Cloning and Analysis of NBS-LRR Type Resistance Gene Analogues in Sweet Potato (Ipomoea batatas)
Zeng HuiCai
Abstract
Zeng HuiCai
Abstract
The degenerate primers were designed based on the conserved NBS-LRR motifs among the known disease-resistance genes. A fragment of about 500 bp was amplified from genomic DNA of sweet potato using the specifically designed degenerate primers. After cloning and sequencing,20 NBS-LRR type of disease-resistance gene analogue (RGAs) in sweet potato were observed. The deduced amino acid sequence of DNA fragment contains the conserved motifs of NBS-LRR type RGAs,such as P-loop,Kinase-2α,Kinase-3α and GLPL domain. The 20 RGAs could be sorted into two subclasses,namely TIR-NBS-LRR type and non-TIR-NBS-LRR type. Compared with the known resistance genes including N,L6 and M,the percentages of homologous amino acid sequence in 10 TIR-NBS-LRR range between 21%-44%. While other 10 non-TIR-NBS-LRR assume 15%-46% homology with the known resistance genes (Prf,RPM1,RPS2,etc.). Consequently the RGAs may further be used as molecular marker for screening the candidate disease-resistance genes in sweet potato.
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The degenerate primers were designed based on the conserved NBS-LRR motifs among the known disease-resistance genes. A fragment of about 500 bp was amplified from genomic DNA of sweet potato using the specifically designed degenerate primers. After cloning and sequencing,20 NBS-LRR type of disease-resistance gene analogue (RGAs) in sweet potato were observed. The deduced amino acid sequence of DNA fragment contains the conserved motifs of NBS-LRR type RGAs,such as P-loop,Kinase-2α,Kinase-3α and GLPL domain. The 20 RGAs could be sorted into two subclasses,namely TIR-NBS-LRR type and non-TIR-NBS-LRR type. Compared with the known resistance genes including N,L6 and M,the percentages of homologous amino acid sequence in 10 TIR-NBS-LRR range between 21%-44%. While other 10 non-TIR-NBS-LRR assume 15%-46% homology with the known resistance genes (Prf,RPM1,RPS2,etc.). Consequently the RGAs may further be used as molecular marker for screening the candidate disease-resistance genes in sweet potato.
Key concepts: Gene, Genetics, Biology, Cloning (programming), Sequence analysis, Molecular cloning, genomic DNA, Plant disease resistance