罗布麻EMS突变体库构建及黄酮变异突变体筛选

罗布麻(Apocynum venetum L.)突变体的获得是人工创新种质资源的有效途径,也是开展品种选育、功能基因组研究的基础。本研究以采自东营野生罗布麻种子作为材料,预试验结果表明0.7%的甲基磺酸乙酯(EMS,ethylmethylsulfone)溶液诱变罗布麻种子13 h能够达到半致死剂量。在此浓度和处理时间下构建了一个1452株的突变体群体,对获得的M1植株进行农艺性状和总黄酮含量性状筛选,初步获得125份农艺性状突变材料,其中包括8株白化植株、23株叶型变异株、16株叶色黄纹突变体、32株苗期分枝植株以及46株叶位变异突变体,突变频率分别为0.55%、1.6%、1.1%、2.2%和3.2%。此外,还筛选到2株总黄酮含量明显高于野生型和1株低于野生型的材料。在突变群体中对黄酮代谢途径中编码黄烷酮-3-羟化酶的基因(F3H,flavanone 3-hydroxylase gene)进行测序鉴定,结果鉴定到3株F3H基因的突变体。本研究构建的罗布麻突变体库表型丰富且突变位点可以鉴定,是罗布麻新种质筛选、品种选育以及基因功能研究的良好材料。

Construction of EMS Mutant Library and Screening of Total Flavonoid Content Mutants of Apocynum venetum L.

The acquisition of Apocynum venetum L.mutant library is an effective way to innovate germplasm resources artificially,and it is also the basis of varieties breeding and functional genome research.In this study,wild A.venetum L.seeds collected in Dongying were used to construct mutant library.Pre-experiment results indicated that 0.7%ethylmethylsulfone(EMS)solution was the LD50 for seeds of A.venetum L.treated for 13 h.A mutant library of 1452 individual plants was constructed with the treatment in this concentration and duration.The agronomic traits and total flavonoid contents of the M1 population were screened.A total of 125 plants with abnormal agronomic traits were obtained,including 8 albinos,23 leaf form variants,16 yellow-striped leaf variants,32 branching seedling variants,and 46 leaf position variants.The mutation frequencies were 0.55%,1.6%,1.1%,2.2%and 3.2%,respectively.Moreover,the total flavonoids in two plants were significantly higher and in one plant was lower than that in the wild type.The flavanone 3-hydroxylase gene(F3H)involved in the flavone metabolism pathway was sequenced in the mutant library.The results showed that one in every 6.7 plants appeared as an F3H gene mutant.The mutant library of A venetum L.constructed in this study showed abundant phenotypic variations,which provided good materials for selection of new germplasm,varieties breeding and gene function research in A.venetum L..