Agracetus, 8520 University Green, Middleton, WI 53562.
Proc Natl Acad Sci U S A. 1989 Oct;86(19):7500-4. doi: 10.1073/pnas.86.19.7500.
DNA-coated gold particles were introduced into meristems of immature soybean seeds using electric discharge particle acceleration to produce transgenic fertile soybean plants. The lineages of integrated foreign DNA in two independently transformed plants were followed in the first (R(1)) and second (R(2)) generation of self-pollinated progeny. One plant (4615) was transformed with the Escherichia coli genes for beta-glucuronidase and neomycin phosphotransferase II; the other (3993) was transformed only with the gene for beta-glucuronidase. Segregation ratios for the introduced gene(s) were approximately 3:1 for plant 4615 and 1:1 for plant 3993 in the R(1) generation. DNA analysis showed 100% concordance between presence of the foreign gene sequences and enzyme activity. Moreover, all copies of the foreign genes are inherited as a unit in each plant. Plant 3993 segregated in a 1:1 ratio in the R(2) generation. R(1) plants derived from plant 4615, which expressed both genes, gave either 100% or 3:1 expression of both genes in the R(2) generation, demonstrating recovery of both homozygous and heterozygous R(1) plants. Our results show that foreign DNA introduced into soybean plants using electric discharge particle acceleration can be inherited in a Mendelian manner. Results also demonstrate cotransformation of tandem markers and show that both markers are inherited as closely linked genes in subsequent generations. These results indicate that whole plants can be derived from single transformed cells by a de novo organogenic pathway.
采用放电粒子加速将 DNA 包覆的金颗粒导入未成熟大豆种子的分生组织中,从而产生转基因可育大豆植株。将整合的外源 DNA 谱系在自花授粉后代的第一(R(1))和第二(R(2))代中进行追踪。一株(4615)转化了大肠杆菌的β-葡萄糖醛酸酶和新霉素磷酸转移酶 II 基因;另一株(3993)仅转化了β-葡萄糖醛酸酶基因。在 R(1)代中,对于 4615 号植物,引入的基因的分离比约为 3:1;对于 3993 号植物,分离比为 1:1。DNA 分析表明,外源基因序列和酶活性之间存在 100%的一致性。此外,在每株植物中外源基因都作为一个单位遗传。在 R(2)代中,3993 号植物以 1:1 的比例分离。从 4615 号植物衍生而来的 R(1)代植物,表达了两种基因,在 R(2)代中均以 100%或 3:1 的比例表达,表明恢复了两种纯合和杂合的 R(1)代植物。我们的结果表明,通过放电粒子加速将外源 DNA 导入大豆植物中,可以按照孟德尔遗传方式进行遗传。结果还表明串联标记的共转化,并表明在随后的世代中,两个标记作为紧密连锁的基因遗传。这些结果表明,整个植物可以通过从头发生的器官发生途径从单个转化细胞中衍生而来。