Regulation of age-dependent expression patterns of five transcription factors in <i>Larix kaempferi</i>

Xiangyi Li; Dongxia Cheng; Liwang Qi; Jinwei Zhan; Wanfeng Li; Xiangyi Li; Dongxia Cheng; Liwang Qi; Jinwei Zhan; Wanfeng Li

doi:10.48130/FR-2023-0018

Figures (4) Tables (1)

Figure 1.
Process of mRNA splicing. RNA splicing involves cutting an intron (gray) from pre-mRNA and joining together the two neighboring exons (black). The spliced exons form the functional RNA, and the intron is usually degraded. Arrows indicate the positions of the primers used to detect the pre-mRNA (gray) and spliced mRNA (black).
Figure 2.
Structure and expression pattern of LaDAL1. (a) Schematic representation of the gene structure of LaDAL1. White indicates exon; gray indicates intron; gray arrows indicate the positions of the primers used to measure the pre-mRNA; and black and horizontal arrows indicate the positions of the primers used to measure the spliced mRNA. After the prediction of DNA methylation, the CpG islands were revealed. (b)–(d) Expression patterns of LaDAL1 pre-mRNA and spliced mRNA during tree aging detected by three different primer pairs. The lateral branches of 3-, 5-, and 13-year-old active Larix kaempferi trees (n ≥ 6, sampled on 4 July 2019) were used to examine the expression patterns, which were assayed by qRT-PCR with LaEF1A1 as the internal control. The capitalized Roman numerals (I–VI) in panels (a)–(d) represent the different primers. The p-values of the differences between 5- and 3-year-old trees were calculated. One-way ANOVA Duncan’s test was used for statistical analysis.
Figure 3.
Structure and expression pattern of LaAGL2-2/3. (a) Schematic representation of the gene structure of LaAGL2-2/3. White indicates exon; gray indicates intron; gray arrows indicate the positions of the primers used to measure the pre-mRNA; and black and horizontal arrows indicate the positions of the primers used to measure the spliced mRNA. After the prediction of DNA methylation, the CpG islands were revealed. (b)–(d) Expression patterns of LaAGL2-2/3 (b, c) and LaAGL2-2 (d) pre-mRNA and spliced mRNA during tree aging detected by three different primer pairs. The lateral branches of 3-, 5-, and 13-year-old active Larix kaempferi trees (n ≥ 6, sampled on 4 July 2019) were used to detect the expression patterns, which were assayed by qRT-PCR with LaEF1A1 as the internal control. The capitalized Roman numerals (I–VI) in panels (a)–(d) represent the different primers. The p-values of the differences between 5- and 3-year-old trees were calculated. One-way ANOVA Duncan’s test was used for statistical analysis.
Figure 4.
Structure and expression pattern of LaAP2-1. (a) Schematic representation of the gene structure of LaAP2-1. White indicates exon; gray indicates intron; gray arrows indicate the positions of the primers used to measure the pre-mRNA; and black and horizontal arrows indicate the positions of the primers used to measure the spliced mRNA. After the prediction of DNA methylation, the CpG islands were revealed. (b)–(d) Expression patterns of LaAP2-1 pre-mRNA and spliced mRNA during tree aging detected by three different primer pairs. The lateral branches of 1-, 3-, and 5-year-old active Larix kaempferi trees (n ≥ 6, sampled on 4 July 2019) were used to detect the expression patterns, which were assayed by qRT-PCR with LaEF1A1 as the internal control. The capitalized Roman numerals (I–VI) in panels (a)–(d) represent the different primers.

Gene (accession: mRNA/DNA)	Primer	Sequence (5'-3')	Position	Size (bp)
LaDAL1 (MN790744/WOXR02001943.1)	I	Forward-CGATGCAGAAGTGGCGCTAA	first exon	94
	I	Reverse-GTCAACAGCGCAAAGAAAGGA	first intron	94
	II	Forward-GCTCTCAGTGCTGTGCGAT	first exon	249
	II	Reverse-CCGAGATCTTCCCCCAACAAA	fourth exon	249
	III	Forward-AGACCAGATTGAGGAGCTTCG	fifth exon	90
	III	Reverse-GGGACGGAATAGCGTGCATTA	fifth intron	90
	IV	Forward-GGTTGAGCTCCTTCAGCGAT	third exon	163
	IV	Reverse-GCGAAGCTCCTCAATCTGGT	fifth exon	163
	V	Forward-GTACTAACGGGCCTTGGGAT	seventh exon	172
	V	Reverse-TTCCAGCTTCAAAAGTGCCAAT	seventh intron	172
	VI	Forward-ACGCAGGTGATGCTAGACCA	fifth exon	165
	VI	Reverse-CCAAGGCCCGTTAGTACCAG	seventh exon	165
LaAGL2-2 (MN790745/WOXR02003181.1)	I	Forward-GAGTTTGCTAGTGCCGGGTA	first exon and first intron	207
	I	Reverse-GTTGGGGGAAGATCTGGGTC	first intron	207
	II	Forward-GGGCTGCTGAAGAAAGCCTA	first exon	110
	II	Reverse-TTCATGCCGGCACTAGCAAA	second exon	110
	III	Forward-GTTGGGGGAAGATCTGGGTC	fourth exon	242
	III	Reverse-TGCGTTGTGTCGTATTTAGGTC	fourth intron	242
	IV	Forward-TGCAGCAACTCGAACATCAAC	fourth exon	76
	IV	Reverse-TGGCCTAGCATAACCTGCG	fifth exon	76
	V	Forward-AATTCAAGCCTCCCGACTGT	sixth exon	207
	V	Reverse-TCCGGGGACTACATATTGGC	sixth intron	207
	VI	Forward-TGCTCTCTTACACCCGCAAC	sixth exon	141
	VI	Reverse-CCACCACCCTTGCACGTAT	seventh exon	141
LaAP2-1 (MN790757/WOXR02007023.1)	I	Forward-CCCCGGAGTTCTGAGGAAAC	first exon	197
	I	Reverse-TTGCTAGAGGCCTCGTGTTC	first intron	197
	II	Forward-GCTCGCAATATCGTGGAGTG	first exon	117
	II	Reverse-TAGCAGCGGCATGAGCAGTA	third exon	117
	III	Forward-CAGCTATCAAGTTTCGAGGCG	fourth exon	200
	III	Reverse-ACTGCCATCCAAATGACTACC	fourth intron	200
	IV	Forward-TTCGAGGCGTTGAAGCTGAT	fourth exon	131
	IV	Reverse-TCCACGAGAGAAACCAGTGC	fifth exon	131
	V	Forward-CTGAAGCTCACATGAGGGAGG	ninth exon	158
	V	Reverse-TCCGCTCAGTCCATCTTTATGC	ninth intron	158
	VI	Forward-CCTGACCATCTGGGTAACTGT	ninth exon	159
	VI	Reverse-TACTGGAGTTGTTGGTCCGC	tenth exon	159

Table 1.

Polymerase chain reaction primers for the amplification of age-related transcription factors.