Transgenic indica rice resistant to sap-sucking insects
Transgenic indica rice resistant to sap-sucking insects
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DOI：
10.1046/j.1467-7652.2003.00022.x
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发表时间：
2003-05-01
影响因子：
13.8
通讯作者：
Rao, KV
中科院分区：
文献类型：
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作者：
Nagadhara, D;Ramesh, S;Rao, KV
农杆菌介导的遗传转化在对吸汁昆虫敏感的籼稻中进行了优化，褐飞虱（BPH）和绿色叶蝉（GLH）。利用水稻韧皮部特异性蔗糖合成酶启动子驱动雪花莲凝集素基因（gna）沿着CaMV 35 S启动子驱动的除草剂抗性基因（bar）对雪花莲进行遗传转化。在含有膦丝菌素的培养基上，选择与携带Ti质粒pSB 111-bar-gna-的农杆菌菌株LBA 4404共培养后的胚性愈伤组织。PCR和Southern杂交分析证实了这两个基因稳定整合到转基因（TO）水稻植株的基因组中。北方和Western印迹分析表明gna在转基因植株中表达。在T-1和T-2代中，gna和bar转基因以3：1的比例显示共分离。在T-1和T-2中表达gna的植物后代表现出对BPH和GLH害虫的显著抗性。这是第一篇关于对两种昆虫都表现出高抗性的转基因籼稻的报道。
Agrobacterium-mediated genetic transformation has been optimized in indica rice susceptible to sap-sucking insects, viz., brown planthopper (BPH) and green leafhopper (GLH). Snowdrop lectin gene (gna) from Galanthus nivalis, driven by phloem-specific rice-sucrose-synthase promoter, along with herbicide resistance gene (bar) driven by CaMV 35S promoter, was employed for genetic transformation. Embryogenic calli - after cocultivation with Agrobacterium strain LBA4404 harbouring Ti plasmid pSB111-bar-gna- were selected on the medium containing phosphinothricin. PCR and Southern blot analyses confirmed the stable integration of both the genes into genomes of transgenic (TO) rice plants. Northern and Western blot analyses revealed the expression of gna in the transgenic plants. In the T-1 and T-2 generations, the gna and bar transgenes showed co-segregation at a ratio of 3 : 1. Plant progenies expressing gna, in T-1 and T-2 exhibited substantial resistance against BPH and GLH pests. This is the first report dealing with transgenic indica rice exhibiting high resistance to both insects.