CAJ | 학술논문

高效遗传转化体系是研究籼稻分子设计育种和功能基因的基础。文章以西南地区4个重要籼稻恢复系乐恢188、万恢88、Q2和蜀恢527成熟胚为外植体，NB（Nutrient Broth）培养基为基本培养基，通过改变激素配比，研究4个籼稻恢复系的愈伤组织诱导、转化和再生体系。结果表明：添加3．0mg/L2，4-D是诱导愈伤组织的最适处理条件，出愈率达75．4％～93．3％；添加3．0g/L山梨醇、5．0mg/LKT、1．0mg/LNAA、3．0mg/L6-BA和3．0mg/L2，4-D可明显提高万恢88、乐恢188的分化率；添加10mg/LKT和0．4mg/LNAA能显著增强Q2的分化能力；以N6、N6D培养基组合后再添加10mg/LKT和0．4mg/LNAA是蜀恢527的最佳分化培养基。采用上述培养基进行遗传转化，4种籼稻材料植株再生率可达到38．5％～65．9％，转基因阳性率70％～90％。在此基础上利用农杆菌介导法将拟南芥热激蛋白基因HSP101导入万恢88，获得了耐热性比其野生型提高的转基因籼稻植株。
고효유전전화체계시연구선도분자설계육충화공능기인적기출。문장이서남지구4개중요선도회복계악회188、만회88、Q2화촉회527성숙배위외식체，NB（Nutrient Broth）배양기위기본배양기，통과개변격소배비，연구4개선도회복계적유상조직유도、전화화재생체계。결과표명：첨가3．0mg/L2，4-D시유도유상조직적최괄처리조건，출유솔체75．4％～93．3％；첨가3．0g/L산리순、5．0mg/LKT、1．0mg/LNAA、3．0mg/L6-BA화3．0mg/L2，4-D가명현제고만회88、악회188적분화솔；첨가10mg/LKT화0．4mg/LNAA능현저증강Q2적분화능력；이N6、N6D배양기조합후재첨가10mg/LKT화0．4mg/LNAA시촉회527적최가분화배양기。채용상술배양기진행유전전화，4충선도재료식주재생솔가체도38．5％～65．9％，전기인양성솔70％～90％。재차기출상이용농간균개도법장의남개열격단백기인HSP101도입만회88，획득료내열성비기야생형제고적전기인선도식주。
High efficiency transformation and regeneration of Indica rice is a vital foundation for molecular breeding and function analysis of Indica-specific genes .In this paper ,embryogenic callus from four popular Indica restorer lines in southwest China ,Lehui 188 ,Wanhui 88 ,Q2 and Shuhui 527 ,were used to explore callus induction ,transformation and regeneration by using different combinations of plant hormones in culture media .It was shown that the callus of four lines were best initiated on Nutrient Broth (NB) medium contai-ning 3.0 mg/L of 2 ,4-D with high induction frequency from 75.4% to 93.3% .The differentiation efficiency of Wanhui 88 and Lehui 188 was significantly improved by the combination of 3.0 g/L sorbitol , 5.0 mg/L KT ,1.0 mg/L NAA ,3.0 mg/L 6-BA and 3.0 mg/L 2 ,4-D ,while the differentiation efficiency of Q2 was effectively promoted by the combination of 10 mg/L KT and 0.4 mg/L NAA .The N6 and N6D medium combination with the additions of 10 mg/L KT and 0.4 mg/L NAA was the optimal differentiation medium for Shuhui 527 .The plantlet regeneration frequency could reach 38.5%-65.9% ,and the observed transformation frequency ranged from 70% to 90% .Together ,the highly efficient transformation and regen-eration system was established from mature embryo-derived callus of Lehui 188 ,Wanhui 88 ,Q2 and Shuhui 527 .According to the transformation system tested ,Arabidopsis heat shock protein gene HSP101 was intro-duced into Wanhui 88 and positive transgenic plants w ere obtained w hich had improved heat resistance com-pared to wide type .