工业大麻CsGASAs基因家族鉴定及表达分析OA

Gibberellin-regulated protein(GASA)plays a significant role in plant growth,development and stress resistance.In this study,we used the industrial hemp(Cannabis sativa)cultivar'Longma 5'as the material to identify and characterize the CsGASAs gene family,aiming to elucidate its regulatory mechanisms in industrial hemp growth,development,and stress physiology.Using Arabidopsis thaliana GASA family sequences as a reference,the transcriptome data of CsGASA genes in industrial hemp were screened via the National Center for Biotechnology Information(NCBI)database and bioinformatics tools such as TBtools.A total of 11 CsGASAs family members were identified.Bioinformatics analysis revealed that all CsGASAs were hydrophilic and were predicted to be located in the Golgi apparatus,cytoplasm,or nucleus.Gene structure analysis indicated that the CsGASAs contained two untranslated regions and two to four coding sequences.Promoter cis-acting element analysis identified various regulatory elements,including light-responsive elements and hormone-responsive elements related to abscisic acid(ABA),auxin and gibberellin A3(GA3).The expression patterns of CsGASAs under exogenous phytohormone treatments[e.g.,GA3,ABA,salicylic acid(SA)]and drought stress were analyzed using quantitative reverse transcription polymerase chain reaction(qRT-PCR).The results showed that this gene family was regulated by drought stress and multiple phytohormones,with the highest expression levels on the 8th day of drought stress or after SA treatment.Further analysis revealed that the expression levels of CsGASA1 and CsGASA8 were significantly upregulated on the 8th day of drought stress,indicating that they may play a role in the stress response.Based on the presence of ABA-responsive elements,it was speculated that CsGASAs may participate in the drought stress response via the ABA signal pathway.Additionally,we successfully cloned the CsGASA6 gene,which had a coding sequence length of 345 bp,encoding 115 amino acids.Sequencing confirmed 99.64%homology with the reference sequence in the NCBI database.An overexpression vector for the CsGASA6 gene was constructed,and transgenic Arabidopsis thaliana plants were successfully obtained via Agrobacterium-mediated transformation.Integrated bioinformatics and qRT-PCR results confirm that the CsGASAs gene family in industrial hemp is highly conserved and may be involved in drought stress response and hormone signal transduction,potentially enhancing plant stress resistance.