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Figure 1.
Techniques and prospects for genetic transformation of conifers. (a) Agrobacterium-mediated genetic transformation. (b) Genetic transformation via biolistic bombardment. (c) Protoplast transformation. (d) Potential strategies for transformation improvement in conifers.
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Tree species Plasmids Strains Genes Promoters Ref. Pinus Pinus pinea p35SGUSint EHA105/LBA4404/
C58GV3850uidA 35S [28] Pinus strobus pGIN/pBIV/pBIVSAR/pBINm-gfp5-ER C58pMP90 GUS 35S/2 × 35S [9] pCAMBIA1301 GV3101 GUS 35S [10] Pinus taeda pAD1289/pToK47/pBISN1/pWWS006 LBA4404/GV3101/EHA105 GUS 35S [51] pPCV6NFHygGUSINT GV3101 GUS 35S [52] pGUS3/pSSLa.3 EHA101/EHA105 GUS 35S/RbcS [53] pCAMBIA1301 EHA105 GUS 35S [54] pCAMBIA1301 GV3101/EHA105/LBA4404 GUS 35S [55] pBIGM LBA4404 Mt1D/GutD 35S [22] Pinus radiata pBI121 LBA4404 GUS 35S [56] pGA643 AGL1 GUS 35S [11] pGUL/pKEA EHA105 NPTII/uidA/Bar 35S [57] pMP2482/pTGUS/ p4CL/pSLJ1111/pLN27/pLUG KYRT1 GFP 35S/CoA ligase 1 [32] Pinus pinaster pPCV6NFGUS C58pMP90 GUS 35S [58] pBINUbiGUSint EHA105/AGL1/LBA4404 GUS ubi1 [59] Pinus patula pAHC25 LBA4404 GUS ubiquitin [12] Pinus elliottii pCAMBIA1301 EHA105/GV3101/LBA4404 GUS 35S [30] Pinus massoniana pBI121 EHA105 CslA2 35S [13] Pinus tabuliformis pBI121 GV3101 GUS 35S [31] Larix Larix decidua pRi11325 Rhizogenes strains 11325 Ri plasmid / [7] pCGN1133/pWB139 strains 11325 Bt/aroA 35S [21] hybrid larch pMRKE70Km C58pMP90 NPTII 35S [60] pCAMBIA1301 GV3101 GUS 35S [61] Larix olgensis pCAMBIA1300/pBI121 GV3101 GUS 35S/PtHCA2-1 [35] VB191103 GV3101 LoHDZ2 35S [25] Larix kaempferi Super1300-GFP GV3101 LaCDKB1;2 Super [24] Picea Picea sitchensis MOG23 LBA4404/strain 1065 GUS 35S [62] Picea abies pAD1289/pToK47/pBISN1/pWWS006 LBA4404/GV3101/EHA105 GUS 35S [51] pBIV10 C58/pMP90 GUS 2 × 35S [63] pET-22b LBA4404 Cry3A 35S [23] Picea mariana pBIV10 C58/pMP90 GUS 2 × 35S [63] Picea glauca pBIV10 C58/pMP90 GUS 2 × 35S [63] pBI121 EHA105/GV3101/
LBA4404GUS 35S [29] pUC19 C58pMP90 WUS/CHAP3A G10 [64] Abies Abies spp. pTS2 AGLO GUS 2 × 35S [65] Abies koreana pBIV10/MP90 C58/pMP90/LBA4404 GUS 2 × 35S [66] Taxus Taxus brevifolia/Taxus baccata / Bo542/C58 / / [8] Chamaecyparis Chamaecyparis obtusa pBin19-sgfp C58/pMP90 GFP 35S [67] Cryptomeria Cryptomeria japonica pIG121-Hm/pUbiP-GFP-Hyg GV3101/pMP90 GFP/GUS 35S/ubiquitin [68] pIG121-Hm GV3101/pMP90 GFP 35S [69] Table 1.
Plant expression vector construction.
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Tree species Acceptor materials Co-culture time OD600nm Results Ref. Pinus Pinus pinea Cotyledons 3 d 1 Cotyledons forming buds [28] Pinus strobus Embryogenic tissues 2 d 0.6 Regenerated plant [9] Mature zygotic embryos 12 h 0.8−1.0 Regenerated plant [10] Pinus taeda Embryogenic tissues 2 d 1 Transient expression [51] Mature zygotic embryos 3−5 d / Regenerated plant [52] Shoot apex 7 d / Transgenic plants [53] Mature zygotic embryos 3−5 d 0.8−1.0 Transgenic plants [54] Mature zygotic embryos 3−5 d 0.8−1.0 Transgenic plants [55] Mature zygotic embryos 3−5 d 0.5−1.0 Improve salt tolerance [22] Pinus radiata Embryogenic tissues 1 d 0.6 Stable transformation [56] Cotyledons 5−60 min OD550nm = 0.4 Transgenic plants [11] Embryogenic tissues 5 d OD550nm = 0.5−0.8 Transgenic plants [57] Micropropagated shoot 3 d OD550nm = 0.35−0.4 Transgenic plants [32] Pinus pinaster Embryogenic tissues 36 h 0.6 Transgenic plants [58] Embryogenic tissues 3 d 0.3 Transgenic plants [59] Pinus patula Embryogenic tissues 2 d 0.5−0.75 Transgenic tissues [12] Pinus elliottii Mature zygotic embryos 3 d 0.9 Transgenic plants [30] Pinus massoniana Mature zygotic embryos 3 d 0.5 Transgenic plants [13] Pinus tabuliformis Callus/hypocotyls/Needles 3 d 0.8 Transient expression [31] Larix Larix decidua Hypocotyls 2−3 d / Regenerated plant [7] Hypocotyls 4 d / Regenerated plant [21] hybrid larch Embryogenic tissues 2 d 0.3 Regenerated plant [60] Embryogenic tissues 2 d 0.5 Regenerated plant [61] Larix olgensis Embryogenic tissues 3 d 0.6 Transgenic plants [35] Embryogenic tissues 2 d 0.5 Enhance stress resistance [25] Larix kaempferi Embryogenic tissues 2 d 0.1 Promotes cell proliferation [24] Picea Picea sitchensis Embryogenic cell lines 3 d 0.8−1.1 Stable transformation [62] Picea abies Embryogenic tissues 2 d 1 Transient expression [51] Embryogenic tissues 2 d 0.6 Transgenic plants [63] Embryogenic tissues 2 d / Transgenic plants [23] Picea mariana Embryogenic tissues 2 d 0.6 Transgenic plants [63] Picea glauca Embryogenic tissues 2 d 0.6 Transgenic plants [63] Embryogenic tissues 2 d 1 Transgenic plants [29] Embryogenic tissues / / Transgenic plants [64] Abies Abies spp. Embryogenic tissues 2 d 0.6 Transgenic plants [65] Abies koreana Embryogenic tissues 3 d 0.6 Transgenic plants [66] Taxus Taxus brevifolia/Taxus baccata Shoot segments 3 d / Gall formation [8] Chamaecyparis Chamaecyparis obtusa Embryogenic tissues 2 d 0.3 Transgenic plants [67] Cryptomeria Cryptomeria japonica Embryogenic tissues 2 d 0.15 Enhance transformation [68] Embryogenic tissues 2 d 0.2−0.6 Transgenic plants [69] Table 2.
Agrobacterium-mediated transformation in conifers.
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Tree species Acceptor materials Plasmids Promoters Genes Results Ref. Pinus Pinus taeda Cotyledons pBI221 35S GUS Transient expression [73] Pinus radiata Suspension cells pBI221 35S GUS Transient expression [87] Embryogenic tissues pCW103/pCWI22 2 × 35S gusA Transient expression [88] Cotyledons pBI121/pCGUΔl/
pAIGusN/pActl-D35S/UbBI/Adhl/Actl gusA Transient expression [89] Embryogenic tissues pRC101/pCW122 35S/Emu uidA Transgenic plants [83] Embryogenic tissues pAHC25/pCW122 maize ubiquitin/35S GUS/Bar Transgenic plants [14] Calli pCW122/pCADsense 35S npt II/Cad Transgenic calli [90] Embryogenic tissues pMYC3425/pAW16/
pCW132/pRN2Emu/ubi Cry1Ac Transgenic plants [15] Pinus concorta/Pinus banksiana Mature pollen pBM113Kp/pRT99GUS/
pAct1-D/pGA98435S/rice actin GUS Transient expression [81] Pinus sylvestris Calli/Vegetative buds/
Suspension cellspBI221 35S GUS Transient expression [91] Pollen pBI221/pRT99/pBI410/
pBI426/pBM11335S/EmP/UbB1 GUS Transient expression [79 ] Pinus strobus Embryonal masses p35S-GFP/mGFP4 35S GFP Transient expression [72] Pinus aristata/Pinus griffithii/Pinus monticola Pollen tubes pBI221 35S GUS Transient expression [92] Pinus patula Embryogenic tissues pAHC25 35S Bar/GUS Somatic embryos [48] Pinus nigra Embryogenic tissues pCW122 2 × 35S GUS Somatic embryos [86] Pinus roxbughii Mature zygotic embryos pAHC25 maize ubiquitin Bar/GUS Transgenic plants [16] Picea Picea glauca Zygotic embryos/Seedlings/
embryogenic calluspUC19 35S GUS Transient expression [82] Somatic embryos pBI426 35S GUS Stable transformation [93] Somatic embryos pTVBT41100 35S GUS/Bt Transgenic plants [49] Embryonal masses p35S-GFP/mGFP4 35S GFP Transient expression [72] Embryogenic tissues pKUB/pBI426 maize ubiquitin/35S cry1Ab Transgenic plants [26] Picea mariana Embryogenic tissues pRT99GUS/pBM113Kp 35S GUS Transient expression [94] Embryogenic tissues pRT99GUS/pGUSInt/
pMON990935S/Em protein of wheat/Rbcs/NOS/
Actin/ArabinGUS Transient expression [76] Mature pollen pBM113Kp/pRT99GUS/
pAct1-D/pGA98435S/rice actin GUS Transient expression [81] Embryonal masses pRT99GUS/pBI426 35S GUS Transgenic plants [84] Pollen/Embryonal masses/ Somatic embryos p35S-GFP/mGFP4 35S GFP Transient expression [72] Mature somatic embryos pBI221.23 35S GUS Transgenic plants [17] Picea abies Somatic embryo pRT99gus 35S GUS Stable transformation [77] Embryogenic tissues pRT99gus/pJIT65/
Dc8gus/pBMI13Kp35S/2 × 35S/
Act1-D/Dc8GUS Transient expression [80] Pollen pBI221/pRT99/pBI410/
pBI426/pBM11335S/EmP/UbB1 GUS Transient expression [79] Embryogenic tissues pCW122 35S GUS Transgenic plants [95] Embryogenic tissues pAHC25 maize ubiquitin Bar Transgenic plants [78] Embryogenic tissues pAHC25/pCW122 maize ubiquitin/35S GUS/Bar Transgenic plants [14] Embryogenic tissues pAHC25 maize ubiquitin CCR Transgenic plants [27] Larix Larix × eurolepis Embryogenic tissues pRT99GUS/pGUSInt/
pMON990935S/Em protein of wheat/Rbcs/NOS/
Actin/ArabinGUS Transient expression [76] Larix laricina Embryonal masses pBI426/pRT99gus/
pRT66gus/pRT55gus35S/2 × 35S GUS Transient expression [75] Larix gmelinii Zygotic embryos pUC-GHG/pBI221-HPT 35S GUS/GFP Transgenic plants [34] Pseudotsuga Pseudotsuga menziesii Cotyledons pTVBTGUS 35S GUS Transient expression [74] Chamaecyparis Chamaecyparis nootkatensis Mature pollen pBM113Kp/pRT99GUS/
pAct1-D/pGA98435S/rice actin GUS Transient expression [81] Tsuga Tsuga heterophylla Mature pollen pBM113Kp/pRT99GUS/
pAct1-D/pGA98435S/rice actin GUS Transient expression [81] Abies Abies nordmanniana Embryogenic tissues pCW122 35S GUS Transgenic plants [85] Table 3.
Biolistic bombardment genetic transformation in conifers.
Figures
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Tables
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