烟草Tubby-like基因家族鉴定及非生物胁迫表达分析OA

The plant Tubby-like protein(TLP)gene family plays a crucial regulatory role in diverse biological processes,including abiotic stress responses and growth and development.To elucidate the molecular mechanisms underlying the response of tobacco TLP genes to abiotic stress,this study employed bioinformatics approaches to identify TLP family members in tobacco.A comprehensive analysis was conducted,encompassing phylogenetic tree construction,cis-acting element prediction,conserved domain and motif characterization,protein-protein interaction(PPI)network prediction,and expression profiling.The results demonstrated that a total of 26 NtTLP family genes were identified across the entire tobacco genome.The encoded proteins varied in length from 88 to 741 amino acids(aa),with molecular weights ranging from 10.14 to 83.27 kDa and isoelectric points(pI)spanning 4.72 to 9.78.Subcellular localization prediction revealed that the majority of NtTLP members(17 out of 26)were localized in the nucleus.Promoter analysis indicated that the cis-acting elements within the NtTLP genes could be categorized into three major classes:hormone-responsive elements,stress-responsive elements,and elements associated with growth,development and biological processes.Among these,hormone-responsive elements were the most abundant,with up to 14 distinct types.Furthermore,tissue expression profiling showed that NtTLP family genes exhibited tissue-and developmental stage-specific expression patterns in tobacco,with generally high expression levels detected in seeds.Protein-protein interaction prediction identified NtTLP02 and NtTLP06 as hub proteins,potentially engaging in extensive interaction networks with other NtTLP members.Quantitative real-time PCR(qRT-PCR)analysis indicated that both NtTLP02 and NtTLP06 responded to heat,salt,cold,and drought stresses to varying degrees;Notably,NtTLP02 displayed a more sensitive response to cold and drought stresses.Subsequent subcellular localization assays confirmed that both NtTLP02 and NtTLP06 proteins were localized in the nucleus.Collectively,these findings suggest that NtTLP02 and NtTLP06 are important candidate genes mediating tobacco responses to abiotic stresses.This study provides valuable genetic resources for further elucidating the regulatory mechanisms underlying abiotic stress tolerance in tobacco.