Family Characteristics Of Chalcone Synthetase Gene Of H276A And PPR Gene Of H268 In Gossypium Barbadense

Cotton is an important fiber producing plant,which shows the Heterosis of increasing production in a specific hybrid combination.Cytoplasmic male sterility(CMS)is an important way to utilize plant heterosis.However,the utilization of Heterosis in cotton is limited because there are few types of cytoplasmic male sterility.Therefore,it is important to study the molecular mechanism of cytoplasmic male sterility for cotton germplasm innovation and heterosis utilization.Chalcone synthetase(CHS)is a key enzyme,it is closely related to the development of plant pollen and CMS.As an important source of restorer genes,PPR(pentatricoptide repeat)gene family plays an important role in plant growth and development,expression and regulation of organelle genes and other biological processes.In cytoplasmic male sterility system,CMS induced gene can express CMS specific polypeptide and RF fertility restorer of Fertility)can encode and produce PPR protein.PPR protein can regulate the expression level of CMS induced gene,reduce the level of toxic protein by reducing or inhibiting the transcription of sterility related genes,so as to restore fertility of CMS plants.However,the systematic and comprehensive analysis of CHS Gene and PPR gene in G.barbadense,especially the response to CMS,needs to be improved.In order to fill this knowledge gap,the whole genome characteristics of CHS and PPR Gene in Gossypium hirsutum were studied by using the whole genome information ofGossypium hirsutum.In addition,the gene expression of CHS family in different organs and different developmental stages of pollen abortion of sterile line H276 A was analyzed.The results will provide an important theoretical basis for the study of pollen development and the molecular mechanism of pollen abortion.In addition,the PPR protein family genes of h276 A and H268 were compared and analyzed,and the PPR protein coding genes were obtained.To provide a theoretical basis for the discovery of subsequent fertility restoration genes.The main results are as follows:(1)In order to explore the characteristics of chalcone synthetase family genes in Gossypium hirsutum,we searched and analyzed the protein database of Gossypium hirsutum,identified 20 Gb CHS genes in total,and classified all Gb CHSs into five categories according to the rootless phylogenetic tree of Gb CHS protein sequence;gene structure analysis showed that most Gb CHSs genes were composed of two exons and one intron;Blast It is found that the Gb CHS sequence is highly similar to the Gb CHS sequence,indicating that the CHS family is conservative among species;20 motifs have been identified,among which 1,3,5 and 6 belong to the Chal-sti-syntu-N domain,2,4 and 7belong to the Chal-sti-syntu-C domain;the amplification mechanism of the Gb CHS gene family has been discussed,and 25 repetitive events(12 Gb CHS genes)have been identified,including 23 fragments The amplification of Gb CHS gene family mainly occurs in the fragment repeat event,and the evolution is slow.(2)The tissue-specific expression of 20 Gb CHS genes was analyzed by q RT PCR.The results showed that the expression patterns of Gb CHS family genes in different organs of cotton showed diversity.Combined with functional prediction and gene expression,the abnormal expression of Gb CHS06,10,16 and 19 may be related to pollen abortion of Gossypium hirsutum.(3)Using H276 A c DNA of Gossypium barbadense as template,thefull-length CDs sequence of Gb CHS16 was cloned.The length of Gb CHS16 amplification fragment was 1158 bp,and p BI121-Gb CHS16-EGFP overexpression vector gene was constructed.(4)The PPR protein coding gene of Gossypium hirsutum was identified by bioinformatics,subcellular location prediction and gene function analysis were carried out,and the PPR protein family genes of H276 A and H268 were further identified by high-throughput sequencing technology.The PPR sequences obtained were homologous compared with the amino acid sequences with fertility restoration and phylogenetic tree was constructed.912 PPR protein coding genes were identified,among which 846 had sequence differences between H276 A and H268,7226 SNPs and 301 In Del heterotopias were obtained,among which 2143 SNPs and 134 In Del heterotopias were involved in mitochondria.The results showed that: XP＿016708712(LOC107923023),XP＿016710013(LOC107924193)has more than33% homology with PPR amino acid sequence with fertility restoration function identified by many plants,and has a relatively close relationship.