不同抗倒性甜荞茎秆木质素合成关键酶基因的表达分析

doi:10.3864/j.issn.0578-1752.2015.09.20

Abstract

Abstract: 【Objective】The objective of this study is to explore the critical period and key enzyme genes in lignin synthesis of stem in common buckwheat and to provide a theoretical basis for common lodging resistance breeding cultivation by determining the lignin content and related genes expression characteristics at different stages of different lodging resistance cultivars. 【Method】Degenerate primers were designed to amplify part of the CDS sequences of CAD, CCR, F5H, COMT and CCOAOMT genes and blast was conducted in the NCBI database. The lignin content at the 2nd internode from the bottom were analyzed at branching stage, full bloom stage and milk ripe stage among four different lodging resistances of common buckwheat by UV spectrophotometry. Quantitative real-time polymerase chain reaction was used to analyze the expression characteristics of nine related genes (PAL, 4CL, C4H, C3H, CAD, CCR, F5H, COMT, and CCOAOMT), and the relative expression level was defined as 2^-^△△CT. Microsoft Excel 2003 and SPSS 19 were used for sorting data, drawing and analysis of variance, correlation analysis, respectively.【Result】Amplified CAD, CCR, F5H, COMT and CCOAOMT genes sequences were used to design the subsequent qRT-PCR primers. Lignin content of stem gradually increased from branching stage to milk ripe stage among different lodging resistances of common buckwheat, and lodging resistance cultivar was significantly higher than that of lodging susceptible cultivar. The expression levels of four genes (PAL, 4CL, C4H, CCR) varied significantly or extremely significantly among different cultivars and stages, CCOAOMT, F5H varied significantly among stages and cultivars, respectively. The remaining three genes (C3H, CAD, COMT) showed no significant difference among the cultivars and stages. The expressions of genes PAL, 4CL, C4H, CAD, CCOAOMT, F5H and CCR increased and then decreased, and the maximum appeared at full bloom stage, and lodging resistance cultivar was significantly higher than that of lodging susceptible cultivar. For COMT, the expression level decreased gradually from branching stage to full bloom stage, compared with the front seven genes, it showed an opposite expression pattern between lodging resistance and lodging susceptible cultivars at the first two stages. There was no obvious changing rule in gene C3H from branching stage to milk ripe stage. The lignin content was significantly and positively correlated with expressions of genes PAL, 4CL, C4H, CCOAOMT, CAD, and CCR, negatively correlated with C3H, and negatively correlated at branching stage and full bloom stage with COMT, while positively correlated at milk ripe stage with COMT.【Conclusion】The common buckwheat cultivar with lodging resistance showed high lignin content in the stem. The PAL, 4CL, C4H, CAD, CCOAOMT and CCR genes play a crucial role in the pathway of lignin synthesis. Full bloom stage is the critical period in lignin synthesis of stem in common buckwheat.

Key words: common buckwheat, lignin, key enzyme gene, qRT-PCR, lodging resistance

HU Dan, LIU Xing-bei, WANG Can, YANG Hao, LI He-xin, RUAN Ren-wu, YUAN Xiao-hui, YI Ze-lin. Expression Analysis of Key Enzyme Genes in Lignin Synthesis of Culm Among Different Lodging Resistances of Common Buckwheat (Fagopyrum esculentum Moench)[J].Scientia Agricultura Sinica, 2015, 48(9): 1864-1872.

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Expression Analysis of Key Enzyme Genes in Lignin Synthesis of Culm Among Different Lodging Resistances of Common Buckwheat (Fagopyrum esculentum Moench)

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