Transformation of Pea Lectin Gene and Parasponia Haemoglobin Gene into Rice and Their Expressions  

豌豆凝集素和血红蛋白基因对水稻的转化和表达(英文)

作　　者：张静娴[1] 王忆平[2] 沈世华[1] 王逸群[1] 高越峰[1] 单雪琴[1] 荆玉祥[1] 

机构地区：[1]中国科学院植物研究所,北京100093 [2]北京大学生命科学学院,北京100871

出　　处：《Acta Botanica Sinica》2001年第3期267-274,共8页Acta Botanica Sinica（植物学报：英文版）

基　　金：NationalNaturalScienceFoundationofChina (3 95 70 194)

摘　　要：Lectin and leghemoglobin in legumes play the important roles, respectively, in recognition of host plants to their rhizobial bacteria, and lowering the oxygen partial pressure around bacteroids and protecting nitrogenase from oxygen in symbiotic nitrogen-fixing nodules. In order to extend the host range of the rhizobial bacteria and to make them fix nitrogen in non-legumes, pea lectin gene (pl) and Parasponia hemoglobin gene ( phl,) have been constructed into a plant expression vector (pCBHUL) and the vector pCBHUL was introduced into rice calli from immature young embryos by particle bombardment. After the calli were regenerated into plantlets on the resistant-selecting media containing hygromycin, they were identified by PCR and Southern blot hybridization. It was indicated that the pi and phb genes were integrated into nucleic genome of the transformed rice plants. GUS activity and the product of the pi gene were determined by GUS staining, Western blot and in situ hybridization at translational level. Eighteen out of 40 plants resistant to hygromycin were positively identified by PCR analysis with the rate of 45%. The pi gene was expressed in 3 out of 18 plants with 17% and 7.5% in 40 plants. The results may provide a clue for exploring whether Rhizobium leguminosarum by. viceae could extend its host range and make the transgenic rice plants have the possibility of being symbiotic, or associative to nitrogen fixation.为了扩大根瘤菌的宿主范围和试探根瘤菌在非豆科植物上的固氮作用 ,将豌豆凝集素基因 (pl)和Parasponiaandersonii血红蛋白基因 (phb)构建在同一个植物表达载体上 ,用基因枪法将其导入水稻 (OryzasativaL .ssp .japonica)。经PCR扩增和Southern杂交分析 ,证明外源目的基因已整合到水稻基因组中。GUS组织化学染色及豌豆凝集素基因的Western印迹实验和表达产物的原位杂交 ,证实外源基因在转基因水稻中表达。在 40个转化植株中 18株有pl和phb基因的PCR产物 ,得率为 45 %。再用 18株植物做pl基因的Westernblot检测 ,有 3株有翻译表达 ,占 40株的 7.5 % ,18株的 17%。

关 键 词：Parasponia hemoglobin gene pea lectin gene particle bombardment transformation gene expression transgenic rice 

分 类 号：Q943.2[生物学—植物学]