甘薯肌醇-1-磷酸合成酶基因的克隆及序列分析  

Cloning and Sequence Analysis of Myo Inositol-1-phosphate Synthase Gene in Sweetpotato*

作　　者：柳哲胜[1] 刘庆昌[1] 翟红[1] 王玉萍[1] 

机构地区：[1]中国农业大学北京市作物遗传改良重点实验室,北京100094

出　　处：《农业生物技术学报》2006年第2期219-225,共7页Journal of Agricultural Biotechnology

基　　金：国家杰出青年科学基金(No.30225028);教育部教学科研奖励计划和国家高技术研究与发展计划(863)项目(No.2003AA207140)资助

摘　　要：甘薯(Ipomoeabatatas(L.)Lam.)新品系农大603是从感茎线虫(Ditylenchusdestructor)病品种徐薯18的辐照后代中获得的一个抗茎线虫病的突变体。以农大603和徐薯18块根的mRNA为模板,根据植物抗线虫病基因NBS保守氨基酸序列设计引物,进行RT-PCR分析,发现农大603的肌醇-1-磷酸合成酶(Myoinositol-1-phosphatesynthase,MIPS)基因的表达量高于徐薯18。采用3'RACE技术扩增出MIPS基因的3'末端cDNA。根据植物MIPS基因5'端一段保守的氨基酸序列设计兼并引物,并与3'端的特异引物组合,扩增出该基因的5'端cDNA序列。DNA序列比对表明,甘薯MIPS基因与大豆(Glycinemax)、番茄(Lycopersiconesculentum)的MIPS基因同源性较高,分别达83.63%和83.89%。甘薯MIPS基因全长cDNA的克隆,有利于进一步研究该基因与抗甘薯茎线虫病的关系。Nongda 603, a mutant resistant to sweetpotato stem nematode （Ditylenchus destructor）, was obtained fi＇om the gamma-irradiated progenies of sweetpotato （Ipomoea batatas （L.） Lam.） cv. Xushu 18 susceptible to the stem nematode. Primers were designed fi＇om the NBS （nucleotide-binding site） conserved amino acid sequence of plant nematode resistance genes. The mRNAs of storage roots of Nongda 603 and Xushu 18 were used as temple. RT-PCR analysis indicated that the mRNA abundance of Myo inositol- l-phosphate synthase （M/PS） gene in Nongda 603 was higher than that in Xushu 18. The 3＇ cDNA of MIPS gene was amplified using 3＇ RACE and the 5＇ cDNA of MIPS gene was amplified by PCR using the specific primer of 3＇ cDNA and the degenerate primer designed based on the conserved amino acid sequence of the 5＇ end of plant MIPS genes. The DNA sequence alignment showed that sweetpotato MIPS gene had high homology to MIPS genes of soybean （ Glycine max） and tomato （Lycopersicon esculentum ） with the identities of 83.63 % and 83.89 %, respectively. The cloning of sweetpotato MIPS gene is useful for the further research of the relationship between sweetpotato MIPS gene and sweetpotato stem nematode resistance.

关 键 词：甘薯 肌醇-1-磷酸合成酶基因 茎线虫病 克隆 

分 类 号：S188[农业科学—农业基础科学]