Agrobacterium rhizogenes-mediated transformation of Superroot-derived Lotus corniculatus plants: a valuable tool for functional genomics.
Agrobacterium rhizogenes-mediated transformation of Superroot-derived Lotus corniculatus plants: a valuable tool for functional genomics.
复制标题
发根农杆菌介导的超级根衍生百脉根植物的转化：功能基因组学的一个有价值的工具
DOI：
10.1186/1471-2229-9-78
复制
发表时间：
2009-06-25
影响因子：
5.3
通讯作者：
Han T
中科院分区：
文献类型：
--
作者：
Jian B;Hou W;Wu C;Liu B;Liu W;Song S;Bi Y;Han T
背景
转基因技术为植物基因功能研究提供了有力的工具。然而，一些豆科植物仍然是目前的转化技术，限制了功能基因组研究的程度，可以进行on. Superroot百脉根是一个连续的根克隆系统，允许直接体细胞胚胎发生和大量再生的植物。近年来，获得转基因L。corniculatus植株从Superroot-derived叶到A.描述了根癌农杆菌介导的转化。但转化效率较低，从基因转移到PCR鉴定需要6个月左右。
结果
在本研究中，我们开发了一个A。发根农杆菌介导的超根L. corniculatus进行基因功能研究，结合高效的A.发根农杆菌介导的转化及超级根的快速再生体系。利用A.通过对影响转化频率的一些参数的验证，对携带pGFPGUSPlus的发根农杆菌K599进行了改良。以单节茎段为外植体，预培养2天，接种OD 600 = 0.6的K599，在pH5.4的培养基上22 ℃共培养2天，可显著提高转化率。作为概念证明，超根导出的L.使用所述系统，用来自小麦的编码Na+/H+反向转运蛋白（TaNHX 2）的基因转化小Corniculatus。获得了转基因超级根植物，并具有增加的耐盐性，如从TaNHX 2的表达所预期的。
结论
这是一种快速、有效的基因功能研究工具。本研究结合超根再生体系的简单高效和A.发根农杆菌介导的转化通过对影响转化率的一些参数的验证，对该体系进行了改进，在GUS检测的基础上，转化率可达92%。超根的高效转化和再生体系的建立为L.具小角的
Background
Transgenic approaches provide a powerful tool for gene function investigations in plants. However, some legumes are still recalcitrant to current transformation technologies, limiting the extent to which functional genomic studies can be performed on. Superroot of Lotus corniculatus is a continuous root cloning system allowing direct somatic embryogenesis and mass regeneration of plants. Recently, a technique to obtain transgenic L. corniculatus plants from Superroot-derived leaves through A. tumefaciens-mediated transformation was described. However, transformation efficiency was low and it took about six months from gene transfer to PCR identification.
Results
In the present study, we developed an A. rhizogenes-mediated transformation of Superroot-derived L. corniculatus for gene function investigation, combining the efficient A. rhizogenes-mediated transformation and the rapid regeneration system of Superroot. The transformation system using A. rhizogenes K599 harbouring pGFPGUSPlus was improved by validating some parameters which may influence the transformation frequency. Using stem sections with one node as explants, a 2-day pre-culture of explants, infection with K599 at OD600 = 0.6, and co-cultivation on medium (pH 5.4) at 22°C for 2 days enhanced the transformation frequency significantly. As proof of concept, Superroot-derived L. corniculatus was transformed with a gene from wheat encoding an Na+/H+ antiporter (TaNHX2) using the described system. Transgenic Superroot plants were obtained and had increased salt tolerance, as expected from the expression of TaNHX2.
Conclusion
A rapid and efficient tool for gene function investigation in L. corniculatus was developed, combining the simplicity and high efficiency of the Superroot regeneration system and the availability of A. rhizogenes-mediated transformation. This system was improved by validating some parameters influencing the transformation frequency, which could reach 92% based on GUS detection. The combination of the highly efficient transformation and the regeneration system of Superroot provides a valuable tool for functional genomics studies in L. corniculatus.
登录
查看更多内容
影响因子：
7.2
作者：
Collier, R;Fuchs, B;Taylor, CG
通讯作者：
Taylor, CG
影响因子：
6.2
作者：
Chabaud, M;de Carvalho-Niebel, F;Barker, DG
通讯作者：
Barker, DG
影响因子：
6.2
作者：
Liu, Sheng-Jun;Wei, Zhi-Ming;Huang, Jian-Qiu
通讯作者：
Huang, Jian-Qiu
影响因子：
6.2
作者：
Lee, MH;Yoon, ES;Choi, YE
通讯作者：
Choi, YE
影响因子：
7.4
作者：
Graham, PH;Vance, CP
通讯作者：
Vance, CP