悬浮细胞再生表达外壳蛋白的转基因Indica水稻对水稻条纹病毒的抗病性(英文)  

Resistance to Rice Stripe Virus Conferred by Expression of Coat Protein in Transgenic Indica Rice Plants Regenerated from Bombarded Suspension Culture

作　　者：燕义唐[1] 王晋芳[1] 邱并生[1] 田波[1] 

机构地区：[1]中国科学院微生物研究所,北京100080

出　　处：《中国病毒学》1997年第3期260-269,共10页Virologica Sinica

摘　　要：合成、克隆了水稻条纹病毒中国株的外壳蛋白基因并进行了序列分析，由Indica水稻成熟胚的愈伤组织形成胚性运浮细胞。用含有CP基因的pROK2表达载体的DNA包被1.09μm直径钨粉颗粒轰击培养细胞。被轰击的培养物在含有G418（40mg／mL，）的培养基中进行选择培养，由对G418抵抗的愈伤组织中获得10株再生株。用32P－dCTP标记的CP基因作为探针，以Southernblot测定其转化特性。由抗病的和对照的植株抽提基因组DNA用EcoRI和BamHI进行酶切，其中两个植株显示出0.6kh和0.7kb两条条交带．其大小与CP基因相对应。Westernblot和ELISA测定进步证明CP（32kDa）在转基因水稻中表达。16株转基因植株和100株对照植株用带毒的叶蝉接种，接种病毒后24d只有37.5％的CP转基因植株产生病毒症状，而对照植株为96％。进一步证明转基因水稻植株具有对RSV的抗病性。转基因植株T1代CP的表达分离比例为3.6:1。The coat prolein (CP) gene of a Chinese isolate of rice stripe virus (RSV) was synthesized,cloned and sequenced. Embryogenic suspension cultures were initialed from calli which derived from Indiea rice mature embryo. For bombardment of cell suspension culture, tungsten particles of diameter of 1.09 μm were coated with DNA of the expression vector pRoK Ⅱ harbouring CP coding sequence downstream CaMV 35S promoter. Bombarded cultures were selected by growing in the medium containing G418 (40mg/mL).Ten plantlets regenerated from G418 resistant calli were analysed by Southem blot Date of receiption: 1996 -09-13, date of correction: 1997-02- 26 This work was supported by grant RF 95001 # 309 from the Rockfeller FoundationTo whom correspondence should be addressedlLsing 32P - dtryP 1abelled CP gene as probe. The genomic DNAs extrac1cd frotn resistant as well as th`>control plants were digested with EcoRI and BamHI. Samples from two plants showed tw0 hyl,ridlzat1onbands with the sizes 0f 0. 6 kb and 0. 7 kb corresponding t0 the RSV CP gene, Western blot and E!xlSAfurther demonstrated that RSV CP (32 kDa) was cxl'I-essed in tranSgenic rlce plante. SIxteen transgenicplants and l00 control pIanLs were fed by pIanthoPpers carrying RSV. Only 37. 5% of the CP genetransformed plants develoPed viral sytnptome compared lo 96 % of contro plants 24 days after 1noctIlati0nwith RSV, indicating that 1hc tranSgenic rice pIants were engincercd with rcslstance to RSV. Tlle CPexpression of Tl progenies from the tranSgenic rice plants Segregated into 3. 6: 1 ratio.

关 键 词：转基因籼稻 水稻 外壳蛋白抗病性 水稻条纹病毒 

分 类 号：S511.034[农业科学—作物学] S432.41