Cis-acting Research Articles

Genome-wide identification, characterization and expression profiles of FORMIN gene family in cotton (Gossypium Raimondii L.)

BackgroundGossypium raimondii serves as a widely used genomic model cotton species. Its genetic influence to enhance fiber quality and ability to adapt to challenging environments both contribute to increasing cotton production. The formins are a large protein family that predominately consists of FH1 and FH2 domains. The presence of the formin domains highly regulates the actin and microtubule filament in the cytoskeleton dynamics confronting various abiotic stresses such as drought, salinity, and cold temperatures.ResultsIn this study, 26 formin genes were analyzed and characterized in G. raimondii and mostly were found in the nucleus and chloroplast. According to the evolutionary phylogenetic relationship, GrFH were dispersed and classified into seven different groups and shared an ancestry relationship with MtFH. The GrFH gene structure prediction revealed diverse intron-exon arrangements between groups. The FH2 conserved domain was found in all the GrFH distributed on 12 different chromosomes. Moreover, 11 pairs of GrFH transpired segmental duplication. Among them, GrFH4-GrFH7 evolved 35 million years ago (MYA) according to the evolutionary divergence time. Besides, 57 cis-acting regulatory elements (CAREs) motifs were found to play a potential role in plant growth, development, and in response to various abiotic stresses, including cold stress. The GrFH genes mostly exhibited biological processes resulting in the regulation of actin polymerization. The ERF, GATA, MYB, and LBD, major transcription factors (TFs) families in GrFH, regulated expression in abiotic stress specifically salt as well as defense against certain pathogens. The microRNA of GrFH unveiled the regulatory mechanism to regulate their gene expression in abiotic stresses such as salt and cold. One of the most economic aspects of cotton (G.raimondii) is the production of lint due to its use in manufacturing fabrics and other industrial applications. The expression profiles of GrFH in different tissues particularly during the conversion from ovule to fiber (lint), and the increased levels (up-regulation) of GrFH4, GrFH6, GrFH12, GrFH14, and GrFH26 under cold conditions, along with GrFH19 and GrFH26 in response to salt stress, indicated their potential involvement in combating these environmental challenges. Moreover, these stress-tolerant GrFH linked to cytoskeleton dynamics are essential in producing high-quality lint.ConclusionsThe findings from this study can contribute to elucidating the evolutionary and functional characterizations of formin genes and deciphering their potential role in abiotic stress such as cold and salt as well as in the future implications in wet lab.

Genome-wide characterization of Solanum tuberosum UGT gene family and functional analysis of StUGT178 in salt tolerance

UDP-glycosyltransferases (UGTs) widely exist in plants and play essential roles in catalyzing the glycosylation reaction associated with metabolic processes. UGT gene family has been identified in many species to date. However, the comprehensive identification and systematic analysis have not been documented yet in the latest potato genome. In this study, a total of 295 UGT members (StUGT) were identified and found to be unevenly distributed on twelve chromosomes of potato. All StUGT genes were classified into 17 groups (A-P, R) and the UGT genes within the same groups have similar structural characterization. Tandem duplication was the major driving force for the StUGT gene expansion. The prediction of cis-acting elements showed that the development process, light, phytohormone, and abiotic stress-responsive elements generally existed in StUGT promoter regions. Analysis of spatial and temporal expression patterns demonstrated that StUGT genes were widely and differentially expressed in various tissues. Additionally, to investigate the salt stress-responsive genes, we analyzed the expression profiles of the StUGT genes under salt treatment. A total of 50 and 20 StUGT genes were continuously up- and down-regulated, respectively, implicating that these genes were involved in the regulation of salt tolerance. Among them, the StUGT178 gene, which was significantly induced by salt stress and contains salt-responsive element, was considered as one of the most relevant candidate genes. Transient transformation of the StUGT178 promoter in tobacco revealed that the transcriptional activation activity of the StUGT178 gene was strengthened under salt treatment. Furthermore, the heterologous expressions of the promoter and coding protein of the StUGT178 gene in Arabidopsis further demonstrated that the StUGT178 gene significantly responds to salt treatment, and enhanced salinity tolerance by regulating antioxidant enzyme activity and H2O2 accumulation. These results provide comprehensive information for a better understanding of the StUGT genes and offer a foundation for uncovering their function associated with salt stress in potato.

Genome-Wide Identification and Evolutionary Analysis of Functional BBM-like Genes in Plant Species

Background/Objectives: BABY BOOM (BBM), a transcription factor from the APETALA2 (AP2) protein family, plays a critical role in somatic embryo induction and apomixis. BBM has now been widely applied to induce apomixis or enhance plant transformation and regeneration efficiency through overexpression or ectopic expression. However, the structural and functional evolutionary history of BBM genes in plants is still not well understood. Methods: The protein sequences of 10 selected plant species were used to locate the branch of BBM-Like by key domain identification and phylogenetic tree construction. The identified BBML genes were used for further conserved motif identification, gene structural analysis, miRNA binding site prediction, cis-acting element prediction, collinear analysis, protein–protein interaction network construction, three-dimensional structure modeling, molecular docking, and expression pattern analysis. Results: A total of 24 BBML proteins were identified from 10 representative plant species. Phylogenetic relationship analysis displayed that BBML proteins from eudicots and monocots were divided into two clusters, with monocots exhibiting a higher number of BBMLs. Gene duplication events indicated that whole genome/segmental duplication were the primary drivers of BBML genes’ evolution in the tested species, with purifying selection playing a key role during evolution processes. Comparative analysis of motif, domains, and gene structures revealed that most BBMLs were highly evolutionarily conserved. The expression patterns of BBML genes revealed significant tissue specificity, particularly in the root and embryo. We also constructed protein–protein interaction networks and molecular docking models to identify functional pathways and key amino acid residues of BBML proteins. The functions of BBMLs may differ between monocots and eudicots, as suggested by the functional enrichment of interacting proteins. Conclusions: Our research delved into the molecular mechanism, evolutionary relationships, functional differentiation, and expression patterns of BBML genes across plants, laying the groundwork for further investigations into the molecular properties and biological roles of BBMLs.

Genome-Wide Identification of the ALMT Gene Family in Nine Rosaceae Species and Functional Analysis Associated with Organic Acid Accumulation in Prunus mume

ALMT (aluminum-activated malate transporter) proteins play crucial roles in the transport of organic acids and have significant implications for plant stress responses and development. While extensively studied in some plants, the characteristics and functional divergence of the ALMT gene family have not yet been thoroughly explored in Prunus mume and some other Rosaceae species. In this study, we systematically analyzed the ALMT gene family across nine Rosaceae species to explore their evolutionary relationships, structural characteristics, and functional roles. A total of 138 ALMT genes were identified and categorized into four groups based on a phylogenetic analysis. The motif analysis confirmed the accuracy of the phylogenetic grouping. The collinearity analysis indicated that whole-genome duplication events were the primary drivers of ALMT gene expansion in these species. Furthermore, the cis-acting element analysis revealed diverse regulatory elements associated with environmental responses, including abscisic acid, light, and jasmonic acid. The gene expression correlation analysis showed that PmALMT1 is primarily associated with malic acid accumulation, whereas PmALMT8 is related to citric acid accumulation. Further transient expression in Nicotiana benthamiana validated the above conclusion. This comprehensive analysis provides valuable insights into the evolution, function, and regulation of the ALMT gene family in Rosaceae species.

An integrative TAD catalog in lymphoblastoid cell lines discloses the functional impact of deletions and insertions in human genomes.

The human genome is packaged within a three-dimensional (3D) nucleus and organized into structural units known as compartments, topologically associating domains (TADs), and loops. TAD boundaries, separating adjacent TADs, have been found to be well conserved across mammalian species and more evolutionarily constrained than TADs themselves. Recent studies show that structural variants (SVs) can modify 3D genomes through the disruption of TADs, which play an essential role in insulating genes from outside regulatory elements' aberrant regulation. However, how SV affects the 3D genome structure and their association among different aspects of gene regulation and candidate cis-regulatory elements (cCREs) have rarely been studied systematically. Here, we assess the impact of SVs intersecting with TAD boundaries by developing an integrative Hi-C analysis pipeline, which enables the generation of an in-depth catalog of TADs and TAD boundaries in human lymphoblastoid cell lines (LCLs) to fill the gap of limited resources. Our catalog contains 18,865 TADs, including 4596 sub-TADs, with 185 SVs (TAD-SVs) that alter chromatin architecture. By leveraging the ENCODE registry of cCREs in humans, we determine that 34 of 185 TAD-SVs intersect with cCREs and observe significant enrichment of TAD-SVs within cCREs. This study provides a database of TADs and TAD-SVs in the human genome that will facilitate future investigations of the impact of SVs on chromatin structure and gene regulation in health and disease.

Genome-Wide Identification and Expression Analysis of Pseudo-Response Regulators (PRRs) in the Tea Plant Camellia sinensis (L.) O. Kuntze

The circadian clock plays a vital role in facilitating plant adaptation to rhythmically changing environmental factors. Pseudo-response regulators (PRRs) are key components regulating the plant circadian clock and have been extensively characterized in model plants. However, the PRRs in the tea plant have not been comprehensively studied. In this study, seven CsPRRs were cloned from the tea plant. Domain, phylogenetic evolution, gene structure, motifs, and cis-acting element analysis revealed their sequence characters and suggested that the first subgroup members, CsPRR1a, 1b, 5a, 5b, 7, and 37, may be responsible for circadian rhythm regulation and abiotic stress responses, while the second subgroup member, CsPRR2, may be involved in development and chloroplast function regulation. Most CsPRRs showed relatively higher expression in flowers, implying their potential roles in photoperiod-regulated flower induction. Moreover, rhythmic expression of CsPRR7, 5b, 5a, 37, 1b, and 1a was observed under long-day conditions in a sequential manner. Additionally, CsPRRs were differently induced/inhibited by cold, heat, and drought stresses in tissue-specific and photoperiod-related manners. A stronger cold induction of CsPRRs was observed under long-day conditions than under short-day conditions. And, among the two tested tissues, changes in the expression of CsPRRs caused by various stresses were more obvious in young shoots. Studies using a floriferous cultivar (FDDB) and an oliganthous cultivar (PYTZ) implied that CsPRRs also played crucial roles in tea-plant flower induction. This study presents the first comprehensive analysis of CsPRRs in the tea plant, providing vital information for further elucidation of CsPRR functions. It also suggests that tissue type and photoperiod conditions should be taken into consideration when conducting gene function studies in the tea plant.

Dynamic O-GlcNAcylation governs long-range chromatin interactions in V(D)J recombination during early B-cell development.

V(D)J recombination secures the production of functional immunoglobulin (Ig) genes and antibody diversity during the early stages of B-cell development through long-distance interactions mediated by cis-regulatory elements and trans-acting factors. O-GlcNAcylation is a dynamic and reversible posttranslational modification of nuclear and cytoplasmic proteins that regulates various protein functions, including DNA-binding affinity and protein-protein interactions. However, the effects of O-GlcNAcylation on proteins involved in V(D)J recombination remain largely unknown. To elucidate this relationship, we downregulated O-GlcNAcylation in a mouse model by administering an O-GlcNAc inhibitor or restricting the consumption of a regular diet. Interestingly, the inhibition of O-GlcNAcylation in mice severely impaired Ig heavy-chain (IgH) gene rearrangement. We identified several factors crucial for V(D)J recombination, including YY1, CTCF, SMC1, and SMC3, as direct targets of O-GlcNAc modification. Importantly, O-GlcNAcylation regulates the physical interaction between SMC1 and SMC3 and the DNA-binding patterns of YY1 at the IgH gene locus. Moreover, O-GlcNAc inhibition downregulated DDX5 protein expression, affecting the functional association of CTCF with its DNA-binding sites at the IgH locus. Our results showed that locus contraction and long-range interactions throughout the IgH locus are disrupted in a manner dependent on the cellular O-GlcNAc level. In this study, we established that V(D)J recombination relies on the O-GlcNAc status of stage-specific proteins during early B-cell development and identified O-GlcNAc-dependent mechanisms as new regulatory components for the development of a diverse antibody repertoire.

GH3 Gene Family Identification in Chinese White Pear (Pyrus bretschneideri) and the Functional Analysis of PbrGH3.5 in Fe Deficiency Responses in Tomato.

Iron (Fe) deficiency poses a major threat to pear (Pyrus spp.) fruit yield and quality. The Gretchen Hagen 3 (GH3) plays a vital part in plant stress responses. However, the GH3 gene family is yet to be characterized, and little focus has been given to the function of the GH3 gene in Fe deficiency responses. Here, we identified 15 GH3 proteins from the proteome of Chinese white pear (Pyrus bretschneideri) and analyzed their features using bioinformatics approaches. Structure domain and motif analyses showed that these PbrGH3s were relatively conserved, and phylogenetic investigation displayed that they were clustered into two groups (GH3 I and GH3 II). Meanwhile, cis-acting regulatory element searches of the corresponding promoters revealed that these PbrGH3s might be involved in ABA- and drought-mediated responses. Moreover, the analysis of gene expression patterns exhibited that most of the PbrGH3s were highly expressed in the calyxes, ovaries, and stems of pear plants, and some genes were significantly differentially expressed in normal and Fe-deficient pear leaves, especially for PbrGH3.5. Subsequently, the sequence of PbrGH3.5 was isolated from the pear, and the transgenic tomato plants with PbrGH3.5 overexpression (OE) were generated to investigate its role in Fe deficiency responses. It was found that the OE plants were more sensitive to Fe deficiency stress. Compared with wild-type (WT) plants, the rhizosphere acidification and ferric reductase activities were markedly weakened, and the capacity to scavenge reactive oxygen species was prominently impaired in OE plants under Fe starvation conditions. Moreover, the expressions of Fe-acquisition-associated genes, such as SlAHA4, SlFRO1, SlIRT1, and SlFER, were all greatly repressed in OE leaves under Fe depravation stress, and the free IAA level was dramatically reduced, while the conjugated IAA contents were notably escalated. Combined, our findings suggest that pear PbrGH3.5 negatively regulates Fe deficiency responses in tomato plants, and might help enrich the molecular basis of Fe deficiency responses in woody plants.

Genome-wide identification and expression analysis of the AS2/LOB gene family in physic nut.

AS2/LOB genes, a class of transcription factors ubiquitously existing in plants, are vital for plant growth, development, and stress tolerance. Despite the availability of the physic nut genome, information regarding the expression profiles and evolutionary histories of its AS2/LOB genes remains scarce. An elaborate exploration of the AS2/LOB gene family was conducted, including phylogeny, exon-intron structure, chromosomal location, conserved domain characteristics, conserved motifs, promoter cis-acting elements, protein interaction, and expression profiles under normal growth and abiotic stress conditions. In this study, 28 AS2/LOB genes (JcASLs) were identified in the physic nut genome. Phylogenetic analysis, based on homologs from Arabidopsis, classified the 28 JcASLs genes into two groups (calss I and II). Chromosome localization indicated that the 28 JcASLs genes were unevenly distributed across nine chromosomes. RNA-seq and qRT-PCR results revealed that the majority of the 28 JcASLs genes exhibited differential expression in tissues such as roots, cortex stems, leaves, and seeds. Notably, JcASL8 and JcASL13 were exclusively expressed in seeds, and 16 JcASLs genes responded to drought and salt stress at least at one time point under at least one treatment condition. These results establish a basis for future investigations into the molecular mechanism by which the JcASLs genes regulate physic nut's response to drought and salt stress and their role in modulating the growth and development of physic nut.

Genome-wide analysis and expression profiling of the polyamine oxidase gene family in Solanum tuberosum L.

BackgroundPolyamine oxidase (PAO) is a crucial enzyme involved in the breakdown of polyamines (PAs) in plants. It not only regulates the levels of PAs, but also plays a role in the oxidative decomposition of PAs and the release of stress-related signals, contributing to the plant’s response and resistance to various adversities. While there have been numerous studies on the response of PAO to stress in other crops, there is a lack of research on this topic in potatoes, a major food crop.ResultsIn this study, we aimed to explore the biological function of the StPAO gene in potato growth and development, as well as its expression patterns under stress. Using bioinformatics methods, we identified 14 StPAO genes in the potato genome. Protein sequence comparisons revealed a high similarity between the PAO proteins of potato and Arabidopsis. Chromosomal mapping and gene structure analysis showed that the StPAO genes were not evenly distributed on the chromosome and all contained an amino-oxidase domain. Furthermore, analysis of the promoters of these genes revealed the presence of abiotic and stress-related cis-acting elements, indicating their potential role in responding to different stresses. To investigate the expression patterns of these genes under stress, we used qRT-PCR to study their response to high temperature, drought, and ABA stress. Our results showed that StPAO6 and StPAO10 were significantly up-regulated under high temperature stress, indicating that they were involved in the process of potato resistance to high temperatures. Similarly, StPAO1, StPAO3, and StPAO4 were significantly up-regulated under drought stress, indicating their potential role in potatoes’ responses to drought. After ABA treatment, the expression levels of StPAO4, StPAO5, StPAO7, and StPAO14 were significantly up-regulated, suggesting their involvement in chemical defense mechanisms. Interestingly, the expression of StPAO11–13 was inhibited by all three stresses.ConclusionsIn conclusion, our study highlights the multifunctional nature of the StPAO gene family in potatoes, which plays a crucial role in coping with various stresses. This research deepens our understanding of the potato StPAO gene family and provides a reference for future studies on its function. It also serves as a theoretical basis for breeding stress-resistant potato varieties in the future.

Identification of ANS gene regulating fruit color in jujube (Ziziphus jujuba Mill. cv. Lingwuchangzao) and analysis of its expression under elevated temperature and drought stress

BackgroundThe anthocyanin synthase (ANS) gene is pivotal in plant anthocyanin biosynthesis, significantly influencing fruit coloration. Despite extensive on various studies, there remains a notable absence of research on the regulatory functions of the ANS gene in the ‘Lingwuchangzao’ variety under elevated temperature and drought stress.ResultsIn this study, 107 ZjANS genes were identified. Analyses of their physicochemical properties and gene structures revealed that most are localized in the cytosol and nucleus, with only seven ZjANS genes exhibiting basic characteristics. All genes contained the conserved domains 2OG-FeII_Oxy and DIOX-N. Chromosomal localization and Ka/Ks analysis indicated that these genes are distributed across 12 chromosomes, with 22 tandem gene pairs, suggesting purifying selection. Phylogenetic and synteny analysis demonstrated that ZjANS genes are evolutionarily conserved, forming collinear gene pairs with four species. Cis-acting element analysis identified elements related to anthocyanin synthesis and environmental stress, such as LTR and MBS. Predictions of protein interaction indicated that certain ZjANS proteins interact with MYB, CHI, and GA proteins, potentially involved in the anthocyanin synthesis pathway. Transcriptome and qRT-PCR analysis revealed that thirteen ZjANS genes positively respond to coloring in jujube under elevated temperature and drought stress, with ZjANS17 identified as particularly significant.ConclusionThis study identified 107 ZjANS genes and systematically analyzed their evolutionary conservation. Thirteen ZjANS genes were found to positively influence fruit coloring in ‘Lingwuchangzao’ under elevated temperature and drought stress. These findings provide a foundation for further investigations into ZjANS genes and the breeding new jujube varieties.

Comparative genomics analysis of the MYB gene family in barley: preliminary insights into evolution and biological function in Blue Qingke.

The Myeloblastosis related (MYB) family is one of the most widely distributed transcription factor families in plants and plays a significant role in plant growth and development, hormone signal transduction, and stress response. There are many reports on MYB family species, but the research on Qingke is still limited. This study used comparative genomics methods to analyze gene and protein structure, protein physicochemical properties, chromosome localization, and evolution. A bioinformatics approach was used to systematically analyze the HvMYB gene family. At the milk stage, soft dough stage, and mature stage, White and Blue Qingke grains were selected for RNA sequencing (RNA-seq), among which two proteins interacted (HvMYB and HvMYC). The expression of this gene family was analyzed through RNA-seq, and the expression levels of HvMYB and HvMYC in the grains of two different color varieties were analyzed by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Finally, the interaction between HvMYB and HvMYC was verified by bimolecular fluorescence complementation (BiFC) experiments. A total of 92 Qingke HvMYB genes were identified and analyzed, and 92 HvMYB proteins were classified into five categories. Cis-acting elements associated with abscisic acid response, light response, and methyl jasmonate (MeJA) response were found in the promoter regions of most MYB genes. Using qRT-PCR combined with RNA-seq analysis showed that MYB gene was highly expressed in the soft dough stage and was varietal specific. Subcellular localization indicated that HvMYB was located in the nucleus and cell membrane, HvMYC was located in the nucleus, cell membrane, and cytoplasm. Through BiFC analysis, it has been proven that HvMYB in the MYB family and HvMYC in the basic helix-loop-helix (bHLH) family can interact. This study provides a preliminary theoretical basis for understanding the function and role of the Qingke MYB gene family and provides a reference for the molecular mechanism of Qingke gene evolution.

An enhancer-promoter-transcription factor module orchestrates plant immune homeostasis by constraining camalexin biosynthesis

Effective plant defense against pathogens relies on highly coordinated regulation of immune gene expression. Enhancers, as cis-regulatory elements, are indispensable determinants of dynamic gene regulation, but the molecular functions in plant immunity are not well understood. In this study, we identified a novel enhancer, CORE PATTERN-INDUCED ENHANCER 35 (CPIE35), which is rapidly activated upon pathogenic elicitation and negatively regulates anti-fungal resistance through modulating WRKY15 expression. During immune activation, CPIE35 activates the transcription of WRKY15 by forming chromatin loops with the promoter of WRKY15 in a WRKY18/40/60-, WRKY33-, and MYC2-dependent manner. WRKY15 directly binds to the promoters of PAD3 and GSTU4, suppressing their expression and leading to the reduced camalexin synthesis and resistance. Interestingly, CPIE35 region is evolutionarily conserved among Brassicaceae plants, and the CPIE35-WRKY15 module exerts similar functions in Brassica napus to negatively regulate anti-fungal resistance. Our work reveals the “enhancer-promoter-transcription factor” regulatory mechanism in maintenance of immune homeostasis, highlighting the importance and conserved role of enhancers in fine-tuning immune gene expression in plants.

Inferring causal relationships among histone modifications in exon skipping event

Alternative splicing is a crucial processes of gene expression. Over 90% multi-exonic genes in human genome undergo alternative splicing. Although the splicing code has been proposed, it still couldn’t satisfactorily explain the tissue-specific alternative splicing. Results of co-transcriptional RNA processing analysis demonstrated that, except for trans- and cis-acting elements, histone modifications also play a role alternative splicing. In the present work, we analyzed the associations among 27 kinds of histone modifications in H1 human embryonic stem cell. In order to illustrate the casual relationships between histone modification and alternative splicing, we built the Bayesian network and validated its robustness by using cross validation test. In addition to the combinatorial patterns, distinct histone modification patterns were also observed in the alternative spliced exons and surrounding intron regions, indicating that histone modifications could substantially mark alternative splicing.

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.

Evolutionary Insights and Expression Dynamics of the Glutathione Peroxidase (GPX) Gene Family in Water lily (Nymphaea colorata) in Response to Multiple Abiotic Stresses

The glutathione peroxidase (GPX) family in plants contains various isoenzymes, each with distinct subcellular localizations, tissue-specific expression patterns, and responses to environmental stress. In current study, we conducted genome-wide bioinformatics analyses of the GPX gene family in Nymphaea colorata, identifying five putative NcGPX genes dispersed across five of the fourteen chromosomes. Examination of the protein sequences revealed that NcGPXs comprehend three conserved cysteine residues, with predicted in-silico subcellular localizations in chloroplast, extracellular, or cytoplasm. We reported that NcGPX genes protein lengths and molecular weight (MW) ranged from 171-248 amino acids (aa) and 18.798-27.076 kDa, respectively. A comparative neighbor-joining phylogenetic study revealed that GPX genes were clustered into four main groups (Group A– D) from water lily and two closely related plant species, alongside with Arabidopsis thaliana. The universal analysis showed that segmental duplications occurred within the NcGPX gene family. Investigation of gene structure and motifs suggested that most NcGPX genes have relatively conserved exon-intron structures and motif arrangements. Promoter region exploration of NcGPX genes revealed numerous cis-acting regulatory elements associated with development, stress, and hormone responses. The identified 3D protein structures showed that water lily GPXs form conserved dimeric protein signatures. Finally, gene expression and enzymes accumulation of NcGPXs in response to different abiotic stresses, in leaves of N. colorata were analysed using real-time RT-qPCR and spectrophotometry. The results indicate that different members of the GPX gene family are co-ordinately regulated under specific environmental stress conditions, establishing a foundation for studying their gene functions and improving abiotic stress tolerance in water lily genetic breeding.

Deciphering genetic mechanisms of Al toxicity tolerance through meta-QTL analysis in rice (Oryza sativa L.)

Rice is known for its tolerance to high aluminum (Al) concentrations in soil. However, the precise genetic and physiological mechanisms are yet to be fully understood. Recent research has identified several candidate genes (CGs) and quantitative trait loci (QTLs) associated with Al toxicity tolerance in rice. Nevertheless, many more QTLs/genes are yet to be precisely mapped. We employed meta-QTL (M-QTL) analysis, integrating 12 independent mapping studies and 5 Genome-Wide Association Studies (GWAS). The meta-analysis identified 53 M-QTLs from 157 projected QTLs, which were further narrowed down to 28 M-QTLs based on the number of overlapping QTLs on a consensus map. Gene identification through batch retrieval from the RAP database yielded 2,765 non-redundant genes within the 28 M-QTL regions. Comparison of M-QTL CGs with six expression datasets associated with Al toxicity tolerance in rice resulted in the identification of 219 CGs with significant differential expression. Notably, 34 CGs were identified to be common across at least 2 studies. Further downstream analyses of CGs revealed the presence of cis-regulatory elements, transcription factors, and transporter proteins related to the Al toxicity tolerance response. Additionally, we analyzed the expression patterns of the four CGs, namely NRT2.3, ALMT4, MT1, and MTP11, which showed significant upregulation in the Al toxicity-tolerant rice genotype, Anjali. Conversely, in the sensitive genotype Swarna, only NRT2.3 exhibited upregulation, while ALMT4, MT1, and MTP11 were downregulated. Our study highlights significant meta-regions that hold the potential for improving rice genotypes for enhanced tolerance to Al toxicity in acidic soils.

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.

Genome‑wide identifcation and expression analysis of growth-regulating factors under drought in Brassica juncea

Growth-regulating factors (GRFs) are plant-specific transcription factors involved in the regulation of plant growth, development, and abiotic stress processes. However, the function of Brassica juncea (L.) Czern & Coss GRFs remain largely unknown. In this study, 34 BjGRF genes were identified in B. juncea. BjGRF members of the same subfamily were found to share a similar motif composition and gene structure. In total, 663 cis-acting elements were found in the promoter regions of BjGRF genes, which were related to light response, hormone response, environmental stresses, and plant growth/development. Additionally, 48 pairs of segmental duplication genes were identified during gene duplication events, and no tandemly duplicated genes were identified. qRT-PCR analysis showed that the 34 BjGRF genes were primarily expressed in the roots, followed by the leaves. Furthermore, the 10 BjGRF genes were screened in response to drought stress, and the expression patterns of the genes were relatively consistent, with a maximum expression level at 3 or 24 h, and the selected BjGRF03 and BjGRF32 might be involved in drought stress. This preliminary study clarifies the response of the BjGRF gene family to drought stress and provides ideas for further analyses of the biological functions of BjGRF genes.

Integrative transcriptome and metabolome analyses reveal the mechanism of melatonin in delaying postharvest senescence in cowpeas

Rapid postharvest senescence and quality deterioration severely limit logistics of cowpeas. Melatonin (MEL) is a pivotal bioactive molecule that can modulate multiple physiological attributes in plants. In this study, physiological, transcriptomic and metabolomic analyses were conducted to explore the effects of exogenous MEL on cowpea senescence and its underlying mechanisms. Physiological results showed that 100 μM MEL treatment maintained sensory quality (greeness, firmness and soluble solids content), reduced weight loss as well as inhibited the degradation of chlorophyll (Chl) and protopectin. Preservation of color and firmness in cowpeas was greatly attributed to inhibition of expression of genes related to Chl and cell wall metabolism, which was based on a transcriptomic analysis. Integrated transcriptomic and metabolomic analyses revealed that MEL promoted transcription of genes associated with amino acid and carbohydrate metabolism, resulting in the accumulation of amino acid and sugar metabolites. Additionally, by integrating transcription factor-binding site analysis with cis-acting element analysis, we constructed a regulatory network of transcription factors underlying MEL-mediated antisenescence. The present study found a series of potential candidate genes and metabolites involved in regulating senescence process and provided an insight into improving postharvest quality of cowpeas.