Peng J, Wen F, Lister R L, Hodges T K
Department of Botany and Plant Pathology, Purdue University West Lafayette, IN 47907.
Plant Mol Biol. 1995 Jan;27(1):91-104. doi: 10.1007/BF00019181.
Inheritance of foreign genes neo and gusA in rice (Oryza sativa L. cv. IR54 and Radon) has been investigated in three different primary (T0) transformants and their progeny plants. T0 plants were obtained by co-transforming protoplasts from two different rice suspension cultures with the neomycin phosphotransferase II gene [neo or aph (3') II] and the beta-glucuronidase gene (uidA or gusA) residing on separate chimeric plasmid constructs. The suspension cultures were derived from callus of immature embryos of indica variety IR54 and japonica variety Radon. One transgenic line of Radon (AR2) contained neo driven by the CaMV 35S promoter and gusA driven by the rice actin promoter. A second Radon line (R3) contained neo driven by the CaMV 35S promoter and gusA driven by a promoter of the rice tungro bacilliform virus. The third transgenic line, IR54-1, contained neo driven by the CaMV 35S promoter and gusA driven by the CaMV 35S. Inheritance of the transgenes in progeny of the transgenic rice was investigated by Southern blot analysis and enzyme assays. Southern blot analysis of genomic DNA showed that, regardless of copy numbers of the transgenes in the plant genome and the fact that the two transgenes resided on two different plasmids before transformation, the introduced gusA and neo genes were stably transmitted from one generation to another and co-inherited together in transgenic rice progeny plants derived from self-pollination. Analysis of GUS and NPT II activities in T1 to T2 plants provided evidence that inheritance of the gusA and neo genes was in a Mendelian fashion in one plant line (AR2), and in an irregular fashion in the two other plant lines (R3 and IR54-1). Homozygous progeny plants expressing the gusA and neo genes were obtained in the T2 generation of AR2, but the homozygous state was not found in the other two lines of transgenic rice.
在三个不同的初级（T0）转基因植株及其子代植株中，研究了水稻（Oryza sativa L. cv. IR54和Radon）中外源基因neo和gusA的遗传情况。T0植株是通过将来自两种不同水稻悬浮培养物的原生质体与新霉素磷酸转移酶II基因[neo或aph(3')II]和位于单独嵌合质粒构建体上的β-葡萄糖醛酸酶基因（uidA或gusA）共转化获得的。悬浮培养物源自籼稻品种IR54和粳稻品种Radon未成熟胚的愈伤组织。Radon的一个转基因株系（AR2）包含由CaMV 35S启动子驱动的neo和由水稻肌动蛋白启动子驱动的gusA。第二个Radon株系（R3）包含由CaMV 35S启动子驱动的neo和由水稻东格鲁杆状病毒启动子驱动的gusA。第三个转基因株系IR54-1包含由CaMV 35S启动子驱动的neo和由CaMV 35S驱动的gusA。通过Southern杂交分析和酶活性测定研究了转基因水稻子代中转基因的遗传情况。基因组DNA的Southern杂交分析表明，无论植物基因组中转基因的拷贝数以及转化前两个转基因位于两个不同质粒上的事实，导入的gusA和neo基因都能稳定地从一代传递到另一代，并在自花授粉产生的转基因水稻子代植株中共同遗传。对T1至T2植株中GUS和NPT II活性的分析提供了证据，表明gusA和neo基因在一个株系（AR2）中以孟德尔方式遗传，而在另外两个株系（R3和IR54-1）中以不规则方式遗传。在AR2的T2代中获得了表达gusA和neo基因的纯合子代植株，但在另外两个转基因水稻株系中未发现纯合状态。
