利用cDNA-AFLP技术鉴定菊花品种‘紫荷’的抗白锈病相关基因

doi:10.3864/j.issn.0578-1752.2012.05.013

Abstract

Abstract: 【Objective】An experiment was conducted to study the molecular mechanism of Chrysanthemum morifolium ‘Zihe’ response to the white rust, and to screen the white rust resistance genes. 【Method】 Infections were conducted by spraying the C. morifolium ‘Zihe’ leaves with suspension of 106 white rust sporangia per mL pure water for 3, 6, 12, 24, 48, 72, 96 h. cDNA-AFLP was used to moniter the gene expression under the white rust infections, and RT-PCR was used to validate the gene expression patterns. RACE method was used to get two fully cDNA sequences encoding 14-3-3 proteins, and phylogenetic analysis was also performed. 【Result】 Seventy-six primer combinations were used to investigate 4 950 cDNA fragments. After sequencing of 80 ESTs, the nucleic acid sequences of 51 ESTs were obtained, and among them, 18 ESTs were obtained, and showed homologous to the resistance-related genes of other species. These 18 ESTs were sorted into six functional categories: disease resistance protein, signal transduction, photosynthesis and photorespiration, retroelement polyprotein-like, senescence and metabolism pathways. RT-PCR showed that the cDNA-AFLP results was accurate and believable. Furthermore, two fully cDNA sequences of genes encoding 14-3-3 proteins were obtained: CmGFR01, CmGFR02. These two genes were 1 062 and 1 098 bp, encoded ORFs of 260 and 271 amino-acid residues. Phylogenetic analysis showed that the CmGFR01 and CmGFR02 might belong to a new subfamily of the higher plants 14-3-3 genes family. 【Conclusion】Gene expression profiling in response to white rust and differentially expressed genes of C. morifolium ‘Zihe’ were identified via cDNA-AFLP analysis. These genes could help us to understand the mechanism of response of plants to white rust. Furthermore, the isolated two genes, CmGFR01 and CmGFR02, could be used for the gene enigeering of the cultivated chrysanthemum resistance to the biotic stresses.

Key words: Chrysanthemum morifolium, white rust, cDNA-AFLP, 14-3-3 gene

HUANG He, WANG Shun-Li, DAI Si-Lan. cDNA-AFLP Analysis of White Rust Response Genes in Chrysanthemum morifolium ‘Zihe’[J].Scientia Agricultura Sinica, 2012, 45(5): 926-935.

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cDNA-AFLP Analysis of White Rust Response Genes in Chrysanthemum morifolium ‘Zihe’

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