Cloning And Expression Analysisi Of Jasmonic Acid Biosynthesis Pathway Genes In Solanum Melongena L.

Male sterility has great economic and social benefit in breeding production. Using the male sterile lines in Solanum melongena breeding can solve seed production problem, reduce cost and improve quality. Functional male sterility is a special type of plant male sterility. Its non dehiscent anthers cannot release pollen and cause sterility, but the pollen is fertile. Anther dehiscence mechanism of male sterile hasn’t been clear yet. So far, based on mutants research in Jasmonic acid(JA) biosynthetic pathway structural gene, signal transduction gene and transcription factors, it general believed that Jasmonic acid plays an important role in regulating anther dehiscence, filament elongation and pollen development.This study use Solanum melongena functional male sterile line F13-1-7 and T4, and fertile line 142 as experimental materials. Cloning the JA biosynthetic pathway key structural genes:DAD1, LOX, AOS, AOC, and OPR3. Analysis the expression of these genes in different flower buds development period in fertile line and functional male stertile line with Real-Time quantitative PCR technology. Try to explore the possible reason for functional male sterility in Solanum melongena and to provide some foundation for revealing the mechanism of functional male sterility and for Solanum melongena functional male sterile resources application. The main results as follows:1、Amplify LOX gene from Solanum melongena flower buds with cDNA as template. The gene fragment was 2557bp. Its open reading frame cotained 2508bp and estimated encoding 836 amino acid residues. The Solanum melongena LOX gene has high homology with that from other species. Protein analysis showed that the amino acid sequences contained the LH2 domain and the lipoxygenase superfamily conserved domain.2、Amplify AOC gene from Solanum melongena flower buds with cDNA as template. The gene fragment was 757bp. Its open reading frame cotained 744bp and estimated encoding 248 amino acid residues. The Solanum melongena AOC gene has high homology with that from other species. Protein analysisi showed that the amino acid sequences contained the Allence_ox_cyc superfamily conserved domain.3、Amplify OPR3 gene from Solanum melongena flower buds with cDNA as template. The gene fragment was 1227bp. Its open reading frame contained 1191bp and estimated encoding 397 amino acid residues. The Solanum melongena OPR3 gene has high homology with that from other species. Protein analysis showed that the amino acid sequences contained the OYE_like_FMN and TIM_phosphate_binding superfamily conserved domain.4、Amplify DAD1 gene from Solanum melongena flower buds with DNA as template. The gene fragment was 1052bp,with no intron. Its open reading frame cotained 1032bp and estimated encoding 244 amino acid residues. The Solanum melongena DAD1gene has high homology with that from other species. Protein analysis showed that the amino acid sequences contained the Esreraselipase superfamily conserved domain.5、In the process of flower bud development, the variation tendency of LOX expression level in fertile line and male sterile line was same. Before blossom, the accumulation of LOX in male fertile line was earlier than in male sterile line. At early period of flower bud development, the expression of AOS、AOC and OPR3 in fertile line and in sterile line was little different. But from 2 days before blossom to 2 days after blossom, the variation tendency of expression level of these 3 gene in fertile line and male sterile line was opposite.From 2 days before blossom to the blossom day, the key structure gene of JA biosynthetic pathway, AOS, AOC, LOX and OPR3 were largely accumulated in the fertile line, and were much higher than that in male sterile line. This period is important period for JA synthesis and anther dehiscence. Low expression level of these genes in male sterile line may relate to its anther disdehiscence.The accumulation of LOX gene in male sterile line was 2 period earlier than in male sterile line. At 1 day after blossom, expression level of LOX increased and OPR3 gene maximum expressed in male sterile line. These may relate to anther disdehiscence.