Cheng Ling, Huang Fugang, Jiang Zhe, Lu Baiyi, Zhong Xiaohui, Qiu Yongfu
College of Agriculture, Yangtze University, Jingzhou, 434025, Hubei, China.
Agricultural College, Guangxi University, Nanning, 530005, Guangxi, China.
Plant Methods. 2021 Nov 29;17(1):121. doi: 10.1186/s13007-021-00823-5.
The rice gall midge (RGM, Orseolia oryzae, Wood-Mason), an important stem-feeding pest worldwide, has caused serious production losses over the past decades. Rice production practices indicate that the most reliable method for managing RGM is the deployment of cultivars that incorporate host resistance. However, the conventional phenotypic screening method of rice resistance to RGM suggested by the International Rice Research Institute (IRRI) has been used for approximately 30 years, and only 12 rice varieties/lines (including controls) can be evaluated in one tray. It is not suitable for high-throughput phenotyping of rice germplasm. Moreover, a suitable method to prepare samples for molecular biological studies of rice resistance against RGM is imperative with the rapid development of modern molecular techniques.
The proper density of seedlings/RGM was determined for four seeding arrangements. A high-throughput phenotyping method (HTPM) for 60 lines/varieties infested with 36 female RGM adults in one tray, as described by method 4-3 (seeded 60 lines/varieties), was developed and verified using mutant screening. Furthermore, one RGM resistance gene flanked by markers 12RM28346 and 12RM28739 on chromosome 12 was simultaneously detected using method 2-2 (seeded 30 lines/varieties in one tray) treated with 24 RGM and analyzed using conventional and simplified grading systems. Genetic analysis of the RGM resistance gene was confirmed using a method identical to that suggested by IRRI. Finally, one bucket with 24 seedlings treated with at least five female RGM adults was efficacious and could offer adequate samples for insect development observation or molecular biological studies.
A highly efficient and reliable procedure for evaluation of resistance in rice to RGM was developed and improved, and was verified through mutant screening, gene mapping, genetic analysis, and insect growth and development observations.
稻瘿蚊（RGM，Orseolia oryzae，伍德 - 梅森）是一种在全球范围内重要的蛀茎害虫，在过去几十年中已造成严重的产量损失。水稻生产实践表明，管理稻瘿蚊最可靠的方法是部署具有寄主抗性的品种。然而，国际水稻研究所（IRRI）建议的传统水稻对稻瘿蚊抗性的表型筛选方法已经使用了约30年，且一个托盘只能评估12个水稻品种/品系（包括对照）。它不适用于水稻种质的高通量表型分析。此外，随着现代分子技术的快速发展，迫切需要一种合适的方法来制备用于水稻抗稻瘿蚊分子生物学研究的样本。
确定了四种播种方式下合适的秧苗/稻瘿蚊密度。开发了一种高通量表型分析方法（HTPM），该方法用于在一个托盘中用36只雌性稻瘿蚊成虫侵染60个品系/品种，如方法4 - 3（播种60个品系/品种）所述，并通过突变体筛选进行了验证。此外，使用方法2 - 2（一个托盘播种30个品系/品种），用24只稻瘿蚊处理，并用传统和简化分级系统进行分析，同时检测到位于第12号染色体上标记12RM28346和12RM28739侧翼的一个稻瘿蚊抗性基因。使用与IRRI建议的方法相同的方法对稻瘿蚊抗性基因进行了遗传分析。最后，一个装有24株用至少五只雌性稻瘿蚊成虫处理过的秧苗的桶效果良好，可为昆虫发育观察或分子生物学研究提供足够的样本。
开发并改进了一种高效、可靠的水稻对稻瘿蚊抗性评估程序，并通过突变体筛选、基因定位、遗传分析以及昆虫生长发育观察进行了验证。
