CAJ | 학술논문

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Parasponia andersonii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%。为水稻与根瘤菌的相互作用和固氮作用的可能性研究奠定了一定的基础。
위료확대근류균적숙주범위화시탐근류균재비두과식물상적고담작용,장완두응집소기인(pl)화Parasponia andersonii혈홍단백기인(phb)구건재동일개식물표체재체상,용기인창법장기도입수도(OryzasativaL.ssp.japonica)。경PCR확증화Southern잡교분석，증명외원목적기인이정합도수도기인조중。GUS조직화학염색급완두응집소기인적Western인적실험화표체산물적원위잡교，증실외원기인재전기인수도중표체。재40개전화식주중18주유pl화phb기인적PCR산물,득솔위45%。재용18주식물주pl기인적Westernblot검측,유3주유번역표체,점40주적7.5%，18주적17%。위수도여근류균적상호작용화고담작용적가능성연구전정료일정적기출。
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 (phb) 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 pl and phb genes were integrated into nucleic genome of the transformed rice plants. GUS activity and the product of the pl 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 pl 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 bv. viceae could extend its host range and make the transgenic rice plants have the possibility of being symbiotic, or associative to nitrogen fixation.