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为了建立黑龙江粳稻组织培养高效再生体系,以东农427和东农428的成熟胚为材料,探讨预先浸胚、低温处理、培养基种类、凝固剂种类、ABA、谷氨酰胺、活性炭、硝酸银、2,4-D浓度以及6-BA浓度等因素对水稻再生体系建立的影响。结果表明,东农427经预先浸胚和低温处理可使培养力分别提高6%和15%,最适培养基为NB;ABA和谷氨酰胺分别适合在分化和诱导阶段添加,分化时添加活性炭和硝酸银分别使分化率提高38%和9%;最适2,4-D和6-BA分别为1mg·L~(-1)和4mg·L~(-1)。东农428经预先浸胚和低温处理可使培养力分别提高14%和9%,最适培养基为MS;ABA适合在诱导阶段添加,谷氨酰胺适合在整个培养阶段添加;分化时添加活性炭和硝酸银分别使分化率提高1%和9%;最适2,4-D和6-BA分别为1mg·L~(-1)和3mg·L~(-1)。2个品种的凝固剂在诱导阶段宜采用琼脂,分化阶段宜采用Phytagel。本研究为进一步优化水稻再生体系提供了依据,为后续水稻遗传转化奠定了基础。
In order to establish an efficient regeneration system for tissue culture of Japonica rice in Heilongjiang Province, the mature embryos of Dongnong 427 and Dongnong 428 were used as material to investigate the effects of pre-soaking, low temperature treatment, medium types, coagulant types, ABA, glutamine, activated carbon, , 2,4-D concentration and 6-BA concentration on the establishment of rice regeneration system. The results showed that the cultivating ability of Dongnong 427 increased by 6% and 15% respectively with pre-soaking and low temperature treatment, and the optimum medium was NB. ABA and glutamine were suitable for differentiation and induction stage, respectively. Activated carbon And silver nitrate increased the differentiation rate by 38% and 9%, respectively. The optimal 2,4-D and 6-BA concentrations were 1 mg · L -1 and 4 mg · L -1, respectively. Dongnong 428 could enhance the culture ability by 14% and 9%, respectively, with pre-soaking embryos and low temperature treatment. The optimal medium was MS. ABA was suitable for the induction stage and glutamine was suitable for the whole culture stage. Activated carbon And silver nitrate increased the rate of differentiation by 1% and 9%, respectively. The optimal 2,4-D and 6-BA were 1 mg · L -1 and 3 mg · L -1, respectively. Two varieties of coagulant should be used in the induction stage of agar, differentiation stage should adopt Phytagel. This study provided the basis for further optimization of rice regeneration system and laid the foundation for subsequent rice genetic transformation.