北玄参毛状根诱导及其植株再生

doi:10.11983/CBB16246

摘要/Abstract

摘要： 利用发根农杆菌A4和R1601感染北玄参(Scrophularia buergeriana)叶片外植体, 诱导产生毛状根, 产生的毛状根可在无激素的液体和固体MS培养基上快速生长。rolB基因的PCR检测表明, Ri质粒中的T-DNA片段整合到了北玄参毛状根的基因组中。毛状根在附加0.5 mg·L^-16-BA及0.1 mg·L^-1NAA的MS固体培养基上形成绿色愈伤组织, 之后形成不定芽, 并获得再生植株; 毛状根在附加0.5 mg·L^-16-BA及0.02 mg·L^-1NAA的MS固体培养基上培养约15-20天可直接形成不定芽, 且不定芽的诱导率达85%。

关键词: 北玄参, 发根农杆菌, 毛状根, 植株再生

Abstract: Hairy roots were induced by infecting leaf explants of Scrophularia buergeriana with Agrobacterium rhizo- genes strains A4 and R1601. Hairy roots could grow rapidly and autonomously in liquid or solid plant-growth regulator-free MS medium. The transformation was confirmed by PCR amplification of the rolB gene of the Ri plasmid from S. buergeriana hairy roots. Hairy roots could form green callus in MS medium with 0.5 mg·L^-1 6-BA and 0.1 mg·L^-1 NAA, then regenerated plantlets were induced from the callus. A whole transgenic plantlet was obtained. Adventitious buds could be directly induced from hairy roots in MS medium with 0.5 mg·L^-1 6-BA and 0.02 mg·L^-1 NAA after about 15-20 d, with the induction rate of buds up to 85%.

Key words: Scrophularia buergeriana, Agrobacterium rhizogenes, hairy root, plant regeneration

任如意, 薛巨坤, 国会艳, 魏继承. 北玄参毛状根诱导及其植株再生. 植物学报, 2017, 52(6): 783-787.

Ren Ruyi, Xue Jukun, Guo Huiyan, Wei Jicheng. Induction of Hairy Roots of Scrophularia buergeriana and Its Plant Regeneration. Chinese Bulletin of Botany, 2017, 52(6): 783-787.

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链接本文: https://www.chinbullbotany.com/CN/10.11983/CBB16246

https://www.chinbullbotany.com/CN/Y2017/V52/I6/783

图/表 3

图1 北玄参毛状根的诱导、离体培养与植株再生(A) 发根农杆菌A4侵染16天后叶片外植体产生的毛状根; (B) MS+0.2 mg·L-1 6-BA+0.1 mg·L-1 NAA培养基上毛状根形成的绿色愈伤组织; (C) MS+0.5 mg·L-1 6-BA+0.1 mg·L-1 NAA培养基上毛状根形成的绿色愈伤组织并有不定芽生成; (D) MS+0.5 mg·L-1 6-BA+ 0.02 mg·L-1 NAA培养基上毛状根直接长出不定芽; (E) 不定芽形成的芽丛; (F) 再生植株

Figure 1 Induction, in vitro culture and plants regeneration of hairy root in Scrophularia buergeriana(A) Hairy root formation from leaf explants 16 days after infection with the strain of Agrobacterium rhizogenes A4; (B) Green callus induction on hairy roots in medium MS+0.2 mg·L-1 6-BA+0.1 mg·L-1 NAA; (C) Green callus and adventitious buds in- duction on medium MS+0.5 mg·L-1 6-BA+0.1 mg·L-1 NAA; (D) Adventitious shoots formation on medium MS+0.5 mg·L-1 6- BA+0.02 mg·L-1 NAA; (E) Adventitious buds; (F) Transplanted plant

图2 北玄参毛状根(A4) rolB基因PCR扩增产物的凝胶电泳1-4: 毛状根DNA扩增片段; 5, 6: 非转化根DNA扩增结果; 7: 发根农杆菌菌落扩增的片段; M: DL2000 marker

Figure 2 Gel electrophoresis analysis of PCR fragments of rolB gene amplified from the genome DNA of hairy roots (A4)1-4: Fragments amplified from genome DNA of hairy roots; 5, 6: Amplification products from genome DNA of untransformed roots; 7: Fragments amplified from the colony of Agrobacte- rium rhizogenes; M: DL2000 marker

表1 6-BA和NAA组合对北玄参毛状根愈伤组织诱导及植株再生的影响

Table 1 Effect of 6-BA and NAA on callus induction and adventitious shoots formation of hairy roots in Scrophularia buergeriana

Med- ium	6-BA (mg·L^-1)	NAA (mg·L^-1)	Change of hairy roots
1	0.1	0.02	Few adventitious buds induced
2	0.1	0.1	Formation of light yellow callus
3	0.2	0.02	Few adventitious buds induced
4	0.2	0.1	Formation of green callus
5	0.5	0.02	More adventitious buds developed
6	0.5	0.1	Firstly formed green callus, then developed more adventitious buds

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