›› 2013, Vol. 25 ›› Issue (6): 0-1188.

• 作物科学 •    

影响大豆遗传转化主要因素及相关条件优化

马晓丽1,2,冯小锋3,杨清华2,郁晓敏2,朱丹华2,*
  

  1. 1浙江师范大学 化学与生命科学学院,浙江 金华 321004;2 浙江省农业科学院 作物与核技术利用研究所,浙江 杭州 310021;3 长江大学 园艺园林学院,湖北 荆州 434023
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2013-11-25 发布日期:2013-11-25

Optimization of main factors and relevant conditions affecting soybean Agrobacterium\|mediated transformation system

MA Xiao\|li;FENG Xiao\|feng;YANG Qing\|hua;YU Xiao\|min;ZHU Dan\|hua;*   

  1. (1 College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China;2 Instit
  • Received:1900-01-01 Revised:1900-01-01 Online:2013-11-25 Published:2013-11-25

摘要: 以浙春3号,华春6号,0428三个大豆基因型的胚尖、子叶节和下胚轴为受体材料,运用农杆菌介导法转化GUS基因,通过对GUS基因瞬时表达率的比较,探讨大豆遗传转化的主要影响因素(基因型、外植体等),并进一步对遗传转化条件进行优化。结果表明:以胚尖为受体时,3个基因型的GUS基因瞬时表达率都很低,在丛生芽的再生部位没有瞬时表达。以下胚轴为受体时,萌发5 d,共培养3 d后,浙春3号、华春6号、0428在丛生芽再生部位的GUS瞬时表达率达到最高,分别为4392%,5445%,5847%。而以子叶节为受体时,按照常规方法(萌发5 d),浙春3号、华春6号、0428分别在共培养5,4,4 d时,其丛生芽再生部位的GUS瞬时表达率最高,分别为2305%,1379%,478%;利用浙春 3号对子叶节法的萌发时间进行了进一步的优化,发现萌发时间为3 d时在丛生芽再生部位的GUS瞬时表达率、芽诱导率较萌发5 d都有较大幅度的提高。

关键词: 大豆基因型, 胚尖, 子叶节, 下胚轴, GUS瞬时表达率, 芽诱导率

Abstract: In this study, we optimized main factors (genotypes, explants, etc) affecting soybean Agrobacterium\|mediated transformation system by comparing GUS transient expression rate with embryonic tip, cotyledonary node and hypocotyl of Zhechun No.3, Huachun No.6 and 0428. The results showed GUS transient expression rate of the three soybean genotypes was low in the embryonic tip transformation system and tufty shoots showed no expression. Using the hypocotyl transformation system, the maximum expression rate of Zhechun No.3, Huachun No.6 and 0428 in tufty shoots was 4392%, 5445%, and 5847%, respectively, after 5 days of germination and 3 co\|culture days. Using the cotyledonary node transformation system(5\|day germination),the optimal co\|culture time for Zhechun No.3, Huachun No.6 and 0428 were 5, 4, and 4 days respectively and the maximum expression rate of Zhechun No.3, Huachun No.6 and 0428 in tufty shoots was 2305%, 1379%, and 478%, respectively. The germination time of cotyledonary node transformation system was further optimized using Zhechun No.3. The results showed that both gene expression and shoot induction increased dramatically in 3 germination days compared to 5 germination days.

Key words: soybean, embryonic tip, cotyledonary node, hypocotyl, GUS transient expression rate, bud induction rate