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

EMS-assisted Mutagenesis of Tartary Buckwheat and Expression Analysis of Rutin Biosynthesis Genes in Selected Mutants

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M3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。

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,FtF3 H,Ft4 CL,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.