Gene Family Research Articles

Molecular identification and characterization of the superoxide dismutase (SOD) gene family in Tetranychus urticae (Acari: Tetranychidae) and the role of TuSOD2 gene under short-term heat stress

Superoxide dismutases (SODs) are key enzymes for scavenging insects' excess reactive oxygen species (ROS) and play a crucial role in resisting a variety of stresses. The deletion of SOD genes can alter the response of insects to abiotic stresses. In this study, four genes of the superoxide dismutase family in Tetranychus urticae were identified and analyzed, among which three copper/zinc SODs and a manganese SOD. The result of quantitative real-time PCR showed that TuSOD2 expression level was up-regulated significantly under short-term heat stress, but TuSOD1, TuSOD2, and TuSOD4 were down-regulated or no significant difference, which indicated that T. urticae responded to oxidative stress induced by heat stress by raising the expression level of superoxide dismutase gene. Among them, TuSOD2 gene plays a key role in response to heat stress in T. urticae, and other SOD genes may be responsible for other oxidative stresses. RNA interference experiments showed that the knocking down of TuSOD2 reduced the activities of various antioxidant enzymes, resulting in a decline in the tolerance to high temperature and an increase in the mortality of T. urticae under short-term heat stress. To summarize, SOD, especially TuSOD2 plays an important role in treating short-term heat stress in T. urticae.

Comprehensive identification of PIN and PILS in crape myrtle genomes reveals their putative functions in bud-to-branch development and callus generation

Polar auxin transport in plants is facilitated by influx and efflux transporters encoded by PIN-FORMED (PIN) and PIN-like (PILS) genes, respectively. While the auxin transporter gene families have been extensively studied in various monocot and dicot species, a comprehensive genome-wide analysis of PIN and PILS gene families in Lagerstroemia indica is currently lacking. In this study, we identified 22 LiPIN and LiPILS genes in L. indica genome, distributed across 17 chromosomes. Gene structure and conserved motif analyses revealed relative conservation within the same group. Additionally, we identified 16 syntenic gene pairs in LiPIN and LiPILS genes, with Ka/Ks values below 1 indicating purifying selection during evolutionary processes. Expression profiling indicated that several genes, notably LiPIN3a and LiPILS5b/6a/6b/6c, responded to salt stress. LiPIN1d, LiPIN5, and LiPILS3a were potentially linked to bud-to-branch development in L. indica. Moreover, the expression levels of LiPILS3a and LiPILS5a exhibited significant differences during the callus formation process, indicating their potential as key regulatory factors in this developmental stage. These findings offered new insights into auxin transporter genes in L. indica and enhanced our understanding of their roles in stress tolerance, growth, and development.

Genome-wide identification and expression analysis of genes encoding late embryogenesis proteins in Cicer arietinum

Late embryogenesis abundant (LEA) proteins play defensive roles during seed maturation and seed germination processes. However, there is no such investigation was carried out in chickpea. In present study, genome wide identification and characterization of LEA encoding genes has been investigated, and identified 65 and 74 LEA encoding genes in desi and kabuli cultivar of chickpea, respectively. All these genes have been classified into eight subfamilies on the bases of their phylogenetic analysis and conserved domain. Maximum members of LEA encoding genes were found to be a part of the LEA_2 gene family. The analysis of physicochemical properties of LEAs was also conducted. LEA encoding genes have been found to be located in all chromosomes (8 chr) of chickpea and identified as involved in response to stimulus, biological processes, molecular functions and cellular components based upon gene ontology analysis. Gene expression analysis of randomly selected 8 LEA encoding genes has been carried out during different seed developmental stages which revealed the higher expression of LEA encoding genes during later stage of seed development in chickpea and proved their potential role in desiccation process during seed maturation. During seed germination, expression analysis of LEA encoding genes was found to be higher during the initial stages of seed germination. In conclusion, this work highlights the genome wide identification and characterization of LEA encoding genes in chickpea and proposed potential roles during seed developmental processes. This information could also be useful as a reference investigation for molecular breeding of chickpea for recalcitrant behaviour of seed.

GmSTOP1-3 regulates flavonoid synthesis to reduce ROS accumulation and enhance aluminum tolerance in soybean

Aluminum (Al) toxicity is a significant limiting factor for crop production in acid soils. The functions and regulatory mechanisms of transcription factor STOP1 (Sensitive to Proton Rhizotoxicity 1) family genes in Al-tolerance have been widely studied in many plant species, except for soybean. Here, expression of GmSTOP1-3 was significantly enhanced by Al stress in soybean roots. Overexpression of GmSTOP1-3 resulted in enhanced root elongation and decreased Al content, which was accompanied by increased antioxidant capacity under Al treatment. Furthermore, RNA-seq identified 498 downstream genes of GmSTOP1-3, including genes involved in flavonoid biosynthesis. Among them, the expression of chalcone synthase (GmCHS) and isoflavone synthase (GmIFS) were highly enhanced by GmSTOP1-3 overexpression. Further quantitative flavonoid metabolome analysis showed that overexpression of GmSTOP1-3 significantly increased the content of naringenin chalcone, naringenin, and genistein in soybean roots under Al treatment, which positively correlated with the expression level of the genes relative to flavonoid biosynthesis. Notably, genistein had a significant positive correlation with the expression levels of GmIFS. Combination of Dual Luciferase Complementation (LUC) and Electrophoretic Mobility Shift Assays (EMSA) revealed that GmSTOP1-3 directly bound to the promoters of GmCHS/GmIFS and activated both genes’ transcription. Taken together, these results suggest that GmSTOP1-3 enhances soybean Al tolerance partially through regulating the flavonoid synthesis.

Genome-wide analysis of the R2R3-MYB gene family and identification of candidate genes that regulate isoflavone biosynthesis in red clover (Trifolium pratense)

Red clover (Trifolium pratense) is a perennial legume with high feeding and medicinal value attributed to its abundant isoflavone content. Previous studies reported that R2R3-MYB transcription factors are involved in the biosynthesis of isoflavones; however, their specific role in red clover remains poorly understood. Through comprehensive genome-wide and transcriptome analyses, a total of 138 TpR2R3-MYB genes were identified and classified into 30 distinct subgroups within a phylogenetic tree. Importantly, six of these subgroups showed associations with isoflavone biosynthesis in red clover. The majority of segmental duplication events (Ka/Ks < 1) indicated that the TpR2R3-MYB gene underwent strong purifying selection during evolution. The qRT-PCR analysis demonstrated high expression levels of TpMYB79 and TpMYB53 in Minshan red clover at full flowering stage, consistent with the trend for isoflavone content determination, suggesting that TpMYB79 and TpMYB53 might be important regulators of isoflavone biosynthesis in red clover. Additionally, we observed nucleus and vacuole membrane localization of TpMYB53 and TpMYB79, with TpMYB53 primarily exerting transcriptional activation through its C-terminal activation motifs while TpMYB79 exhibited no transcriptional activity. These findings provided a foundation for the study of the biological function of R2R3-MYB transcription factors in red clover.

GhLCYε-3 characterized as a lycopene cyclase gene responding to drought stress in cotton

Drought stress significantly affects crop productivity. Carotenoids are essential photosynthetic pigment for plants, bacteria, and algae, with signaling and antioxidant functions. Lutein is a crucial branch product in the carotenoid synthesis pathway, which effectively improves the stress tolerance of higher plants. lycopene cyclase, a central enzyme for lutein synthesis, holds great significance in regulating lutein production. This research establishes a correlation between lutein content and stress resistance by measuring the drought resistance and lutein content of various cotton materials. To identify which crucial genes are associated with lutein, the lycopene cyclase family (LCYs) was analyzed. The research found that LCYs form a highly conserved family divided into two subfamilies, LCY-ε (lycopene ε-cyclase) and LCY-β (lycopene β-cyclase). Most members of the LCY family contain photoresponsive elements and abscisic acid elements. qRT-PCR demonstrates showed that most genes responded positively to drought stress, and GhLCYε-3 was expressed significantly differently under drought stress. Virus-induced gene silencing (VIGS) assay showed that the content of GhLCYε-3 was significantly increased with MDA and PRO, and the contents of chlorophyll and lutein were significantly decreased in pYL156 plants. The decrease in GhLCYε-3 expression is speculated to lead to reduced lutein content in vivo, resulting in the accumulation of reactive oxygen species (ROS) and decreased drought tolerance. This research enriched the understanding of LCY gene family and lutein function, and provided a new reference for cotton planting in arid areas. SynopsisLycopene cyclase plays an important role in enhancing the ability of scavenging ROS and drought resistance of plants.

Genome-wide identification of HIPP and mechanism of SlHIPP4/7/9/21/26/32 mediated phytohormones response to Cd, osmotic, and salt stresses in tomato

Heavy-metal-associated isoprenylated plant proteins (HIPPs) contributed to abiotic tolerance in vascular plants. Up to now, the HIPP gene family of tomato (Solanum lycopersicum L.) had not been thoroughly understood. In the present study, 34 SlHIPP genes were identified from the tomato genome using the Hidden Markov Model (HMM). The phylogenetic analysis revealed that the evolution of SlHIPPs was highly conserved. The cis-acting element analysis indicated that SlHIPP genes might be involved in phytohormones and abiotic stresses. We constructed venn diagram with 17 genes containing stress-related motifs as well as 15 genes and 19 genes expressing in leaves and roots in RNA-seq data, suggesting that SlHIPP4/7/9/21/26/32 were selected as candidate genes for study. The quantitative real-time PCR (qRT-PCR) analysis showed that 6 candidate genes were indicated to be involved in osmotic and salt stress tolerance and SlHIPP7/21/26/32 responded to cadmium (Cd) tolerance. The virus-induced silencing of 6 candidate genes caused growth inhibition in stress conditions, further illustrating that 6 candidate genes played a positive role in abiotic conditions. Importantly, the phytohormone analysis implied that 6 candidate genes mediated abscisic acid (ABA), salicylic acid (SA), gibberellin (GA3), auxin (IAA), or methyl jasmonate (MeJA) responses to Cd, osmotic, or salt stress tolerance. These findings indicated that SlHIPP4/7/9/21/26/32 were key regulators of abiotic stress responses in tomato seedlings, functioning through multiple phytohormone pathways.

Comprehensive analysis of Capsicum annuum CaLhcs uncovered the roles of CaLhca5.1 and CaLhcb1.7 in photosynthesis and stress tolerance

The light-harvesting chlorophyll a/b-binding proteins (Lhcs) are integral to plants' capture and transfer of light energy during photosynthesis. However, the Lhc gene family remains unexplored in pepper. In this study, 37 CaLhcs (Capsicum annuum Lhc) were identified from the reference genome and classified into five subfamilies (Lhca, Lhcb, CP24, CP26, and CP29) based on phylogenetic relationships and conserved domains, with members of each subfamily displaying similar conserved motifs and gene structures. cis-element analysis revealed an enrichment of light-responsive elements within CaLhcs (46.1 %). Transcriptome analysis showed that most CaLhcs are specifically expressed in leaves, flowers, and pericarp and are responsive to stressors, including NaCl, cold, heat, H2O2, and d-mannitol. Post-transcriptional regulation analysis identified 11 miRNAs that target nine CaLhcs through cleavage. RT-qPCR analysis validated the involvement of CaLhcs in response to NaCl stress. Localization studies confirmed that CaLhca4.1, CaLhcb1.1, CaLhca1.7, CaLhcb1.11, and CaLhcb6.1 are chloroplast-localized, whereas CaLhca5.1 localizes in the nucleus. Overexpression of CaLhcb1.7 and CaLhca5.1 increased chlorophyll content and net photosynthetic rate, enhancing photosynthesis. Additionally, CaLhcb1.7 and CaLhca5.1 reduced ROS accumulation, bolstering the plant's resistance to pathogens and salt stress. These findings provide a foundation for further exploration of CaLhcs in photosynthesis and stress tolerance mechanisms.

Identification of the FBN gene family in tomato and functional analysis of SlFBN11 in the electron transport under low night temperature

FBNs are lipid-associated proteins that play a critical role in plant growth and stress response. However, the mechanisms of how FBNs proteins participate in the low night temperature response in tomato still unclear. Here we conducted a comprehensive genome-wide analysis of the FBN gene family in Solanum lycopersicum. In total, 14 SlFBN genes were identified, and information on their gene structures, protein motifs, phylogenetic relationships, and stress-related cis-regulatory elements (CREs) was provided. Among these, SlFBN11 was selected as a promising candidate for further functional characterization. The silencing of SlFBN11 destroys the redox balance of the PSI reaction center under low night temperature (LNT) stress, which led to increased ROS accumulation. Surprisingly, we found that the silencing of SlFNR2 also displayed an imbalance in electron transport of the PSI under LNT stress. Further experiments showed SlFBN11 can interact with SlFNR2 to positively response electron transport low night temperature. Collectively, the study provides a comprehensive analysis of the FBN genes family in Solanum lycopersicum and provides a theoretical basis for our understanding of the function of FBN genes in adaptation to LNT stresses.

GhBZR15 improved the tolerance of cotton to cold and salt stress revealed by genome-wide characterization of BZR genes family

Brassinazole resistant (BZR) transcription factors play important roles in brassinosteroids (BRs) signaling pathway of plants, which are involved in many developmental processes and the responses to abiotic stress by regulating the expression of related genes. In order to explore the molecular mechanism of BZR transcription factors in cotton in response to abiotic stress, the BZR transcription factors in four cotton genomes were identified and analyzed. A total of 67 BZR transcription factors were identified and divided into three subfamilies according to their genetic relationship. The expression levels of BZR under cold, heat, salt, and drought treatments were analyzed to predict their function under abiotic stress. Promoter analysis of BZR proteins showed that light signaling can enhance the BZR transcriptional activity. The subcellular localization experiment showed that GhBZR15 protein was mainly distributed in the nucleus. Arabidopsis thaliana L. overexpressing GhBZR15 gene showed stronger cold tolerance and salt tolerance than wild type. These results, combined with the GhBZR protein-interaction network, suggest that the interaction of GhBZR with phytochrome interacting factors (PIF4) plays an important role in abiotic stress responses. These findings provide a basis for further understanding the function of BZR in cotton.

Genome-wide identification of gap junction gene family and their expression profiles under low temperature stress in noble scallop Chlamys nobilis

Gap junctions, formed by gap junction proteins (GJ), play crucial roles in cell signaling and immune responses. The structure and function of the GJ from vertebrates (called connexins) have been extensively studied. However, little is known about the proteins forming gap junctions in invertebrates (called innexins). In this study, 14 GJ genes of Chlamys nobilis were identified. GJ proteins are mainly distributed on the plasma membrane, and all proteins are hydrophilic Phylogenetic tree analysis showed that the GJ proteins in C. nobilis were distantly related to those in vertebrates but closely related to those in invertebrates. Conserved motifs analysis of these GJ proteins in C. nobilis identified to have 10 conserved motifs, similar to gap junction proteins in other bivalves. Moreover, expression profiles of CnGJ genes under chronic and acute low temperature stress were also investigated. Results showed that chronic low temperature stress had a significant effect on the expression levels of CnGJ genes, and the expression profiles of CnGJ genes showed significantly variation under acute low temperature stress. All these results indicated that CnGJ genes play important roles in environmental adaptation in scallops. The present study initially elucidated the function of gap junction genes in noble scallop C. nobilis, which provides new insights into the GJ genes in mollusks and will help us better understand their roles in environmental stress in scallops.

Genome-wide identification and expression analysis of the GT8 gene family in tomato(Solanum lycopersicum) and the functional of SlGolS1 under cold stress

Glycosyltransferases (GTs) are a diverse superfamily of enzymes involved in glycosylation reactions, with the GT8 (glycosyltransferase 8) playing a crucial role in plant growth, development, and abiotic stress responses. Tomato, widely cultivated and is a thermophilic plant. So it is significant to study how GT8 regulates cold resistance in tomato for plant growth. In this study, we screened the whole genome of tomato by using bioinformatics methods and identified 40 members of the GT8 gene family. Analysis of cold stress transcriptome data and qRT-PCR experiments revealed the potential significance of SlGolS1 in responding to cold stress. SlGolS1 was highly expressed in the stems and flowers of tomato, with its mature protein localized in the chloroplast. Used the VIGS method to transiently silence the SlGolS1 gene, the SlGolS1-silenced plants (pTRV2-SlGolS1) rendered tomato more sensitive to cold stress compared with the control (pTRV2) tomato plant phenotype after cold treatment; enzyme activity assays showed that oxidative damage was more severe in the pTRV2-SlGolS1. In summary, SlGolS1 positively regulates cold resistance in tomato.

Comprehensive analysis of the oligopeptide transporter gene family in maize: Genome-wide identification, structural characterization, and stress-responsive expression

Oligopeptide transporter (OPT) proteins are integral proteins in cell membranes that perform a crucial role in cellular metal homeostasis, mineral uptake, transport of small peptides and abiotic stress tolerance. Hence, in-depth evolutionary, structural, molecular and expression analysis could guide in exploring the mechanism of how these transporters contribute to nutrient balance and abiotic stress tolerance in maize. Following a series of bioinformatic analyses, eight OPTs in maize were identified. These transporters showed significant variations in molecular weight, and have one to five introns, 11 to 15 motifs, 11 to 18 transmembrane domains, multiple beta-strands, multiple alpha and transmembrane helices. ZmOPT1, 5, and 6 are plasma membranes and the rest are endoplasmic reticulum localized. All ZmOPTs are distributed in six different chromosomes having significant phytohormone-responsive cis-elements might be due to their great roles in hormonal regulation in maize. The significant transport activity of these transporters plays a great role in biological processes, cellular components and molecular functions in maize during gene ontology analysis. Co-expression of five genes among eight OPTs with 56 genes might be due to their great role in regulating other genes in multiple signaling pathways or vice-versa. Significant tissue-specific and drought, salinity, nitrogen starvation, and heat stress induced in silico expression following real time PCR validation of ZmOPT1, 7, and 8 genes might guide to elucidate the potential roles of these transporters in specific tissues and abiotic stress tolerance. Hence, the findings of the research might guide plant biologists to develop new varieties of maize having higher efficiency of peptide and metal ion transport, balance, and abiotic stress tolerance. Findings of these structural and expression analyzes might guide the plant biologist for single transgenesis, or developing and transforming programming-based synthetic genetic circuits in maize for advancing research in peptide transport, metal hemostasis, and increasing abiotic stress tolerance in maize.

Identification and Immunological Characterization of Annexin B8 and Annexin E1 from Spirometra Erinaceieuropaei Spargana.

Sparganosis is a parasitic zoonotic disease that poses a serious threat to public hygiene and human health. Annexin is a phospholipid-binding protein with calcium ion binding activity, serving various important functions, including interaction with the parasite and regulation of the host's immune response. In this study, two annexin (ANX) family genes, Spirometra erinaceieuropaei (S. erinaceieuropaei) Annexin B8 (SeANXB8) and E1 (SeANXE1), isolated from spargana, were cloned and immunologically characterized. Both recombinant S. erinaceieuropaei Annexin B8 (rSeANXB8) and E1 (rSeANXE1) were specifically recognized by serum from rats immunized with the recombinant proteins, displaying strong immunoreactivity. They are also among the major components of sparganum excretion/secretion products (ESPs). SeANXE1 was identified in the parasite's tegument, testis, genital pore, ovary, and eggs, while SeANXB8 was found in the parasite's tegument and eggs. Plasminogen (PLG)-binding assays revealed that the two annexins could bind to human PLG in a concentration-dependent manner, which was blocked by the corresponding antibodies. These findings suggest that SeANXB8 and SeANXE1 may be involved in host-parasite interaction and may influence the host's immune response during sparganosis. They could be potential diagnosis and vaccination targets for sparganosis.

In Silico Genome-wide Identification and Expression Analysis of the Pin-2 Serine Protease Inhibitor Gene Family in Eggplant (Solanum melongena L.)

In Silico Genome-wide Identification and Expression Analysis of the Pin-2 Serine Protease Inhibitor Gene Family in Eggplant (Solanum melongena L.)

TaGPAT6 enhances salt tolerance in wheat by synthesizing cutin and suberin monomers to form a diffusion barrier.

One mechanism plants use to tolerate high salinity is the deposition of cutin and suberin to form apoplastic barriers that limit the influx of ions. However, the mechanism underlying barrier formation under salt stress is unclear. Here, we characterized the glycerol-3-phosphate acyltransferase (GPAT) family gene TaGPAT6, encoding a protein involved in cutin and suberin biosynthesis for apoplastic barrier formation in wheat (Triticum aestivum). TaGPAT6 has both acyltransferase and phosphatase activities, which are responsible for the synthesis of sn-2-monoacylglycerol (sn-2 MAG), the precursor of cutin and suberin. Overexpressing TaGPAT6 promoted the deposition of cutin and suberin in the seed coat and the outside layers of root tip cells and enhanced salt tolerance by reducing sodium ion accumulation within cells. By contrast, TaGPAT6 knockout mutants showed increased sensitivity to salt stress due to reduced cutin and suberin deposition and enhanced sodium ion accumulation. Yeast-one-hybrid and electrophoretic mobility shift assays identified TaABI5 as the upstream regulator of TaGPAT6. TaABI5 knockout mutants showed suppressed expression of TaGPAT6 and decreased barrier formation in the seed coat. These results indicate that TaGPAT6 is involved in cutin and suberin biosynthesis and the resulting formation of an apoplastic barrier that enhances salt tolerance in wheat.

Analysis of Alzheimer's disease associated deleterious non-synonymous single nucleotide polymorphisms and their impacts on protein structure and function by performing in-silico methods.

Alzheimer's disease (AD) is a neurodegenerative disorder that is presented with a progressive loss of memory, a decline in cognitive abilities and multiple changes in behavior. Its pathogenicity has been linked to genetic factors in approximately 60-80% of the cases specifically APOE gene family and as well as other gene families. This study utilized advanced computational biology methods to analyze AD-associated nsSNPs extracted from the NHGRI-EBI GWAS Catalog. Ensembl Variant Effect Predictor (VEP) is used to annotate the variants associated with AD. Annotated missense variants were subjected to PolyPhen-2, SNPs&Go, PredictSNP servers which were used to predict pathogenicity of selected missense variants by protein sequence information. DynaMut and DUET servers were applied to determine protein stability due to the amino acid change by integrating protein structure information. Missense variations associated with AD were annotated to 26 proteins and further analyzed in our study. Following rigorous data filtration steps, 15 candidate variants (13 proteins) were identified and subjected to sequence and structure-based analysis. Finally in this in-silico study, five deleterious non-synonymous single nucleotide polymorphisms (nsSNPs) were identified in ACKR2(V41A), APOE(R176C), ATP8B4(G395S), LAMB2(E987K), and TOMM40(R239W), and these findings were subsequently backed-up by existing in-vivo and in-vitro literature. This study not only provides invaluable insight into the intricate pathogenic mechanisms underlying AD but also offers a distinctive perspective that paves the way for future, more comprehensive investigations aimed at unraveling the molecular intricacies responsible for the development and progression of AD. Nonetheless, it is imperative that further rigorous in vivo and in vitro experiments are conducted to validate and expand upon the findings presented here.

Utilizing the mutant library to investigate the functional characterization of GhGLR3.4 regulating jasmonic acid to defense pest infestation.

The glutamate receptor (GLR) serves as a ligand-gated ion channel that plays a vital role in plant growth, development, and stress response. Nevertheless, research on GLRs in cotton is still very limited. The present study conducted a comprehensive analysis of GLRs gene family in cotton. In total, 41 members of the GLR family were identified in cotton unveiling distinct subgroups in comparison to Arabidopsis. Among these members, the third subgroup highlights its pivotal role in cotton's defense against insect infestation. Furthermore, the CRISPR/Cas9 system was utilized to create a mutant library of GLR members, which consisted of a total of 135 independent mutant lines, resulting in the production of novel cotton materials with valuable breeding potential for pest control. Further, this study elucidates the influence of GhGLR3.4 on jasmonic acid (JA) pathway signal transduction and demonstrated its participation in the influx of intracellular Ca2+, which regulates "calcium transients" following stimulation, thereby influencing multiple intracellular reactions. The study also found that GhGLR3.4 influences the synthesis of the JA pathway and actively partakes in long-distance signal transmission among plants, facilitating the transfer of defense signals to neighbor leaves and thereby triggering systemic defense. Consequently, this research advances our knowledge of plants' comprehensive defense mechanism against insect pest infestation.

Identification of the EXORDIUM gene family and functional analysis of VaEXO2a during cold stress response in Vitis amurensis

Identification of the EXORDIUM gene family and functional analysis of VaEXO2a during cold stress response in Vitis amurensis

Identification analysis of GAD gene family, and the role of BoGAD 5 in GABA enrichment in broccoli sprouts

Identification analysis of GAD gene family, and the role of BoGAD 5 in GABA enrichment in broccoli sprouts