Kim Hee Jin
Cotton Fiber Bioscience Research Unit, USDA-ARS, Southern Regional Research Center, New Orleans, LA, USA.
Methods Mol Biol. 2013;958:179-87. doi: 10.1007/978-1-62703-212-4_15.
Although well-accepted as the ultimate method for cotton functional genomics, Agrobacterium tumefaciens-mediated cotton transformation is not widely used for functional analyses of cotton genes and their promoters since regeneration of cotton in tissue culture is lengthy and labor intensive. In certain cases, A. rhizogenes-induced hairy root culture has been a suitable molecular tool for functional analyses of genes and promoters for plants that are difficult to regenerate by A. tumefaciens-mediated transformation. Similarly, A. rhizogenes-induced hairy root cultures are an alternative tool for cotton functional genomics. In this chapter, the advantages and disadvantages of using A. rhizogenes-induced cotton hairy root culture over A. tumefaciens-mediated cotton transformation are discussed. The procedures for transformation, generation, selection, and molecular analyses of transgenic cotton hairy roots are introduced by describing the functional analysis of a cotton promoter in cotton hairy roots generated by A. rhizogenes-mediated transformation.
尽管根癌农杆菌介导的棉花转化作为棉花功能基因组学的最终方法已被广泛接受,但由于棉花组织培养再生过程漫长且劳动强度大,该方法并未广泛用于棉花基因及其启动子的功能分析。在某些情况下,发根农杆菌诱导的毛状根培养已成为一种合适的分子工具,用于对难以通过根癌农杆菌介导的转化进行再生的植物的基因和启动子进行功能分析。同样,发根农杆菌诱导的毛状根培养也是棉花功能基因组学的一种替代工具。在本章中,将讨论使用发根农杆菌诱导的棉花毛状根培养相对于根癌农杆菌介导的棉花转化的优缺点。通过描述由发根农杆菌介导的转化产生的棉花毛状根中棉花启动子的功能分析,介绍了转基因棉花毛状根的转化、产生、选择和分子分析程序。