Selection of Maize Inbred Lines with High Regeneration and Susceptibility to Agrobacterium tumifacien  

具有高再生性且对农杆菌敏感玉米自交系的筛选(英文)

作　　者：王钰[1] 付绍红 文颖[3] 张志明[1] 夏燕莉[1] 刘玉贞[1] 荣廷昭[1] 潘光堂[1] 

机构地区：[1]四川农业大学玉米研究所,雅安625014 [2]成都市第二农业科学研究所,温江611130 [3]四川省制糖糖料工业研究所,资中641200

出　　处：《Journal of Genetics and Genomics》2007年第8期749-755,共7页遗传学报（英文版）

基　　金：This work was supported by the National Natural Sciences Foundation of China (No. 30370889);the Program for Changjiang Scholars and Innovative Research Team in University of China (No. IRT0453);Beijing Agricultural Innovative Platform-Beijing Natural Science Fund Program;the National High-tech R&D Program of China (No. 2006 AA100103);the National Key Technolo-gies R&D Program (No. 2006 BAD01A03);the Program of the National Ministry of Agriculture (No. 2003-Q03)

摘　　要：Ten-maize inbred lines of maize （Zea mays L.） with high-induction rate and proliferation ability of embryonic calli were selected from 70-maize inbred lines by immature embryo culturing. Some of the embryonic calli were transferred onto regeneration medium to examine the ability of regeneration, some were transformed via Agrobacterium tumifaciens C58 carrying intron-β-glucuronidase （gus） gene, and GV3301 carrying the green fluorescent protein （gfp） gene to study the susceptibility of different genotypes in maize to A. tumifaciens. All embryonic calli initiated from 10-maize inbred lines were able to regenerate into plantlets, and the regeneration frequencies of inbred lines 6010, 6038, 6015, 6051, and 6060 were 61.11%, 31.94%, 45%, 33.33%, and 56.94%, respectively, which were higher than that of other lines. Analysis of variance indicated that the susceptibility of the various genotypes in maize to A. tumifacien C58 showed a significant difference among each other, and the inbred lines 6010, 6015, 6051, 6050, 6058, 6060, 6069, 6077 were susceptible to A. tumifacien C58, of which frequency of gus expression were over 70%. Expression of GFP was observed in six-inbred lines （6050, 6015, 6051, 6058, 6069, 6077）. The inbred lines 6051, 6010, 6015, 6060, and 6050 had the high regeneration and the susceptibility to A. tumifaciens C58; and the inbred lines 6051, 6015, and 6060 had the high regeneration and the susceptibility to Agrobacterium tumifaciens GV3301.以 70 个高代玉米自交系的幼胚为材料在 NB 培养基上进行离体培养,通过多次继代选择,从中筛选出了胚性愈伤组织诱导率高、克隆能力强的 10 个自交系,将这 10 个自交系的一部分胚性愈伤组织用作绿苗再分化,研究其再生能力;另一部分用农杆菌 C58 和 GV3301 转化,利用植物载体中携带的 gus 和 gfp 报告基因的瞬时表达为指标研究玉米基因型对农杆菌的敏感性。绿苗再分化结果表明,10 个自交系都具有绿苗再分化能力,其中自交系 6010、6038、6015、6051 和 6060的绿苗再分化力相对较高,分别为 61.11%、31.94%、45%、33.33%和 56.94%。gus 瞬时表达率方差分析结果表明:玉米基因型对农杆菌 C58 的敏感性存在极显著差异,在自交系 6034、6038 的胚性愈伤组织上没有检测到 gus 瞬时表达,即这两个基因型对农杆菌 C58 不敏感,不能被其转化;其他 7 个基因型的平均 gus 瞬时表达率均大于 70%,说明这 7 个基因型对农杆菌 C58 敏感。GFP 荧光检测结果表明,在自交系 6034、6038、6069 和 6010 的胚性愈伤组织上没有检测到 gfp 瞬时表达,在其他 6 个自交系上检测到绿色荧光蛋白的表达。因此,认为自交系 6015、6051 和 6060 是对农杆菌 GV3301 敏感且具有高再生能力的玉米转基因受体材料;自交系 6051、6010、6015、6060 和 6050 是再生能力强且对农杆菌 C58 敏感的玉米转基因受体材料。

关 键 词：maize inbred lines embryogenic calli Agrobacterium tumifacien GFP GUS 

分 类 号：S513[农业科学—作物学]