CAJ | 학술논문

根据大麦MLa基因的保守区域设计了4对家族性引物.通过用家族性引物对小麦(Triticum aestivum L.)抗白粉病品系TAM104R在接种和未接种两种条件下的基因差异表达进行RT-PCR分析,获得了一个在接种条件下特异表达的基因片段RJ-3-3L,并用RACE方法获得了其cDNA全长,命名为TaMla1.序列比对显示:TaMlal与大麦MLa位点的基因家族成员具有高度同源性,TaMla1编码的氨基酸功能基序扫描表明其为一个CC-NBS-LRR型抗病蛋白.用一套中国春缺-四体材料将TaMla1定位到了小麦的1A染色体上,这正是大麦MLa基因位点在小麦中的同源区段所在的染色体.这些结果表明,TaMla1为一个类MLa抗白粉病基因.同时我们还获得了一个在不接种条件下特异表达的基因片段RW-2-3L,序列分析表明它与MLa基因也高度同源,推测其可能是一个小麦白粉病的敏感基因或抗性负调控因子.
근거대맥MLa기인적보수구역설계료4대가족성인물.통과용가족성인물대소맥(Triticum aestivum L.)항백분병품계TAM104R재접충화미접충량충조건하적기인차이표체진행RT-PCR분석,획득료일개재접충조건하특이표체적기인편단RJ-3-3L,병용RACE방법획득료기cDNA전장,명명위TaMla1.서렬비대현시:TaMlal여대맥MLa위점적기인가족성원구유고도동원성,TaMla1편마적안기산공능기서소묘표명기위일개CC-NBS-LRR형항병단백.용일투중국춘결-사체재료장TaMla1정위도료소맥적1A염색체상,저정시대맥MLa기인위점재소맥중적동원구단소재적염색체.저사결과표명,TaMla1위일개류MLa항백분병기인.동시아문환획득료일개재불접충조건하특이표체적기인편단RW-2-3L,서렬분석표명타여MLa기인야고도동원,추측기가능시일개소맥백분병적민감기인혹항성부조공인자.
According to the conservative region of resistance gene homologue (RGH) family of the barley Mla locus, we synthesized four pairs of family primers. RT-PCR with the series of family-specific primers was used to analyze the differential expression of genes between the inoculated seedlings and the uninoculated seedlings of a wheat ( Triticum aestivum L.) line which showed high resistance to powdery mildew. As a result, a putative resistance gene fragment, RJ-3-3L, expressed specifically in the inoculated condition, was isolated and then, its full-length cDNA, designated as TaMla1, was obtained using rapid amplification of cDNA ends (RACE). Ey alignment analysis and functional-motif scanning, the TaMla1 shared high similarity to Mla alleles in barley and coded a typical CC-NBS-LRR (coiled-coil, nucleotide binding site,Leu-rich repeat) resistance protein. Subsequently, by Southern blot using a series of nulli-tetrasomics of wheat, the TaMla1 was located on wheat chromosome 1A, where the ortholog of the barley Mla locus lies.These findings showed that TaMla 1 is one of the Mla-like alleles in wheat. In addition, another gene fragment, RW-2-3L, its transcription remarkably inhibited in the inoculated seedlings, was isolated and homology analysis showed that it was also highly similar to Mla members in barley. We inferred that the gene derived RW-2-3L was also a Mla-like allele and probably acted as a powdery mildew susceptible gene or a negative factor for resistance to powdery mildew.