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维持良好的愈伤组织状态和提高分化率是栽培稻(Oryza sativa L.)转基因成功的重要因素。本实验研究了培养基中添加山梨醇和添加不同比例的蔗糖与山梨醇对粳型旱稻(Oryza sativa L.ssp.japonica)297和籼型水稻(Oryza sativa L.ssp.indica)9311两品种成熟胚愈伤组织分化的影响,分别比较了愈伤组织诱导、继代和分化阶段添加不同浓度山梨醇和分化阶段不同蔗糖浓度与山梨醇配比对愈伤组织分化率和转基因效率的影响,结果发现,两个品种都以3个阶段分别添加山梨醇10、20和40 g/L即10-20-40的处理获得最高分化率,分别为88.9%和86.1%,比对照(3个阶段都不加山梨醇)提高了45.1%和32.6%。此结果也在旱稻297 Activator(Ac)基因转化中得到验证。此外,不管愈伤组织诱导和继代阶段是否添加山梨醇,分化阶段蔗糖为5.5 g/L和山梨醇为40 g/L的配比都使愈伤组织分化率高达82.7%~95.2%,愈伤组织呈紧实鲜黄,比其他处理有更多的绿芽点。同时该处理还使9311愈伤组织褐化得到抑制。这些结果表明,山梨醇能够使愈伤组织保持良好的状态、抑制籼稻愈伤组织褐化、提高分化率和转基因效率,这一结果对栽培稻乃至其他作物的组织培养和转基因绿苗再生具有一定参考价值。
Maintaining good callus status and increasing the rate of differentiation is an important factor in the success of transgenic rice in Oryza sativa L. In this study, we studied the effects of adding sorbitol and adding different proportions of sucrose and sorbitol on the mature embryos of two varieties of Oryza sativa L.ssp. Japonica 297 and Oryza sativa L.ssp.indica 9311 The effects of different concentrations of sorbitol and differentiation stages of sucrose and sorbitol on callus differentiation rate and transgene efficiency were compared among callus induction, subculture and differentiation stages, respectively. The results showed that, The two cultivars were all treated with sorbitol at 10, 20 and 40 g / L in 10 stages, ie, 10-20-40 in 3 stages, respectively, with the highest differentiation rates of 88.9% and 86.1%, respectively. Compared with the control Sorbitol) increased by 45.1% and 32.6%. This result was also validated in Upland 297 Activator (Ac) gene transformation. In addition, the rate of callus differentiation reached as high as 82.7% ~ 95.2% with the sucrose concentration of 5.5 g / L and the sorbitol concentration of 40 g / L at the differentiation stage irrespective of whether sorbitol was added during callus induction and subculture Wound tissue was firm yellow, more green shoots than other treatments. At the same time, the treatment also inhibited browning of 9311 callus. These results show that sorbitol can maintain callus in good condition, inhibit browning of indica rice, improve differentiation rate and transgene efficiency, which has certain effects on tissue culture and regeneration of transgenic green shoots in cultivated rice and other crops Reference value.