首页|Cloning of Resistance Gene Analogs (RGAs) from Red Tangerine (Citrus reticulata Blanco) by Candidate Gene Approach and Overlapping Extension Strategy
Cloning of Resistance Gene Analogs (RGAs) from Red Tangerine (Citrus reticulata Blanco) by Candidate Gene Approach and Overlapping Extension Strategy
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Citrus is one of the most important fruit crop globally because of its fresh consumption， juice producation， and many additional minor uses including medicinal utilization。 However， diseases cause substantial economic losses to citrus industry。 Disease-resistance breeding is one of the most important aims for citrus breeding program。 In this study， red tangerine (Citrus reticulate Blanco) ， which is resistant to Citrus exocortis viroid， were used for resistant gene homolog cloning by both candidate gene approach and overlap extension strategy。 In the candidate gene approach， the degenerate primers were designed based on the conserved P-loop and GLPL motifs of NBS-LRR (nucleotide binding site-Leucine-rich repeats) type disease-resistance genes。 While in the overlap extension strategy， two additional primers， which targeted to Kinase2 motif， were designed;and two rounds of PCR were used to produce the expected products。 The PCR products obtained were about 500bp。 Sequence analysis revealed that 27 of the 50 clones are resistance gene analogs (RGA)。 These sequences exhibited 23%～99% nucleotide identities with each other。 The deduced amino acid sequences of the DNA fragments contained the conserved motifs of NBS-LRR type RGAs， such as P-loop， kinase-2a， kinase-3a and GLPL domains。 Phylogenetic analysis classified these RGAs into 4 evolutionary clades， named I ， II ， ffl， IV type respectively， and the IV type is TIR-NBS-LRR type。 The cloned genes from this study have potential utilization in the development of molecular markers associated with disease resistance， and the phylogenetic analysis of the resistance genes analogs provides evolutionary mechanism of resistance genes in citrus。
Red tangerine (Citrus reticulata Blanco)NBS-LRRResistance gene analogs
Xu Q、Deng XX
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National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, Wuhan 430070, P. R.China
International citrus congress
Wuhan(CN)
Proceedings of the International Society of Citriculture.;vol. 1.
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