转Bt基因海岛棉抗虫性鉴定及新品系筛选

发布时间：2018-12-19 16:02

【摘要】：海岛棉作为新疆特有的资源,其优良的纤维品质和商业价值是其他作物不可替代的。本研究以K222、新海25、新海30为受体,利用农杆菌介导技术导入Bt基因,通过PCR检测、Southern杂交、Bt-Cry1Ab/1Ac试纸条检测和抗虫性鉴定,证明Bt基因整合到海岛棉基因组中并得到表达。在分子检测基础上,对转Bt基因海岛棉品系的农艺性状、产量性状和纤维品质进行考察,结果表明：(1)PCR检测结果得出9个转Bt基因品系都含有目的基因,并且阳性率都在90%以上；Southern杂交结果可以看出7个转Bt基因品系出现了杂交条带,3个品系为单拷贝,3个品系为双拷贝,1个品系三拷贝。表明Bt基因已经成功整合到海岛棉基因组中。(2)用Bt免疫试纸条检测转Bt基因品系及对照,转Bt基因品系的试纸条上出现两条紫红色条带,而对照的试纸条上只出现一条紫红色条带,K2-ZKC66的阳性率达到了90%。(3)抗虫性鉴定结果表明转Bt基因品系的叶片危害较轻,叶片较为完整,对照叶片危害较重,叶片不完整,7个转Bt基因抗虫棉幼虫校正死亡率都在60%以上。(4)Bt基因的导入对9个海岛棉品系的农艺性状、产量性状和纤维品质产生不同程度的变化,当受体为K222时,转基因品系农艺性状(除果枝数外)与对照之间差异极显著(P0.01)；当受体为XH25和XH30时,转基因品系农艺性状与对照之间差异不显著。(5)9个转基因品系之间株高、始节高、始节数和铃数差异极显著(P0.01),有效果枝数和有效铃数差异显著(P0.05)。(6)9个转基因品系的产量性状与对照之间差异不显著；农艺性状与产量呈显著相关。(7)综合考虑9个转Bt基因抗虫棉在各方面的表现,筛选出K2-ZKC66,K2-ZKC31,K2-ZKC70三个表现优良的新品系。
[Abstract]:As a unique resource in Xinjiang, island cotton has excellent fiber quality and commercial value that can not be replaced by other crops. In this study, using K222, Xinhai 25 and Xinhai 30 as receptors, the Bt gene was introduced by Agrobacterium tumefaciens. The gene was detected by PCR, Southern hybridization, Bt-Cry1Ab/1Ac strip detection and insecticidal identification. The results showed that Bt gene was integrated into the genome and expressed in sea island cotton. On the basis of molecular detection, the agronomic characters, yield characters and fiber quality of island cotton lines with Bt gene were investigated. The results showed that: (1) the results of PCR analysis showed that all 9 lines with Bt gene had target genes. The positive rate was above 90%. The results of Southern hybridization showed that there were hybridization bands in 7 lines with Bt gene, 3 lines with single copy, 3 lines with double copy and 1 strain with three copies. The results showed that the Bt gene had been successfully integrated into the genome of island cotton. (2) two purplish red bands were found on the test strip of the transgenic Bt gene strain, and the Bt gene lines and the control were detected by Bt immunological test strip. However, there was only one purplish red band on the strip of the control, and the positive rate of K2-ZKC66 reached 90%. (3) the results of insect-resistance identification showed that the leaf damage of the transgenic strain with Bt gene was lighter, the leaf of the transgenic strain was relatively intact, and the damage of the control leaf was more serious. The corrected larval mortality of 7 transgenic cotton lines with Bt gene was above 60%. (4) the agronomic characters, yield characters and fiber quality of 9 island cotton lines were changed in different degree by introducing Bt gene. When the receptor was K222, the agronomic characters of transgenic lines (except the number of fruit branches) were significantly different from those of the control (P0.01). When the receptors were XH25 and XH30, there was no significant difference between the agronomic characters of the transgenic lines and the control. (5) the plant height, the number of initial nodes and the number of bolls were significantly different among the 9 transgenic lines (P0.01). There was no significant difference in the number of effective branches and effective bolls (P0.05). (6) between the yield characters of 9 transgenic lines and the control. Agronomic traits were significantly correlated with yield. (7) considering the performance of 9 transgenic cotton lines with Bt gene in various aspects, three new lines, K2-ZKC66, K2-ZKC31and K2-ZKC70, were selected.
【学位授予单位】：新疆农业大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：S562

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