激光生物学报
激光生物學報
격광생물학보
ACTA LASER BIOLOGY SINICA
2009年
2期
264-269
,共6页
玉米自交系%淀粉分支酶sbeIIb基因%遗传转化%RNAi表达载体
玉米自交繫%澱粉分支酶sbeIIb基因%遺傳轉化%RNAi錶達載體
옥미자교계%정분분지매sbeIIb기인%유전전화%RNAi표체재체
maize elite inbred line%starch branching enzyme (sbeIIb) gene%genetic transformation%RNAi expression vector
玉米高直链淀粉育种是玉米分子育种的一个重要研究方向.本实验中,首先研究了不同诱导愈伤培养基对再生体系的影响,确定了LS+2,4-D 2.0 mg/L+L-pro 700 mg/L+CH 500 mg/L+3 %蔗糖为诱导培养基.同时,构建并验证了含有淀粉分支酶sbeIIb基因双干涉片段载体和胚乳特异性启动子的表达载体pCAMBIA 1301+Glu+1620,并转入根癌农杆菌EHA105,以农杆菌转化法转化玉米自交系178.通过PCR检测,5株转化株表现阳性,初步证明了干涉片段已整合入玉米基因组中.
玉米高直鏈澱粉育種是玉米分子育種的一箇重要研究方嚮.本實驗中,首先研究瞭不同誘導愈傷培養基對再生體繫的影響,確定瞭LS+2,4-D 2.0 mg/L+L-pro 700 mg/L+CH 500 mg/L+3 %蔗糖為誘導培養基.同時,構建併驗證瞭含有澱粉分支酶sbeIIb基因雙榦涉片段載體和胚乳特異性啟動子的錶達載體pCAMBIA 1301+Glu+1620,併轉入根癌農桿菌EHA105,以農桿菌轉化法轉化玉米自交繫178.通過PCR檢測,5株轉化株錶現暘性,初步證明瞭榦涉片段已整閤入玉米基因組中.
옥미고직련정분육충시옥미분자육충적일개중요연구방향.본실험중,수선연구료불동유도유상배양기대재생체계적영향,학정료LS+2,4-D 2.0 mg/L+L-pro 700 mg/L+CH 500 mg/L+3 %자당위유도배양기.동시,구건병험증료함유정분분지매sbeIIb기인쌍간섭편단재체화배유특이성계동자적표체재체pCAMBIA 1301+Glu+1620,병전입근암농간균EHA105,이농간균전화법전화옥미자교계178.통과PCR검측,5주전화주표현양성,초보증명료간섭편단이정합입옥미기인조중.
High amylose maize (Zea mays L.) breeding is an important field of molecular maize breeding. In this study, through research of the effect of inducing culture, the optimal inducing culture was fixed as LS+ 2,4-D 2.0 mg/L+L-pro 700 mg/L+CH 500 mg/L+3 % sucrose. Then, we constructed and detected the expression vector pCAMBIA 1 301+Glu+1 620 which carries endosperm specific promoter (Glu promoter from wheat glutenin) and two interferential fragment of the 16 and 20 exon in maize starch branching sbeIIb gene, transferred into Agrobacterium tumefaciens strain EHA105, and transformed the maize elite inbred line 178 with the Agrobacterium-protocol. Five transgenic plants which were detected by PCR reaction showed positive signal, indicating that interferential fragment was transformed preliminarily into genome of maize.