高粱茎尖再生体系及其遗传转化影响因子的研究

本文对高粱茎尖再生体系及其转化影响因子进行了研究。不同基因型的茎尖再生频率有很大差异,说明基因型是影响茎尖再生的一个重要因素。在9种不同的激素组合中,以0.25MG/L KT、0.5MG/LBAP和0.25MG/L IAA对茎尖的再生效果最好。在MS培养基的基础上,采用B5培养基维生素并添加200MG/L L-ASN,10MG/L抗坏血酸和100MG/L PVP可提高茎尖再生频率。对农杆菌介导高粱茎尖转化的因子进行了优化,获得较高转化频率的条件为:茎尖预培养时间3D,在OD600值为0.5农杆菌菌液中感染10MIN,共培养3D。

REGENERATION SYSTEM AND INFLUENCING FACTORS FOR GENETIC TRANSFORMATION OF SORGHUM SHOOT APEX

Regeneration system and influencing factors for genetic transformation of sorghum shoot apex were studied.Regeneration frequency varied obviously with genotypes,suggesting that the genotype is one of the important factors to the regeneration of shoot apex.Among 9 different combination of regulator tested,the 0.25mg/L KT combined with 0.5mg/L BAP and 0.25mg/L IAA was the best for the generation.The regeneration frequency of sorghum shoot apex was also increased by the supplement of 10mg/L ascorbic acid,200mg/L PVP,100mg/L L-Asn and vitamins as listed in B_5 to the MS medium.Factors affecting transformation of shoot apex mediated by Agrobacterium tumefacien were also optimized.The highest transformation frequency indicated by transient expression of gus gene was detected under the conditions of pre-culture for 3 days and subsequently infection for about 10 minutes in Agrobacterium suspension with OD_(600) value about 0.5 followed by co-culture for 3 days.