杨树农杆菌遗传转化研究30年

doi:10.11707/j.1001-7488.20221211

摘要/Abstract

摘要：

杨树是我国最为重要的国土绿化和工业用材树种之一，精准高效培育速生、优质和高抗的杨树新品系是今后杨树育种理论和技术发展的重要方向。以农杆菌介导的遗传转化体系为基础的基因工程育种已成为缩短林木育种周期、提升育种效率的核心技术。自20世纪80年代证实杨树是农杆菌的天然寄主以来，国内外开展了大量构建农杆菌介导杨树稳定遗传转化体系的研究。本文综合分析了已有研究报道，详细回顾了不同派系杨树树种遗传转化体系的构建历程，系统整理了影响杨树遗传转化体系的主要因素，并对杨树基因工程育种的研究前景进行了展望，以期为进一步提升杨树遗传转化效率提供信息依据，为开发顽拗性木本植物和生产推广良种的遗传转化体系提供参考方案。

关键词: 杨树, 遗传转化, 根癌农杆菌, 发根农杆菌, 组培再生

Abstract:

Poplar (Populus spp.) is one of the most important tree species used for land greening and industrial timber production in China, and efficiently and precisely breeding new cultivars with rapid growth, high quality and strong stress tolerance will be an important direction for the future theoretical and technological development in poplar breeding. The Agrobacterium-mediated genetic transformation system has been the critical technology for shortening breeding cycle and increasing breeding efficiency. A large number of researches have been conducted on developing the Agrobacterium-mediated genetic stable transformation system for poplar since it was proved to be a native host of Agrobacterium in 1980s. We performed a comprehensive analysis of existing literatures, reviewed the historical process of constructing Agrobacterium-mediated genetic transformation system in different poplar species, summarized the main factors affecting transformation efficiency, and also discussed some future perspectives in this area. This review will provide useful information for further improving the poplar transformation efficiency and present reference protocols for establishing transformation systems in recalcitrant plant species and elite cultivars widely applied in production.

Key words: Populus, genetic transformation, Agrobacterium tumefaciens, A. rhizogenes, tissue culture regeneration

中图分类号:

S722.3

陈赢男,胡传景,诸葛强,胡建军,尹佟明. 杨树农杆菌遗传转化研究30年[J]. 林业科学, 2022, 58(12): 114-129.

Yingnan Chen,Chuanjing Hu,Qiang Zhuge,Jianjun Hu,Tongming Yin. Thirty Years of Agrobacterium-Mediated Genetic Transformation of Populus[J]. Scientia Silvae Sinicae, 2022, 58(12): 114-129.

图/表 3

表1

目前已建立农杆菌介导遗传转化体系的杨树"

派系 Section	杨树树种或杂交种 Populus species/hybrid	外植体 Explant	农杆菌菌种 Agrobacterium strain	侵染液OD值 OD value	筛选剂及其浓度 Selective agent and its concentration	参考文献 Reference
白杨组 Leuce	Populus alba (clone Villafranca)	茎节间 Stem internode	A. tumefaciens EHA105	0.6	100 mg·L^-1卡那霉素 Kanamycine	Confalonieri et al., 2000
	P. alba	茎尖 Shoot tip	A. tumefaciens LBA 4404	0.8	100 mg·L^-1卡那霉素 Kanamycine	Soliman et al., 2017
	P. alba var. pyramidalis (= P. bolleana)	叶盘 Leaf disk	A. tumefaciens LBA4404	0.4	15~20 mg·L^-1卡那霉素 Kanamycine	诸葛强等，2003 Zhuge Qiang et al., 2003
	P. alba var. pyramidalis (= P. bolleana)	叶盘 Leaf disk	A. tumefaciens GV3101	0.6~0.8	90 mg·L^-1潮霉素 Hygromycin	Ma et al., 2019
	P. tomentosa	叶盘 Leaf disk	A. rhizogenes R1000 (pTVK85)	对数生长期 Logarithmic growth phase	无 None	卜学贤等，1991 Bu Xuexian et al., 1991
	P. tomentosa	叶盘 Leaf disk	A. tumefaciens LBA4404	0.6	30~50 mg·L^-1卡那霉素 Kanamycine	Hu et al., 2005
	P. tremula	茎节间 Stem internode	A. rhizogenes LBA9402 A. tumefaciens EHA101	0.1~0.2	50 mg·L^-1卡那霉素 Kanamycine	Tzfira et al., 1996; 1997a
	P. tremula	茎段 Stem segment	A. tumefaciens EHA105	0.15~0.30	100 mg·L^-1卡那霉素Kanamycine；0.2 units·mL^-1潮霉素Hygromycin	Tzfira et al., 1997b
	P. tremuloides(clone 271)	叶盘 Leaf disk	A. tumefaciens C58	未明确说明 Unspecified	40~100 mg·L^-1卡那霉素 Kanamycine	Tsai et al., 1994
	P. tremuloides	下胚轴 Hypocotyl	A. tumefaciens C58	农杆菌克隆直接侵染 Agrobacterium colony	100 mg·L^-1卡那霉素 Kanamycine	Cseke et al., 2007
白杨派杂种 White poplar hybrid	P. alba ×P. grandidentata (clone NC-5339)	叶盘 Leaf disk	A. tumefaciens C58	2×10⁸ cfu·mL^-1	60 mg·L^-1卡那霉素 Kanamycine	Fillatti et al., 1987
	P. alba × P. tremula (clone 357)	节间 Internode	A. tumefaciens C58-C1 Rif (pMP90)	2~4×10⁶ bacteria·mL^-1	100 mg·L^-1卡那霉素 Kanamycine	De Block, 1990
	P. alba × P. grandidentata (clone Hansen)	叶盘 Leaf disk	A. tumefaciens A281	0.5	20~40 mg·L^-1卡那霉素 Kanamycine	Klopfenstein et al., 1991
	P. alba × P. glandulosa cv. 84K	叶盘 Leaf disk	A. tumefaciens LBA4404	0.4~0.6	50~60 mg·L^-1卡那霉素 Kanamycine	张冰玉等，2005 Zhang Bingyu et al., 2005
		愈伤组织 Callus	A. tumefaciens GV3101	0.6	1.5 mg·L^-1潮霉素 Hygromycin	Wen et al., 2022
	P. alba × P. berolinensis	叶盘 Leaf disk	A. tumefaciens EHA105	0.6	25~50 mg·L^-1卡那霉素 Kanamycine	Wang et al., 2011
	P. davidiana × P. bolleana	叶盘 Leaf disk	A. tumefaciens EHA105	0.6	50 mg·L^-1卡那霉素 Kanamycine	Wang et al., 2011
	P. davidiana × P. bollena	叶盘 Leaf disk	A. tumefaciens EHA105	0.8~1.0	50 mg·L^-1卡那霉素 Kanamycine	Han et al., 2013
	P. canescens × P. grandidentata (clone 36-67-S-1)	叶盘 Leaf disk	A. tumefaciens EHA105	1.0	80 mg·L^-1卡那霉素 Kanamycine	Dai et al., 2003
	P. kitakamiensis (P. sieboldii×P. gradidentata)	叶盘 Leaf disk	A. tumefaciens LBA4404	未明确说明 Unspecified	100 mg·L^-1卡那霉素 Kanamycine	Zhan et al., 1997
	P. tremula× P. alba (clone INRA 717. 1B4)	茎节间 Stem internode	A. tumefacien 82.139与C58/pMP90 混合接种Co-inoculation	82.139 OD=0.1 C58/pMP90 OD= 1.0	50 mg·L^-1卡那霉素 Kanamycine	Brasileiro et al., 1991
		茎或叶 Stem or leaf	A. tumefacien C58/pMP90	0.3	50 mg·L^-1卡那霉素 Kanamycine	Brasileiro et al., 1992; Leple et al., 1992
		茎节间 Stem internode	A. tumefacien C58/pMP90	0.3~0.5	100 mg·L^-1卡那霉素 Kanamycine	Han et al., 1997
		叶盘 Leaf disk	A. tumefacien LBA4404	0.3	50 mg·L^-1卡那霉素 Kanamycine	Gallardo et al., 1999
	P. tremula × P. tremuloides	茎段 Stem segment	A. tumefacien GV3101 (pMP90RK)	0.2~0.5	60 mg·L^-1卡那霉素 Kanamycine	Nilsson et al., 1992
	P. tremula ×P. tremuloides (clone T89)	完整无菌苗 Sterile seedling	A. tumefacien GV3101 (pMP90)	1.0	未使用抗生素	Takata et al., 2012
	P. tremuloides × P. davidiana (clone 18-59-S-11)	叶盘 Leaf disk	A. tumefacien EHA105	1.0	60 mg·L^-1卡那霉素 Kanamycine	Dai et al., 2003
	Poplar 741[P. alba×(P. davidiana+P. simonii)× P. tomentosa]	叶片 Leaf	A. tumefacien LBA4404	未明确说明 Unspecified	50 mg·L^-1卡那霉素 Kanamycine	田颖川等，2000 Tian Yingchuan et al., 2000
青杨组 Tacamahaca	P. trichocarpa (clone Nisqually-1)	茎节间或叶盘 Stem internode or leaf disk	A. tumefacien C58/pMP90	0.5~0.6	25~100 mg·L^-1卡那霉素 Kanamycine	Ma et al., 2004
		茎节间(温室苗) Stem internode (Greenhouse plant)	A. tumefacien C58	未明确说明 Unspecified	50~100 mg·L^-1卡那霉素 Kanamycine	Song et al., 2006
		茎节间或叶柄 Stem internode or petiole	A. tumefacien GV3101	0.4	20~30 mg·L^-1卡那霉素 Kanamycine	Li et al., 2017
	P. ciliata	叶柄(温室苗) Petiole (Greenhouse plant)	A. tumefacien LBA4404	10⁸ cells·mL^-1	50 mg·L^-1卡那霉素 Kanamycine	Thakur et al., 2005
	P. cathayana cv. Zhonglinmeihe	顶芽和腋芽(原位转化) Apical and axillary bud (in situ transformation)	A. tumefacien LBA4404	0.4~1.0	150 mg·L^-1卡那霉素 Kanamycine	Yang et al., 2010
	P. angustifolia P. balsamifera	茎节间或腋芽 Stem internode or axillary bud	A. tumefacien GV3101	0.5	—	Maheshwari et al., 2016
黑杨组 Aigeiros	P. nigra (clone San Giorgio and Jean Pourtet)	叶盘 Leaf disk	A. tumefacien GV2260	6×10⁸ cells·mL^-1	100 mg·L^-1卡那霉素 Kanamycine	Confalonieri et al., 1994; 1995
	P. nigra var. italica	茎段 Stem Segment	A. tumefacien LBA4404	5×10⁸ cells·mL^-1	150 mg·L^-1卡那霉素 Kanamycine	Mohri et al., 1996
	P. nigra var. italica	茎节间(生长箱植株) Stem internode (Plants in a growth chamber)	A. tumefacien EHA105	0.5	10~100 mg·L^-1卡那霉素 Kanamycine	Nishiguchi et al., 2006
	P. deltoides (PE68-022× P. deltoides)	叶盘 Leaf disk	A. tumefacien A281、GV2260、C58、GV3850、EHA105	7×10⁸或1.2× 10⁹ bacteria·mL^-1	100 mg·L^-1卡那霉素 Kanamycine	Confalonieri et al., 1997
	P. deltoides (clone 172-2)	茎节间或叶盘 Stem internode or leaf disk	Agrobacterium strain ABI	0.3~0.4	25~100 mg·L^-1卡那霉素 Kanamycine	Han et al., 2000
	P. deltoides cv. Juba (P. deltoides cv. 50 × P. deltoides cv. 36)	叶片 Leaf	A. tumefacien EHA105	0.5	5 mg·L^-1潮霉素 Hygromycin	王桂英 2012 Wang Guiying 2012
黑杨派杂种 Black poplar hybrid	P. × euramericana (Neva, PE68-022 × P. nigra, 71-060 × P. nigra)	叶盘 Leaf disk	A. tumefacien A281、GV2260、C58、GV3850、EHA105	7×10⁸或1.2×10⁹ bacteria·mL^-1	100 mg·L^-1卡那霉素 Kanamycine	Confalonieri et al., 1997
	P. × euramericana cv. ‘74/76’	叶片 Leaf	A. tumefacien EHA105	0.4~0.6	30 mg·L^-1卡那霉素 Kanamycine	任亚超 2013 Ren Yachao 2013
	P. nigra var. betulifolia × P. trichocarpa (clone NC5331)	茎节间或叶盘 Stem internode or leaf disk	A. tumefaciens ASE9749	1-2×10⁸ cells·mL^-1	60 mg·L^-1卡那霉素 Kanamycine	Li et al., 1997
	P. deltoides × P. nigra (OP-367)	茎节间或叶盘 Stem internode or leaf disk	Agrobacterium strain ABI	0.3~0.4	25~100 mg·L^-1卡那霉素 Kanamycine	Han et al., 2000
	P. deltoides × P. euramericana (clone Nanlin895)	叶盘 Leaf disk	A. tumefacien EHA105	0.7	50 mg·L^-1卡那霉素 Kanamycine	Movahedi et al., 2014
黑杨派与青杨派杂种	P. trichocarpa × P. deltoides (clone H11)	茎段 Stem segment	A. tumefaciens A281、A348	5×10⁸ cells·mL^-1	—	Parsons et al., 1986
	P. trichocarpa × P. deltoides (clone H11)	茎段 Stem segment	A. rhizogenes R1000	5×10⁸ cells·mL^-1	—	Pythoud et al., 1987
	P. trichocarpa × P. deltoides (clone 064)	茎节间 Stem internode	A. tumefaciens C58-C1 Rif (pMP90)	2×10⁸ bacteria·mL^-1.	200 mg·L^-1卡那霉素 Kanamycine	De Block, 1990
	P. trichocarpa × P. deltoides (clone 24-305-242-246\H11)	嫩枝茎节间 Greenwood stem internode	A. rhizogenes R1000、R1240、R1600、R1601	1.0	100 mg·L^-1卡那霉素 Kanamycine	Han et al., 1996
	P. trichocarpa × P. deltoides (clone 184-402)	茎节间 Stem internode	A. tumefaciens C58/pMP90	0.3~0.5	100 mg·L^-1卡那霉素 Kanamycine	Han et al., 1997
	P. trichocarpa × P. deltoides (clone 24-305\ 184-4029-434)	茎节间或叶盘 Stem internode or leaf disk	Agrobacterium strain ABI	0.3~0.4	25~100 mg·L^-1卡那霉素 Kanamycine	Han et al., 2000
	P. nigra × P. maximowiczii (clone NM6)	茎节间、叶柄或叶盘 Stem internode, petiole or leaf disk	A. tumefaciens MP90	0.4~0.5	25~100 mg·L^-1卡那霉素 Kanamycine	Yevtushenko et al., 2010

表1

图1

表2

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基因名称 Gene name	基因功能 Gene function	物种 Species	参考文献 Reference
4CL1	木质素含量 Lignin content	杨树(P. trichocarpa)	Min et al., 2012
AGO1	横向器官发育 Lateral organ development	拟南芥(Arabidopsis thaliana)	Yang et al., 2006
REVOLUTA	茎和叶发育 Stem and leaf development	拟南芥(A. thaliana)	Talbert et al., 1995
LS	侧枝发育 Shoot branching	番茄(Lycopersicon esculentum)	Schumacher et al., 1999
Tb1	分蘖 Tiller	玉米(Zea mays)	Doebley et al., 1995
SD1	株高 Plant height	水稻(Oryza sativa)	Ashikari et al., 2002
PIN2	株型 Plant morphogenesis	水稻(O. sativa)	Chen et al., 2012

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