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中国农学通报 ›› 2017, Vol. 33 ›› Issue (36): 131-138.doi: 10.11924/j.issn.1000-6850.casb17040038

所属专题: 生物技术 水稻

• 植物保护 农药 • 上一篇    下一篇

转cry1C基因抗虫超级粳稻田间目的基因表达及抗螟虫性

李冬雪,田崇兵,刘长华,李荣田   

  1. 黑龙江大学农业资源与环境学院,黑龙江大学生命科学学院/分子生物学省高校重点实验室,黑龙江大学农业资源与环境学院,黑龙江大学生命科学学院/分子生物学省高校重点实验室
  • 收稿日期:2017-04-08 修回日期:2017-12-19 接受日期:2017-05-25 出版日期:2017-12-29 发布日期:2017-12-29
  • 通讯作者: 刘长华
  • 基金资助:
    国家高技术研究发展计划(863 计划)“寒区水稻绿色性状基因聚合与种质创新”(2014AA10A603-12);转基因生物新品种培育重大专项“粳 稻种植区抗虫水稻新品种的培育”(2016ZX08001001-001-007);黑龙江省博士后资助项目“抗稻瘟病水稻空育131 多系品种的培育”(LBH-Z12230); 黑龙江省大学生创新创业训练计划项目“转Bt 基因水稻HD3 品系目的基因整合及表达”(2016102127294);黑龙江省大学生创新创业训练计划项目 “黑龙江大学与黑龙江省龙娃粮油进出口有限公司“物联网+农业”对接服务项目”(20161021301)。

Target gene expression and stem borer resistance of insect-resistant super japonica rice transformed with cry1C gene in field

李冬雪,田崇兵,刘长华 and 李荣田   

  • Received:2017-04-08 Revised:2017-12-19 Accepted:2017-05-25 Online:2017-12-29 Published:2017-12-29

摘要: 为了研究转基因抗虫超级粳稻田间目的基因表达及抗螟虫性,将10个来自于不同独立抗性愈伤组织的转cry1C*基因抗虫超级粳稻品系种植于田间,利用实时荧光定量PCR方法检测孕穗期叶片、茎鞘和幼穗等器官目的基因mRNA,利用酶联免疫吸附(ELISA)法检测孕穗期叶片、茎鞘和幼穗及收获后糙米的Cry1C蛋白,利用田间目测调查二化螟危害的白穗率。结果及分析显示,转基因超级粳稻不同品系及不同器官cry1C*基因田间表达量明显不同,cry1C*基因mRNA表达量叶片>茎鞘>幼穗,蛋白质表达量叶片>茎鞘>幼穗>糙米。转基因超级粳稻田间目的基因表达,在mRNA水平和蛋白质水平,不同器官间存在正相关关系,各器官Cry1C蛋白质含量和糙米Bt蛋白质含量呈正相关。在本研究范围内,不论转基因粳稻植株Cry1C蛋白质含量高或低的品系,田间均表现为高抗二化螟。培育转基因抗虫粳稻品种时,注意对目的基因适量表达的抗虫基因型的选择。

关键词: 黑蒜, 黑蒜, 加工, 组分变化, 生理功能

Abstract: To study the target gene expression and stem borer resistance in transgenic insect-resistant super Japonica rice in field, 10 super rice strains with insect-resistant genes Japonica rice transformed with cry1C* from different independent resistant callus were grown in field. The mRNA of target gene in rice leaf, stem sheath and young panicle at booting stage were detected by real-time PCR. The Cry1C protein in rice leaf, stem sheath, young panicle at booting stage and in brown rice after harvest were detected by enzyme-linked immunosorbent assay (ELISA). The white head plants damage of Chilo suppressalis was investigated by visual inspection in field. The results showed that the expression of cry1C* in different organs and different transgenic super Japonica rice strains was obviously different. The quantities of mRNA expression of cry1C* gene followed an order: leaf>stem sheath>young panicle, and the quantities of cry1C* protein followed an order: leaf>stem sheath>young panicle> brown rice. The positive correlation of target gene expression in transgenic super Japonica rice existed in different organs at mRNA and protein levels. The positive correlation existed between Cry1C protein content in organs and Bt protein content in brown rice. The stains, no matter with high or low Cry1C protein contents, had high resistance to Chilo suppressalis in filed in our study. The insect resistant genotype which properly expressed target genes should be paid attention in breeding transgenic insect-resistant Japonica varieties.