《植物生理学报》 2015, 51(12): 2201-2206

大麦单株小孢子植株再生及胁迫培养研究

郭桂梅^1,2,*, 何婷^1,2,*, 刘成洪^1,2, 高润红^1,2, 陈志伟^1,2, 徐红卫^1,2, 李颖波^1,2, 王亦菲^1,2, 陆瑞菊^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

通信作者：郭桂梅；E-mail: cs7@saas.sh.cn, sw1@saas.sh.cn；Tel: 021-62201032, 021-62202965

摘　要：

为了建立单株起源的小孢子高频再生体系用于突变体库创制和筛选研究, 以人工气候室种植的大麦为供体材料, 研究了不同培养批次和不同单株起源对小孢子愈伤诱导和植株再生频率的影响, 以及盐胁迫(500 mg•L^-1 NaCl)对单株起源小孢子培养阶段的筛选效果。结果表明, 大麦单株来源的小孢子的愈伤诱导产量在不同培养批次中出现较大的波动性, 抽穗进入第15周取材其愈伤产量获得最高, 通过连续培养单株小孢子平均成苗2 035株; 盐胁迫对单株小孢子的愈伤诱导和植株再生存在显著的抑制作用, 单株平均愈伤产量和绿苗再生率与对照相比分别下降了46%和67%, 不同单株之间的小孢子再生植株频率存在较大差异。结论: 利用人工可控条件种植的大麦单株, 通过连续小孢子培养可以获得高频率纯合再生植株; 盐胁迫可以在小孢子培养阶段实现有效筛选。

关键词：大麦; 单株; 小孢子培养; 盐胁迫; 愈伤诱导; 植株再生

收稿：2015-09-22   修定：2015-11-12

资助：上海市基础研究项目(14JC1405300)、上海市种业发展项目[沪农科种字(2015)第3号]和大麦青稞产业技术体系(CARS-05)。

Regeneration from Single Plant via Microspore Culture and Culture Response to Salt Stress

GUO Gui-Mei^1,2,*, HE Ting^1,2,*, LIU Cheng-Hong^1,2, GAO Run-Hong^1,2, CHEN Zhi-Wei^1,2, XU Hong-Wei^1,2, LI Ying-Bo^1,2, WANG Yi-Fei^1,2, LU Rui-Ju^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

Corresponding author: GUO Gui-Mei; E-mail: cs7@saas.sh.cn, sw1@saas.sh.cn; Tel: 021-62201032, 021-62202965

Abstract:

In order to establish an efficiency regeneration system via isolated microspore culture from single donor plant for future application in creation and screening of mutant population, each barley plant grown in climate chamber was used as single donor for isolated microspore culture, and the effects of different culture batches and different donor origins on the callus induction and plant regeneration, as well as the role of salt stress (500 mg•L^-1 NaCl) in the microspore culture were studied. The results showed that the callus production fluctuated between the culture batches in continuous microspore culture from single donor plant, which reached the summit at the fifteenth week, and in average 2 035 plants was regenerated from single donor plant. In addition, salt stress severely repressed the callus induction and plant regeneration, at 46% and 67% decline comparing to control respectively, and the plant regeneration frequency varied between donor plants. Conclusion: It is applicable to produce homogenous plants with high efficiency from a single donor plant grown in climate chamber via continuous microspore culture, and salt stress is effective for screening during microspore culture.

Key words: barley; single plant; microspore culture; salt stress; callus induction; plant regeneration