小麦矮腥黑粉菌导致小麦花药细胞异常发育的侵染过程及相关基因研究

Tilletia controversa Kühn (TCK) was one of the international quarantine diseases as the most difficult to control in wheat smut, which lead to dwarf bunt of wheat and abnormal development of wheat anther cells. We already found the infection lead to the abnormal of the cells in leaves by scanning electron microscopy and found the teliospores not only in kernel, but also in root, stem and leaves. The results mentioned above showed a big difference from the references with the conclusion that the teliospores were only existed in the kernel and the infection of TCK will not affect the wheat physiology based on the section of wheat tissues. These new findings broaden the traditional opinions and maybe showed new infection process of TCK in wheat. Based on the results which mentioned above and the development of the inoculation method which got the kernel with teliospores successfully, we will firstly track the fungi with confocal microscope in three-dimensional, which was a big improvement and could provide much more information than the section technology. Secondly, we will detect the wheat anther cells’ development (epidermal cell, endothecial cell, middle layer cell, tapetal cell, archesporial cell) under infection, combined with gene sequencing, RNA-Seq, HIGS and genetic transformation system of the fungi, we will analyze, forecast and verificate the function of the genes which related to the abnormal development of wheat anthers. These results will contribute to its effective prevention and resistance breeding, and will also provide an example of the train of thought and the method for the infection process and interaction mechanism of smut disease with their hosts.

小麦矮腥黑粉菌（Tilletia controversa Kühn, TCK）是麦类黑粉菌中危害最大且极难防治的国际重要检疫性病菌，其侵染导致小麦花药细胞异常发育。申请人前期研究发现（如该病菌侵染可导致叶肉细胞变形及冬孢子亦分布于除穗部外的根、茎、叶中）拓宽了组织切片技术所获传统观点（该病菌侵染不影响小麦正常生理及冬孢子仅分布于小麦穗部），预示存在新的侵染过程。基于上述发现及已建立的TCK人工接种体系，本项目将采用激光共聚焦显微技术继续观察其侵染过程，特别关注其侵染对小麦花药细胞（药壁表皮、药室内壁、中层、绒毡层及孢子囊）不同发育阶段的异常影响；进而基于TCK基因组信息，结合RNA-Seq、HIGS技术及该病菌遗传转化体系，预测导致花药异常发育相关基因，并进行其功能分析，为深入研究TCK的侵染机制、有效防控及抗性育种奠定理论依据，亦为其它黑粉菌对禾谷类寄主的侵染研究提供可借鉴的思路和方法。

小麦矮腥黑粉菌（Tilletia controversa Kühn, TCK）是麦类黑粉菌中危害最大且极 难防治的国际重要检疫性病菌，其侵染导致小麦花药细胞异常发育。申请人前期研究发现 （如该病菌侵染可导致叶肉细胞变形及冬孢子亦分布于除穗部外的根、茎、叶中）拓宽了 组织切片技术所获传统观点（该病菌侵染不影响小麦正常生理及冬孢子仅分布于小麦穗部 ），预示存在新的侵染过程。基于上述发现及已建立的小麦矮腥黑粉菌人工接种体系，本项目采用激光共聚焦显微技术建立了小麦花药细胞的发育历程，研究了该病菌侵染后对小麦花药细胞（药壁表皮、 药室内壁、中层、绒毡层及孢子囊）不同发育阶段的异常影响；基于小麦矮腥黑粉菌的基因组信息，结合该病菌的遗传转化体系，我们采用RNA-seq的方法预测了导致花药异常发育关键阶段的相关基因，为深入研究该病菌的侵染机制、有效防控及抗性育种奠定理论依据，亦为其它黑粉菌对禾谷类寄主的侵染研究提供可借鉴的思路和方法。