Cloning and analysis of a NBS-LRR disease resistance gene candidate PnAG1 from peanut (Arachis hypogaea L.)
Vol. 14 No. 6 (2011), Research Articles
Vol. 14 No. 6 (2011)

Cloning and analysis of a NBS-LRR disease resistance gene candidate PnAG1 from peanut (Arachis hypogaea L.)

Research Articles
Published 2011-10-24
Shi-hua Shan
Shandong Peanut Research Institute
Ting-ting Zhang
Shandong Peanut Research Institute
Chun-Juan Li
Shandong Peanut Research Institute
Chen Yang
Shandong Peanut Research Institute
Cai-xia Yan
Shandong Peanut Research Institute
Shu-bo Wan
Shandong Academy of Agricultural Sciences
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Supplementary Files

Phylogenetic tree of deduced amino acid sequence of PnAG1 gene
Comparison of the deduced amino acid sequence of PnAG1 with10 known disease resistance proteins Sequences were aligned using the CLUSTAL W program.
The amplication curves of PnAG1 gene by real-time fluorescence quantitative PCR
The standard curve of PnAG1 gene
The amplification curves of DF-actin gene by real-time fluorescence quantitative PCR
The standard curve of DF-actin gene
The amplification curves of PnAG1 gene and Control Gene by real-time qPCR
The melting curves of PnAG1 gene and Control Gene by real-time qPCR
The changes of PnAG1 gene expression during the pod-maturing phase in testa of JH1012 and J11.
The changes of PnAG1 gene expression during the pod-maturing phase in embryo of JH1012 and J11
Table.1 Degenerate primers used in amplification
Table.2 Primers used in real-time fluorescence quantitative PCR
Table 3 Copy number of DF-actin gene
Table 4 Copy number of AG1 gene

Keywords

Peanut
NBS-LRR
Real-time fluorescence quantitative PCR
Bioinformatics

How to Cite

1.
Shan S- hua, Zhang T- ting, Li C-J, Yang C, Yan C- xia, Wan S- bo. Cloning and analysis of a NBS-LRR disease resistance gene candidate PnAG1 from peanut (Arachis hypogaea L.). Electron. J. Biotechnol. [Internet]. 2011 Oct. 24 [cited 2024 Nov. 23];14(6). Available from: https://www.ejbiotechnology.info/index.php/ejbiotechnology/article/view/v14n6-3
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Abstract

Background: Based on the conserved sequences of a known NBS resistance gene, a pair of degenerate primers was designed to amplify the NBS-LRR resistance gene from peanut using PCR and RACE methods. Results: Analyzing the amino acid sequence by BLAST on NCBI, which was deduced from the 1088bp-long gene named PnAG1-2, showed that it had a certain homology with some resistance proteins, among which Arachis cardenasii resistance protein gene had the highest homology (66%). Relative quantification PCR analysis indicated that PnAG1-2 gene expresses more in J11 (an A. flavus-resistant variety) than in JH1012 (an A. flavus-susceptible variety) when the harvest time was coming. Conclusions: In this study, the NBS-LRR resistance sequence was successfully cloned from peanut and prokaryotic expression was done on the gene, which provided a foundation for cultivating anti-A. flavus peanut varieties.

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