Establishment and Application of Genetic Transformation System for Populus simonii×P. nigra ‘Baicheng’

doi:10.7525/j.issn.1673-5102.2023.05.004

Abstract

Abstract:

Stems of Populus simonii×P. nigra ‘Baicheng’ in vitro plants were selected as explants and MS medium was used to establish tissue culture system by adjusting the hormone concentrations of 6-BA， NAA， TDZ and IBA. Based on the tissue culture system， the optimal concentration of kanamycin and infection time were confirmed to establish Agrobacterium tumefaciens-mediated genetic transformation system for P. simonii×P. nigra ‘Baicheng’. By using this system， the transgenic plants overexpressed a key tension wood formation regulator LBD39 （Lateral Organ Boundaries Domain） were created successfully. The results showed that the tissue culture system consisted of three stages， including adventitious bud differentiation induction （MS+ 0.5 mg·L^-1 6-BA+0.1 mg·L^-1 NAA+0.001 mg·L^-1 TDZ， shoot differentiation rate=92.6%）， stem induction （MS+0.2 mg·L^-1 6-BA+0.05 mg·L^-1 NAA， multiplication coefficient=6.5） and rooting induction （1/2 MS+0.4 mg·L^-1 IBA， rooting percentage=100%）. The optimal kanamycin concentration for genetic transformation was 30 mg·L^-1 and the optimal infection time was 20 min， and the transgenic plants were obtained successfully after 30 d of adventitious bud differentiation induction， 15-30 d of stem induction and 25 d of rooting induction with 2% transformation efficiency respectively. By using this system， five overexpressed plants of LBD39 were obtained， and the transformation efficiency was 3.3%.

Key words: Populus simonii×P. nigra ‘Baicheng’, tissue culture, genetic transformation, LBD39

CLC Number:

S792.11

Xu HE, Yuan GAO, Qunye ZHANG, Chenguang ZHOU, Wei LI, Shuang LI. Establishment and Application of Genetic Transformation System for Populus simonii×P. nigra ‘Baicheng’[J]. Bulletin of Botanical Research, 2023, 43(5): 667-678.

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处理 Treatment	植物生长调节剂 Plant growth regulators/（mg·L^-1）			不定芽分化率 Adventitious bud differentiation rate/%
处理 Treatment	6-BA	NAA	TDZ	不定芽分化率 Adventitious bud differentiation rate/%
1	0.1	0.1	—	63.0±2.14 e
2	0.2	0.1	—	71.6±1.64 d
3	0.3	0.1	—	79.0±2.67 c
4	0.4	0.1	—	82.7±2.47 bc
5	0.5	0.1	—	86.4±1.23 ab
6	0.5	0.1	0.001	92.6±2.11 a

处理 Treatment	植物生长调节剂 Plant growth regulators/（mg·L^-1）		抽茎增殖系数 Stem proliferation coefficient
处理 Treatment	6-BA	NAA	抽茎增殖系数 Stem proliferation coefficient
1	0.1	0.05	4.0±0.08 b
2	0.1	0.10	2.8±0.12 d
3	0.2	0.05	6.5±0.10 a
4	0.2	0.10	3.1±0.10 c

处理 Treatment	植物生长调节剂 Plant growth regulators/（mg·L^-1）		生根率 Rooting percentage/%
处理 Treatment	IBA	NAA	生根率 Rooting percentage/%
1	0.1	0.02	72.5±1.44 f
2	0.1	—	78.3±0.83 e
3	0.2	0.02	79.2±2.20 de
4	0.2	—	82.5±2.89 de
5	0.3	0.02	84.2±2.20 cd
6	0.3	—	88.3±1.67 bc
7	0.4	0.02	92.5±1.44 b
8	0.4	—	100.0±0.00 a

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