《植物生理学报》 2012, 48(11): 1069-1078

源于小孢子培养的大麦耐盐变异体获取

陆瑞菊^1,2, 徐红卫^1,2,*, 陈志伟^1,2, 何婷^1,2, 杜志钊^1,2, 高润红^1,2, 王亦菲^1,2, 邹磊^1,2, 郭桂梅³,卜姝明³,刘成洪^1,2,**, 黄剑华^1,2,**

1上海市农业科学院生物技术研究所, 上海201106; 2上海市农业遗传育种重点实验室, 上海201106; 3上海海洋大学水产与生 命学院, 上海201306

通信作者：徐红卫；E-mail: dainney@163.com, sw1@saas.sh.cn；Tel: 021-62202965;021-62201032

摘　要：

以大麦品种‘花30’作为供试材料, 比较了甲基磺酸乙酯(EMS)和平阳霉素处理小孢子, 60Co γ-射线辐照处理离体穗和 干种子, 对300 mg•L-1 NaCl胁迫培养下游离小孢子的愈伤组织产量和愈伤组织在0.3% NaCl胁迫筛选下的绿苗产量的影 响。结果表明, EMS处理离体小孢子和60Co γ-射线辐照干种子的愈伤组织产量和绿苗产量明显优于平阳霉素处理小孢子 和60Co γ-射线辐照离体穗。以16份源于种子辐照处理的再生植株自交一代种子为供试材料, 比较了在0.3% NaCl胁迫下种 子的发芽率和幼苗的成活率以及植株的分蘖数、株高和单株产量。结果表明, ‘花30’发芽率为0, 供试的16份耐盐变异体 中, 有14份材料在NaCl胁迫下的发芽率优于‘花30’, 鉴定出4份耐盐性明显优于‘花30’的变异体材料。选择耐盐变异体作为 供试材料, 测定了变异体中Na+/H+逆向转运蛋白基因NHX1、NHX2和NHX3和编码甜菜碱醛脱氢酶(BADH)的两个同工酶 基因BBD1和BBD2的表达模式和表达量, 结果表明变异体耐盐性的提高与这些基因的表达量存在联系。

关键词：大麦; 小孢子离体诱变和胁迫培养; 耐盐变异体; 单株产量; Na+/H+逆向转运蛋白; 甜菜碱醛脱氢酶

收稿：2012-09-24   修定：2012-11-05

资助：大麦现代产业技术体系项目(CARS-05)和上海市科技兴农重 点攻关项目[沪农科攻字(2009)第2-1号, 农青年科技2012 (15)]。

Screening of Variants Derived from Barley (Hordeum vulgare L.) Microspore Culture for Salt Tolerance

LU Rui-Ju^1,2, XU Hong-Wei^1,2,*, CHEN Zhi-Wei^1,2, HE Ting^1,2, DU Zhi-Zhao^1,2, GAO Run-Hong^1,2, WANG Yi-Fei^1,2, ZOU Lei^1,2, GUO Gui-Mei³, BU Shu-Ming³, LIU Cheng-Hong^1,2,**, HUANG Jian-Hua^1,2,**

¹Biotech Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai 201106, China; ²Shanghai Key Laboratory of Agricultural Genetics and Breeding, Shanghai 201106, China; ³College of Fisheries and Life Science, Shanghai Ocean University, Shanghai 201306, China

Corresponding author: XU Hong-Wei; E-mail: dainney@163.com, sw1@saas.sh.cn; Tel: 021-62202965;021-62201032

Abstract:

Using barley (Hordeum vulgare) variety ‘Hua 30’ as donor material, effects on callus yield under 300 mg·L^-1 NaCl stress and the green plantlet differentiated rate under 0.3% NaCl stress from the microspores treated with ethyl methane sulfonate (EMS) and bleomycin or the microspores from irradiated spikes and dry seeds were investigated. The results showed that callus yield and green plantlet from microspores with EMS treatment and microspore derived from irradiated dry seeds were obviously superior to the microspores treated by bleomycin and microspore derived from irradiated spikes. A series of indexes such as seed germination rate, seedling survival rate, plant tiller number, plant height and yield per plant were compared on seeds from 16 regenerated plants by seed irradiation under 0.3% NaCl stress. The germination rate of ‘Hua 30’ is 0, while that of 14 of the 16 salt tolerance mutants has an advantage over ‘Hua 30’, moreover, 4 materials were obviously better than ‘Hua 30. Based on these salt tolerant mutants, the expression patterns and quantity of Na⁺/H⁺ antiporter genes (NHX1, NHX2 and NHX3), and BBD1 and BBD2 encoding two betaine aldehyde dehydrogenase (BADH) isozymes were measured. As a result, the improvement in salt tolerance of variants had corresponding relation with these genes.

Key words: barley; microspore in vitro mutation and stress culture; salt-tolerant variant; yield per plant; Na⁺/H⁺antiporter; betaine aldehyde dehydrogenase