Fast, efficient and reproducible genetic transformation of Phaseolus spp. by Agrobacterium rhizogenes
Fast, efficient and reproducible genetic transformation of Phaseolus spp. by Agrobacterium rhizogenes
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DOI：
10.1038/nprot.2007.259
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发表时间：
2007-01-01
期刊：
影响因子：
14.8
通讯作者：
Sanchez, Federico
中科院分区：
文献类型：
--
作者：
Estrada-Navarrete, Georgina;Alvarado-Affantranger, Xochitl;Sanchez, Federico
该转化程序在菜豆和其他菜豆品种、地方品种和材料上高效地产生了毛状根(70-90%)。发根农杆菌伤害幼苗子叶节，并使植株处于高湿度条件下后，毛状根迅速发育。毛状根的发生总是先于毛状根的形成，15天后，当根在伤处发育时，具有正常根的茎在毛状根区下方裂开。使用报告基因可以很容易地识别与双元载体共转化的转基因根和根瘤。该方法除了诱导出对根瘤菌结瘤敏感的转基因毛状根和有效固氮外，还为高通量功能基因组学方法研究根生物学和根-微生物相互作用奠定了基础。该方案可在30天内完成。
This transformation procedure generates, with high efficiency (70-90%), hairy roots in cultivars, landraces and accessions of Phaseolus vulgaris ( common bean) and other Phaseolus spp. Hairy roots rapidly develop after wounding young plantlets with Agrobacterium rhizogenes, at the cotyledon node, and keeping the plants in high-humidity conditions. Callogenesis always precedes hairy-root formation, and after 15 days, when roots develop at wounded sites, the stem with the normal root is cleaved below the hairy root zone. Transgenic roots and nodules co-transformed with a binary vector can be easily identified using a reporter gene. This procedure, in addition to inducing robust transgenic hairy roots that are susceptible to being nodulated by rhizobia and to fixing nitrogen efficiently, sets the foundation for a high-throughput functional genomics approach on the study of root biology and root-microbe interactions. This protocol can be completed within 30 days.