西瓜未成熟胚高效再生体系的建立
2014-04-29董焱张洁张海英宫国义郭绍贵任毅许勇
董焱 张洁 张海英 宫国义 郭绍贵 任毅 许勇
摘 要:为了建立西瓜高效遗传转化的再生体系,以西瓜杂交品种‘秀玲、自交系‘97103和野生种质‘PI296341的未成熟胚为外植体,分析了基因型、未成熟胚发育时间和植物生长调节剂等因素对体外再生率的影响,获得了西瓜离体培养再生植株。结果表明,以授粉后24 d的西瓜未成熟胚为外植体,在MS+SH维生素+6-BA 2.2 mg·L-1培养基上,‘秀玲、‘97103和‘PI296341的未成熟胚外植体能够诱导出不定芽,不定芽诱导率为87.78%、82.78% 和86.11%,不定芽在MS+SH维生素+6-BA 0.2 mg·L-1+IAA 0.02 mg·L-1培养基上伸长、展叶,在MS+IBA 1.0 mg·L-1的培养基上诱导生根得到完整的再生植株。以未成熟胚为外植体获得再生植株的途径,可为西瓜遗传转化提供有效的受体系统。
关键词: 西瓜; 未成熟胚; 再生
Plant Regeneration from Immature Embryos of Watermelon
DONG Yan1,2, ZHANG Jie2, ZHANG Hai-ying2, GONG Guo-yi2, GUO Shao-gui2, REN Yi2, XU Yong1,2
(1.Plant Science and Technology College, Beijing University of Agriculture, Beijing 102206 China; 2. Beijing Vegetable Research Center, Beijing Academy of Agriculture and Forestry Sciences, Beijing 100097 China)
Abstract:In order to obtain watermelon regeneration system for genetic transformation,the immature embryos of watermelon ‘Feeling,‘97103and‘PI296341were taken as explants in this experiment. The impact the immature embryo development time,genotypes and plant growth regulators on the rate of regeneration were explored and regenerated plantlets were obtained. Adventitious buds were regenerated in MS+SH Vitamin+6-BA 2.2 mg·L-1,and the regeneration rates were 87.78%,82.78% and 86.11% for 3 genotypes tested. The regenerated buds grew into shoots in the MS medium with 6-BA 0.2 mg·L-1+IAA 0.02 mg·L-1,and rooted in MS+IBA 1.0 mg·L-1. The result indicated that the plantlet regeneration system from immature embryos can be a proper system for genetic transformation in watermelon.
Key words: Watermelon; Immature embryos; Regeneration
西瓜(Citrullus lanatus)为葫芦科(Cucurbitaceae)西瓜属一年生草本蔓生植物,是一种重要的园艺作物。西瓜遗传基础狭窄,种质资源缺乏,很多重要的性状无法通过常规育种方法进行改良。基因工程技术就成为对常规育种的有效补充手段。近年来,植物基因工程的理论和实践逐步走向成熟和完善,并成为现代生物技术的核心领域,亦是农作物品种改良的重要手段。转基因能够定向改良西瓜性状,并且是基因功能验证的重要手段,而开展这些工作的前提是建立西瓜高效体外再生系统。
国内外许多学者已经相继采用顶芽[1]、子叶[2]、下胚轴[3]、花药[4]和原生质体[5]等进行西瓜组织培养和遗传转化技术。但是与番茄等园艺植物相比,西瓜体外再生能力弱、不容易通过愈伤组织途径获得再生植株,存在着高效基因型少、再生频率低、分化芽伸长慢、易出现玻璃化苗等问题[6-8],此外,不同研究者所得的结论存在很大的差异,重复性差,目前还缺乏对西瓜体外再生技术的系统研究。……
