Based on both cDNA sequence of barley powdery mildew resistance control element Mlo and DNA sequence of the known putative disease resistance gene from Triticum monococcum L., we designed some primers to amplify resis...Based on both cDNA sequence of barley powdery mildew resistance control element Mlo and DNA sequence of the known putative disease resistance gene from Triticum monococcum L., we designed some primers to amplify resistant homologous sequences in the near isogenic lines (NILs) of powdery mildew resistance using RT-PCR method. Two expressed cDNA fragments were isolated from wheat genome. One showed 83% homology to the Mlo gene of barley. The other contained two possible open reading frames (ORFs). NBS conservative domains 2, 3 of disease resistance gene and 13 LRR structures similar to rice Pib protein terminal were found respectively in the two ORFs. It indicated that the latter fragment belongs to NBS-LRR-like genes. The obvious difference of RT-PCR products was observed between the before challenged and the challenged for 72 h by Blumeria graminis f. sp. tritici, which, implied that this sequence could be associated with disease resistance of wheat. Using nulli-tetrasomic lines of 'Chinese Spring', the NBS-LRR-like gene had been located on chromosome 1D.展开更多
One resistance gene analog fragment named RGA-CIN14 was isolated from TcLr19 wheat,which contains kinase-2,kinase-3a,and the GLPL motif of the NBS-spanning region,using degenerated primers according to the nucleotide ...One resistance gene analog fragment named RGA-CIN14 was isolated from TcLr19 wheat,which contains kinase-2,kinase-3a,and the GLPL motif of the NBS-spanning region,using degenerated primers according to the nucleotide binding site （NBS） conserved domain.Based on the RGA-CIN14,a full-length cDNA,CIN14,which was 2 987 bp encoding 880 amino acids,was obtained by using the method of the rapid amplification cDNA ends （RACE）.Bioinformatics analysis showed that the deduced amino acids of CIN14 protein consisted of a NB-ARC conserved domain and many leucine-rich repeats （LRR） domains.The phylogenetic tree analysis indicated a considerable identity of the protein encoded by CIN14 with that of wheat leaf rust resistance gene Lr1,but a lower similarity with Lr21.The expression profile of the CIN14 gene detected by semi-quantitative RT-PCR showed that the CIN14 gene was not induced by Puccinia triticina and it was a constitutive gene with low abundance in the wheat leaf tissue.The resistance homology sequence was successfully obtained,which provides the shortcut for cloning of the resistance gene in TcLr19 wheat.展开更多
文摘Based on both cDNA sequence of barley powdery mildew resistance control element Mlo and DNA sequence of the known putative disease resistance gene from Triticum monococcum L., we designed some primers to amplify resistant homologous sequences in the near isogenic lines (NILs) of powdery mildew resistance using RT-PCR method. Two expressed cDNA fragments were isolated from wheat genome. One showed 83% homology to the Mlo gene of barley. The other contained two possible open reading frames (ORFs). NBS conservative domains 2, 3 of disease resistance gene and 13 LRR structures similar to rice Pib protein terminal were found respectively in the two ORFs. It indicated that the latter fragment belongs to NBS-LRR-like genes. The obvious difference of RT-PCR products was observed between the before challenged and the challenged for 72 h by Blumeria graminis f. sp. tritici, which, implied that this sequence could be associated with disease resistance of wheat. Using nulli-tetrasomic lines of 'Chinese Spring', the NBS-LRR-like gene had been located on chromosome 1D.
基金funded by the National Natural Science Foundation of China (30771391,30700505)the Natural Science Foundation of Hebei Province,China (C2008000281)
文摘One resistance gene analog fragment named RGA-CIN14 was isolated from TcLr19 wheat,which contains kinase-2,kinase-3a,and the GLPL motif of the NBS-spanning region,using degenerated primers according to the nucleotide binding site （NBS） conserved domain.Based on the RGA-CIN14,a full-length cDNA,CIN14,which was 2 987 bp encoding 880 amino acids,was obtained by using the method of the rapid amplification cDNA ends （RACE）.Bioinformatics analysis showed that the deduced amino acids of CIN14 protein consisted of a NB-ARC conserved domain and many leucine-rich repeats （LRR） domains.The phylogenetic tree analysis indicated a considerable identity of the protein encoded by CIN14 with that of wheat leaf rust resistance gene Lr1,but a lower similarity with Lr21.The expression profile of the CIN14 gene detected by semi-quantitative RT-PCR showed that the CIN14 gene was not induced by Puccinia triticina and it was a constitutive gene with low abundance in the wheat leaf tissue.The resistance homology sequence was successfully obtained,which provides the shortcut for cloning of the resistance gene in TcLr19 wheat.
