Cao Bihao, Meng Chengmin, Lei Jianjun, Chen Guoju
Horticulture Biotechnology Research Institute, Genetic and Quality Improved Center of Vegetable, South-China Agricultural University, Guangzhou 510642, China.
Sheng Wu Gong Cheng Xue Bao. 2008 May;24(5):881-6.
In order to induce male sterility of Brassica campestris L. subsp. chinensis Makino var. parachinensis, we introduced the chimeric pTA29-barnase gene into it by Agrobacteriumtume faciens transformation. We obtained the transgenic plants, and determined them by PCR, Southern blotting and RT-PCR analysis. Results indicated that the RNase (barnase) gene had been transferred into genome of plant, and its expression level was different among transformation plants. All transgenic plants were male sterile; there was no vigor or a little pollen without fertility in the anther of transgenic plants. The transgenic plants failed to produce seeds under the condition self-control pollination, but hybrid seeds set were obtained when these transgenic plants were cross-pollinated artificially with normal pollen from untransformed plants. Progeny from cross-pollinated maintainer line with transgenic plants segregated in the 1:1 for male sterility and male fertility, and these phenotypes corresponded directly to the presence or absence of the chimeri TA29-barnase gene. The male fertile plants of co-separated progenies could die by spraying 10 mg/L PPT in cotyledon seedling stage. The hybrid F1 between male sterility and other varieties showed heterosis in yield and growth. All these show that it is an efficient method to induce male sterility in Brassica campestris L. subsp. chinensis Makino var. parachinensis by TA29-barnase ene, there is potential on heterosis breeding of Brassica campestris L. subsp. chinensis Makino var. parachinensis.
为了诱导不结球白菜雄性不育,通过农杆菌介导法将嵌合的pTA29 - barnase基因导入不结球白菜中。获得了转基因植株,并通过PCR、Southern杂交和RT - PCR分析对其进行鉴定。结果表明,核糖核酸酶(barnase)基因已转入植物基因组中,且其在不同转化植株中的表达水平存在差异。所有转基因植株均表现为雄性不育;转基因植株的花药无活力或仅有少量无育性的花粉。转基因植株在自花授粉条件下不能产生种子,但当用未转化植株的正常花粉对这些转基因植株进行人工异花授粉时,可获得杂交种子。转基因植株与保持系杂交后代中,雄性不育和雄性可育的分离比例为1:1,这些表型与嵌合的TA29 - barnase基因的存在与否直接相关。共分离后代中的雄性可育植株在子叶期喷施10 mg/L的PPT后会死亡。雄性不育系与其他品种的杂交F1代在产量和生长方面表现出杂种优势。所有这些表明,利用TA29 - barnase基因诱导不结球白菜雄性不育是一种有效的方法,在不结球白菜杂种优势育种方面具有潜力。