苯丙氨酸与UV-C对苦荞芦丁含量影响及相关基因表达分析

doi:10.3864/j.issn.0578-1752.2011.23.002

摘要/Abstract

摘要： 【目的】克隆与苦荞芦丁生物合成相关基因序列，探讨外源前体物质及UV-C辐射条件下芦丁含量及基因表达水平的影响，为分子选育高芦丁含量荞麦品种奠定基础。【方法】基于简并引物结合3′RACE的方法，克隆苦荞芦丁合成途径相关基因；高效液相色谱（HPLC）测定持续添加外源苯丙氨酸（L-phenylalanine，Phe）前体（1、2、4 mg•L-1）1—5 d和辐射时间（1、3、5、7 h）UV-C处理条件下叶片芦丁含量的变化，qRT-PCR分析相关基因表达水平。【结果】Phe（4 mg•L-1）处理1 d，芦丁含量（26.15 mg•gFW-1）为未处理对照（12.55 mg•gFW-1）的2倍；UV-C辐射3 h，芦丁含量（17.36 mg•gFW-1）明显高于对照；克隆到3个基因FtCHS，FtF3H和FtFLS-like，qRT-PCR分析表明Phe（1 mg•L-1）处理4 d，FtCHS相对表达量为对照的4.84倍，Phe（4 mg•L-1）处理5 d，FtF3H相对表达量为对照的1.06倍，Phe（2 mg•L-1）处理5 d，FtFLS-like相对表达量为对照的3.90倍；UV-C辐射1—3 h，FtCHS、FtF3H和FtFLS-like相对表达量分别高于对照。而FtFLS-like基因，仅在UV-C辐射1 h，相对表达量升高为对照的127.71倍。【结论】添加不同浓度Phe前体及不同时间UV-C辐射，芦丁含量变化及与芦丁合成相关基因表达量存在差异，为进一步探讨芦丁生物合成途径及影响因素奠定基础。

关键词: 苦荞, 芦丁, 苯丙氨酸, UV-C, 基因表达

Abstract: 【Objective】 The objective of this study is to investigate the content of rutin and the expression of the related genes in response to L-phentermine (Phe) and UV-C treatment, and to provide molecular information for high rutin content buckwheat breeding. 【Method】 Rutin biosynthesis-related genes were cloned from the leaves of F. tataricum by 3′RACE. Rutin content in leaves under Phe treatment (1, 2 and 4 mg?L-1, 1 to 5 d) was compared with that of untreated buckwheat by high performance liquid chromatography (HPLC). The gene expression level was analyzed by qRT-PCR. 【Result】 Three partial CDS of rutin biosynthesis-related genes were cloned. They were chalcone synthase (FtCHS), flavonol synthase-like (FtFLS-like) and flavone 3-hydroxylase (FtF3H). Seedlings were cultured in solution containing Phe (4 mg×L-1) for 1 d, rutin content was two times more than the control (12.55 mg×gFW-1), reached 26.15 mg×gFW-1. The higher rutin content (17.36 mg×gFW-1) was found in leaves at 3 h under UV-C treatment. Differential expression patterns of FtCHS, FtFLS-like and FtF3H were observed in different Phe concentration treatments in leaves of F. tartaricum. Seedlings were cultured in solution containing Phe (1 mg×L-1) for 4 d, FtCHS expression level was 4.84 fold higher than the control. The higher expression levels of FtF3H and FtFLS-like were observed after 4 and 2 mg×L-1 Phe treatment for 5 d, they were 1.06 and 3.90 fold higher than the control, respectively. UV-C radiation for 1-3 h, the expression of FtCHS, FtFLS-like and FtF3H was higher than control. In particular, the expression level of FtFLS-like at 1 h under UV-C radiation was 127.71 fold higher than the control. 【Conclusion】 Rutin content could be enhanced to a great extent by Phe treatment and UV-C radiation, and gene expression patterns of rutin biosysthesis-related genes were regulated by UV-C radiation. It can be used to analyze the pathway of rutin biosynthesis. Further study should be focused on the influential factors of rutin metabolic process.

Key words: Fagopyrum tartaricum, rutin, L-phenylalanine, UV-C, gene expression

孙朝霞, 侯思宇, 杨武德. 苯丙氨酸与UV-C对苦荞芦丁含量影响及相关基因表达分析[J]. 中国农业科学, 2011, 44(23): 4772-4780.

SUN Chao-Xia, HOU Si-Yu, YANG Wu-De. Effect of Exogenous L-Phentermine and UV-C on the Accumulation of Rutin Compounds and the Expression of Rutin Biosynthesis Genes in Fagopyrum tartaricum[J]. Scientia Agricultura Sinica, 2011, 44(23): 4772-4780.

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