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苦荞突变体库构建与突变体中芦丁合成相关基因表达分析
孙朝霞1, 侯思宇1, 郝彦蓉1, 王东航1, 高豪1, 韩渊怀1, 李红英1, 刘龙龙2, 周美亮3
0
(1.山西农业大学农学院;2.山西省农业科学院种质与资源研究所;3.中国农业科学院作物科学研究所)
摘要:
摘要:苦荞是重要的小杂粮作物之一,营养物质丰富,是天然芦丁的重要来源。突破苦荞育种难题,创制苦荞新种质是目前研究的重要方面。本试验利用甲基磺酸乙酯(EMS)构建了黑丰1号苦荞突变体库,明确了当EMS浓度为1.2%时,诱变效果较好。通过对M1代 突变株表型观察统计,共获得叶色、叶型、株型、粒型变异单株102株,突变率为3.85%;高效液相色谱技术(HPLC)测定1000株M23代 材料,获得高芦丁含量突变株系2个和低芦丁突变体株系5个;qRT-PCR对芦丁含量突变体株系中芦丁代谢关键酶基因(CHS、F3H、4CL、FLS、UFGT)进行表达量分析,发现不同株系中上述基因的表达量与芦丁含量相关性不明显,但个别基因如FtFLS基因表达量在高芦丁含量突变体中达到对照的4.55倍。通过突变体的筛选丰富了苦荞基因资源,创新了苦荞新种质,也为苦荞芦丁代谢的分子基础研究提供了材料保证与技术支持。
关键词:  苦荞  突变体  芦丁  基因表达
DOI:10.13430/j.cnki.jpgr.20190425002
投稿时间:2019-04-25修订日期:2019-12-22
基金项目:国家重点研发计划中欧政府间合作项目(2017YFE0117600);国家燕荞麦产业体系(CARS-07-A1);山西省应用基础研究项目(201801D221296); 山西省回国留学人员科研资助项目(2017-069);国家自然基金青年项目(31301385)
EMS-assisted Mutagenesis of Tartary Buckwheat and Expression Analysis of Rutin Biosynthesis Genes in Selected Mutants
SUN Zhao-xia1, HOU Si-yu1, HAO Yan-rong1, WANG Dong-hang1, GAO Hao1, HAN Yuan-huai1, LI Hong-ying1, LIU Long-long2, ZHOU Mei-liang3
(1.School of Agronomy,Shanxi Agricultural University;2.Institute of Germplasm Resources, Shanxi Academy of Agriculture Science;3.Institute of Crop Science, Chinese Academy of Agricultural Sciences)
Abstract:
Tartary buckwheat is one of minor grain crops in China, which has rich nutrition and is an important source of natural rutin. It is an important aspect of present research to break through of breeding and create new germplasm in tartary buckwheat. The mutation library was constructed in “Heifengyihao” by EMS. The half lethal treatment of EMS mutagenesis was 1.2%, the effect of plentiful phenotype was recorded in M1 generation. 102 plants with obvious phenotypic variation, including leaf color, leaf shape, plant type and grain sizes, were obtained from M3 materials and the mutation rate was 3.85%. The rutin content of 1000 plants in mutants’ library was determined by HPLC, 2 mutant lines with high (rutin content >16 mg/g) and 5 lines of low rutin (rutin content <10 mg/g) were obtained respectively. Comparison of rutin synthesis-related genes through qRT-PCR showed a relative weaker relationship between FtCHS, FtF3H, Ft4CL, FtUFGT transcripts and rutin content mutation lines. However, it also found that FtFLS showed the higher expression level in line with higher rutin content, which reached 4.55 times of control. The screening of mutants enriched the resources of tartary buckwheat, innovated the new germplasm of tartary buckwheat, and provided material guarantee and technical support for the molecular basis research of tartary buckwheat metabolism.
Key words:  tartary buckwheat  mutant  rutin  gene expression

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