外源钙对巴西橡胶树易碎愈伤组织中根癌农杆菌介导的基因转移的影响。
Effect of exogenous calcium on Agrobacterium tumefaciens-mediated gene transfer in Hevea brasiliensis (rubber tree) friable calli.
作者信息
Montoro P, Teinseree N, Rattana W, Kongsawadworakul P, Michaux-Ferriere N
机构信息
Rubber Project, CIRAD/KAPI, Kasetsart University, Bangkok 10900, Thailand Fax: +662-9428684 e-mail:
Department of Agriculture, Chatuchak, Bangkok 10900, Thailand, , , , , , TH.
出版信息
Plant Cell Rep. 2000 Sep;19(9):851-855. doi: 10.1007/s002990000208.
The influence of CaCl was investigated on Agrobacterium tumefaciens-mediated gene transfer in Hevea brasiliensis friable calli which are usually proliferated on maintenance medium (MM) containing 9 mM CaCl.Five A. tumefaciens strains (C58pMP90, C58pGV2260, AGL1, LBA4404 and EHA 105) and two binary vectors (pGIN and pCAMBIA2301) were tested and the strain EHA105pC2301 was selected to conduct further experiments. The calli were precultured on MM containing a range of CaCl concentrations, then inoculated with Agrobacterium suspension. Transfer of friable calli from MM containing 9 mM CaCl to calcium-free medium significantly enhanced the transient β-glucuronidase activity. Interestingly, the use of calcium-free Agrobacterium resuspension medium to inoculate friable calli again dramatically increased the transformation efficiency. Induction of Agrobacterium's virulence with acetosyringone remained an important factor to stimulate transformation.
研究了CaCl对巴西橡胶树易碎愈伤组织中根癌农杆菌介导的基因转移的影响，这些易碎愈伤组织通常在含有9 mM CaCl的继代培养基（MM）上增殖。测试了5种根癌农杆菌菌株（C58pMP90、C58pGV2260、AGL1、LBA4404和EHA 105）和2种双元载体（pGIN和pCAMBIA2）选择EHA105pC2301菌株进行进一步实验。将愈伤组织在含有一系列CaCl浓度的MM上预培养，然后用农杆菌悬液接种。将含有9 mM CaCl的MM上的易碎愈伤组织转移到无钙培养基中显著提高了瞬时β-葡萄糖醛酸酶活性。有趣的是，再次使用无钙农杆菌重悬培养基接种易碎愈伤组织显著提高了转化效率。用乙酰丁香酮诱导农杆菌的毒性仍然是刺激转化的一个重要因素。
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