Mutagenesis and Creation of Photoperiod/Thermo-sensitive Genic Male Sterile Lines in Rice

doi:10.16819/j.1001-7216.2026.250613

Abstract

Abstract:

【Objective】The two-line hybrid rice breeding technology relies on thermo-sensitive genic male sterile (TGMS) lines carrying the tms5 locus. This locus originates from a natural mutation and exhibits varying sterility onset temperatures across different genetic backgrounds. Additionally, genetic drift may occur in subsequent generations. Thus, elevated sterility onset temperatures and genetic drift are key bottlenecks restricting the development of two-line hybrid rice technology.【Method】Using the japonica rice variety Zhonghua 11(ZH11) as material, we established a method for creating P/TGMS lines. Rice seeds were treated with ethylmethanesulfonate (EMS), and sterile individuals were screened from the M₂ population under high-temperature conditions. After ratooning, these fertility-restored plants were obtained under low-temperature conditions.【Result】Through years of experimentation, approximately 10% of the sterile plants were able to self-fertilize under low temperatures. Seeds from these fertility-restored lines were collected and further validated for sterility under high temperatures and fertility under low temperatures. These validated P/TGMS lines were subjected to genetic population construction combined with BSA-seq (Bulked Segregant Analysis sequencing) for gene mapping, resulting in the identification of 18 new P/TGMS lines in the ZH11 background. Among these, ostms15, ostms16, ostms18, and ostms19 have been published. Notably, the fertility transition temperature of ostms15 is significantly lower than that of the widely used tms5 locus in the same genetic background. Currently, more than 20 research institutions have utilized this sterile line for hybrid rice breeding.【Conclusion】The P/TGMS line creation technology proposed in this study can generate a series of novel P/TGMS lines, potentially overcoming the bottleneck of P/TGMS line scarcity in two-line hybrid rice breeding and supporting the sustainable development of hybrid rice breeding.

Key words: rice, EMS mutagenesis, photoperiod/thermo-sensitive genic male sterility, two-line hybrid breeding

摘要：

【目的】水稻两系杂交育种技术高度依赖携带 tms5 位点的温敏雄性核不育系。该位点源自自然突变，在不同遗传背景中具有不同的不育起点温度，而且在后代中可能存在遗传漂变，所以不育起点温度偏高和遗传漂变是制约水稻两系杂交技术发展的关键瓶颈。【方法】本研究以粳稻中花 11 种子为材料，建立了光温敏核不育系的创制方法。水稻种子经过化学诱变剂甲基磺酸乙酯(EMS)诱变处理(M₁)自交收种后，在高温环境下的 M₂群体中筛选出不育单株。将这些不育单株割蔸后继续生长，此时环境温度逐渐降低，部分新抽穗的植株育性得到恢复。【结果】经多年实验，约10%的不育株在低温条件下能够自交结实。收集这些育性恢复株系的种子，进一步进行高温不育和低温可育的验证。这些经过验证的光温敏不育系，采用构建遗传群体结合 BSA-seq (混池分离分析)基因定位方法，已获得18个新的中花11背景的光温敏雄性不育位点，其中ostms15，ostms16，ostms18，ostms19已经发表。在这些位点中，ostms15 不育系的育性转换温度显著低于同样遗传背景下在生产上广泛应用的 tms5不育系，目前已有20多家科研单位将该不育系用于水稻杂交育种。【结论】本研究提出的水稻光温敏不育系的创制技术，可以获得一系列新型光温敏不育系，有望彻底突破水稻两系杂交中光温敏不育系缺乏的瓶颈，助力水稻杂交育种的持续发展。

关键词: 水稻, EMS诱变, 光/温敏核雄性不育, 两系杂交法

ZHU Jun, YANG Yanming, YANG Zhongnan. Mutagenesis and Creation of Photoperiod/Thermo-sensitive Genic Male Sterile Lines in Rice[J]. Chinese Journal OF Rice Science, 2026, 40(4): 460-468.

朱骏, 杨延铭, 杨仲南. 水稻光温敏核不育系的诱变创制[J]. 中国水稻科学, 2026, 40(4): 460-468.

Figures/Tables 4

Fig. 1. EMS mutagenesis process for P/TGMS lines in rice A, Schematic diagram of the EMS mutagenesis process in rice; B, Illustration of rice plants with different fertility during the screening process; C, Summary data of screening across different batches. HT, High temperature; LT, Low temperature.

Fig. 2. Fertility validation of rice P/TGMS lines A, P/TGMS lines planted at different times; B, Seed-setting rate of three different batches of P/TGMS lines under high-temperature conditions; C, Seed-setting rate of three different batches of P/TGMS lines under low-temperature conditions; ostms15, ostms18 and ostms19, Three PGMS lines screened in this study; tms5, tms5 sterile line in the ZH11 genetic background; AT, Ambient temperature.

Fig. 3. Mapping process of the candidate gene for P/TGMS lines through BSA-seq A, Structure of VCF format files from raw data of BSA-seq and annotation of key information; B, Genetic basis of sequencing analysis; C, Distribution map of filtered SNP sites, where purple circles mark the candidate gene linkage regions, and purple arrows indicate the specific locus of candidate gene confirmed through further analysis; D, Complementation process and phenotypic performance of complemented plants. HT, High temperature.

Fig. 4. New P/TGMS loci identified using this technology The localization information of identified P/TGMS loci using the EMS mutagenesis technique described in this study across the 12 chromosomes of rice. Blue markers indicate previously reported P/TGMS loci, while red markers represent new P/TGMS loci cloned in this study.

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