Abstract

The nucleotide-binding site (NBS)-Leucine-rich repeat (LRR) gene family accounts for the largest number of known disease resistance genes, and is one of the largest gene families in plant genomes. In the present study, based on the NBS domain, resistance gene analogues (RGAs) have been isolated from peanut, which named PnAG₃. A full-length cDNA, PnAG₃ was obtained by rapid amplification of cDNA ends (RACE) method. Sequence analysis indicated that the length of PnAG₃ was 1 882 bp, including a complete open reading frame of 1 335 bp encoding PnAG₃ protein of 444 amino acids. Multiple analysis showed that it had a certain homology with known resistance proteins, among which Arachis cardenasii resistance protein had the highest homology (48.01%). The polypeptide has a typical structure of nonTIR-NBS-LRR genes. Real-time fluorescence quantitative PCR analysis showed that after A. flavus infection, the expression of PnAG₃ gene in J11 (A. flavus resistance species) has increased 16.68, 11.16 and 25.96 times in seed coat, kernel and pericarp, respectively. But it only increased 2-3 times in JH1012 (A. flavus sensitive species). The cloning of putative resistance gene from peanut provides a basis for studying the structure and function of peanut disease-resistance relating genes and disease resistant genetic breeding in peanut.

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