CAJ | 학술논문

小麦是世界主要粮食作物之一，但籽粒赖氨酸含量低，影响其营养品质。为了通过生物技术提高小麦籽粒中赖氨酸含量，选用小麦LKR-SDH基因靠近3′端385 bp的保守序列，以水稻pGluB为启动子，拟南芥FAD2第一内含子为间隔序列，pUC18为中间栽体，构建LKR-SDH基因的hpRNA表达盒。然后将hpRNA表达盒插入到pAHC25的Hind Ⅲ酶切位点上，成功构建小麦LKR-SDH基因RNAi植物表达载体pAHC25-TaLKR/SDH-RNAi。通过限制性内切酶酶切鉴定，表明pAHC25-TaLKR/SDH-RNAi载体构建正确。进一步采用基因枪法将其转入到扬麦158中，获得了部分小麦转化植株。旨在为小麦高赖氨酸品种的培育提供候选材料。
소맥시세계주요양식작물지일，단자립뢰안산함량저，영향기영양품질。위료통과생물기술제고소맥자립중뢰안산함량，선용소맥LKR-SDH기인고근3′단385 bp적보수서렬，이수도pGluB위계동자，의남개FAD2제일내함자위간격서렬，pUC18위중간재체，구건LKR-SDH기인적hpRNA표체합。연후장hpRNA표체합삽입도pAHC25적Hind Ⅲ매절위점상，성공구건소맥LKR-SDH기인RNAi식물표체재체pAHC25-TaLKR/SDH-RNAi。통과한제성내절매매절감정，표명pAHC25-TaLKR/SDH-RNAi재체구건정학。진일보채용기인창법장기전입도양맥158중，획득료부분소맥전화식주。지재위소맥고뢰안산품충적배육제공후선재료。
Wheat is one of main food crops in the world , but its nutritional value is limited by its low lysine content .In order to improve lysine content in wheat grains by biotechnology , we selected and used the 385-bp consered sequence fragment of wheat LKR-SDH gene, and constructed the RNAi hairpin expression cassette of LKR-SDH gene by using rice pGluB as promoter , the first intron of Arabidopsis thaliana FAD2 as interval sequence , and the pUC18 vector as intermediate vector .Then the HindⅢfrag-ments containing the RNAi hairpin expression cassette were sub -cloned into the pAHC25 vector, and the constructed new RNAi ex-pression vector of wheat LKR-SDH gene was named as pAHC25-TaLKR/SDH-RNAi.Enzyme digestion showed that these re-combined plasmids were successfully constructed .The pAHC25-TaLKR/SDH-RNAi was used to transfer the immature embryos of wheat cultivar Yangmai 158 by gold particle bombardment , and some transgenic wheat seedlings were obtained .Our research re-sults might provide candidate materials for high -lysine wheat breeding .