LI Shuang,YUAN Lin,JIA Gui-xia.Study on the Pollen-tube Pathway Transformation in Lily[J].HEILONGJIANG AGRICULTURAL SCIENCES,2011,(08):15-18.
百合花粉管通道法遗传转化的研究
- Title:
- Study on the Pollen-tube Pathway Transformation in Lily
- 文章编号:
- 1002-2767（2011）08-0015-04
- 关键词:
- 花粉管通道途径; 绿色荧光蛋白（GFP）; 百合
- Keywords:
- pollen-tube pathway; GFP; lily
- 分类号:
- S644
- 文献标志码:
- A
- 摘要:
- 为了提高百合花粉管通道法遗传转化效率，采用pBI121-GFP-FT载体，对外源基因携带形式、导入方式和导入时间进行了研究。对质粒和农杆菌两种形式携带的外源基因的研究结果表明：菌液形式携带外源基因有利于百合花粉管通道法遗传转化。采用切割柱头授粉后涂抹外源基因、常规授粉后涂抹外源基因以及子房注射外源基因3种导入方式进行百合遗传转化，结果表明常规授粉后涂抹外源基因更适宜百合花粉管通道法遗传转化。对授粉后0 h涂抹外源基因和授粉后延迟3 h涂抹外源基因2种处理进行比较研究，结果表明授粉后0 h涂抹外源基因更适宜于百合花粉管通道法遗传转化。将试验所得种子进行荧光观察，转基因植株有绿色荧光。PCR检测结果表明GFP-FT基因已经整合到百合基因组中。
- Abstract:
- In order to improve transgenic efficiency,the expression vector pBI121-GFP-FT was transformed into lily using pollen-tube pathway method and the effects of exogenous vector type,introduction way and introduction time（after pollination）were discussed.The effects of Agrobacterium and plasmid were compared.The results showed that exogenous vector type of Agrobacterium was better for lily transformation.Three introduction methods were carried out,including cutting stigma pollination and conventional pollination followed by exogenous vector and injecting exogenous vector into ovary.The results indicated that smearing exogenous vector after conventional pollination was suitable for lily transformation.Different introduction times after pollination for 0 h and 3 h were considered,results indicated that 0 h was the optimization introduction time.The green fluorescence was observed in the seeds under fluorescent microscope.Results of PCR also proved that the GFP-FT gene had integrated into lily genome.
参考文献/References:
[1]杜捷.兰州百合外源DNA花粉管通道法导入的初步研究[D].兰州：西北师范大学，2003. [2]崔红,李朝霞,时向东,等.花粉管通道法及其在烟草育种中的应用综述[J].烟草科技/栽培与调制,2003（3）:34-36. [3]Luo Z X,Wu R.A simple method for the transformation of rice via the pollen tube pathway[J].Plant Mol. Biol. Rep.,1988,6（3）:165-174. [4]Ishii M,Kunimura J S,Penna T C V,et al.Study on the thermal stability of green fluorescent protein（GFP）in glucose parenteral formulations[J].International journal of Pharmaceutics,2007,7（1-2）:109-117. [5]张宁,侯喜林,余丽芸,等.重组GFP干酪乳杆菌的构建及其在小鼠肠道内的定植分布[J].微生物学报,2010,50（9）:1232-1238. [6]黄国存,董越梅,孙敬三.以GFP为标记用花粉管通道途径导入外源DNA[J].科学通报,1998,43（23）:2531-2534. [7]曾君祉,吴有强,王东江,等.花粉管通道（或运载）法转化的植株后代遗传表现及转化机理的探讨[J].科学通报,1998,43（6）:561-566. [8]谢道昕,范云六,倪万潮,等.苏云金杆菌（Bacillus thuringiensis）杀虫晶体蛋白基因导入棉花获得转基因植株[J].中国科学（B辑）,1991（4）:367-373. [9]刘芳,袁英,高树仁,等.花粉管通道导入技术在农作物分子育种中的应用[J].农业与技术,2007,27（4）:42-43. [10]李卉,武天龙.CaCl2诱导大豆花粉管通道农杆菌转基因研究[J].大豆科学,2007,26（1）:55-59. [11]刘春,穆鼎,明军,等.百合种间杂交受精前障碍的研究[J].园艺学报,2006,33（3）:653-656.
备注/Memo
基金项目：国家“863”计划资助项目（2008AA10Z156）；“十一五”国家科技支撑计划资助项目（2009BADB8B04）；北京市教育委员会产学研合作资助项目