啤酒大麦品种甘啤4号成熟胚再生体系的建立  

Establishing Regeneration System from Mature Embryos of Beer Barley Variety Ganpi 4

作　　者：马玲珑[1,2] 任盼荣 汪军成[1,2] 孟亚雄[1,2] 马小乐[1,2] 李葆春[3] 王化俊[1,2] 

机构地区：[1]甘肃省作物遗传改良与种质创新重点实验室,甘肃省干旱生境作物学重点实验室,兰州730070 [2]甘肃农业大学农学院,兰州730070 [3]甘肃农业大学生命科学技术学院,兰州730070

出　　处：《分子植物育种》2015年第3期663-669,共7页Molecular Plant Breeding

基　　金：国家自然科学基金面上项目(30771331;1171558);国家大麦青稞产业技术体系(CARS-05)共同资助

摘　　要：建立啤酒大麦品种甘啤4号成熟胚再生体系可为其遗传转化奠定基础。本研究以甘啤4号成熟胚为材料,研究不同诱导培养基对其愈伤组织诱导及分化的影响,并通过添加不同浓度梯度的Ag NO3(0,2.5 mg/L,5.0 mg/L,7.5 mg/L,10.0 mg/L,15.0 mg/L)和ABA(0,0.1 mg/L,0.3 mg/L,0.5 mg/L,0.7 mg/L,1.0 mg/L),观察其愈伤组织形态变化,统计出愈率、绿点率,探索适宜甘啤4号成熟胚组织培养的条件。同时在0、7 d、14 d、21 d和二叶期分别测定成熟胚再生过程中SOD、POD、CAT活性、MAD、可溶性蛋白及内源激素(IAA,ABA,GA3)的含量,为优化成熟胚再生体系准备条件。研究结果表明:培养基NMB较适宜甘啤4号成熟胚再生培养,发现添加适量Ag NO3和ABA有助于愈伤组织的诱导及分化,但Ag NO3处理的效果较ABA明显,其诱导甘啤4号愈伤组织的最适浓度分别为7.5 mg/L和0.5 mg/L;观察到不同时期酶活性大致均呈现先上升后下降的趋势,内源激素含量也随着培养时期的不同有所差异。研究结果可为添加外源激素、进一步优化培养条件奠定了基础。This study aimed to build regeneration system from mature embryos of Ganpi 4, a cultivar of beer barley and to lay a foundation for genetic transformation. We employed the mature embryos of Ganpi 4 as the explant material to study the effects of different induction media on callus regeneration by adjusting different levels of Ag NO3concentration（0, 2.5 mg/L, 5.0 mg/L, 7.5 mg/L, 10.0 mg/L, 15.0 mg/L） and ABA concentration（0, 0.1 mg/L,0.3 mg/L, 0.5 mg/L, 0.7 mg/L, 1.0 mg/L） in the culture media to observe the callus morphological changes and to calculate the induction rate and differentiation rate, and further to explore the best tissue culture c onditions of mature embryos of Ganpi 4. By monitoring the activities of SOD, POD, CAT and the content of MAD, soluble protein and endogenous hormones（IAA, ABA, GA3）, we attempted to make a preparation for optimizing regeneration condition of mature embryos. The results indicated that NMB should be the better medium for culturing mature embryos of Ganpi 4, and the callus regeneration be improved by adding Ag NO3 and ABA, which the most suitable concentrations for inducting callus were 7.5 mg/L and 5.0 mg/L, respectively. Whereas the treating effect of Ag NO3 was better than that of ABA. We also confirmed that the activities of enzyme exhibits a rise-up and then fall-down, and there were also some difference among the content of endogenous hormones in different stages, which might be very helpful to provide a guideline for further adding exogenous hormones and optimizing tissue culture situation.

关 键 词：啤酒大麦 甘啤4号 成熟胚 再生体系 愈伤组织诱导与分化 

分 类 号：S512.31[农业科学—作物学]