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为研究辣椒雄性不育的机理,在花期,采用常规的石蜡切片法,对新型辣椒雄性不育材料H9A及其保持系H9B小孢子不同发育时期进行细胞形态学观察;采用DDRT-PCR技术对H9A与H9B小孢子发育过程进行mRNA差异显示,筛选差异表达的基因。切片观察发现:供试雄性不育材料的小孢子败育发生在单核前期,此时花蕾纵径0.526～0.572 cm,横径0.318～0.372 cm,花冠和花萼之比约为1∶4;通过DDRT-PCR,共获得27条差异表达的cDNA片段,其中在H9A中获得12条,在H9B中获得15条。结果说明新型辣椒雄性不育材料H9A的小孢子败育发生在单核前期,败育原因可能是绒毡层细胞程序性死亡异常并极度液泡化膨胀,四分体受挤压后破裂降解,不能形成正常的单核花粉粒。败育时期与花蕾外部形态相关。在败育期表达的27条差异cDNA片段为研究辣椒雄性不育机理的候选基因奠定了基础。
In order to study the mechanism of male sterility in pepper, morphological changes of H9A and its maintainer H9B microspore at different developmental stages were observed by conventional paraffin section method at flowering stage. DDRT-PCR was used to detect H9A And H9B microspore development mRNA differences show that the screening of differentially expressed genes. The microscopic observation showed that the microspore abortion of the male sterile material occurred in the early stage of mononuclear stage, at the time of 0.526 ~ 0.572 cm in length and 0.318 ~ 0.372 cm in diameter. The ratio of corolla to calyx was about 1: 4. DDRT-PCR, a total of 27 differentially expressed cDNA fragments were obtained, of which 12 were obtained in H9A and 15 in H9B. The results showed that the microspore abortion of the new type of pepper male sterile material H9A occurred in the early stage of mononuclear stage. The reason of abortion may be abnormal apoptosis of tapetum cells and extremely vacuolar expansion. The tetrad is degraded by rupture after extrusion and can not The formation of normal mononuclear pollen grains. Abortion period and buds related to the external form. The 27 cDNA fragments expressed in the abortive period laid the foundation for the study of the candidate gene of pepper male sterility mechanism.