Construction of cDNA Library from Fertility Related Genes of Male Sterile Wheat with Aegilops kotschyi Cytoplasm by SSH

Abstract Male sterility with Aegilops kotschyi cytoplasm has a great potential in hybrid wheat ( Triticum aestivum L.) breeding for its stable sterility and broad-spectrum of restoring gene resources. To further study the genetic mechanism of male sterility with Ae. kotschyi cytoplasm, a male sterile line ms(Kots)-90-110(A) and its near isogenic line BC 4 F 1 (fertility restored by rk5451) were used to construct sterility and fertility cDNA libraries, using the suppression subtractive hybridization (SSH) method. Wheat anthers with pollen mother cells at the dikaryophase were sampled for RNA extraction via microscopic examination. Both cDNA libraries were good in quality with high efficiency of SSH. A total of 120 positive clones were randomly selected and sequenced from the 2 libraries, and 100 high quality sequences were obtained. According to BLAST screening and functional annotation, energy-related genes were identified with high frequencies in the fertility cDNA library, whereas, MADS-box transcriptional factor TaAGL7, ubiquitin-conjugating enzyme, and adenosine diphosphate glucose pyrophosphatase were detected in the sterility cDNA library. The MADS-box transcription factor TaAGL7 played an important role in regulating flower development. The immature spike ubiquitin-conjugating enzyme would probably relate to apoptosis. The expression of adenosine diphosphate glucose pyrophosphatase directly affected starch synthase and the normal supply of energy. The genes detected in the present study are probably important genes associated with the fertility in wheat with Ae. kotschyi cytoplasm.