Characterization and Comparative Analysis of RWP-RK Proteins from Arachis duranensis, Arachis ipaensis, and Arachis hypogaea

RWP-RK proteins are important factors involved in nitrate response and gametophyte development in plants, and the functions of RWP-RK proteins have been analyzed in many species. However, the characterization of peanut RWP-RK proteins is limited. In this study, we identified 16, 19, and 32 RWP-RK members from Arachis duranensis, Arachis ipaensis, and Arachis hypogaea, respectively, and investigated their evolution relationships. The RWP-RK proteins were classified into two groups, RWP-RK domain proteins and NODULE-INCEPTION-like proteins. Chromosomal distributions, gene structures, and conserved motifs of RWP-RK genes were compared among wild and cultivated peanuts. In addition, we identified 12 orthologous gene pairs from the two wild peanut species, 13 from A. duranensis and A. hypogaea, and 13 from A. ipaensis and A. hypogaea. One, one, and seventeen duplicated gene pairs were identified within the A. duranensis, A. ipaensis, and A. hypogaea genomes, respectively. Moreover, different numbers of cis-acting elements in the RWP-RK promoters were found in wild and cultivated species (87 in A. duranensis, 89 in A. ipaensis, and 92 in A. hypogaea), and as a result, many RWP-RK genes showed distinct expression patterns in different tissues. Our study will provide useful information for further functional and evolutionary analysis of the RWP-RK genes.

The Housekeeping Transporter SLC25A44 Bridges between the Secondary Metabolism and Mitochondrial Electron Transfer Chains

The solute carrier family 25 (SLC25) participates in the transport of metabolites and cofactors across the membranes of mitochondria, plastids, peroxisomes, and endoplasmic reticulum. By calling for genomic blocks involved in adjacent metabolic reactions, this report introduces gene clusters of the Slc25 subfamily 44, stilbene and chalcone synthases, and subunits of the mitochondrial electron transfer complexes. The Slc25A44 gene was found ubiquitously expressed and transcriptionally co-regulated with energy metabolism genes in human, mouse, and Arabidopsis thaliana. The Slc25A44s also had no homozygous missense mutation and were highly conserved at intra-species level with the majority of polymorphism present in the non-coding regions. When expressed in oocytes, AdSlc25A44 from Arachis duranensis showed transport activity for the common precursors of flavonoids, stilbenoids, and ubiquinone. Accordingly, AdSLC25A44 and its human orthologue HsSLC25A44 elevated the production of para-coumaric, 4-aminobenzoic, and 4-hydroxybenzoic acids in Saccharomyces cerevisiae strains designed to produce para-coumaric acid via different pathways. Moreover, the engineered SLC25 subfamily specific signature, i.e., AdSLC25A44LWW206IQF, had a stronger effect on para-coumaric acid secretion than the native variant. Importantly, the aerobic growth-rate of S. cerevisiae was significantly higher when expressing the AdSLC25A44, HsSLC25A44, or AdSLC25A44LWW206IQF. These results suggest that SLC25A44 is an essential mitochondrion-ER-nucleus zone transporter associated with metabolism of secondary metabolites and energy.

Genome-wide identification and characterization of non-specific lipid transfer protein (nsLTP) genes in Arachis duranensis

Background: Non-specific lipid transfer proteins (nsLTPs) are known to transfer various lipid molecules between lipid bilayers in plants. In Arachis duranensis, however, little is known about the nsLTPs and their responses to biotic and abiotic stresses. Results: In this study, we identified 64 nsLTP family members (AdLTPs) in A. duranensis. These AdLTPs were classified into six subfamilies (Types 1, 2, C, D, E, and G) and were randomly distributed along nine chromosomes. The Ks value and Ka/Ks value significantly differed between Type 1 and Type D subfamilies. Among paralogs, eight AdLTPs were under positive selection, indicating that these AdLTPs might have different functions in the evolutionary history of A. duranensis. qRT-PCR results showed that the expression of AdLTPs changed in response to abiotic stresses, including salinity, PEG, low temperature, and ABA. Using RNA-seq data, we also found three AdLTPs (AdLTP1.14, AdLTPd8, and AdLTPe2) that were possibly associated with resistance to nematode infection. Among them, AdLTP1.14, which belongs to the Type 1 subfamily, was up-regulated at three time points after nematode infection. Co-expression analysis indicated that DOF and WRI1 transcription factors may regulate the AdLTP response to nematode infection. Conclusions: We identified AdLTPs in A. duranensis. Based on both RNA-seq and qRT-PCR datasets, we found that AdLTPs are involved in responses to biotic and abiotic stresses. Our results could provide valuable genomic information for the breeding of peanut cultivars that are resistant to biotic and abiotic stresses.

B-box Proteins in Arachis duranensis: Genome-Wide Characterization and Expression Profiles Analysis

B-box (BBX) proteins are important factors involved in plant growth and developmental regulation, and they have been identified in many species. However, information on the characteristics and transcription patterns of BBX genes in wild peanut are limited. In this study, we identified and characterized 24 BBX genes from a wild peanut, Arachis duranensis. Many characteristics were analyzed, including chromosomal locations, phylogenetic relationships, and gene structures. Arachis duranensis B-box (AdBBX) proteins were grouped into five classes based on the diversity of their conserved domains: I (3 genes), II (4 genes), III (4 genes), IV (9 genes), and V (4 genes). Fifteen distinct motifs were found in the 24 AdBBX proteins. Duplication analysis revealed the presence of two interchromosomal duplicated gene pairs, from group II and IV. In addition, 95 kinds of cis-acting elements were found in the genes’ promoter regions, 53 of which received putative functional predictions. The numbers and types of cis-acting elements varied among different AdBBX promoters, and, as a result, AdBBX genes exhibited distinct expression patterns in different tissues. Transcriptional profiling combined with synteny analysis suggests that AdBBX8 may be a key factor involved in flowering time regulation. Our study will provide essential information for further functional investigation of AdBBX genes.

Defining the function of SUMO system in pod development and abiotic stresses in Peanut

Background Posttranslational modification of proteins by small ubiquitin like modifier (SUMO) proteins play an important role during the developmental process and in response to abiotic stresses in plants. However, little is known about SUMOylation in peanut (Arachis hypogaea L.), one of the world’s major food legume crops. In this study, we characterized the SUMOylation system from the diploid progenitor genomes of peanut, Arachis duranensis (AA) and Arachis ipaensis (BB). Results Genome-wide analysis revealed the presence of 40 SUMO system genes in A. duranensis and A. ipaensis. Our results showed that peanut also encodes a novel class II isotype of the SCE1, which was previously reported to be uniquely present in cereals. RNA-seq data showed that the core components of the SUMOylation cascade SUMO1/2 and SCE1 genes exhibited pod-specific expression patterns, implying coordinated regulation during pod development. Furthermore, both transcripts and conjugate profiles revealed that SUMOylation has significant roles during the pod development. Moreover, dynamic changes in the SUMO conjugates were observed in response to abiotic stresses. Conclusions The identification and organization of peanut SUMO system revealed SUMOylation has important roles during stress defense and pod development. The present study will serve as a resource for providing new strategies to enhance agronomic yield and reveal the mechanism of peanut pod development.

Molecular and transcriptional characterization of phosphatidyl ethanolamine-binding proteins in wild peanuts Arachis duranensis and Arachis ipaensis

Background Phosphatidyl ethanolamine-binding proteins (PEBPs) are involved in the regulation of plant architecture and flowering time. The functions of PEBP genes have been studied in many plant species. However, little is known about the characteristics and expression profiles of PEBP genes in wild peanut species, Arachis duranensis and Arachis ipaensis, the diploid ancestors of cultivated peanuts. Results In this study, genome-wide identification methods were used to identify and characterize a total of 32 peanut PEBP genes, 16 from each of the two wild peanut species, A. duranensis and A. ipaensis. These PEBP genes were classified into 3 groups (TERMINAL FLOWER1-like, FLOWERING LOCUS T-like, and MOTHER OF FT AND TFL1-like) based on their phylogenetic relationships. The gene structures, motifs, and chromosomal locations for each of these PEBPs were analyzed. In addition, 4 interchromosomal duplications and 1 tandem duplication were identified in A. duranensis, and 2 interchromosomal paralogs and 1 tandem paralog were identified in A. ipaensis. Ninety-five different cis-acting elements were identified in the PEBP gene promoter regions and most genes had different numbers and types of cis-elements. As a result, the transcription patterns of these PEBP genes varied in different tissues and under long day and short day conditions during different growth phases, indicating the functional diversities of PEBPs in different tissues and their potential functions in plant photoperiod dependent developmental pathways. Moreover, our analysis revealed that AraduF950M/AraduWY2NX in A. duranensis, and Araip344D4/Araip4V81G in A. ipaensis are good candidates for regulating plant architecture, and that Aradu80YRY, AraduYY72S, and AraduEHZ9Y in A. duranensis and AraipVEP8T in A. ipaensis may be key factors regulating flowering time. Conclusion Sixteen PEBP genes were identified and characterized from each of the two diploid wild peanut genomes, A. duranensis and A. ipaensis. Genetic characterization and spatio-temporal expression analysis support their importance in plant growth and development. These findings further our understanding of PEBP gene functions in plant species.

Characterization of B-box proteins and their contribution to plant development in Arachis duranensis

Background B-box (BBX) proteins are important factors involving in the regulation of plant growth and development, and have been identified in many plant species. However, the characteristics and transcription patterns of BBX genes in wild peanut are limited. Results In the present study, we identified and characterized 24 BBX genes in a wild peanut Arachis duranensis. The AdBBX members distributed on 9 of the 10 chromosomes and chromosome 3 contained the most AdBBX members, with 6 AdBBXs. 16 AdBBX proteins had two distinct BBX domains, 11 members contained one CCT domain, and 7 genes had both BBX and CCT domains. Protein structure analysis revealed that AdBBX were classified into five clades: I (3 genes), II (4 genes), III (4 genes), IV (9 genes) and V (4 genes), on the basis of the diversity of conserved BBX and CCT domains. Moreover, 15 distinct motifs were found in these 24 AdBBX proteins and motif 1 and 5 existed in all the AdBBX proteins. Duplication analysis revealed that 4 interchromosomal duplicated gene pairs were obtained and all of them belonged to group IV. In addition, 95 kinds of cis-acting elements were found in the promoter regions of AdBBXs and 53 types were predicted to have putative functions. The numbers and types of cis-acting elements varied in these AdBBX promoters, as a result, AdBBX genes exhibited distinct expression levels in different tissues. The transcription investigation combined with synteny analysis suggested AdBBX8 might be the key factor involving in flowering time regulation in Arachis duranensis. Conclusion Overall, this study provides a genome-wide identification of BBX genes in a wild peanut Arachis duranensis. Characteristic and transcription pattern analysis revealed their critical roles in plant growth and development. Our study will provide essential information for further functional characteristic investigation of AdBBX genes.