Abstract
Based on conserved regions of a sequence of a previously isolated powdery mildew (Pm) resistance gene, Pm3b, from hexaploid wheat (Triticum aestivum L.), a pair of primers was designed, and 11 resistance gene analogs (RGAs) were obtained using a polymerase chain reaction-based cloning approach. Three RGAs were deemed as pseudogenes, while the remaining eight corresponded to protein-encoding genes. At the nucleotide level, all these RGAs shared a sequence identity of 99% and showed 86% sequence identity with Pm3b. Phylogenetic analysis revealed that the eight protein-encoding genes were paraphyletic with Pm3 alleles. Positively selected sites were identified using the Selecton 2.1 program, and these were located on secondary structures. Based on these findings, the following two inferences could be made on the mode of evolution of the nucleotide-binding site and leucine-rich repeat (LRR) class of resistance genes. First, the majority of evolution events must have occurred primarily in the LRR domain, and this might have contributed to an increase in the proportion of evolution events in other domains. Second, these evolutionary events in the LRR domain must have occurred initially in secondary structures and then in the β-sheet. The crystalline structure models of RGAs were constructed.
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Abbreviations
- BLAST:
-
Basic local alignment and search tool
- CC:
-
Coiled coil
- DNA:
-
Deoxyribonucleic acid
- InDel:
-
Insertion/deletion
- LRR:
-
Leucine-rich repeat
- NBS:
-
Nucleotide-binding site
- PCR:
-
Polymerase chain reaction
- Pm:
-
Powdery mildew
- RGAs:
-
Resistance gene analogs
- TIR:
-
Toll-interleukin receptor-like
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Acknowledgments
The authors thank Prof. Mark E. Sorrels (College of Agriculture and Life Sciences, Cornell University) for the critical review of the manuscript. We also wish to thank two anonymous reviewers for helpful comments that improved the manuscript. This project was supported by the Early Stage of China Key Development Project for Basic Research (973) (2007CB116208).
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De-Shun Feng and Yan Li contributed equally to this paper.
Sequence data of 2Q2, 2Q3, 2Q4, 2Q7, 2Q9, 2Q11, 2Q12, 12Q11, 15Q1, and 15Q7 from this article have been deposited at GenBank under accession numbers EF157980, EF157981, EF157982, EF157983, EF157984, EF157985, EF157986, EF157987, EF157988, EF157989, and EF157990.
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Feng, DS., Li, Y., Wang, HG. et al. Isolation and Evolution Mode Analysis of NBS–LRR Resistance Gene Analogs from Hexaploid Wheat. Plant Mol Biol Rep 27, 266–274 (2009). https://doi.org/10.1007/s11105-008-0080-1
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DOI: https://doi.org/10.1007/s11105-008-0080-1