li Q, Fu X, et al. 2012. Integrated effects of light intensity and fertilization on growth and flavonoid accumulation in Cyclocarya paliurus. Journal of Agricultural and Food Chemistry 60:6286−92

    doi: 10.1021/jf301525s

    CrossRef   Google Scholar

    [22]

    Liu Y, Qian C, Ding S, Shang X, Yang W, et al. 2016. Effect of light regime and provenance on leaf characteristics, growth and flavonoid accumulation in Cyclocarya paliurus (Batal) Iljinskaja coppices. Botanical Studies 57:28

    doi: 10.1186/s40529-016-0145-7

    CrossRef   Google Scholar

    [23]

    Yang W, Liu Y, Fang S, Ding H, Zhou M, et al. 2017. Variation in growth, photosynthesis and water-soluble polysaccharide of Cyclocarya paliurus under different light regimes. iForest - Biogeosciences and Forestry 10:468−74

    doi: 10.3832/ifor2185-010

    CrossRef   Google Scholar

    [24]

    Liu Y, Wang TL, Fang S, Zhou M, Qin J. 2018. Responses of morphology, gas exchange, photochemical activity of photosystem II, and antioxicant balance in Cyclocarya paliurus to light spectra. Frontiers in Plant Science 9:1704

    doi: 10.3389/fpls.2018.01704

    CrossRef   Google Scholar

    [25]

    Feng Y, Lin X, Qian L, Hu N, Kuang C, et al. 2020. Morphological and physiological variations of Cyclocarya paliurus under different soil water capacities. Physiology and Molecular Biology of Plants 26:1663−74

    doi: 10.1007/s12298-020-00849-4

    CrossRef   Google Scholar

    [26]

    Stein WI, Edwards JL, Tinus RW. 1975. Outlook for container-grown seedling use in reforestation. Journal of Forestry 73:337−41

    Google Scholar

    [27]

    Li H. 2000. Principles and techniques of plant physiological biochemical experiment. Beijing: Higher Education Press. pp. 195−97.

    [28]

    Ahmed S, Raza MA, Zhou T, Hussain S, Khalid M, et al. 2018. Responses of soybean dry matter production, phosphorus accumulation, and seed yield to sowing time under relay intercropping with maize. Agronomy 8:282

    doi: 10.3390/agronomy8120282

    CrossRef   Google Scholar

    [29]

    Raza MA, Feng L, Iqbal N, Manaf A, Khalid M, et al. 2018. Effect of sulphur application on photosynthesis and biomass accumulation of sesame varieties under rainfed conditions. Agronomy 8:149

    doi: 10.3390/agronomy8080149

    CrossRef   Google Scholar

    [30]

    Raza AM, Feng L, Iqbal N, Imran K, Meraj AT, et al. 2020. Effects of contrasting shade treatments on the carbon production and antioxidant activities of soybean plants. Functional Plant Biology 47:342−54

    doi: 10.1071/FP19213

    CrossRef   Google Scholar

    [31]

    Lee GS, Choi SC, Lee GJ, Jang Y, Lee JH, et al. 2014. Influence of shading and irrigation on the growth and development of leaves tissue in hot pepper. Korean Journal of Horticultural Science and Technology 32:448−53

    doi: 10.7235/hort.2014.14015

    CrossRef   Google Scholar

    [32]

    Saeidi M, Abdoli M. 2015. Effect of drought stress during grain filling on yield and its components, gas exchange variables, and some physiological traits of wheat cultivars. Journal of Agricultural Science and Technology 17:885−98

    Google Scholar

    [33]

    Fang L, Ma Z, Wang Q, Nian H, Ma Q, et al. 2021. Plant growth and photosynthetic characteristics of soybean seedlings under different LED lighting quality conditions. Journal of Plant Growth Regulation 40:668−78

    doi: 10.1007/s00344-020-10131-2

    CrossRef   Google Scholar

    [34]

    Stagnari F, Galieni A, Pisante M. 2015. Shading and nitrogen management affect quality, safety and yield of greenhouse-grown leaf lettuce. Scientia Horticulturae 192:70−79

    doi: 10.1016/j.scienta.2015.05.003

    CrossRef   Google Scholar

    [35]

    Sui X, Mao S, Wang L, Zhang B, Zhang Z. 2012. Effect of low light on the characteristics of photosynthesis and chlorophyll a fluorescence during leaf development of sweet pepper. Journal of Integrative Agriculture 11:1633−43

    doi: 10.1016/S2095-3119(12)60166-X

    CrossRef   Google Scholar

    [36]

    Fu W, Li P, Wu Y. 2012. Effects of different light intensities on chlorophyll fluo-rescence characteristics and yield in lettuce. Scientia Horticulturae 135:45−51

    doi: 10.1016/j.scienta.2011.12.004

    CrossRef   Google Scholar

    [37]

    Gupta A, Singh M, Laxmi A. 2015. Interaction between glucose and brassinosteroid during the regulation of lateral root development in Arabidopsis. Plant Physiology 168:307−20

    doi: 10.1104/pp.114.256313

    CrossRef   Google Scholar

    [38]

    Rolland F, Baena-Gonzalez E, Sheen J. 2006. Sugar sensing and signaling in plants: conserved and novel mechanisms. Annual Review of Plant Biology 57:675−709

    doi: 10.1146/annurev.arplant.57.032905.105441

    CrossRef   Google Scholar

    [39]

    Sofo A, Dichio B, Xiloyannis C, Masia A. 2004. Lipoxygenase activity and proline accumulation in leaves and roots of olive trees in response to drought stress. Physiologia Plantarum 121:58−65

    doi: 10.1111/j.0031-9317.2004.00294.x

    CrossRef   Google Scholar

    [40]

    Ben Abdallah M, Methenni K, Nouairi I, Zarrouk M, Ben Youssef N. 2017. Drought priming improves subsequent more severe drought in a drought-sensitive cultivar of olive cv. Chétoui. Scientia Horticulturae 221:43−52

    doi: 10.1016/j.scienta.2017.04.021

    CrossRef   Google Scholar

    [41]

    Sulpice R, Pyl ET, Ishihara H, Trenkamp S, Steinfath M, et al. 2009. Starch as a major integrator in the regulation of plant growth. Proceedings of the National Academy of Sciences of the United States of America 106:10348−53

    doi: 10.1073/pnas.0903478106

    CrossRef   Google Scholar

    [42]

    Cakmak I. 2005. The role of potassium in alleviating detrimental effects of abiotic stresses in plants. Journal of Plant Nutrition and Soil Science 168:521−30

    doi: 10.1002/jpln.200420485

    CrossRef   Google Scholar

    [43]

    Hafez E, Farig M. 2019. Efficacy of salicylic acid as a cofactor for ameliorating effects of water stress and enhancing wheat yield and water use efficiency in saline soil. International Journal of Plant Production 13:163−76

    doi: 10.1007/s42106-019-00036-w

    CrossRef   Google Scholar

    [44]

    Chow WS, Ball MC, Anderson JM. 1990. Growth and photosynthetic responses of spinach to salinity: implications of K+ nutrition for salt tolerance. Australian Journal of Plant Physiology 17:563−78

    doi: 10.1071/PP9900563

    CrossRef   Google Scholar

    [45]

    Schachtman DP, Kumar R, Schroeder JI, Marsh EL. 1997. Molecular and functional characterization of a novel low-affinity cation transporter (LCTI) in higher plants. Proceedings of the National Academy of Sciences of the United States of America 94:11079−84

    doi: 10.1073/pnas.94.20.11079

    CrossRef   Google Scholar

    [46]

    Castaňo CAM, Morales LCS, Obando MFH. 2008. Evaluation of the nutritional deficiencies in the blackberry crop (Rubus glaucus) in controlled conditions for low mountainous forest. Agronomía Tropical 16:75−88

    Google Scholar

    [47]

    Cramer GR, Lynch J, Läuchli A, Epstein E. 1987. Influx of Na+, K+ and Ca2+ into roots of salt-stressed cotton seedlings: effects of supplemental Ca2+. Plant Physiology 83:510−16

    doi: 10.1104/pp.83.3.510

    CrossRef   Google Scholar

    [48]

    Mansfield TA, Hetherington AM, Atkinson CJ. 1990. Some aspects of stomatal physiology. Annual Review of Plant Physiology and Plant Molecular Biology 41:55−75

    doi: 10.1146/annurev.pp.41.060190.000415

    CrossRef   Google Scholar

    [49]

    Wakeel A, Farooq M, Qadir M, Schubert S. 2011. Potassium substitution by sodium in plants. Critical Reviews in Plant Sciences 30:401−13

    doi: 10.1080/07352689.2011.587728

    CrossRef   Google Scholar

    [50]

    Gill SS, Tuteja N. 2010. Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiology and Biochemistry 48:909−30

    doi: 10.1016/j.plaphy.2010.08.016

    CrossRef   Google Scholar

  • Cite this article

    Feng Y, Zhi L, Pan H, Chen Y, Xu J. 2023. Shade improves seedling quality of ornamental Cyclocarya species under plastic greenhouse cultivation. Ornamental Plant Research 3:13 doi: 10.48130/OPR-2023-0013
    Feng Y, Zhi L, Pan H, Chen Y, Xu J. 2023. Shade improves seedling quality of ornamental Cyclocarya species under plastic greenhouse cultivation. Ornamental Plant Research 3:13 doi: 10.48130/OPR-2023-0013

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Shade improves seedling quality of ornamental Cyclocarya species under plastic greenhouse cultivation

Ornamental Plant Research  3 Article number: 13  (2023)  |  Cite this article

Abstract: Plastic greenhouse cultivation is a widespread and convenient way of cultivating high-quality seedlings, which are often damaged by higher temperatures during summer. Cyclocarya paliurus, a medical and ornamental species with low-quality seedlings, was investigated using three shading levels (treatment with no shade net (SH0), treatment with one layer of shade net (SH1), and treatment with two layers of shade net (SH2)). The growth and physiological responses of seedlings under plastic greenhouse cultivation were investigated from June to September. The results showed that the survival rate of seedlings reached 100%, and seedling growth and biomass were the best under SH2, with higher plant height and leaf area than that under other treatments. Water content of seedlings exhibit not difference between three shading levels, and leaves had the highest water content and total soluble sugar content. The chlorophyll content in the leaf increased, but malondialdehyde content decreased with increasing shading layers. Mineral content were in the following order: calcium > potassium > magnesium > sodium, and the translocation factor decreased with increasing shading layers. Antioxidant enzyme activities were in the following order: SOD > PPO > POD > CAT; their activities decreased with increasing shading layers, except for that of PPO. Various correlation existed between physiological response and seedling growth. Shade improved seedling quality through a series of physiological responses under plastic greenhouse conditions. This study provides a solid foundation for greenhouse cultivation of Cyclocarya species.

    • Greenhouse cultivation is a widespread method of cultivating high-quality plants to improve survival and adaptability. However, with the impacts of global climate change, abiotic stresses, such as higher temperature or stronger light intensity due to higher solar radiation, are becoming a great threat to plants especially in tropical, subtropical and temperate regions, consequently retarding plant growth or causing plant death under greenhouse cultivation[14]. To date, shade practice has been a common and effective method that alleviates or prevents the damage on plant cultivation under greenhouse conditions[2, 57]. Aied et al.[2] reported that eggplant grown under greenhouse conditions with 30%–70% shading, performed better than that grown in the open field.

      Light is an important factor that affects plant growth[7, 8]. Shade practice reduces light intensity under greenhouse cultivation, and excessive or slight shading causes unfavorable light intensity and further affects plant growth, hence, it is necessary to determine the optimal shading levels for plant growth. In practice, shading levels in greenhouse cultivation depend on plant species and genotypes. Slight shade (25%–50% of full sunlight) was suitable for Azolla growth[9]. Two dwarf Lisianthus species showed the best growth under 20% shade condition in a study performed by Rezai et al.[10].

      Plants respond to shading conditions through morphological changes and physiological behaviors. With regard to morphology, suitable shading conditions benefited the morphological development of Anthurium andreanum[7] and increased the plant height, internode length, leaf number, and leaf area[11]. Meanwhile, a well-developed root system is formed to increase nutrient absorption capacity[12,13]. In contrast, many plants under light-limiting or excessive light conditions showed undesirable morphological traits and reduced plant quality[11]. Regarding physiological behaviors, the biosynthesis and degradation of chlorophyll is actively regulated by light[14,15], and suitable shading levels upregulate chlorophyll content[8, 11]. In addition, relative water content (RWC) and soluble sugar content are also related to shading levels[11,16]. Moreover, the massive production of reactive oxygen species (ROS) owing to abiotic stress leads to accelerated activity of antioxidant enzymes, such as superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) in plant cells to minimize the negative impacts[11]. In all, various physiological activities of plants are in order when plants are cultivated under greenhouse cultivation with suitable shading conditions.

      Cyclocarya paliurus (Batal) Iljinskaja (C. paliurus), belonging to the family Juglandaceae, is a native, medicinal and ornamental tree distributed in the highlands of southern China[17]. In recent years, the demand for its leaf production has increased with the exploration and exploitation of its medical value because the leaf contains many bioactive compounds, such as triterpenoids, polysaccharides, and phenolic compounds etc., which benefit human health[1820]. The harvesting of its leaf was mainly from natural forests of Cyclocarya species, resulting in serious damage to the natural forests. Moreover, the shape of Cyclocarya species can be used as ornamental trees in urban parks due to its graceful shape and copper-like fruits. Thus, it is necessary to develop artificial cultivation of Cyclocarya species.

      Seedlings are the only way to artificially cultivate Cyclocarya species, and seedling cultivation has an important effect on the quality of artificial cultivation. Previous reports on seedling cultivation of C. paliurus only focused on improving leaf bioactive content using light and fertilization treatments under laboratory conditions[2124]. For example, Deng et al.[21] reported that leaf production per seedling increased under intermediate shade and fertilization treatments, and Yang et al.[23] investigated the effect of differential light quality and intensity on the growth and water-soluble polysaccharides of C. paliurus. However, in the actual process of forestry breeding, seedling is often cultivated in the open field. Considering the difference in experimental controllability under laboratory conditions, changeable environmental factors in the open field directly or indirectly lead to the uneven growth and low quality of seedling. This reduced the survival and adaptability of seedlings after transplantation. Therefore, it is necessary and urgent to cultivate high-quality seedlings of Cyclocarya species.

      Plastic greenhouse cultivation has been used to cultivate seedling growth of Cyclocarya species[25]. Feng et al.[25] reported that 50%–60% soil water capacity in plastic greenhouses significantly improved seedling quality. However, during the process of seedling cultivation, seedling growth was inhibited or seedlings died which was found to be caused by higher temperatures, especially in summer. To avoid or alleviate this problem, we hypothesized that shade could improve the seedling quality of C. paliurus under plastic greenhouse conditions. Therefore, the objective of this study was to determine the optimal shading level for C. paliurus seedlings from June to September and obtain high-quality seedlings based on their growth performances and physiological behaviors under different shading levels.

    • Seeds were collected from the natural forest of Jinggangshan (Jiangxi province, China, named JX), and the collected seeds were treated using the method of Fang et al.[17]. After stratification treatment for five months, the germinated seedlings were first transplanted into cultivation bags (10.8 cm in diameter and 14.5 cm in height) filled with a medium (a mixture of nutritional soil: perlite (10:1, g/g)) and then cultured in an incubation room at 15 ± 2 °C (day) and 10 ± 2 °C (night), with natural light environment (about 15 ± 1 h photoperiod). Each bag contained one plant.

      Seedlings with even growth (height, marked H1) were selected and exposed to different shading levels in a plastic greenhouse under natural condition. Shade nets were set at 1.8 m height from the ground with different layers of shade net: no layer of shade net (marked SH0, as control, 60%–70% of full sunlight), one layer of shade net (marked SH1, 30%–40% of full sunlight) and two layers of shade net (marked SH2, 10%–20% of full sunlight); and plastic sheeting were not fitted around the plastic greenhouse. Soil water capacity was maintained at 50%–60%, and the lost water was supplemented at approximately 6:00 AM every day, and the position of the seedling was adjusted every 5 d. The experiment was established in a randomized complete block design, and each shading treatment contained three replicates and 15 seedlings per replicate.

      After 100 d of shading treatment (from June to September), the surviving seedlings were observed and investigated for growth analysis. Three seedlings were randomly selected and separated into leaves, shoots, and roots to determine and analyze their water content (WC) and mineral content, and the fourth compound leaf from the top to bottom with the same orientation were obtained from other seedlings and then immediately frozen in liquid nitrogen to determine and analyze their total soluble sugar (TSS) content, antioxidant enzyme activity, chlorophyll content and malondialdehyde (MDA) content.

    • Seedling height (the final height, marked H2), ground diameter (DH), and leaf area (LA) of the seedlings under each shading treatment were measured. After washing, three parts of seedling were weighed (marked FW), and then heated at 105 °C for 15 min and dried to a constant weight (marked DW) at 85 °C. Finally, the growth and biomass of seedlings were assessed and the growth index was calculated according to the method described by Feng et al.[25].

    • Based on the measurement of FW and DW, WC was calculated with the formula 1[26]:

      WC(%)=FWDWFW×100 (1)
    • According to the method described by Feng et al.[25], dried samples were digested and extraction was obtained using the electric-heating digestion method by the Block Digestion System (AIM600, Clontarf Qld 4019 Australia), then the mineral content was measured using Inductively Coupled Plasma - Optical Emission Spectrometer (ICP-OES, optima 7000DV, PerkinElmer Co., USA), and finally the mineral content (potassium (K), calcium (Ca), magnesium (Mg), sodium (Na)) and translocation factor (TF) were calculated according to Eqns 2 & 3:

      C(mg/g)=Cm×VDW (2)
      TF=Cshoot+CleafCroot (3)

      Cm is the mineral content measured by ICP-OES, mg/L; V is the total volume of extraction, L; DW is the dried weight of the sample, g; Cshoot, Cleaf, and Croot are the mineral content in the shoots, leaves and roots, respectively.

    • Total soluble sugar (TSS) was extracted and measured according to the method described by Li[27]. First, fresh leaves were ground and added to 10 mL distilled water, and then heated at 100 °C for 30 min, finally filtered to a volume of 25 mL. Next, 0.5 mL extraction was added to a mixture of 1.5 mL of distilled water, 0.5 mL of Anthrone and ethyl acetate mixture (Anthrone:ethyl acetate = 1:50 g/mL) and 5 mL of H2SO4, and then kept at 100 °C for 1 min. Finally, the absorbance was measured at 630 nm using a UV-visible spectrophotometer (TU-1900, Beijing Purkinje General Instrument Co., Ltd, China). TSS content was calculated using Eqn 4:

      C(%)=Cs×VW×0.5×1000000×100 (4)

      Cs was obtained from the standard curve in which sugar was used as the standard curve, mg/L; V is the total volume of extraction, mL; W indicates the fresh weight of the sample, g.

    • Chlorophyll content was determined using the method explained by Li[27]. Briefly, fresh leaves were ground and added to 10 mL 80% acetone, and then filtered to a volume of 25 mL. Then, the absorbance was measured using a UV-visible spectrophotometer (TU-1900, Beijing Purkinje General Instrument Co., Ltd, China) at 470 nm (A470), 663 nm (A663) and 645 nm (A645). Finally, chlorophyll content and carotenoid content were calculated using Eqns 5, 6, 7 & 8 respectively.

      Ca=13.95A6656.88A649 (5)
      Cb=24.96A6497.32A665 (6)
      Cx.c=1000A4702.05Ca114.8Cb245 (7)
      Chlorophyllcontent(mg/g)=(Ca+Cb)×VW (8)

      V is the total volume of extraction, mL; W indicates the fresh weight of the sample, g.

    • MDA content was extracted from fresh leaves using an improved thiobarbituric acid-malondialdehyde (TBA-MDA) assay method[27], and then measured at 450 nm, 532 nm and 600 nm by a UV-visible spectrophotometer (TU-1900, Beijing Purkinje General Instrument Co., Ltd, China), respectively. Finally, MDA content was calculated using Eqns 9 & 10, respectively:

      C(umoLL)=6.45(A532A600)0.56A450 (9)
      MDAcontent(umoL/g)=C×VW (10)

      V is the total volume of extraction, L; W indicates the weight of the fresh sample, g.

    • Fresh leaves were ground and added to 15 mL 0.05 mol/L phosphate buffer (PBS, pH 7.8), then kept at 4 °C for 15 min, finally centrifuged at 4 °C, 11,000 ×g for 10 min (TGL-16M, Hunan Xiangyi Laboratory Instrument Development Co., Ltd, China). Enzyme extraction (labeled E) was obtained and antioxidant enzyme activity was immediately determined.

      SOD activity was determined using the method explained by Li[27] with slight modifications. Reaction mixture including 0.1 mL enzyme extraction (labeled E1), 3.0 mL 0.05 mol/L PBS (pH 7.8), 0.6 mL 130 mol/L methionine, 0.6 mL 0.1 mmol/L ethylene diamine tetraacetic acid disodium, 0.4 mL distilled water, 0.6 mL 0.02 mmol/L riboflavin and 0.6 mL 0.75 mmol/L nitro-blue tetrazolium was reacted at 4,000 lx for 20 min, but the control was kept in the dark. After that, the absorbance was measured at 560 nm by a UV-visible spectrophotometer (TU-1900, Beijing Purkinje General Instrument Co., Ltd, China).

      POD activity was measured according to the method described by Li[27] with slight modification. Reaction mixture containing 2.9 mL 0.05 mol/L PBS (pH 7.8), 1 mL 2% H2O2 (v/v), 1 mL 50 mmol/L guaiacol and 0.1 mL enzyme extraction (labeled E2) was heated at 34 °C for 3 min. Then, the absorbance was immediately measured at 470 nm, 6 times with an interval of 1 min by a UV-visible spectrophotometer (TU-1900, Beijing Purkinje General Instrument Co., Ltd, China).

      Polyphenol oxidase (PPO) activity was determined using the method described by Li[27] with slight modification. Reaction mixture containing 3.5 mL 0.05 mol/L PBS (pH 7.8), 1 mL 0.1 mol/L catechol and 0.5 mL enzyme extraction (labeled E3) was heated at 37 °C for 10 min. Then, 2 mL 20% trichloroacetic acid (w/v) was quickly added to the reaction mixture and centrifuged at 4 °C, 11,000 ×g for 10 min (TGL-16M, Hunan Xiangyi Laboratory Instrument Development Co., Ltd, China). Finally, the absorbance was measured immediately at 420 nm, 6 times with an interval of 1 min by a UV-visible spectrophotometer (TU-1900, Beijing Purkinje General Instrument Co., Ltd, China).

      CAT activity was measured using the method described by Li[27] with slight modification. Reaction mixture was prepared using 2 mL 0.05 mol/L PBS (pH 7.8), 0.5 mL 0.18% H2O2 (v/v), 1 mL distilled water and 0.5 mL enzyme extraction (labeled E4). The absorbance was immediately measured at 240 nm, 6 times with an interval of 1 min by a UV-visible spectrophotometer (TU-1900, Beijing Purkinje General Instrument Co., Ltd, China).

      Antioxidant enzyme activity was calculated with the following equations:

      SODactivity(U/gFW)=(AckAe)×Ve0.5×Ack×W×Ve1 (11)
      PODactivity(U/gmin)=D470×Ve0.01×W×Ve2×T (12)
      PPOactivity(U/gmin)=D420×Ve(0.01×W×Ve3×T) (13)
      CATactivity(U/gmin)=D240×Ve0.1×W×Ve4×T (14)

      Ack and Ae are the absorbance of the control and samples at 560 nm, respectively; D470 is the change of absorbance measured at 470 nm; D420 is the change of absorbance measured at 420 nm; D240 is the change of absorbance measured at 240 nm; Ve is the total volume of E, mL; Ve is the total volume of E, mL; Ve1 is the volume of E1, mL; Ve2 is the volume of E2, mL; Ve3 is the volume of E3, mL; Ve4 is the volume of E4; T is the reaction time, min; W is the weight of fresh leaf, g.

    • One-way analysis of variance was conducted to evaluate the effect of shading treatment on growth and physiological characteristics, followed by Tukey's Highly Significant Differences (HSD), and the standard error of differences between means was calculated with p set to 0.05. All statistical analyses were performed using SPSS Statistics 18 version 16.0 for Windows (SPSS Inc., Chicago, IL, USA). Correlation analysis was performed to identify the relationship between the growth index of seedlings and physiological index using the software package Origin 9.1 (Northampton, MA01060, USA). Network analysis was performed using Gephi (version 0.9.2, WebAtlas, France) to analyze the correlation between various physiological index.

    • In the treatment without shade net (SH0, as a control), the survival rate of seedlings was 84.44%, and seedling growth was significantly inhibited. Specifically, seedlings had the lowest height and growth biomass. Leaves were seriously burnt, but RSR was the highest (Fig. 1, Table 1). Under shading conditions, the survival rate of seedlings increased significantly, but there was no difference under SH1 and under SH2. Seedling growth and biomass significantly increased with increasing layers of shade net. In particular, seedlings under SH2 showed favorable growth with the largest leaf area, highest plant height, and highest plant weight, but RSR decreased with increasing layers of shade net (Table 1, Fig. 1).

      Figure 1. 

      Growth of Cyclocarya paliurus seedlings under different shading levels. (a), (c) Seedling growth in plastic greenhouse under three shading levels; (b), (d) seedling growth in plastic greenhouse under SH1 and SH0, respectively; SH0 indicates seedling growth with no shade net (about 60%−70% of full sunlight); SH1 indicates seedling growth with one layer of shade net (about 30%−40% of full sunlight); SH2 indicates seedling growth with two layers of shade net (about 10%−20% of full sunlight). Seedlings were treated in plastic greenhouses with different shading levels for 100 d (from June to September), and plastic sheeting was not fitted around the plastic greenhouse and the shade net was set at 1.8 m height from the ground.

      Table 1.  Variations in seedling growth of Cyclocarya paliurus with different shading treatments

      Treatment Survival rate
      (%)
      The fourth leaf area
      (cm2)
      DH
      (mm)
      The growth rate of stem
      (cm/d)
      DM:FM
      (g/plant)
      RSR
      SH0 84.44 ± 3.85b 3.46 ± 0.42c 1.48 ± 0.24c 0.02 ± 0c 4.04 ± 0.72b 0.70 ± 0.19a
      SH1 95.56 ± 3.85a 9.19 ± 1.045b 2.76 ± 0.42b 0.08 ± 0.02b 6.34 ± 2.79b 0.58 ± 0.13ab
      SH2 100 ± 0a 33.19 ± 3.09a 5.29 ± 0.76a 0.31 ± 0.03a 18.84 ± 3.72a 0.41 ± 0.08b
      SH0 means seedling growth with no shade net; SH1 means seedling grwoth with one layer of shade net; SH2 means seedling growth with two layers of shade net. Different letters (a,b,--) indicate significant differences between shading treatments by Tukey's Highly Significant Differences at p set to 0.05. DH represents ground diameter; DM:FM represents dry mass: fresh mass; RSR represents the ratio of underground weight to aboveground weight.
    • WC of seedlings under SH1 and SH2 was slightly higher than that under SH0, but no differences exist between SH0, SH1, and SH2 (Fig. 2a). However, WC varied in the three parts (root, shoot, and leaf) under different shading levels. Specifically, WC in the root increased, but WC in the shoot decreased with increasing layers of shade net, and no difference was observed between SH0, SH1, and SH2. WC in the leaf under SH1 and SH2 significantly increased by 12.54% and 12.78%, respectively, in comparison with that under SH0 (Fig. 2a).

      Figure 2. 

      (a) Water content and (b) total soluble sugar content in Cyclocarya paliurus seedlings under different shading levels. SH0 indicates seedling growth with no shade net; SH1 indicates seedling growth with one layer of shade net; SH2 indicates seedling growth with two layers of shade net. * means significant differences (p < 0.05 according to Tukey's HSD) between SH0, SH1 and SH2.

    • Leaves under SH2 showed the highest TSS content, TSS content under SH0 and SH1 decreased by 13.93% and 14.67%, respectively, but there was no difference between three shading levels (Fig. 2b).

    • Chlorophyll (Chl) content of leaves under SH2 was 2.52 times and 1.67 times higher than that under SH0 and SH1, respectively (Fig. 3a). Chl a content was much higher than Chl b content and carotenoid content, but both of them showed a similar trend (Fig. 3b, c, d).

      Figure 3. 

      Chlorophyll content and carotenoid content in Cyclocarya paliurus leaves under different shading levels. (a) Chlorophyll content; (b) chlorophyll a content; (c) chlorophyll b content; and (d) carotenoid content. SH0 indicates seedling growth with no shade net; SH1 indicates seedling growth with one layer of shade net; SH2 indicates seedling growth with two layers of shade net. * means significant differences (p < 0.05 according to Tukey's HSD) between SH0, SH1 and SH2.

    • Under the three shading levels, MDA content in leaf under SH2 was the lowest, and its content under SH0 and SH1 significantly increased by 59.15% and 60.25%, respectively (Fig. 4). But there was no difference between under SH0 and SH1.

      Figure 4. 

      MDA content in Cyclocarya paliurus leaves under different shading levels. SH0 indicates seedling growth with no shade net; SH1 indicates seedling growth with one layer of shade net; SH2 indicates seedling growth with two layers of shade net. * means significant differences (p < 0.05 according to Tukey's HSD) between SH0, SH1 and SH2.

    • Four kinds of mineral (K, Ca, Mg, and Na) were detected in seedlings, and their contents were in the order of Ca>K>Mg>Na (Fig. 5a). The translation factor of the four minerals decreased with the increasing layers of shade net, and there was a significant difference between under SH0, SH1, and SH2 (Fig. 5b).

      Figure 5. 

      Mineral contents in Cyclocarya paliurus seedling under different shading levels. (a) mineral contents in seedlings; (b) translocation factor; (c) K content; (d) Ca content; (e) Mg content; (f) Na content. SH0 indicates seedling growth with no shade net; SH1 indicates seedling growth with one layer of shade net; SH2 indicates seedling growth with two layers of shade net. * means significant differences (p < 0.05 according to Tukey's HSD) between SH0, SH1 and SH2.

      K content of seedlings was significantly decreased with the increasing layers of shade net (Fig. 5a), K content under SH0 and SH1 was rich in the leaf (Fig. 5c). K content in the root under SH1 was higher than that under SH0 and SH2, but no change was observed between under SH0, SH1 and SH2. K content in the shoot and leaf decreased with increasing layers of shade net (Fig. 5c).

      Ca content of seedlings under SH1 significantly decreased, but increased significantly under SH2 in comparison with that under SH0 (Fig. 5a). Ca content in the leaf was higher than that in the root and shoot (Fig. 5d). Ca content in the root and shoot had a similar trend with that in seedlings. Leaves showed a decreasing Ca content with increasing layers of shade net.

      Seedling under SH1 had the highest Mg content (Fig. 5a), Mg content was also rich in leaf. Mg content in the root slightly increased, but Mg content in the shoot and leaf significantly decreased with increasing layers of shade net (Fig. 5e).

      Na content of seedlings reached significant difference under SH2 (Fig. 5a), Na content in the root was higher than that in the leaf and in shoot. Na content in the root and shoot significantly increased, but decreased in the leaf with increasing layers of shade net (Fig. 5f).

    • Four kinds of antioxidant enzymes (SOD, PPO, POD, and CAT) were detected in the leaf and their activities were in the following order: SOD>PPO>POD>CAT (Fig. 6). However, their activities varied with different shading levels. Leaf under SH0 had the highest SOD activity, and SOD activity decreased significantly with increasing layers of shade net. The change of POD and CAT activities were similar to that of SOD activity. However, PPO activity increased significantly with increasing layers of shade net, and its activity under SH2 was 25.67 and 23.03 times higher than that under SH0 and SH1, respectively.

      Figure 6. 

      Variations in antioxidant enzyme activities in Cyclocarya paliurus leaves under different shading levels. (a) CAT activity; (b) PPO activity; (c) POD activity; and (d) SOD activity. SH0 indicates seedling growth with no shade net; SH1 indicates seedling growth with one layer of shade net; SH2 indicates seedling growth with two layers of shade net. * means significant differences (p < 0.05 according to Tukey's HSD) between SH0, SH1 and SH2.

    • The growth index of seedlings had positive or negative relationships with various physiological indices under three shading levels (Fig. 7a). And various relationships exist between them. For example, seedling height and leaf area had a negative relation with growth biomass (DM:FM, RSR); PPO activity had a significant positive relation with TSS and Chl content. Antioxidant enzyme activity (SOD, POD, CAT) had a positive relation with mineral content (K, Ca, Mg, Na); but physiological index had a negative relation with antioxidant enzyme activity and mineral content (Fig. 7b).

      Figure 7. 

      (a) Correlation analysis and (b) network analysis between different studied factors. LA, DH, RH, DF, RSR indicates leaf area, ground diameter, the growth rate of shoot, the ratio of dry mass to fresh mass, the ratio of underground weight to aboveground weight, respectively; WC, TSS, Chl, PPO indicate water content, total soluble sugar content, Chlorophyll content, polyphenol oxidase activity;* represents significant differences (p < 0.05 according to Tukey's HSD) between SH0, SH1, and SH2. antioxidant enzyme; mineral content; physiological index; MDA content. Blue line means a positive relation between each other; red line means a negative relation between each other.

    • Greenhouse cultivation is an effective method of cultivating high-quality seedlings. However, higher temperatures or stronger light intensity impairs the successful growth of many plants under greenhouse cultivation, especially plastic greenhouse cultivation. The morphological characteristics of many seedlings were the increase of leaf thickness, the reduction of leaf area and plant height, and the serious sunburn of leaves[2,3,14]. In accordance with previous morphological changes, our findings also showed that plastic greenhouse cultivation with no shade net (SH0) inhibited plant height and leaf area and caused serious leaf sunburn (Table 1, Fig. 1). Our finding of MDA content under SH0 further inferred that seedling growth and metabolism were seriously inhibited or damaged even at the cellular level (Figs 26), resulting in higher activity of antioxidant enzymes and higher content of minerals to alleviate this damage according to network analysis (Fig. 7). This result suggested that plastic greenhouse cultivation without shading condition was unsuitable to cultivate seedlings of C. paliurus.

      Shading practices effectively alleviate or prevent damages in plants under greenhouse cultivation[2,58,14]. Our findings further confirmed that suitable shading conditions favor the growth of seedlings. For one reason, growth index and biomass serve as a direct indicator of plant response to environmental conditions[2830]. Our findings showed that both shading levels (SH1 and SH2) benefited seedling growth, especially two layers of shade net (SH2), where growth index and biomass of seedlings were much better than that under SH0 (Fig. 1, Table 1). Similar findings were reported by Lee et al.[31], who showed that 30% shade promoted plant growth. However, Liu et al.[22] reported that full light intensity conditions resulted in the highest growth and total biomass production of one-year-old C. paliurus plants in an open field, probably because cultivation conditions were different from our study. For another reason, chlorophyll takes part in light utilization and promotes plant growth by improving light interceptions and absorptions and accelerating photosynthetic metabolism[3234]. In corroboration with previous findings[2,35], our finding further revealed that the chlorophyll content in leaves had a significant positive relationship with plant height and TSS content or PPO activity (Fig. 7). However, the significant decrease of chlorophyll content under SH0 and SH1 may be caused by the serious damage to the cellular membrane, leading to the inhibition of chlorophyll synthesis, the promotion of chlorophyll degradation, or the prevention of the conversion of Chl a to Chl b to a large extent. This was further illustrated by the negative relation between Chl content and MDA content (Fig. 7b). This was in corroboration with the results for rose[8], Azolla microphylla[11] and lettuce[34, 36]. Finally, the change of MDA content under different shading levels further inferred that shading condition was suitable for seedling cultivation by alleviating the damage of adverse conditions. This was shown from the network analysis between MDA and other physiological index (Fig. 7b).

      Sugar is a substrate that participates in many biosynthetic processes and energy production in plants, including the regulation of lateral root emergence and development and biosynthesis and degradation of auxin via an interaction with phytochrome[37]. In addition, sugar is also a substrate for sensing and signaling plant systems under biotic or abiotic stress[38]. Sugar can protect olive trees during drought conditions[39,40] and improve C. paliurus seedlings during drought conditions[25]. In the present study, total sugar content in the leaves under three shading levels had no difference (Fig. 2), but TSS content positively correlated with morphological traits (plant height, leaf area) and other physiological components (Chl content, PPO activity), and negatively correlated with RSR (Fig. 7), suggesting that sugar acts as a major integrator for osmotic response to abiotic stress and influences root development by promoting auxin degradation[25,33,37,38,41].

      Minerals are indispensable nutrients for the growth and development of plants, and as non-enzymatic antioxidant defense systems, minerals respond to adverse conditions by regulating many physiological activities[42,43]. In our study, the content of four minerals varied in the three parts and had different relationships with physiological index (Figs 5 & 7b). K is essentially for stomatal function, cell expansion, osmoregulation, and cellular or whole-plant homeostasis[4446]. Mg is the central atom of chlorophyll and enzyme activator for photosynthesis. High K and Mg contents were observed in the leaf under SH0 and SH1, and they had a positive relationship with antioxidant enzymes, but had a negative relationship with Chl content, TSS content or WC (Figs 5 & 7b). This result can be attributed to the transfer of high content of K and Mg from roots to leaves to protect seedlings from high solar radiation. Ca is important for preserving membrane integrity, signaling osmoregulation, and influencing K/Na selectivity[47,48]. This was further inferred from the change of Ca content under different shading levels (Fig. 5). Na improves plant water balance and water use efficiency by modifying stomatal control and contributes to the maintenance of cell turgor and expansion[49]. A decreasing tendency of Na transfer ability from root to stem or leaf was observed with increasing shading levels in our study (Fig. 5).

      Enzymatic defense mechanisms (superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD)) have been developed to reduce the negative effects of reactive oxygen species (ROS), which affect plants at the cellular level under stress[50]. In accordance with Feng et al.[25], who reported that C. paliurus seedlings had increased antioxidant enzyme activities under different soil water capacities, higher activities of SOD, CAT and POD were observed in leaf under SH0 in comparison with that under SH2, highlighting the effectiveness of antioxidant enzyme systems in protecting cellular apparatuses under unsuitable conditions. Similar result was also reported in rose[8]. Moreover, SOD activity was much higher than CAT activity and POD activity, and they had positive correlation with MDA and mineral content (Figs 6 & 7b). This indicates that SOD is the main antioxidant enzyme in the enzymatic defense mechanism, and three antioxidant enzymes (SOD, CAT, and POD) participated in alleviating excessive ROS at the cellular level via a cascade mechanism[50] and coordinated non-enzymatic mechanisms (K, Ca, Mg, and Na) to maintain the healthy growth of plants. However, our findings showed that PPO activity under SH0 and SH1 had a lower level compared to that under SH2 and positively correlated with growth index except for RSR (Figs 4 & 7b), inferring that PPO may have another role in seedling growth; however, further experimental investigation is needed to confirm this inference.

    • Cyclocarya paliurus seedlings were cultivated in a plastic greenhouse under three shading levels. The results showed that (1) shading conditions with two layers of shade net (SH2) improved the survival rate and growth of Cyclocarya paliurus seedlings, and seedlings significantly increased plant height, leaf area, and biomass accumulation; (2) in comparison with that under SH0 and SH1, chlorophyll content increased and MDA content reduced significantly under SH2, indicating that suitable shading conditions was beneficial to seedling growth and normal metabolism; and (3) the physiological responses of seedlings varied with shading levels and showed various correlation with seedling growth, indicating that mineral and antioxidant enzymes could coordinate to alleviate or protect seedling from damage under SH0 and SH1, and SOD was the main enzymatic mechanism.

      • This work was supported by Natural Science Foundation of Fujian province of China (No. 2022J011107), Talent project of Quanzhou city of China (2021C043R) and The college students innovations special project of Fujian province (S202110399065; S202210399049).

      • The authors declare that they have no conflict of interest.

      • Copyright: © 2023 by the author(s). Published by Maximum Academic Press, Fayetteville, GA. This article is an open access article distributed under Creative Commons Attribution License (CC BY 4.0), visit https://creativecommons.org/licenses/by/4.0/.
    Figure (7)  Table (1) References (50)
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    Feng Y, Zhi L, Pan H, Chen Y, Xu J. 2023. Shade improves seedling quality of ornamental Cyclocarya species under plastic greenhouse cultivation. Ornamental Plant Research 3:13 doi: 10.48130/OPR-2023-0013
    Feng Y, Zhi L, Pan H, Chen Y, Xu J. 2023. Shade improves seedling quality of ornamental Cyclocarya species under plastic greenhouse cultivation. Ornamental Plant Research 3:13 doi: 10.48130/OPR-2023-0013

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