《植物生理学报》 2017, 53(9): 1813-1818

粳稻F₁小孢子培养条件的优化

刘成洪^*, 何婷^*, 郭桂梅, 高润红, 陈志伟, 徐红卫, 李颖波, 周龙华, 陆瑞菊^**, 黄剑华^**

上海市农业科学院生物技术研究所, 上海市农业遗传育种重点实验室, 上海201106

通信作者：刘成洪；E-mail: luruiju62@163.com; sw1@saas.sh.cn

摘　要：

为了提高水稻小孢子培养效率, 加速小孢子培养技术在水稻育种中的应用, 本研究以上海本地早熟杂交粳稻品种‘申优17’为材料, 对F₁小孢子培养中前期幼穗低温预处理时间(12和16 d)、诱导胚性愈伤转分化培养时间(4周、7周和13周)以及自发加倍频率等技术环节进行了研究。结果表明: 低温(4°C)处理水稻幼穗12 d在诱导愈伤组织产量和分化绿苗产量上都要优于16 d处理, 且在3个转分化培养时间上都显现出这一优势效果; 比较3个转分化培养时间, 其中7周的转分化培养时间可以获得较高的分化绿苗数; 小孢子再生植株中自发加倍成二倍体频率约为50%, 而单倍体约占40%。上述结果表明, 选择合适的幼穗低温预处理时间(1~2周)和转分化培养时间(7周)有利于提高水稻小孢子培养中的绿苗分化频率。

关键词：粳稻; F₁小孢子; 低温预处理; 绿苗

收稿：2017-06-27   修定：2017-08-18

资助：上海市科技兴农重点攻关项目(应用基础类) (沪农科攻字(2015)第6-1-4号)和上海市种业发展项目(沪农科种字(2015)第2号)。

Optimization of the culture procedures for F1 microspores from japonica rice (Oryza sativa L.)

LIU Cheng-Hong^*, HE Ting^*, GUO Gui-Mei, GAO Run-Hong, CHEN Zhi-Wei, XU Hong-Wei, LI Ying-Bo, ZHOU Long-Hua, LU Rui-Ju^**, HUANG Jian-Hua^**

Biotech Research Institute, Shanghai Academy of Agricultural Sciences, Shanghai Key Laboratory of Agricultural Genetics and Breeding, Shanghai 201106, China

Corresponding author: LIU Cheng-Hong; E-mail: luruiju62@163.com; sw1@saas.sh.cn

Abstract:

In order to improve the efficiency of microspore culture in rice and explore the application of the culture technology in rice breeding, the local variety ‘Shenyou 17’ in Shanghai zone, an early maturing japonica rice hybrids was used as materials to improve the culture procedures for microspores isolated from F1 plants. The low-temperature pretreatment period on young spikes (12 and 16 d), the time of induced embryonic calli to transfer to differentiation medium (4, 7 and 13 weeks) and the frequency of spontaneous chromosome doubling were studies in this study. The results showed that the low-temperature (4°C) pretreatment on young spikes for 12 days achieved higher yields of both induced callus and regenerated green plants than the pretreatment for 16 days, which is consistent no matter what the time of transferring for differentiation. However, the highest yield of green plants was obtained in 7 weeks of induced calli transferred for differentiation when comparing three periods of transferring for differentiation. In addition, half of microspore-derived plants were diploids by spontaneous chromosome doubling, and the percent of haploids is forty or so. In conclusion, the optimized periods of low-temperature pretreatment on young spikes (1–2 weeks) and induced calli to transfer for differentiation (7 weeks) are preferred to improve the yield of green plants in microspore culture for japonica rice.

Key words: japonica rice; F1 microspores; low-temperature pretreatment; green plants