Abstract
The Negative on TATA-less (NOT) 2_3_5 domain proteins play key roles in mRNA metabolism and transcription regulation, but few comprehensive studies have focused on this protein family in plants. In our study, a total of 30 NOT2_3_5 genes were identified in four cotton genomes: Gossypium. arboretum, G. raimondii, G. hirsutum and G. barbadense. Phylogenetic analysis showed that all the NOT2_3_5 domain proteins were divided into two classes. The NOT2_3_5 genes were expanded frequently, and segmental duplication had significant effects in their expansion process. The cis-regulatory elements analysis of NOT2_3_5 promoter regions indicated that NOT2_3_5 domain proteins might participate in plant growth and development processes and responds to exogenous stimuli. Expression patterns demonstrated that all of the GhNOT2_3_5 genes were expressed in the majority of tissues and fiber development stages, and that these genes were induced by multiple stresses. Quantitative real-time PCR showed that GbNOT2_3_5 genes were up-regulated in response to verticillium wilt and the silencing of GbNOT2_3_5-3/8 and GbNOT2_3_5-4/9 led to more susceptibility to verticillium wilt than controls. Identification and analysis of the NOT2_3_5 protein family will be beneficial for further research on their biological functions.
Highlights
Negative on TATA-less (NOT) proteins are important constitutive components of the carbon catabolite repression 4 (CCR4)-NOT complex, which exists in all eukaryotes and plays a key role in mRNA metabolism and transcription regulation [1]
Several kinds of NOT proteins have been isolated from yeasts, flies, humans and Arabidopsis thaliana, including NOT1, NOT2, NOT3, NOT4, NOT5, NOT9, NOT10 and NOT11 [5]
Trypan blue assay showed that leaves from silenced cotton were severely dyed sbhlouwe, einddtihcaattinsgilemnocriengdeGadbNceOllTs2in_3th_5e-l3e/a8veasnodfGTRbNV:OGTbN2_O3T_25_-34_/95 gpelannetss acfotmerpVr.odmahilsiaeed cotton riensfiescttaionnce(FdiguurriengS4V)..Adallholifatehienaobcouvleatrieosnu.lts showed that silencing GbNOT2_3_5-3/8 and GbNOT2_3_5-4/9 genes compromised cotton resistance during V. dahliae inoculation
Summary
Negative on TATA-less (NOT) proteins are important constitutive components of the carbon catabolite repression 4 (CCR4)-NOT complex, which exists in all eukaryotes and plays a key role in mRNA metabolism and transcription regulation [1]. Many previous studies indicated that NOT2, NOT3 and NOT5 function together and play key roles in the vegetative growth and transcription machinery of yeast [12]. Previous studies indicated that NOT2 played a key role in miRNA biogenesis and miRNA regulated expression level of target genes involving in plant growth, development and response to abiotic and biotic stresses [18]. We characterize NOT2_3_5 domain proteins from diploid and tetraploid cotton species and examine their expression patterns during normal growth and development and after abiotic and biotic stresses. Two of these genes were induced in a verticillium-resistant cotton variety after pathogen inoculation. Verticillium wilt, a soil-borne vascular disease, causes devastating losses of cotton yield and quality every year [25,26].The results will provide new approaches to improve resistance to verticillium wilt in cotton and provide new inspiration on further research on transcriptional regulation of NOT proteins
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