Analysis Of SPL8-like Genes In Arabidopsis And Beyond

The SBP-Box genes(also known as the SPL genes)encode plant–specific transcription factors.These transcription factors bind to DNA through the GTAC core sequence and regulate the expression of downstream genes.They play an important role in plant development,disease resistance,and increasing crop yield.SPL8 is a gene that is not regulated transcriptinally by miR156/7 and plays an important role in the development of anthers in Arabidopsis thaliana.The SPL8 knockout leads to infertility of several early arisingflowers.The main reason for the infertility is the decreased number of pollen sacs and pollen in anthers.Through sequence alignment analysis,it was found that SPL8-like genes are also present in other plants,but their function has not been reported much.We isolated and identified the SPL8-like genes in Solanum lycopersicum and Brachypodium distachyon and transformed them into Arabidopsis spl8-1 mutants to observe whether the phenotype of spl8-1 mutant can be rescued and further analyzed its molecular function.Through the amino acid sequence alignment,we found that the N-terminus of the protein encoded by the SPL8-like gene has a conserved region consisting of 12 amino acids.This conserved region is located upstream of the SBP-box domain,but whether it affects the function of the SPL8 is not clear.We used point mutations to mutate five highly conserved amino acids and transformed the mutated AtSPL8 gene into the spl8-1 mutant for phenotypic rescue experiments and analyzed the importance of this conservative region for normal functioning of SPL8.This paper studied a set of methods for establishing Arabidopsis growth chambers,explored the factors affecting the growth of Arabidopsis,and optimized the culture conditions of Arabidopsis thaliana.And the SPL8 gene of Arabidopsis thaliana,two SPL8-like genes(SlySPL8-1,SlySPL8-2)of Solanum lycopersicum,and two SPL8-like genes(BdSPL8-1,BdSPL8-2)of Brachypodium distachyon were cloned.The results showed that AtSPL8 can completely complement the spl8-1 mutant phenotype,whereas the SPL8-Like genes of Solanum lycopersicum and Brachypodium distachyon cannot complement the Arabidopsis spl8-1 mutant phenotype.In addition,our results also indicate that mutations in glycine at positions 148 and 152 all resulted in the loss of SPL8 function of the complementary spl8-1 mutant sterility phenotype.In summary,this article established a management system for Arabidopsis growth chambers,provided examples and practical experience for the Arabidopsis growth platform.To explore the function of SPL8-Like genes in Solanum lycopersicum and Brachypodium distachyon,and to lay a foundation for the future research on the function of other SPL8-like genes,using the importance of conserved amino acids on the function of SPL8,to construct male sterile lines for production and provide the new genetic resources for Crossbreeding.