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与菌根真菌共生的兰科福建金线莲差异表达基因的筛选

, PP. 218-225

Keywords: 福建金线莲,兰科菌根共生,瘤菌属,差异显示PCR,基因筛选

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Abstract:

研究菌根真菌促进药用植物生长发育的分子作用机制在农业上有着十分重要的理论意义和实践意义.本研究从野生福建金线莲(Anoectochilusroxburghii)根中分离出一株内生真菌AR-18,经鉴定为瘤菌属(Epulorhizasp.)真菌;将AR-18菌与福建金线莲组培苗接菌培养60天,发现其根中能形成内生菌根并能显著地促进福建金线莲苗的生长.为从与AR-18菌共生的福建金线莲中分离出差异表达基因,本实验采用差异显示PCR(DDRT-PCR)方法,使用3条锚定引物和5条随机引物设计15个引物对,从接菌组和对照组中共分离出52条差异显示条带,其中有9条带能通过双引物重扩增显现.再运用反式Northern点杂交进一步筛选,发现其中5条上调表达基因,包括:编码尿磷酸核苷转移酶基因(UPRTs;EC2.4.2.9)和1条假蛋白基因;1条下调表达基因,编码氨基酸跨膜转运子;1条在接菌组中特异表达基因,编码突变酶K(matK).同时,讨论了上述基因可能的功能,尤其是UPRTs基因和matK基因与菌根真菌互作的功能.本研究是福建金线莲植物在该领域的首次报道.

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