Pyramiding Bt genes for increasing resistance of cotton to two major lepidopteran pests: Spodoptera litura and Heliothis armigera
Pyramiding Bt genes for increasing resistance of cotton to two major lepidopteran pests: Spodoptera litura and Heliothis armigera
复制标题
金字塔化 Bt 基因可提高棉花对两种主要鳞翅目害虫的抗性：斜纹夜蛾和棉铃虫
DOI：
10.1007/s11738-014-1642-5
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发表时间：
2014
影响因子：
2.6
通讯作者：
Zhang Xianlong
中科院分区：
文献类型：
--
作者：
Li Lebin;Zhu Yi;Jin Shuangxia;Zhang Xianlong
为了更有效地控制棉铃虫和斜纹夜蛾这两种主要棉花害虫，提高抗虫性管理的效果，培育了表达苏云金芽孢杆菌Cry9C基因的新型转基因植株，并引入了基因堆积策略。首先，通过农杆菌介导法将含有Cry9C基因的双质粒载体导入棉花优良品种四棉3号。通过聚合酶链式反应和逆转录-聚合酶链式反应证实了Cry9C基因在3个转基因株系中的整合和表达。在这些转基因品系中，通过ELISA法筛选出了表型正常的品系16(L-16)的T0代，其Cry9C的表达水平最高。在L-16的T1群体中，Cry9C的表达水平在29~45μg/g鲜叶之间。昆虫生物测定表明，转S3-35S：：Cry9C基因棉株对棉铃虫有中等毒力，但对棉铃虫有较强的毒力。与表达Cry 1Acc基因的转基因植株进行了比较。采用基因定位策略，对Cry9C基因和Cry 2Aor Cry 1Ac有性杂交分别进行了聚合。在所有F1代中，Cry9C蛋白的表达水平与亲本相近，表明Bt基因在转基因后代中具有较高的遗传力。Cry9C×Cry 2A和Cry9C×Cry 1AC的后代表现出较强的抗逆性。礼拜仪式与他们的父母进行比较。总之，我们的数据表明，我们新产生的转基因植株代表了棉花育种中新的抗虫材料的储备库，而基因聚合技术的成功整合可以为开发多种抗性管理策略提供新的解决方案。
To more effectively control two major cotton insects (cotton bollworm andSpodoptera litura) and improve the efficacy of the pest resistance management, novel transgenic plants expressingBacillus thuringiensis Cry9Cgene were generated, and gene stacking strategy was incorporated. Initially, a binary plasmid vector harboringCry9Cgene was introduced into an elite cotton cultivar Simian-3 byAgrobacterium-mediated transformation. Integration and expression of theCry9Cgenes in three transgenic lines were confirmed by PCR and RT-PCR. Among these transgenic lines, T0 generation of line 16 (L-16) with normal phenotypes were selected by ELISA assays for its highest expression level of Cry9C. In T1 population of L-16, the expression level ofCry9Cranged from 29 to 45 μg/g fresh leaf. The following insect bioassays demonstrated that transgenic S3-35S::Cry9C cotton plants exhibited moderate toxicity toHeliothis armigerabut strong toxicity toS. lituracompared with the transgenic plants expressingCry 1Acgene. For incorporation of gene staking strategy,Cry9Cgene andCry 2AorCry 1Acwere pyramided, respectively by sexual crossing. The expression of Cry9C protein in all F1 progenies had a similar level as the parent plants indicating the high heritability ofBtgenes in transgenic progenies. Progenies from both Cry9C × Cry 2A and Cry9C × Cry 1Ac exhibited higher resistance toS. lituracompared with their parents. Together our data demonstrated that our newly generated transgenic plants represent a reservoir of novel insect-resistant materials in cotton breeding, and the successful incorporation of gene pyramiding technology can provide a new solution of developing multiple resistance management strategies.