Arachis Duranensis Research Articles

Genome-wide analysis of two repeats containing MYB transcription factors in groundnut identifies drought-inducible genes involved as a negative regulator of root nodulation

The MYB gene superfamily encompasses a group of related genes found in all eukaryotes. In contrast to animals, higher plants contain large numbers of two-repeat MYB genes, which have been established in regulating crucial developmental, biotic, and abiotic stress responses that profoundly affect the plant's yield. However, a comprehensive analysis of the two-repeat MYB gene family in groundnut and its progenitors, especially the role of two-repeat MYB genes in response to drought stress, and the effect of those genes in nodulation have not been reported so far. Our recent analysis of the groundnut genome has identified 79 (Arachis duranensis), 84 (Arachis ipaensis), and 161 (Arachis hypogaea) two-repeat MYB genes, which belong to multiple distinct subgroups based on their architecture. Here, we provided a complete overview of the gene structure, protein motif organization, chromosome localization, gene duplication events, and synteny analysis to clarify evolutionary perspectives. Members of the same subgroup showed highly conserved structure and motif compositions. The whole-genome duplication and segmental duplication likely contributed to the expansion of two-repeat MYB genes in Arachis hypogaea. The upstream sequences of most genes contained phytohormone-responsive, stress-responsive, light-responsive, and plant growth-related elements. The genes not only have diverse expression profiles across different development stages but as shown in our experimental findings, most of them were induced by ABA and drought-related stress. Further gene silencing experiments demonstrated that two drought-inducible MYB genes, homologs of Arabidopsis MYB96 and MYB94, function as a negative regulator of root nodulation. The results of this study can serve as a strong foundation for further elucidation of the physiological and molecular function of two-repeat MYB genes in groundnut.

The Stilbene Synthase Family in Arachis: A Genome-Wide Study and Functional Characterization in Response to Stress.

Peanut (Arachis hypogaea) and its wild relatives are among the few species that naturally synthesize resveratrol, a well-known stilbenoid phytoalexin that plays a crucial role in plant defense against biotic and abiotic stresses. Resveratrol has received considerable attention due to its health benefits, such as preventing and treating various human diseases and disorders. Chalcone (CHS) and Stilbene (STS) Synthases are plant-specific type III Polyketide Synthases (PKSs) that share the same substrates and are key branch enzymes in the biosynthesis of flavonoids and stilbenoids, respectively. Although resveratrol accumulation in response to external stimulus has been described in peanut, there are no comprehensive studies of the CHS and STS gene families in the genus Arachis. In the present study, we identified and characterized 6 CHS and 46 STS genes in the tetraploid peanut and an average of 4 CHS and 22 STS genes in three diploid wild species (Arachis duranensis, Arachis ipaënsis and Arachis stenosperma). The CHS and STS gene and protein structures, chromosomal distributions, phylogenetic relationships, conserved amino acid domains, and cis-acting elements in the promoter regions were described for all Arachis species studied. Based on gene expression patterns of wild A. stenosperma STS genes in response to different biotic and abiotic stresses, we selected the candidate AsSTS4 gene, which is strongly induced by ultraviolet (UV) light exposure, for further functional investigation. The AsSTS4 overexpression in peanut hairy roots significantly reduced (47%) root-knot nematode infection, confirming that stilbene synthesis activation in transgenic plants can increase resistance to pathogens. These findings contribute to understanding the role of resveratrol in stress responses in Arachis species and provide the basis for genetic engineering for improved production of valuable secondary metabolites in plants.

The role of microRNAs in responses to drought and heat stress in peanut (Arachis hypogaea).

MicroRNAs (miRNAs) are 21-24nt small RNAs (sRNAs) that negatively regulate protein-coding genes and/or trigger phased small-interfering RNA (phasiRNA) production. Two thousand nine hundred miRNA families, of which ∼40 are deeply conserved, have been identified in ∼80 different plant species genomes. miRNA functions in response to abiotic stresses is less understood than their roles in development. Only seven peanut MIRNA families are documented in miRBase, yet a reference genome assembly is now published and over 480 plant-like MIRNA loci were predicted in the diploid peanut progenitor Arachis duranensis genome. We explored by computational analysis of a leaf sRNA library and publicly available sRNA, degradome, and transcriptome datasets the miRNA and phasiRNA space associated with drought and heat stresses in peanut. We characterized 33 novel candidate and 33 ancient conserved families of MIRNAs and present degradome evidence for their cleavage activities on mRNA targets, including several noncanonical targets and novel phasiRNA-producing noncoding and mRNA loci with validated novel targets such as miR1509 targeting serine/threonine-protein phosphatase7 and miRc20 and ahy-miR3514 targeting penta-tricopeptide repeats (PPRs), in contradistinction to other claims of miR1509/173/7122 superfamily miRNAs indirectly targeting PPRs via TAS-like noncoding RNA loci. We characterized the inverse correlations of significantly differentially expressed drought- and heat-regulated miRNAs, assayed by sRNA blots or transcriptome datasets, with target mRNA expressions in the same datasets. Meta-analysis of an expression atlas and over representation of miRNA target genes in co-expression networks suggest that miRNAs have functions in unique aspects of peanut gynophore development. Genome-wide MIRNA annotation of the published allopolyploid peanut genome can facilitate molecular breeding of value-added traits.

Mapping of QTLs Associated with Biological Nitrogen Fixation Traits in Peanuts (Arachis hypogaea L.) Using an Interspecific Population Derived from the Cross between the Cultivated Species and Its Wild Ancestors.

Peanuts (Arachis hypogaea L.) are an allotetraploid grain legume mainly cultivated by poor farmers in Africa, in degraded soil and with low input systems. Further understanding nodulation genetic mechanisms could be a relevant option to facilitate the improvement of yield and lift up soil without synthetic fertilizers. We used a subset of 83 chromosome segment substitution lines (CSSLs) derived from the cross between a wild synthetic tetraploid AiAd (Arachis ipaensis × Arachis duranensis)4× and the cultivated variety Fleur11, and evaluated them for traits related to BNF under shade-house conditions. Three treatments were tested: without nitrogen; with nitrogen; and without nitrogen, but with added0 Bradyrhizobium vignae strain ISRA400. The leaf chlorophyll content and total biomass were used as surrogate traits for BNF. We found significant variations for both traits specially linked to BNF, and four QTLs (quantitative trait loci) were consistently mapped. At all QTLs, the wild alleles decreased the value of the trait, indicating a negative effect on BNF. A detailed characterization of the lines carrying those QTLs in controlled conditions showed that the QTLs affected the nitrogen fixation efficiency, nodule colonization, and development. Our results provide new insights into peanut nodulation mechanisms and could be used to target BNF traits in peanut breeding programs.

Genome-wide identification of acyl-CoA binding proteins and possible functional prediction in legumes.

Acyl-CoA-binding proteins (ACBPs), members of a vital housekeeping protein family, are present in various animal and plant species. They are divided into four classes: small ACBPs (class I), ankyrin-repeat ACBPs (class II), large ACBPs (class III), and kelch-ACBPs (class IV). Plant ACBPs play a pivotal role in intracellular transport, protection, and pool formation of acyl-CoA esters, promoting plant development and stress response. Even though legume crops are important for vegetable oils, proteins, vegetables and green manure, legume ACBPs are not well investigated. To comprehensively explore the functions of ACBPs in nine legumes (Lotus japonicus, Medicago truncatula, Glycine max, Vigna angularis, Vigna radiata, Phaseolus vulgaris, Arachis hypogaea, Arachis duranensis, and Arachis ipaensis), we conducted genome-wide identification of the ACBP gene family. Our evolutionary analyses included phylogenetics, gene structure, the conserved motif, chromosomal distribution and homology, subcellular localization, cis-elements, and interacting proteins. The results revealed that ACBP Orthologs of nine legumes had a high identity in gene structure and conserved motif. However, subcellular localization, cis-acting elements, and interaction protein analyses revealed potentially different functions from previously reported. The predicted results were also partially verified in Arachis hypogaea. We believe that our findings will help researchers understand the roles of ACBPs in legumes and encourage them to conduct additional research.

Genome-wide exploration of sugar transporter (sweet) family proteins in Fabaceae for Sustainable protein and carbon source.

Sugar transporter proteins (STPs) are membrane proteins required for sugar transport throughout cellular membranes. They plays an imperative role in sugar transmission across the plant and determinants of crop yield. However, the analysis of these important STPs Sugars Will Eventually be Exported Transporters (SWEET) family in legumes is still not well-documented and remains unclear. Therefore, the in-silico analysis of STPs has been performed to unravel their cellular, molecular, and structural composition in legume species. This study conducted a systematic search for STPs in Cajanus cajan using the Blastp algorithm to understand its molecular basis. Here, we performed a comprehensive analysis of 155 identified SWEET proteins across 12 legumes species, namely (Cajanus cajan, Glycine max, Vigna radiate, Vigna angularis, Medicago truncatula, Lupinus angustifolius, Glycine soja, Spatholobus suberectus, Cicer arietinum, Arachis ipaensis, Arachis hypogaea, Arachis duranensis). The amino acid composition and motif analysis revealed that SWEET proteins are rich in essential amino acids such as leucine, valine, isoleucine, phenylalanine, and serine while less profuse in glutamine, tryptophan, cysteine, and histidine. A total of four main conserved motifs of SWEET proteins are also highly abundant in these amino acids. The present study deciphered the details on primary physicochemical properties, secondary, tertiary structure, and phylogenetic analysis of SWEETs protein. Majorities of SWEET proteins (72.26%) are in stable form with an average instability index of 36.5%, and it comprises a higher fraction of positively charged amino acid Arg + Lys residues. Secondary structure analysis shown that these proteins are richer in alpha-helix (40%) than extended strand (30%) and random coil (25%), respectively. Furthermore, to infer their mechanism at a structural and functional level which play an essential roles in growth, development, and stress responses. This study will be useful to examine photosynthetic productivity, embryo sugar content, seed quality, and yield enhancement in Fabaceae for a sustainable source of essential amino acids and carbon source.

Unravelling the treasure trove of drought-responsive genes in wild-type peanut through transcriptomics and physiological analyses of root

Peanut is one of the most valuable legumes, grown mainly in arid and semi-arid regions, where its production may be hindered by the lack of water. Therefore, breeding drought tolerant varieties is of great importance for peanut breeding programs around the world. Unlike cultivated peanuts, wild peanuts have greater genetic diversity and are an important source of alleles conferring tolerance/resistance to abiotic and biotic stresses. To decipher the transcriptome changes under drought stress, transcriptomics of roots of highly tolerant Arachis duranensis (ADU) and moderately susceptible A. stenosperma (AST) genotypes were performed. Transcriptome analysis revealed an aggregate of 1465 differentially expressed genes (DEGs), and among the identified DEGs, there were 366 single nucleotide polymorphisms (SNPs). Gene ontology and Mapman analyses revealed that the ADU genotype had a higher number of transcripts related to DNA methylation or demethylation, phytohormone signal transduction and flavonoid production, transcription factors, and responses to ethylene. The transcriptome analysis was endorsed by qRT-PCR, which showed a strong correlation value (R2 = 0.96). Physio-biochemical analysis showed that the drought-tolerant plants produced more osmolytes, ROS phagocytes, and sugars, but less MDA, thus attenuating the effects of drought stress. In addition, three SNPs of the gene encoding transcription factor NFAY (Aradu.YE2F8), expansin alpha (Aradu.78HGD), and cytokinin dehydrogenase 1-like (Aradu.U999X) exhibited polymorphism in selected different genotypes. Such SNPs could be useful for the selection of drought-tolerant genotypes.

Chloroplast Phylogenomic Analyses Reveal a Maternal Hybridization Event Leading to the Formation of Cultivated Peanuts.

Peanuts (Arachis hypogaea L.) offer numerous healthy benefits, and the production of peanuts has a prominent role in global food security. As a result, it is in the interest of society to improve the productivity and quality of peanuts with transgenic means. However, the lack of a robust phylogeny of cultivated and wild peanut species has limited the utilization of genetic resources in peanut molecular breeding. In this study, a total of 33 complete peanut plastomes were sequenced, analyzed and used for phylogenetic analyses. Our results suggest that sect. Arachis can be subdivided into two lineages. All the cultivated species are contained in Lineage I with AABB and AA are the two predominant genome types present, while species in Lineage II possess diverse genome types, including BB, KK, GG, etc. Phylogenetic studies also indicate that all allotetraploid cultivated peanut species have been derived from a possible maternal hybridization event with one of the diploid Arachis duranensis accessions being a potential AA sub-genome ancestor. In addition, Arachis monticola, a tetraploid wild species, is placed in the same group with all the cultivated peanuts, and it may represent a transitional species, which has been through the recent hybridization event. This research could facilitate a better understanding of the taxonomic status of various Arachis species/accessions and the evolutionary relationship among them, and assists in the correct and efficient use of germplasm resources in breeding efforts to improve peanuts for the benefit of human beings.

A proteomic analysis of peanut seed at different stages of underground development to understand the changes of seed proteins.

In order to obtain more valuable insights into the protein dynamics and accumulation of allergens in seeds during underground development, we performed a proteomic study on developing peanut seeds at seven different stages. A total of 264 proteins with altered abundance and contained at least one unique peptide was detected by matrix-assisted laser desorption ionization time-of-flight/time-of-flight mass spectrometry (MALDI-TOF/TOF MS). All identified proteins were classified into five functional categories as level 1 and 20 secondary functional categories as level 2. Among them, 88 identified proteins (IPs) were related to carbohydrate/ amino acid/ lipid transport and metabolism, indicating that carbohydrate/amino acid/ lipid metabolism played a key role in the underground development of peanut seeds. Hierarchical cluster analysis showed that all IPs could be classified into eight cluster groups according to the abundance profiles, suggesting that the modulatory patterns of these identified proteins were complicated during seed development. The largest group contained 41 IPs, the expression of which decreased at R 2 and reached a maximum at R3 but gradually decreased from R4. A total of 14 IPs were identified as allergen-like proteins by BLAST with A genome (Arachis duranensis) or B genome (Arachis ipaensis) translated allergen sequences. Abundance profile analysis of 14 identified allergens showed that the expression of all allergen proteins was low or undetectable by 2-DE at the early stages (R1 to R4), and began to accumulate from the R5 stage and gradually increased. Network analysis showed that most of the significant proteins were involved in active metabolic pathways in early development. Real time RT-PCR analysis revealed that transcriptional regulation was approximately consistent with expression at the protein level for 8 selected identified proteins. In addition, some amino acid sequences that may be associated with new allergens were also discussed.

Fine-Mapping of a Wild Genomic Region Involved in Pod and Seed Size Reduction on Chromosome A07 in Peanut (Arachis hypogaea L.).

Fruit and seed size are important yield component traits that have been selected during crop domestication. In previous studies, Advanced Backcross Quantitative Trait Loci (AB-QTL) and Chromosome Segment Substitution Line (CSSL) populations were developed in peanut by crossing the cultivated variety Fleur11 and a synthetic wild allotetraploid (Arachis ipaensis × Arachis duranensis)4x. In the AB-QTL population, a major QTL for pod and seed size was detected in a ~5 Mb interval in the proximal region of chromosome A07. In the CSSL population, the line 12CS_091, which carries the QTL region and that produces smaller pods and seeds than Fleur11, was identified. In this study, we used a two-step strategy to fine-map the seed size QTL region on chromosome A07. We developed new SSR and SNP markers, as well as near-isogenic lines (NILs) in the target QTL region. We first located the QTL in ~1 Mb region between two SSR markers, thanks to the genotyping of a large F2 population of 2172 individuals and a single marker analysis approach. We then used nine new SNP markers evenly distributed in the refined QTL region to genotype 490 F3 plants derived from 88 F2, and we selected 10 NILs. The phenotyping of the NILs and marker/trait association allowed us to narrowing down the QTL region to a 168.37 kb chromosome segment, between the SNPs Aradu_A07_1148327 and Aradu_A07_1316694. This region contains 22 predicted genes. Among these genes, Aradu.DN3DB and Aradu.RLZ61, which encode a transcriptional regulator STERILE APETALA-like (SAP) and an F-box SNEEZY (SNE), respectively, were of particular interest. The function of these genes in regulating the variation of fruit and seed size is discussed. This study will contribute to a better knowledge of genes that have been targeted during peanut domestication.

Unraveling the mechanisms of resistance toSclerotium rolfsiiin peanut (Arachis hypogaeaL.) using comparative RNA-Seq analysis of resistant and susceptible genotypes.

Stem rot, a devastating fungal disease of peanut, is caused by Sclerotium rolfsii. RNA-sequencing approaches have been used to unravel the mechanisms of resistance to stem rot in peanut over the course of fungal infection in resistant (NRCG-CS85) and susceptible (TG37A) genotypes under control conditions and during the course of infection. Out of about 290 million reads, nearly 251 million (92.22%) high-quality reads were obtained and aligned to the Arachis duranensis and Arachis ipaensis genomes with the average mapping of 78.91% and 78.61%, respectively. In total, about 48.6% of genes were commonly regulated, while approximately 21.8% and 29.6% of uniquely regulated genes from A. duranensis and A. ipaensis genomes, respectively, were identified. Several annotated transcripts, such as receptor-like kinases, jasmonic acid pathway enzymes, and transcription factors (TFs), including WRKY, Zinc finger protein, and C2-H2 zinc finger, showed higher expression in resistant genotypes upon infection. These transcripts have a known role in channelizing the downstream of pathogen perception. The higher expression of WRKY transcripts might have induced the systemic acquired resistance (SAR) by the activation of the jasmonic acid defense signaling pathway. Furthermore, a set of 30 transcripts involved in the defense mechanisms were validated with quantitative real-time PCR. This study suggested PAMP-triggered immunity as a probable mechanism of resistance, while the jasmonic acid signaling pathway was identified as a possible defense mechanism in peanut. The information generated is of immense importance in developing more effective ways to combat the stem rot disease in peanut.

The Role of the Expansin Gene in the Process of Novel Subterranean Seed Development in Arachis duranensis

Background: Plants in the genus Arachis produce flowers aerially that develop into gynophores that grow into the ground, where they develop into fruits. Although some aerial gynophores develop into pods aboveground, embryo or seed abortion occurs in these individuals. During pod development, the shell wall initially comprises the majority of the fruit volume, but then the seeds expand up to the total pod volume. Expansins are plant cell wall-loosening proteins involved in cell enlargement and a variety of other developmental processes in which cell-wall modification occurs.Methods: In this study, we analyzed the expansin genes during seed development using RNA-seq data by bioinformatic approaches.Results: We identified five expansin genes exhibited up-regulated expression, while four genes were down-regulated expression using Arachis duranensis RAN-seq data. Multiple transcription factors regulate AdEXPs throughout seed development. Genes co-expressed with AdEXPs are involved in metabolic processes throughout pod development into seed development, reframing the current understanding of the novel subterranean peanut fruits.

Assessment of 16 Peanut (Arachis hypogaea L.) CSSLs Derived from an Interspecific Cross for Yield and Yield Component Traits: QTL Validation

Cultivated peanut is an allotetraploid (2n = 4× = 40) with narrow genetic diversity. In previous studies, we developed an advanced backcross quantitative trait loci (AB-QTL) population from the cross between the synthetic allotetraploid ((Arachis ipaensis × Arachis duranensis)4×) and the cultivated variety Fleur11, and mapped several quantitative trait loci (QTLs) involved in yield and yield components. We also developed a chromosome segment substitution line (CSSL) population as a way to mendelize the QTLs and analyzing their effects. In this study, 16 CSSLs were used for assessing the contribution of wild alleles in yield performance and stability across environments, as well as validating QTLs for pod and seed size. The CSSLs and the recurrent parent Fleur11, used as a check, were assessed using an alpha lattice design in three locations during two consecutive rainy seasons in Senegal, totaling six environments. Our results showed that the chromosome segments from the wild species, in general, have no yield disadvantage and contributed positive variation to yield-related traits. Most of the QTLs detected for pod and seed size in the AB-QTL on linkage groups A07, A08, A09, and B06 were also found in the CSSLs, showing that the CSSLs used in this study are accurate material for QTL validation. Several new QTLs have also been identified. Two CSSLs (12CS_031 and 12CS_069) showed consistently higher pod and seed size than Fleur11 in all environments, suggesting that the QTLs were consistent and stable. Our study opens the way for pyramiding these QTLs for peanut improvement.

Phylogenetic analysis and transcriptional profiling of WRKY genes in sunflower (Helianthus annuus L.): Genetic diversity and their responses to different biotic and abiotic stresses

WRKY proteins constitute a large family of transcription factors that play important roles in many aspects of physiological processes and adaptation to environmental challenge. Although previous studies have identified and characterized WRKY genes from sunflower (Helianthus annuus L.) expressed sequence tag and genomic databases, the phylogenetic diversity of WRKY repertoires of oil crop species and the responses of sunflower WRKY proteins to various stressors have not been elucidated. In this study, a total of 860 WRKY members from seven oil crop species, namely, sunflower, sesame, castor bean, soybean, canola, Arachis duranensis, and Arachis ipaensis, were subjected to comparative genomic analysis. Genetic diversity within the WRKY family among these oil crop species was revealed through pairwise comparisons of orthologous groups and phylogenetic analysis. Sequence features and phylogenetic analysis revealed that the WRKY genes of oil crop species belong to well-defined groups or subgroups, and different oil plant species exhibit obvious expansion or loss to different degrees. One hundred nineteen sunflower WRKY (HaWRKY) genes were found to be unevenly distributed across each chromosome. Segmental and tandem duplications contributed to the expansion of group II and III WRKY genes in sunflower, respectively. The duplicated genes generally exhibit divergent expression patterns, which may indicate that neofunctionalization or subfunctionalization of these genes tends to occur after duplication. Moreover, expression profiles derived from transcriptomic data exhibited distinct expression patterns of HaWRKY genes in various tissues and in response to different stress treatments, including hormone treatments, salt and drought stress, infection by the fungus Verticillium dahliae, and colonization by the fungus Rhizoglomus irregulare. According to these results, WRKY genes may be involved in the strong adaptability of sunflower to various environments, thereby contributing to the importance of sunflower as a widely cultivated oil resource. This systematic analysis provides a foundation for further functional characterization of WRKY genes among oil crop species for the improvement of stress resistance.

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

BackgroundPosttranslational 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).ResultsGenome-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.ConclusionsThe 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.

Bioinformatic analysis of globulin gene family in Arachis duranensis

Bioinformatic analysis of globulin gene family in Arachis duranensis

Sequencing of Cultivated Peanut, Arachis hypogaea, Yields Insights into Genome Evolution and Oil Improvement

Cultivated peanut (Arachis hypogaea) is an allotetraploid crop planted in Asia, Africa, and America for edible oil and protein. To explore the origins and consequences of tetraploidy, we sequenced the allotetraploid A. hypogaea genome and compared it with the related diploid Arachis duranensis and Arachis ipaensis genomes. We annotated 39 888 A-subgenome genes and 41 526 B-subgenome genes in allotetraploid peanut. The A. hypogaea subgenomes have evolved asymmetrically, with the B subgenome resembling the ancestral state and the A subgenome undergoing more gene disruption, loss, conversion, and transposable element proliferation, and having reduced gene expression during seed development despite lacking genome-wide expression dominance. Genomic and transcriptomic analyses identified more than 2 500 oil metabolism-related genes and revealed that most of them show altered expression early in seed development while their expression ceases during desiccation, presenting a comprehensive map of peanut lipid biosynthesis. The availability of these genomic resources will facilitate a better understanding of the complex genome architecture, agronomically and economically important genes, and genetic improvement of peanut.

Identification and characterization of aquaporin genes in Arachis duranensis and Arachis ipaensis genomes, the diploid progenitors of peanut

BackgroundAquaporins (AQPs) facilitate transport of water and small solutes across cell membranes and play an important role in different physiological processes in plants. Despite their importance, limited data is available about AQP distribution and function in the economically important oilseed crop peanut, Arachis hypogea (AABB). The present study reports the identification and structural and expression analysis of the AQPs found in the diploid progenitor genomes of A. hypogea i.e. Arachis duranensis (AA) and Arachis ipaensis (BB).ResultsGenome-wide analysis revealed the presence of 32 and 36 AQPs in A. duranensis and A. ipaensis, respectively. Phylogenetic analysis showed similar numbers of AQPs clustered in five distinct subfamilies including the plasma membrane intrinsic proteins (PIPs), the tonoplast intrinsic proteins (TIPs), the nodulin 26-like intrinsic proteins (NIPs), the small basic intrinsic proteins (SIPs), and the uncharacterized intrinsic proteins (XIPs). A notable exception was the XIP subfamily where XIP1 group was observed only in A. ipaensis genome. Protein structure evaluation showed a hydrophilic aromatic/arginine (ar/R) selectivity filter (SF) in PIPs whereas other subfamilies mostly contained a hydrophobic ar/R SF. Both genomes contained one NIP2 with a GSGR SF indicating a conserved ability within the genus to uptake silicon. Analysis of RNA-seq data from A. hypogea revealed a similar expression pattern for the different AQP paralogs of AA and BB genomes. The TIP3s showed seed-specific expression while the NIP1s’ expression was confined to roots and root nodules.ConclusionsThe identification and the phylogenetic analysis of AQPs in both Arachis species revealed the presence of all five sub-families of AQPs. Within the NIP subfamily, the presence of a NIP2 in both genomes supports a conserved ability to absorb Si within plants of the genus. The global expression profile of AQPs in A. hypogea revealed a similar pattern of AQP expression regardless of the subfamilies or the genomes. The tissue-specific expression of AQPs suggests an important role in the development and function of the respective organs. The AQPs identified in the present study will serve as a resource for further characterization and possible exploitation of AQPs to understand their physiological role in A. hypogea.

Cloning and analysis of cellulose synthase genes (CesA) in Acacia mangium

Cellulose synthase (CesA) plays a major regulatory role in the cellulose synthesis pathway in plants and is an important factor in controlling wood fiber quality and yield. In this study, two cellulose synthase genes, AmCesA1 and AmCesA2, were cloned from Acacia mangium using transcriptome de novo sequencing analysis and RACE. In silico analysis revealed that AmCesA1 cDNA was 3793 bp in size, had a 3249 bp ORF, and encoded a 1082 amino acid protein; AmCesA2 cDNA was 3743 bp in size, had a 3228 bp ORF, and encoded a 1075 amino acid protein. AmCesA1 was determined to have six transmembrane regions, and AmCesA2 was determined to have eight transmembrane regions. Cluster analysis showed that AmCesA1 had high degrees of similarity with Glycine max GmCesA1 and Arachis duranensis AdCesA1. AmCesA2 had high degrees of similarity with Leucaena leucocephala LlCesA7 and LlCesA8. Southern blot analysis showed that multiple copies of AmCesA1 were present in the Acacia mangium genome, but only a single copy of AmCesA2 was present. Real-time RT-PCR showed that both genes were widely expressed in roots, stems, and leaves, but AmCesA2 was more highly expressed in stems. The two genes responded to GA3, 6-BA, and MeJA treatments, and the response to GA3 was relatively strong. Here, we presume that the two genes are involved in the formation of primary cell walls and that AmCesA2 is also involved in the formation of secondary cell walls. The expression of both genes was upregulated under different hormone treatments, which indicated that the two genes positively regulate the hormone response.