摘要
以香蕉(Musa spp.)品种‘天宝蕉’(Musa spp.,AAA类群)为试材,对以根癌农杆菌介导法的香蕉遗传转化体系进行较全面的研究,并以该系统进行了ACS反义基因转化香蕉的研究。结果表明:不经预培养的香蕉茎尖横切薄片,侵染前用附加0.1mg/L甘露醇的高渗固体培养基前处理4h,农杆菌重悬液浓度为0D。在1.0左右,重悬液中含100g/L蔗糖,接菌时间为10~15min,于26℃黑暗条件下共培养4d,共培养培养基pH值为5.8是较为适合的转化条件:采用附加100mg/L卡那霉素、2mg/LAgNO,筛选培养基对共培养后的香蕉横切薄片进行筛选,共获得5个转ACS反义基因的抗性芽系;经GUS组织化学法及PCR检测,gus基因已整合进香蕉基因组.
A high efficiency and systematic transgenic procedure mediated by Agrobacterium was developed in Musa spp. cv. Tianbaojiao (AAA group). The introduction of antisense A CS gone to banana was conducted, and the optimized parameters used in Agrobacterium-mediated transformation were obtained. The best transient expression of GUS was obtained under the conditions as follows: the thin cross-sections which were not precultured but were pretreated with 0.1 mol/L mannitol for 4 h, and the Agrobacterium suspension was with OD600 value of 1.0 and supplemented with 100 g/L sucrose, the samples were infected for 10-15 mins, and then they were transferred onto the non-selective medium (pH5.8) supplemented with 0.1 mg/L NAA and 1.0 mg/L BA for co-culture for 4 days at 26 ℃ in the dark. After co-culture, the thin cross-sections were selected on MS medium supplemented with 100 mg/L kanamycin and 2 mg/L AgNO3, which could be used to inhibit Agrobacterium contamination and be good for bud differentiation. At last, 5 resistant bud lines were maintained for further studies. The GUS histochemical and PCR assays proved that the gus gene had integrated into the genome of the 5 resistant bud lines.
出处
《热带作物学报》
CSCD
北大核心
2014年第1期17-23,共7页
Chinese Journal of Tropical Crops
基金
国家香蕉产业体系专项资金(No.CARS-32-11)
福建省农业科技平台(No.2008N2001)
国家科技支撑计划(No.2007BAD07B01)
关键词
香蕉
横切薄片
根癌农杆菌
ACS反义基因
转化
Banana
Thin-cross-section
Agrobacterium-mediated
Antisense A CS gene
Transformation