Agrobacterium rhizogenes-mediated transformation of the parasitic plant Phtheirospermum japonicum.
Agrobacterium rhizogenes-mediated transformation of the parasitic plant Phtheirospermum japonicum.
复制标题
DOI：
10.1371/journal.pone.0025802
复制
发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Shirasu K
中科院分区：
文献类型：
--
作者：
Ishida JK;Yoshida S;Ito M;Namba S;Shirasu K
列当科植物是农业上重要的一类寄生虫，它们攻击经济上重要的作物以从其宿主获得水和营养。尽管其农业的重要性，寄生的分子机制知之甚少。我们建立了列当科兼性寄生植物Phtheirospermum japonicum的瞬时和稳定转化系统。利用发根农杆菌和幼苗，通过超声处理和乙酰丁香酮处理的组合建立了转化方案。从A.发根菌接种通过基因组PCR、Southern blot和RT-PCR方法验证了转基因在日本对虾中的存在和表达。从A.发根农杆菌介导的转化能够在水稻和玉米根上产生吸器。转基因根也形成了明显的感受态吸器响应2，6-二甲氧基-1，4-苯醌（DMBQ），吸器诱导化学品。利用该系统，我们将带有Cyclin B1启动子的报告基因导入日本对虾，并观察了吸器形成过程中的细胞分裂。本研究提供了一种简便有效的毛状根转化方法。转基因标记分析表明，细胞分裂吸器的发展过程中发生24小时后DMBQ处理。这里描述的协议将允许参与植物寄生的基因的功能分析。
Plants within the Orobanchaceae are an agriculturally important group of parasites that attack economically important crops to obtain water and nutrients from their hosts. Despite their agricultural importance, molecular mechanisms of the parasitism are poorly understood. We developed transient and stable transformation systems for Phtheirospermum japonicum, a facultative parasitic plant in the Orobanchaceae. The transformation protocol was established by a combination of sonication and acetosyringone treatments using the hairy-root-inducing bacterium, Agrobacterium rhizogenes and young seedlings. Transgenic hairy roots of P. japonicum were obtained from cotyledons 2 to 3 weeks after A. rhizogenes inoculation. The presence and the expression of transgenes in P. japonicum were verified by genomic PCR, Southern blot and RT-PCR methods. Transgenic roots derived from A. rhizogenes-mediated transformation were able to develop haustoria on rice and maize roots. Transgenic roots also formed apparently competent haustoria in response to 2,6-dimethoxy-1,4-benzoquinone (DMBQ), a haustorium-inducing chemical. Using this system, we introduced a reporter gene with a Cyclin B1 promoter into P. japonicum, and visualized cell division during haustorium formation. We provide an easy and efficient method for hairy-root transformation of P. japonicum. Transgenic marker analysis revealed that cell divisions during haustorium development occur 24 h after DMBQ treatment. The protocols described here will allow functional analysis of genes involved in plant parasitism.
登录
查看更多内容
影响因子：
1.4
作者：
MATERN, U;KNEUSEL, RE
通讯作者：
KNEUSEL, RE
影响因子：
11.6
作者：
Bandaranayake, Pradeepa C. G.;Filappova, Tatiana;Yoder, John I.
通讯作者：
Yoder, John I.
影响因子：
6.9
作者：
Stiller, J;Martirani, L;Gresshoff, PM
通讯作者：
Gresshoff, PM
影响因子：
7.4
作者：
Tomilov, AA;Tomilova, NB;Yoder, JI
通讯作者：
Yoder, JI
影响因子：
10.5
作者：
Kubo, M;Udagawa, M;Demura, T
通讯作者：
Demura, T