黑龙江粳稻再生体系建立的影响因素研究

Study on Factors of Establishment for Heilongjiang Japonica Rice Regeneration System

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。

To establish Heilongiang japonica rice regeneration system,mature embryos of Dongnong 427 and Dongnong428 were selected as experimental materials,and the effect of various factors including pre-soaked embryos,cold treatment,different mediums,different coagulants,ABA,glutamine,activated carbon,silver nitrate,2,4- D and 6-BA concentrations on rice regeneration system were studied. The results showed that the culture ability of Dongnong 427 were increased by 6% and 15%,respectively,after pre-soaked embryos and cold treatment,the optimum medium was NB. The addition of ABA and glutamine were suitable in the stage of differentiation and induction; the differentiation ratewere improved by 38% and 9%,respectively,after the addition of activated carbon and silvernitrate. The optimum concentration of 2. 4- D and 6- BA were 1mg·L^-1 and 4mg·L^-1,respectively. While for Dongnong 428,the culture ability were raised by 14% and 9%,respectively,after pre-soaked embryos and cold treatment,MS was the best medium. ABA was suitably added at induction phase,glutamine was suitably added in the whole culture stage,the addition of activated carbon and silvernitrate during differentiationcould increase the regeneration rate by 1% and 9%,respectively and the most suitable concentration of 2,4- D and 6- BA were 1mg·L^-1 and 3mg·L^-1. Agar was selected as coagulant in the induction stage,while Phytagel was selected in the differentiation stage for both of them. This study would provide a basis for further optimization of regeneration system of rice,and lay the foundation for the subsequent establishment of transformation system of rice.