Cloning And Function Analysis Of AGL24 Gene In Carya Cathayensis

During the growth of hickory,there is a long childhood period,and the male flowers consume a large amount of tree nutrition every year,which seriously affects the yield of the fruit.Studies have shown that AGAMOUS-LIKE24(AGL24)is an important transcription factor that regulates the development of plant floral organs,but how to function in hickory remains to be further studied.We hope to use bioinformatics and molecular biology techniques to study the function of this gene,which can effectively shorten the pecan juvenile period and lay a certain foundation for the research on the regulation mechanism of hickory flower development.The following research results have been obtained:(1)Bioinformatics analysis found two CcAGL24 homologous genes,named CcAGL24 a and CcAGL24 b respectively.They all contain four protein domains: M,I,K,and C,which are typical MADSbox genes.(2)Analysis of the temporal and spatial expression characteristics of CcAGL24 a and CcAGL24 b showed that CcAGL24 a and CcAGL24 b are mainly expressed in the floral organs of hickory,and this gene is highly expressed in the pistil primordium formation period of female flowers,which may play a role in the formation of the pistil primordium.(3)Through heterologous transformation of Arabidopsis thaliana,it was found that CcAGL24 a and CcAGL24 b both promoted the flowering period of the plant and prolonged the cycle from seed germination to death of the plant.The petals of the plant become sepals,the stamens become shorter,and there is a phenomenon that the number of stamens decreases,and the carpel becomes shorter and thicker and develops abnormally.However,the phenotypes of the overexpression plants of the two genes are different.Among them,the overexpression plants of CcAGL24 a have reverse growth from reproductive growth to vegetative growth,and the overexpression plants of CcAGL24 b have multiple sepals.This also indicates that CcAGL24 a and CcAGL24 b may have redundant functions in regulating flower formation,and both have early-flowering phenotypes,but there are differences in floral organ forming functions.(4)Subcellular localization found that CcAGL24 a and CcAGL24 b observed green fluorescence in the nucleus of protoplasts,indicating that the protein of this gene was localized on the nucleus to perform its function.(5)Protein interaction studies have found that CcAGL24 a may interact with CcAGL24 b,Cc SOC1,CcAP1,Cc FLC,and Cc SVP,and the degree of interaction may be different.Among them,CcAGL24 a and CcAP1 may have the strongest degree of interaction,and CcAGL24 a and CcAGL24 a may interact with each other.CcAGL24 b interacts with Cc SOC1 and CcAP1 proteins.(6)Promoter analysis revealed that there are a variety of cis-response elements in the CcAGL24 promoter,including light response,low temperature response,and gibberellin response.It shows that CcAGL24 gene is likely to be affected by light,temperature,low temperature vernalization,gibberellin,etc.in the flowering regulatory network.