摘要
以摩尔多瓦葡萄早期体细胞胚为受体,通过农杆菌介导辅以超声波处理转化hGLP1基因。结果表明,摩尔多瓦葡萄体细胞胚的卡那霉素临界致死质量浓度为10mg/L,农杆菌浸染时辅以超声波处理可显著提高GUS瞬时表达效率(高达69.4%)。采用此体系对摩尔多瓦葡萄进行转化,获得高瞬时表达和稳定表达的转化子,进一步筛选培养获得7株抗性再生植株。利用PCR扩增的方法进行检测,其中2株PCR检测呈阳性,初步证实hGLP1基因已整合到摩尔多瓦葡萄基因组。
ThehGLP1 gene was introduced into grapes through Agrobactrium tumefaciens with soni- cation-assisted treatment using Moldova grapes early somatic embryos as the target tissue. The results showed that Kanamycin critical lethal mass concentration of Moldova grape somatic embryos was 10 mg/L. Transient expression efficiency of GUS was enhanced greatly by Agrobacterium tumefaciens supplemented with ultrasonic treatment, reached up to 69.4%. Moldova grapes were transformed by this system, high transient expression and stable expression transformants were obtained, and seven resistant regenerated plants were obtained with further screening culture. PCR amplification analysis on these resistant shoots showed that there were two PCR positive plants, and confirmed thathGLP1 gene was integrated into the Moldova grape genome. These results will provide useful information for establishment of an efficient transformation method of grape.
出处
《西北农业学报》
CAS
CSCD
北大核心
2013年第2期131-135,共5页
Acta Agriculturae Boreali-occidentalis Sinica
基金
三峡大学人才科研启动基金(KJ2012B023)
国家林业局948项目(2006-4-73)
关键词
葡萄
转基因
农杆菌介导
超声波处理
Grape
Transgene
Agrobacterium-mediated
Ultrasonic treatment
