What role does the Huperzia serpentine COBRA gene family play in response to abiotic stress?
Foreword: Huperzine serpentine is one of the sources of Chinese herbal medicine mille-feuille tower, which has the effects of invigorating blood and stasis, stopping bleeding and building muscles, reducing swelling and pain, clearing heat and detoxifying, reducing fever and dehumidifying, etc., and its main ingredient Huperzine A has a good curative effect on Alzheimer's disease, and the peripheral adverse reactions are mild, and the market prospect is good.
In recent years, the artificial cultivation of Hupera serpenta has made some progress, but because of its slow growth and difficult reproduction, large-scale cultivation has not yet been realized, and the yield is far from meeting the market demand.
A total of 24 base sequences of COBRA gene family were screened from the full-length transcriptome data of Huperzia serpenta, which were named HsCOBRA1~HsCOBRA24, respectively.
The amino acid sequences were obtained based on Huperus serpenta's transcriptome sequencing and ORFfinder, and the physicochemical properties of the proteins encoded by HsCOBRAs were predicted by online software.
Among the 24 HsCOBRAs protein sequences, the number of protein amino acids was 73~663, the molecular weight was between 8494.91~73084.23Da, and the average molecular weight was 26012.36.
The isoelectric points of 9 proteins were between 4.53~6.97, which were acidic proteins, and the isoelectric points of the remaining 15 proteins were between 8.17~10.13, which were basic proteins, 11 proteins were stable proteins, and the remaining 13 proteins were unstable proteins.
HsCOBRAsFamily 成员的Fat Solubility Index 介于61.70~92.37之间,HsCOBRA7、HsCOBRA8、HsCOBRA9、HsCOBRA12、HsCOBRA15为疏水性蛋白,其余蛋白为亲水性蛋白,所有蛋白均带有负电残基3~77个及正电残基6~53个。
The prediction of transmembrane structure showed that each of the 7 proteins had 1 transmembrane structure, and the rest had no transmembrane structure.
Subcellular localization showed that HsCOBRAs family members were widely distributed, with 1 in cell wall, 4 in chloroplast, 10 in nucleus, and the remaining 10 in cell membrane, and the results of protein domain analysis showed that there were 7 domains in HsCOBRAs family.
The secondary structure of Huperzia serpentais COBRA gene family was predicted by software, and the results showed that the secondary structure of HsCOBRAs protein was composed of random coils, α helixes, extended chains, and β folds.
The results showed that there were 19 tertiary structure models for 24 proteins, with the highest proportion of HsCOBRA7 amino acids, and HsCOBRA13, HsCOBRA16, HsCOBRA19 and HsCOBRA24 all had hydrolase structures.
HsCOBRA1、HsCOBRA6、HsCOBRA7均具有水解酶抑制剂结构，HsCOBRA2、HsCOBRA3、HsCOBRA4、HsCOBRA14均具有碳水化合物结构。
HsCOBRA6、HsCOBRA7、HsCOBRA17序列同源性为99.9%,HsCOBRA6、HsCOBRA7蛋白的三级结构相似,可能具有相似的biological 功能,HsCOBRA1、HsCOBRA5序列同源性为100%。
The analysis results of cis-acting elements showed that there were 46 cis-acting elements with functional annotation in HsCOBRAs, and 23 cis-acting elements participated in the optical response.
Some proteins have cis-acting elements that participate in meristem expression and endosperm expression, and respond to abscisic acid (ABA), auxin (IAA), gibberellin (GAs), and salicylic acid, in addition to cis-acting elements that respond to anaerobic, hypoxia, drought, low temperature and other environments.
Among them, ACA can respond to cold, heat, salt stress and ultraviolet stress, ABRE element can specifically bind to BpbZIP1 to respond to salt stress, improve the salt tolerance of the plant by scavenging reactive oxygen species, and LTR retrotransposon can change the transcription level in response to salt, ABA, H2O2 and drought.
The presence of ACA, ABRE, LTR retrotransposons in the promoter region of the HsCOBRAs gene family is abundant, suggesting that the serpentine is indicated
The COBRA gene family plays a role in plant response to abiotic stresses.
According to the transcriptome analysis of the expression pattern of COBRA gene in the leaves, spores, stems and spores of Huperzia serpenta, the expression levels of HsCOBRA2 were the highest in the four tissues, and the expression level of HsCOBRA4 in leaves, spores and stems was second only to HsCOBRA2.
HsCOBRA7, HsCOBRA15 and HsCOBRA20 were almost not expressed in 4 tissues, in addition to 3 genes that were almost not expressed in leaves, 1 gene was almost not expressed in spores, 4 genes were almost not expressed in stems, and 4 genes were almost not expressed in spores.
The expression level of COBRA gene family in Hupera serpenta was leaf > spores> stem > spores, which were one of the main reproductive organs of Hupera serpenta, and the HsCOBRAs gene family may affect the growth and development of its sporangia wall, thus affecting the maturity and germination of spores.
Conclusion:
The COBRA gene family of Hupera serpenta has similar bioinformatics characteristics and rules to other species, and may participate in growth and development, reproduction and respond to adverse environmental stresses.
The study of the molecular bioinformatics characteristics and tissue expression of the COBRA gene family of Hupera serpentii is helpful to elucidate the molecular regulation and growth and development mechanism of Hupera serpentii, provide a new way for the growth of biomass, and provide a theoretical basis for future research on molecular breeding and gene function verification.
