CAJ | 학술논문

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.
이70개고대옥미자교계적유배위재료재NB배양기상진행리체배양,통과다차계대선택,종중사선출료배성유상조직유도솔고、극륭능력강적10개자교계,장저10개자교계적일부분배성유상조직용작록묘재분화,연구기재생능력;령일부분용농간균C58화GV3301전화,이용식물재체중휴대적gus화gfp보고기인적순시표체위지표연구옥미기인형대농간균적민감성.록묘재분화결과표명,10개자교계도구유록묘재분화능력,기중자교계6010、6038、6015、6051화6060적록묘재분화력상대교고,분별위61.11%、31.94%、45%、33.33%화56.94%.gus순시표체솔방차분석결과표명:옥미기인형대농간균C58적민감성존재겁현저차이,재자교계6034、6038적배성유상조직상몰유검측도gus순시표체,즉저량개기인형대농간균C58불민감,불능피기전화;기타7개기인형적평균gus순시표체솔균대우70%,설명저7개기인형대농간균C58민감.GFP형광검측결과표명,재자교계6034、6038、6069화6010적배성유상조직상몰유검측도gfp순시표체,재기타6개자교계상검측도록색형광단백적표체.인차,인위자교계6015、6051화6060시대농간균GV3301민감차구유고재생능력적옥미전기인수체재료;자교계6051、6010、6015、6060화6050시재생능력강차대농간균C58민감적옥미전기인수체재료.
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.