植物学报
植物學報
식물학보
ACTA BOTANICA SINICA
2002年
8期
963-970
,共8页
陆地棉%抗虫棉%转基因植物%根癌土壤杆菌%cry 1Ac3基因%复合启动子
陸地棉%抗蟲棉%轉基因植物%根癌土壤桿菌%cry 1Ac3基因%複閤啟動子
륙지면%항충면%전기인식물%근암토양간균%cry 1Ac3기인%복합계동자
upland cotton%insect-resistant cotton%transgenic plant%Agrobacterium tumefaciens%cry 1Ac3 gene%chimeric promoter
以根癌土壤杆菌( Agrobacterium tumefaciens )介导法分别将植物表达载体pBinMoBc和pBinoBc导入陆地棉( Gossypium hirsutum L.)栽培品种"新陆早1号"、"晋棉7号"、"晋棉12号"和"冀合321".pBinMoBc携带有高效启动子复合OM启动子控制下的 cry 1Ac3基因,pBinoBc携带有35S启动子控制下的 cry 1Ac3基因.经过共培养、卡那霉素筛选抗性愈伤组织及体细胞胚的诱导,得到了再生植株.对T2代的PCR、Southern blotting、ELISA检测及Western blotting证明 cry 1Ac3基因已整合入受体棉花基因组并得到表达.抗虫性检测表明转基因后代对棉铃虫( Heliothis armigera ) 具有良好的抗性,转pBinMoBc T2代与转pBinoBc T2代相比,对棉铃虫具有更快的致死速度.本研究建立了一套高效的陆地棉栽培品种转化体系;进一步的检测结果表明,复合OM启动子可以提高外源基因的表达量从而增强转基因棉的抗虫性.
以根癌土壤桿菌( Agrobacterium tumefaciens )介導法分彆將植物錶達載體pBinMoBc和pBinoBc導入陸地棉( Gossypium hirsutum L.)栽培品種"新陸早1號"、"晉棉7號"、"晉棉12號"和"冀閤321".pBinMoBc攜帶有高效啟動子複閤OM啟動子控製下的 cry 1Ac3基因,pBinoBc攜帶有35S啟動子控製下的 cry 1Ac3基因.經過共培養、卡那黴素篩選抗性愈傷組織及體細胞胚的誘導,得到瞭再生植株.對T2代的PCR、Southern blotting、ELISA檢測及Western blotting證明 cry 1Ac3基因已整閤入受體棉花基因組併得到錶達.抗蟲性檢測錶明轉基因後代對棉鈴蟲( Heliothis armigera ) 具有良好的抗性,轉pBinMoBc T2代與轉pBinoBc T2代相比,對棉鈴蟲具有更快的緻死速度.本研究建立瞭一套高效的陸地棉栽培品種轉化體繫;進一步的檢測結果錶明,複閤OM啟動子可以提高外源基因的錶達量從而增彊轉基因棉的抗蟲性.
이근암토양간균( Agrobacterium tumefaciens )개도법분별장식물표체재체pBinMoBc화pBinoBc도입륙지면( Gossypium hirsutum L.)재배품충"신륙조1호"、"진면7호"、"진면12호"화"기합321".pBinMoBc휴대유고효계동자복합OM계동자공제하적 cry 1Ac3기인,pBinoBc휴대유35S계동자공제하적 cry 1Ac3기인.경과공배양、잡나매소사선항성유상조직급체세포배적유도,득도료재생식주.대T2대적PCR、Southern blotting、ELISA검측급Western blotting증명 cry 1Ac3기인이정합입수체면화기인조병득도표체.항충성검측표명전기인후대대면령충( Heliothis armigera ) 구유량호적항성,전pBinMoBc T2대여전pBinoBc T2대상비,대면령충구유경쾌적치사속도.본연구건립료일투고효적륙지면재배품충전화체계;진일보적검측결과표명,복합OM계동자가이제고외원기인적표체량종이증강전기인면적항충성.
Hypocotyl segments from aseptic seedlings of two important cultivars of upland cotton ( Gossypium hirsutum L.) in Northwest China, "Xinluzao-1", "Jinmian-7", "Jinmian-12" and "Jihe-321" were transformed respectively by two efficient plant expression plasmids pBinMoBc and pBinoBc via Agrobacterium tumefaciens . In pBinMoBc, cry 1Ac3 gene, which encodes the Bt toxin, is under the control of chimeric OM promoter. In pBinoBc, it is under control of CaMV 35S promoter. After co-cultivation with Agrobacterium tumefimpfaciens LBA4404 (containing pBinMoBc or pBinoBc), kanamycin-resistant selection, somatic embryos were induced and regenerated plants were obtained. Then the regenerated plantlets were grafted to untransformed stocks in greenhouse to produce descendants. The integration of cry 1Ac3 gene and its expression in T2 generation of transgenic cotton plants were confirmed by Southern hybridization and Western blotting. The analyses of insect bioassay indicated that the transgenic plants of both constructions have significant resistance to the larvae of cotton bollworm ( Heliothis armigera ) and that cry 1Ac3 gene driven by chimeric OM promoter could endue T2 generation cotton with high pest-resistant ability, implicating that it has a profound application in genetic engineering to breed new pest-resistant cotton varieties.
