REN Ruyi,LI Sixuan,JIN Qiu,et al.Optimization of Hairy Root Induction by Agrobacterium rhizogene in Physalis pubescens L.[J].Northern Horticulture,2017,41(08):115-119.[doi:10.11937/bfyy.201708026]
发根农杆菌诱导毛酸浆毛状根条件的优化
- Title:
- Optimization of Hairy Root Induction by Agrobacterium rhizogene in Physalis pubescens L.
- 文献标志码:
- A
- 摘要:
- 以4种发根农杆菌A4、C58C1、R1601、ATCC15834为诱导菌株侵染毛酸浆叶片、茎段,研究了发根农杆菌种类、外植体类型、不同预培养时间、不同侵染时间、不同共培养时间和乙酰丁香酮对毛酸浆毛状根诱导率的影响,并用PCR检测rolB基因。结果表明:4种发根农杆菌均能诱导毛酸浆外植〖JP2〗体产生毛状根,ATCC15834的诱导率相对较高,叶片优于茎段;最佳侵染时间为6~8 min;预培养和共培养时间均为2~3 d时诱导率较高;乙酰丁香酮(AS)浓度为100 μmol?L-1有利于促进毛状根的产生,经PCR检测证明,rolB基因已整合到毛酸浆毛状根基因组中。
- Abstract:
- Agrobacterium rhizogenes strains of A4,C58C1,R1601,ATCC15834 were used to infect the explants of Physalis pubescens L.,then the influences of different Agrobactrium rhizogenes species,explants types,different preculture times,different infection times and co-culture times and acetosyringone on hairy root induction rate were researched.Using PCR technique to identify the induced hairy roots.The results showed that Agrobacterium rhizogenes strains of A4,C58C1,R1601,ATCC15834 all could induce hairy roots,but inductivity of ATCC15834 was higher.The inductivity of leaf was higher than stem.The best induction conditions of hairy roots were preculturing for 2—3 days,infecting for 6-8 minutes,co-culturing for 2-3 days.Acetosyringone 100 μmol?L-1 would be necessary to upgrade transformation frequencies.PCR detection results indicated that the rolB gene in Ri plasmid of A.rhizogenes had integrated into the genome of P.pubescens and been expressed.
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备注/Memo
第一作者简介:任如意(1968-),女,博士,教授,现主要从事植物基因工程等研究工作。E-mail:swxrry@126.com.