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The B3 transcription factors (TFs) in plants play vital roles in numerous biological processes. Although B3 genes have been broadly identified in many plants, little is known about their potential functions in mediating seed development and material accumulation. Castor bean (Ricinus communis L.) is a non-edible oilseed crop considered an ideal model system for seed biology research. Here, we identified a total of 61 B3 genes in the castor bean genome, which can be classified into five subfamilies, including ABI3/VP1, HSI, ARF, RAV and REM. The expression profiles revealed that RcABI3/VP1 subfamily genes are significantly up-regulated in the middle and later stages of seed development, indicating that these genes may be associated with the accumulation of storage oils. Furthermore, through yeast one-hybrid and tobacco transient expression assays, we detected that ABI3/VP1 subfamily member RcLEC2 directly regulates the transcription of RcOleosin2, which encodes an oil-body structural protein. This finding suggests that RcLEC2, as a seed-specific TF, may be involved in the regulation of storage materials accumulation. This study provides novel insights into the potential roles and molecular basis of B3 family proteins in seed development and material accumulation.

Sunflower occupies the fourth position among oilseed crops the around the world. Eceriferum (CER) is an important gene family that plays critical role in very-long-chain fatty acids elongation and biosynthesis of epicuticular waxes under both biotic and abiotic stress conditions. The aim of present study was to investigate the effect of sunflower CER genes during drought stress condition. Thus, comparative analysis was undertaken for sunflower CER genes with Arabidopsis genome to determine phylogenetic relationship, chromosomal mapping, gene structures, gene ontology and conserved motifs. Furthermore, we subjected the sunflower cultivars under drought stress and used qRT-PCR analysis to explore the expression pattern of CER genes during drought conditions. We identified thirty-seven unevenly distributed CER genes in the sunflower genome. The phylogenetic analysis revealed that CER genes were grouped into seven clades in Arabidopsis, Helianthus annuus, and Gossypium hirsutum. Expression analysis showed that genes CER10 and CER60 were upregulated in sunflower during drought conditions, indicating that these genes are activated during drought stress. The results obtained will serve to characterize the CER gene family in sunflower and exploit the role of these genes in wax biosynthesis under limited water conditions. Key message Cuticular waxes protect the plants from drought stress, so we observed the expression of wax bio synthesis genes in recently sequences genome of Helianthus annuus. We observed that expression of wax biosynthesis genes CER10 and CER60 was upregulated when the plants were subjected to drought stress.

The assessment of the genetic structuring of biodiversity is crucial for management and conservation. This is particularly critical for widely distributed and highly mobile deep‐water teleosts, such as hoki (Macruronus novaezelandiae). This species is significant to Māori people and supports the largest commercial fishery in New Zealand, but uncertainty about its stock structure presents a challenge for management. Here, we apply a comprehensive genomic analysis to shed light on the demographic structure of this species by (1) assembling the genome, (2) generating a catalogue of genome‐wide SNPs to infer the stock structure and (3) identifying regions of the genome under selection. The final genome assembly used short and long reads and is near complete, representing 93.8% of BUSCO genes, and consisting of 566 contigs totalling 501 Mb. Whole‐genome re‐sequencing of 510 hoki sampled from 14 locations around New Zealand and Australia, at a read depth greater than 10×, produced 227,490 filtered SNPs. Analyses of these SNPs were able to resolve the stock structure of hoki into two genetically and geographically distinct clusters, one including the Australian and the other one all New Zealand locations, indicating genetic exchange between these regions is limited. Location differences within New Zealand samples were much more subtle (global F ST = 0.0006), and while small and significant differences could be detected, they did not conclusively identify additional substructures. Ten putative adaptive SNPs were detected within the New Zealand samples, but these also did not geographically partition the dataset further. Contemporary and historical N e estimation suggest the current New Zealand population of hoki is large yet declining. Overall, our study provides the first genomic resources for hoki and provides detailed insights into the fine‐scale population genetic structure to inform the management of this species.

The southernmost stock of longfin smelt (Spirinchus thaleichthys) is approaching extirpation in the San Francisco Estuary (SFE); however, patterns of genetic structure, diversity and gene flow which are vital for management are poorly understood in this species. Here, we use genome-wide data to evaluate population structure of longfin smelt across a broad latitudinal scale across estuaries ranging from the SFE to Yakutat Bay and Lake Washington, and fine scale within the Fraser River and the SFE. Results indicate high genetic structure between major estuaries, fine-scale structure within the Fraser River, and low levels of structure within the SFE. Genetic structure was more pronounced between northern estuaries whereas southern estuaries showed shared ancestry and ongoing gene flow, most notably unidirectional northward migration out of the SFE. Furthermore, we detected signatures of local adaptation within the Fraser River and the Skeena River estuaries. Taken together, our results identify broad patterns of genetic diversity in longfin smelt shaped by co-ancestry, unidirectional migration and local adaptation. Results also suggest that the SFE population is genetically distinct from northernmost populations and an important source for maintaining nearby populations., The authors thank Colin Grant (US Fish and Wildlife Service), who made many of the initial inquiries soliciting longfin smelt samples, and Josh Israel (US Bureau of Reclamation), who performed initial genetic analyses. The authors also thank the following individuals and agencies for help collecting samples: Hobbslab and staff -UC Davis; Matt Dekar and staff -US Fish and Wildlife Service; Kathy Hieb, Jennifer Giannetta, Jeremiah Bautista, Aaron Ng, Rebecca Garwood, James Ray, Justin Garwood, and Mike Wallace -California Department of Fish and Wildlife; Laurie Lloyd, Olaf Langness -Washington Department of Fish and Wildlife, Thomas Quinn, University of Washington; Eric Taylor -The University of British Columbia, John Kelson -Fish BiologistConsultant, Smithers, British Columbia; Mayumi Arimitsu -US Geological Survey, Alaska Science Center. Finally, the authors also thank a number of internal reviewers, Genomic Variation Lab Personnel, and other agency biologists who helped this project, including Matthew Campbell, Daphne Gille, Levi Lewis, Andrea Schreier, Josh Israel, and Alisha Goodbla. Funding for this project was generously provided by the California Department of Water Resources, Agreement Number 4600011196.

Heat shock transcription factor (HSF) is ubiquitous in the whole biological world and plays an important role in regulating growth and development and responses to environment stress. In this study...

Nutrient stress is the most important environmental stress that limits plant growth and development. Although recent evidence highlights the vital functions of long non-coding RNAs (lncRNA) in response to single nutrient stress in some model plants, a comprehensive investigation of the effect of lncRNAs in response to nutrient stress has not been performed in Arabidopsis thaliana. Here, we presented the identification and characterization of lncRNAs under seven nutrient stress conditions. The expression pattern analysis revealed that aberrant expression of lncRNAs is a stress-specific manner under nutrient stress conditions and that lncRNAs are more sensitive to nutrient stress than protein-coding genes (PCGs). Moreover, competing endogenous RNA (ceRNA) network and lncRNA-mRNA co-expression network (CEN) were constructed to explore the potential function of these lncRNAs under nutrient stress conditions. We further combined different expressed lncRNAs with ceRNA network and CEN to select key lncRNAs in response to nutrient stress. Together, our study provides important information for further insights into the role of lncRNAs in response to stress in plants.

Plant lipid transfer proteins (LTPs) are small basic proteins that play important roles in the regulation of various plant biological processes as well as the response to biotic and abiotic stresses. However, knowledge is limited on how this family of proteins is regulated in response to nematode infection in cucumber. In the present study, a total of 39 CsLTP_2 genes were identified by querying databases for cucumber-specific LTP_2 using a Hidden Markov Model approach and manual curation. The family has a five-cysteine motif (5CM) with the basic form CC-Xn-CXC-Xn-C, which differentiates it from typical nsLTPs. The members of CsLTP_2 were grouped into six families according to their structure and their phylogenetic relationships. Expression data of CsLTP_2 genes in 10 cucumber tissues indicated that they were tissue-specific genes. Two genes showed significant expression change in roots of resistant and susceptible lines during nematode infection, indicating their involvement in response to Meloidogyne incognita. This systematic analysis provides a foundation of knowledge for future studies of the biological roles of CsLTP_2 genes in cucumber in response to nematode infection and may help in the efforts to improve M. incognita-resistance breeding in cucumber.

Pseudomonas putida is a micro-organism with great potential for industry due to its stress-endurance traits and easy manipulation of the metabolism. However, optimization is still required to improve production yields. In the last years, manipulation of bacterial small non-coding RNAs (ncRNAs) has been recognized as an effective tool to improve the production of industrial compounds. So far, very few ncRNAs are annotated in P. putida beyond the generally conserved. In the present study, P. putida was cultivated in a two-compartment scale-down bioreactor that simulates large-scale industrial bioreactors. We performed RNA-Seq of samples collected at distinct locations and time-points to predict novel and potentially important ncRNAs for the adaptation of P. putida to bioreactor stress conditions. Instead of using a purely genomic approach, we have rather identified regions of putative ncRNAs with high expression levels using two different programs (Artemis and sRNA detect). Only the regions identified with both approaches were considered for further analysis and, in total, 725 novel ncRNAs were predicted. We also found that their expression was not constant throughout the bioreactor, showing different patterns of expression with time and position. This is the first work focusing on the ncRNAs whose expression is triggered in a bioreactor environment. This information is of great importance for industry, since it provides possible targets to engineer more effective P. putida strains for large-scale production.

Callose deposition occurs during plant growth and development, as well as when plants are under biotic and abiotic stress. Callose synthase is a key enzyme for the synthesis of callose. In this study, 27, 28, 16, and 15 callose synthase family members were identified in Gossypium hirsutum, Gossypium barbadense, Gossypium raimondii, and Gossypium arboreum using the sequence of Arabidopsis callose synthase. The CalSs were divided into five groups by phylogenetic, gene structure, and conservative motif analysis. The conserved motifs and gene structures of CalSs in each group were highly similar. Based on the analysis of cis-acting elements, it is inferred that GhCalSs were regulated by abiotic stress. WGD/Segmental duplication promoted the amplification of the CalS gene in cotton, and purification selection had an important function in the CalS family. The transcriptome data and qRT-PCR under cold, heat, salt, and PEG treatments showed that GhCalSs were involved in abiotic stress. The expression patterns of GhCalSs were different in various tissues. We predicted that GhCalS4, which was highly expressed in fibers, had an important effect on fiber elongation. Hence, these results help us understand the role of GhCalSs in fiber development and stress response.

Background: Germline mutations provide the raw material for all evolutionary processes and contribute to the occurrence of spontaneous human diseases and disorders. Yet despite the daily interactio...

Background: Blueberry (Vaccinium corymbosum L.) is one of the most important commercial fruit tree species. The development of high-quality seedlings is a prerequisite for fruit production. Stem cutting and tissue culture methods are widely applied for propagating blueberry seedlings. Both methods require adventitious roots (ARs), indicating ARs are critical for vegetative propagation. However, the underlying factors and molecular mechanisms regulating blueberry AR formation remain relatively unknown. Results: In this study, the rooting abilities of differentially lignified cuttings from various cultivars or the same cultivars cultured differently were evaluated following an indole-3-butyric acid (IBA) treatment. Field-grown semi-lignified and tissue culture-grown cuttings formed ARs, but the latter had more pericycle and secondary xylem cells and formed ARs more easily and faster. WUSCHEL-related homeobox genes are commonly involved in vascular tissue development and early root meristem maintenance. On the basis of the available Vaccinium corymbosum genome data, 29 putative WOX genes with conserved homeodomains were identified and divided into three major clades (modern/WUS, intermediate, and ancient). These 29 WOX genes were differentially expressed in the root, shoot, leaf, flower bud, and fruit. Additionally, a qRT-PCR analysis revealed that five selected VcWOX genes were responsive to an IBA treatment during AR formation. Accordingly, VcWOX4b was functionally characterized. The overexpression of VcWOX4b in transgenic tobacco inhibited AR formation by altering vascular cell division and differentiation and the indole-3-acetic acid (IAA):cytokinin (CTK) ratio. These observations suggest that VcWOX4b regulates the IAA:CTK ratio to promote primary xylem cell differentiation, thereby inhibiting AR formation. However, an IBA treatment can induce AR formation by inhibiting VcWOX4b expression. Conclusions: Current study elucidates the rooting abilities of various cultivars and the cytological characters of influence on AR formation of blueberry cuttings, which may provide novel insights into the selection of high-quality blueberry cuttings. VcWOX4b, VcWOX8/9a, VcWOX11/12c, and VcWOX13b might regulate blueberry AR formation in an IBA-dependent manner. Ectopic expression of VcWOX4b modulated the IAA:CTK ratio to promotes primary xylem cell differentiation, but inhibit secondary xylem cell differentiation, ultimately leading to decreased AR formation.

The NAC (NAM, ATAF, and CUC) transcription factors (TFs), as plant-specific transcription regulators, play a critical role in various developmental processes and stress responses. However, knowledge of the NAC TFs in rose (Rosa chinensis), one of the most important horticultural crops, is limited. In this study, 116 NAC genes were identified from the rose genome and classified into 16 subfamilies. The RcNACs were unevenly distributed on the chromosomes. Analysis of the stress-related RcNACs based on the transcriptome data showed differences in the expression patterns in different organs and flower developmental stages and during drought and salt stress in rose leaves and roots. Analysis of the cis-regulatory elements (CREs) in the promoter region revealed that stress-related RcNACs possessed three sets of CREs, corresponding to plant growth and development, plant hormonal response, and abiotic and biotic stresses. Reverse transcription-quantitative real-time PCR (RT-qPCR) analysis of 11 representative RcNACs revealed their diverse transcript expression in leaves and roots under abscisic acid (ABA), polyethylene glycol (PEG), and sodium chloride (NaCl) treatments. Furthermore, RcNAC091 silencing verified its role in positively regulating the dehydration stress response. Overall, this study provides helpful insight into stress-related RcNACs and paves the way for stress tolerance via breeding in rose.

Potassium is one of the most essential inorganic cations for plant growth and development. The high affinity K+ (HAK)/K+ uptake (KUP)/K+ transporter (KT) family plays essential roles in the regulation of cellular K+ levels and the maintenance of osmotic balance. However, the roles of these genes in the responses of bananas to low-potassium stress are unclear. In this study, 24 HAK/KUP/KT (MaHAK) genes were identified from banana genomic data. These genes were further classified into four groups based on phylogenetic analysis, gene structure and conserved domain analysis. Segmental duplication events played an important role in the expansion of the MaHAK gene family. Transcriptome analysis revealed the expression patterns of MaHAKs in various tissues under different K+ conditions. MaHAK14b was upregulated under both short- and long-term K+-deficient conditions, suggesting that it plays crucial roles in K+ uptake at low K+ concentrations. Furthermore, MaHAK14b mediated K+ uptake when it was heterologously expressed in the yeast mutant R5421 on low K+ medium. Collectively, these findings provide a foundation for further functional analysis of MaHAK genes, which may be used to improve stress resistance in bananas.

Trypanosoma brucei brucei (T.b.brucei) is an extra-cellular parasite that causes Animal African Trypanosomiasis (AAT) disease in animals. Till day, this disease is more difficult to treat and control due to lack of efficient vaccines and early diagnosis of the parasite infection. T.b.brucei Excretory/Secretory (ES) proteins were involved in pathogenesis and key for understanding the host-parasite interactions. Functions of T.b.brucei's ES proteins were poorly investigated and experimental identification is expensive and time-consuming. Bioinformatics approaches are cost-effective by facilitating the experimental analysis of potential drug targets for parasitic diseases. Here we applied several bioinformatics tools to predict and functionalize the annotation of 1104 ES proteins and immunoinformatics approaches carried out to predict and evaluate the epitopes in T.b.brucei. Secretory information, functional annotations and potential epitopes of each ES proteins were available at http://tbb.insilico.in . This study provides functional information of T.b.brucei for experimental studies to identify potential targets for diagnosis and therapeutics development.

Alternative splicing (AS) is a post-transcriptional regulatory mechanism that expands proteomic diversity. AS often occurs in the context of changing environmental conditions. Nevertheless, it rema...

Thermostable xylanases from thermophilic fungi and bacteria have a wide commercial acceptability in feed, food, paper and pulp and bioconversion of lignocellulosics with an estimated annual market of USD 500 Million. The genome wide analysis of thermophilic fungi clearly shows the presence of elaborate genetic information coding for multiple xylanases primarily coding for GH10, GH11 in addition to GH7 and GH30 xylanases. The transcriptomics and proteome profiling has given insight into the differential expression of these xylanases in some of the thermophilic fungi. Bioprospecting has resulted in identification of novel thermophilic xylanases that have been endorsed by the industrial houses for heterologous over- expression and formulations. The future use of xylanases is expected to increase exponentially for their role in biorefineries. The discovery of new and improvement of existing xylanases using molecular tools such as directed evolution is expected to be the mainstay to meet increasing demand of thermostable xylanases.

Being an essential mineral nutrient, potassium (K+) plays numerous important roles in plant growth and development and determines the yield and quality of crop products. The cellular level of K+ is controlled to a large extent by the K+ transporter, which belongs to the KT/HAK/KUP (HAK) family. However, little is known about these genes in tobacco. In this study, we surveyed the tobacco genome and identified 41 putative NtHAK genes (NtHAKS1–NtHAKS21 and NtHAKT1–NtHAKT20). Investigation of the cis-elements in upstream regions of these NtHAK genes suggests that members of this family respond to environmental cues and phytohormones. Expression data mining reveals that NtHAK genes showed clear sub-genome dominance. In all, these results will provide molecular insights into K+ transporter research in tobacco.

Cotton (Gossypium spp.) is an economically important crop grown for natural fiber and seed oil production. DA1 is a ubiquitin receptor that determines final seed and organ size by restricting the period of cell proliferation. In the present study, we identified 7 DA1-like genes each in cultivated tetraploid (AADD) G. hirsutum and G. barbadense, and 4 and 3 DA1-like genes in their ancestral diploid G. arboreum (A2A2) and G. raimondii (D5D5), respectively. The 7 GhDA1 genes were confirmed to be distributed on four At and three Dt subgenome chromosomes in G. hirsutum. GhDA1-1A showed a high sequence similarity to AtDA1 in Arabidopsis, and they possessed the same functional domains, suggesting conserved functions. The overexpression of GhDA1-1AR301K in Arabidopsis significantly increased seed size and seed weight, indicating that GhDA1-1A is a promising target for cotton improvement. This study provides information on the molecular evolutionary properties of DA1-like genes in cotton, which will be useful for the genetic improvement of cotton.

Over the last decades, identification of RNA-proteins complexes and their binding sites was challenging. Recently, techniques based on crosslinking, immunoprecipitation, and high-throughput sequencing have been developed. An optimized method, called eCLIP-seq, enables to identify precisely the targeted RNAs as well as the transcriptome-wide binding sites at nucleotide resolution. Here we describe the eCLIP-seq protocol in asexual stages of the human malaria parasite, Plasmodium falciparum. This method could facilitate the characterization of RNA-binding proteins in this organism for which few data are currently available.

This chapter is focused on the identification and characterization of long noncoding RNAs (lncRNAs) in Sorghum and their potential role in biological processes related to development and stress. It begins by introducing lncRNAs as key regulators of several biological and molecular functions in different plant species. It then provides a brief about Sorghum research done till date along with studies on lncRNAs. Lastly, the research gaps existing in current knowledge involving lncRNAs have been discussed that can have long-term implications on future research activities in Sorghum, which is widely considered as a smart agriculture crop in the era of climate change and nutritional security.

Background: Black pepper is a prominent spice which is an indispensable ingredient in culinary and traditional medicine. Phytophthora capsici, the causative agent of foot rot disease causes drastic constraint in black pepper cultivation and productivity. To counterattack various biotic and abiotic stresses plants employ a broad array of mechanisms one such includes the accumulation of pathogenesis-related (PR) proteins. Several studies have reported the role of PR-1 proteins in triggering the plant defenses during plant-oomycete interaction.Results: Through the genome-wide survey, eleven PR-1 genes that belongs to a CAP superfamily protein with Caveolin-Binding Motif (CBM) and CAP-derived peptide (CAPE) were identified from P. nigrum. Despite the critical functional domains, PnPR1 homologs differ in their signal peptide motifs, and core amino acid sequence composition in the functional protein domains. The GO, biological function analysis reveals their role in defense responses and response to biotic stimulus whereas the KEGG functional annotation predicted their function in the plant-pathogen interactions. Furthermore, transcriptome-assisted FPKM analysis revealed PnPR-1 genes mapped to P. nigrum - P. capsici interaction pathway. The differentially expressed pathogen-responsive PR-1 gene was validated through qRT-PCR. Subsequent analysis revealed the structural details, phylogenetic relationships, conserved sequence motifs and critical cis-regulatory elements of PnPR-1 genes.Conclusion: This is the first genome-wide study that identified the role of PR-1 genes during P. nigrum - P. capsici interactions. The detailed in silico experimental analysis revealed the vital role of PnPR-1 genes in regulating the first layer of defense towards P. capsici infection in Panniyur-1 plants.

Streptococcus suis (S. suis) is a gram-positive coccus that causes disease in humans and animals. The codon usage pattern of bacteria reveals a range of evolutionary changes that assist them to enhance tolerance to environments. To better understand the genetic features during the evolution of S. suis, we performed codon usage analysis. Nine pathogenic strains of different serotypes and different geographical distribution were analyzed to better understand the differences in their evolutionary process. Nucleotide compositions and relative synonymous codon usage (RSCU) analysis revealed that A/T-ending codons are dominant in S. suis. Neutrality analysis, correspondence analysis and ENC-plot results revealed that natural selection is the predominant element prompting codon usage. Cluster analysis based on RSCU was roughly consistent with the dendrogram rooted genomic BLAST analysis. Comparison of synonymous codon usage pattern between S. suis and susceptible hosts (H. sapiens and S. scrofa) revealed that the codon usage of S. suis is separated from the synonymous codon usage of susceptible hosts. The CAI values implied that S. suis includes a series of predicted highly expressed coding sequences contained in metabolism and transcriptional regulation, revealing the necessity of this pathogen to deal with various environmental conditions. The study of codon usage in S. suis may provide evidence involving the molecular evolution of bacteria and a better understanding of evolutionary relationships between S. suis and its corresponding hosts.

Background: The chloroplast NADH dehydrogenase-like (NDH) complex, homologous to respiratory complex I, participates in photosystem I cyclic electron flow (PSI-CEF) and chlororespiration in photosynthesis. However, little information was available about the ndh genes in Zostera marina which is one of the most productive and wide-distributed seagrasses.Results: The phylogenetic analysis indicated that Z. marina has a complete NDH complex which is rarely observed in marine macrophytes. We identified all 31 ndh genes necessary for the functional NDH complex, with ndhB and pnsb3 occurring as duplication events. Secondary structural analyses of antiporter-like subunits showed that the long amphipathic helix of NdhF was lost in Z. marina, suggesting an alternative mode in the generation of trans-thylakoid proton gradient. The splicing pattern in ndh genes exhibited tissue-specific patterns but no response to photo-regulation. RNA editing in Z. marina presented the ancestral pattern with many of the primitive editing sites and types. The partial editing in ndhF reflected the link between light stress and RNA editing. Moreover, predominant expression in the leaves of most ndh genes implied their major function in photosynthesis. The dynamic profiles of expression in response to light stress suggested that there were two diverse responsive and regulatory mechanisms of the NDH complex in PSI-CEF and chlororespiration.Conclusions: In this study, we performed a systematic analysis of the identification, splicing, editing and pattern of expression of the Z. marina NDH complex. Our study help elucidate the further evolutionary and functional exploration of the NDH complex.

Parkinson’s disease (PD) is primarily characterized by the loss of dopaminergic (DA) neurons in the brain. However, little is known about why DA neurons are selectively vulnerable to PD. To identify genes that are associated with DA neuron loss, we screened through 201 wild-caught populations of Drosophila melanogaster as part of the Drosophila Genetic Reference Panel. Here, we identify the top-associated genes containing single-nucleotide polymorphisms that render DA neurons vulnerable. These genes were further analyzed by using mutant analysis and tissue-specific knockdown for functional validation. We found that this loss of DA neurons caused progressive locomotor dysfunction in mutants and gene knockdown analysis. The identification of genes associated with the progressive loss of DA neurons should help to uncover factors that render these neurons vulnerable in PD, and possibly develop strategies to make these neurons more resilient.

Background: The β amylase (BAM) gene family, known for their property of catalytic ability to hydrolyze starch to maltose units, has been recognized to play critical roles in metabolism and gene regulation. To date, BAM genes have not been characterized in oil crops.Results: In this study, the genome wide survey revealed the identification of 30 BnaBAM genes in Brassica napus (B. napus), 11 BraBAM genes in Brassica rapa (B. rapa), 20 BoBAM genes in Brassica oleracea (B. oleracea), which were divided into 4 subfamilies according to the sequence similarity and phylogenetic relationships. All the BAM genes identified in the allotetraploid genome of B. napus, as well as two parental related species (B. rapa and B. oleracea), were analyzed for the gene structures, chromosomal distribution and collinearity, the sequence alignment of the core glucosyl hydrolase domains was further applied. 30 BnaBAMs, 11 BraBAMs and 17 BoBAMs exhibited uneven distribution on chromosomes of Brassica crops. The similar structural compositions of BAM genes in the same subfamily suggested that they were relatively conserved. Abiotic stresses pose one of the major constraints to plant growth and productivity worldwide. Thus, the responsiveness of BnaBAM genes under abiotic stresses were analyzed in B. napus. The expression patterns revealed a stress responsive behavior of all members, of which BnaBAM3s were more prominent. These differential expression patterns suggested an intricate regulation of BnaBAMs elicited by environmental stimuli. Conclusion: Altogether, the present study provides first insights into the BAM gene family of Brassica crops, which lays the foundation for investigating the roles of stress--responsive BnaBAM candidates in B. napus.

Background Gleditsia sinensis Lam. (Leguminosae), a dioecious perennial arbor, demonstrates important medicinal properties and economic value. These properties can be harnessed depending on the sex of the plant. However, the sex of the plants is difficult to identify accurately through morphological methods before the flowering. Results We used bulked segregant analysis to screen sex-specific simple sequence repeat (SSR) markers in G. sinensis. Five male and five female plants were pooled to form the male and female bulks, respectively, and subjected to whole-genome sequencing. After high-throughput sequencing, 5,350,359 sequences were obtained, in which 2,065,210 SSRs were searched. Among them, the number of duplicated SSRs was the highest. The male plants could reach 857,874, which accounted for 60.86% of the total number of male plants. The female plants could reach 1,447,603, which accounted for 56.25% of the total model of the female plants. Among all the nucleotide repeat types, the A/T-rich motif was the most abundant. A total of 309,516 female strain-specific SSRs were selected by clustering. After designing the primers, the male and female gene pools were amplified, and five pairs of primers (i.e., 27, 34, 36, 39, and 41) were found to amplify the differential bands in the male and female gene pools. Using the five pairs of primers, we performed PCR verification on 10 individuals of known sex, which constructed the gene pool. The female plants amplified a single fragment of lengths (i.e., 186, 305, 266, 203, and 260 bp) and no male plant strip, thereby completing the identification of the male and female sexes of the G. sinensis. Conclusions This study provides accurate sex identification strategies between female and male plants, thus improving the utilization rate of G. sinensis resources.

Background: Heat shock transcription factors (HSFs) play an important role as transcriptional regulatory proteins against heat stress by controlling the expression of heat responsive genes. French bean is a highly thermosensitive crop and therefore, its genome sequence information segregated, characterized here in terms of heat shock transcription factors alongwith its evolutionary significance. Results: In this study, a total comprehensive set of 29 non-redundant full length HSF genes were identified and characterized from Phaseolus vulgaris L. (PvHSF) genome sequence. Detailed gene information such as chromosomal localization, domain position, motif organization and exon- intron identification were analyzed. All the 29 PvHSF genes were mapped on 8 out of 11 chromosomes indicating the gene duplication occurred in french bean genome. Motif analysis and exon- intron structure found to be conserved in each group which showed conserved structure of PvHSFs and heat induced response is highly influenced by the cytoplasmic proteins. Based on structural features and phylogenetic relationship, the HSF genes were grouped into three classes i.e. A to C and 14 groups. Presence of only one pair of paralog sequence suggests that it may be derived from the duplication event during evolution. Comparative genomics study indicated the influence of whole genome duplication and purifying selection on french bean genome during evolution. In silico expression analysis indicated active role of classs A and B family during abiotic stress condition and higher expression in floral organs. Conclusion: A comprehensive analysis of Heat shock transcription factors resulted 29 non-redundant full length PvHSF genes, which were characterized for their occurrence in genome and functional importance through In silico approach.

Cowpea [Vigna unguiculata (L.)Walp.] is an essential food crop that is cultivated in many important arid and semi-arid regions of the world. In this study the genome-wide database of cowpea genes was accessed in search of genomic sequences coding for globulins, specifically members of the cupin superfamily, a well-documented multigenic family belonging to the globulin protein class. A total of seventy-seven genes belonging to the cupin superfamily were found and divided into six families. We classify V. unguiculata genes into two subgroups: classical cupins with one cupin domain (fifty-nine proteins) and bicupins with two cupin domains (eighteen members). In addition, a search for cupin members in other closely related species of the fabaceae family [V. angularis, V. radiatam and Phaseolus vulgaris (common bean)] was performed. Based on those data, a detailed characterization and comparison of the cupin genes on these species was performed with the aim to better understand the connection and functions of cupin proteins from different, but related, plant species. This study was the first attempt to investigate the cupin superfamily in V. unguiculata, allowing the identification of six cupins families and better understand the structural features of those proteins, such as number of domains alternative splicing.

Multidrug and Toxic Compound Extrusion (MATEs) is one of the characteristic transporter families, which plays a key role in the detoxication of endogenous secondary metabolites and exogenous agents in both animal and plant cells. In this study, we identified a total of 67 MATE genes (CcMATEs) from the pigeonpea genome, on which we performed bioinformatics analysis and we group them by phylogenetic analysis. Finally, eight represented CcMATE genes were selected for further qRT-PCR analysis of tissue specificity and response to metal stress in pigeonpea. The results showed that both CcMATE34 and 45 genes were significantly up-regulated and the CcMATE4 gene was only up-regulated in the roots under the stress of Al, Mn and Zn. We speculated that the function of CcMATE34 and 45 might be related to the transport of alkaloids and harmful substances and the function of CcMATE4 might be related to the delivery of flavonoids.

Background: APETALA2/ETHYLENE RESPONSIVE FACTOR (AP2/ERF) transcription factors play important roles in plant growth, development, metabolism, as well as in biotic and abiotic stress responses. However, there are few studies concerning AP2/ERF genes in sugarcane, which is the most critical sugar and energy crop worldwide. Results: A total of 218 AP2/ERF genes were identified in the Saccharum spontaneum genome. Phylogenetic analysis showed that these genes could be divided into four groups, including 43 AP2s, 160 ERFs, and Dehydration-responsive element-binding (DREB) factors, 11 ABI3/VPs (RAV) and 4 Soloist genes. These genes were unevenly distributed on 32 chromosomes. Analysis of the structural of SsAP2/ERF genes showed that 91 SsAP2/ERFs lacked introns. Sugarcane and sorghum have a collinear relationship between 168 SsAP2/ERF genes and sorghum AP2/ERF genes that reflects their similarity. Multiple cis-regulatory elements (CREs) are present in the SsAP2/ERF promoter, and many are related to abiotic stresses, suggesting that SsAP2/ERF activity could contribute to the adaptation of sugarcane crops to environmental changes. The tissue-specific analysis showed spatiotemporal expression of SsAP2/ERF in the stems and leaves of sugarcane at different stages of development. In 10 sugarcane samples, 39 SsAP2/ERFs were not expressed at all, whereas 58 SsAP2/ERFs were expressed in all samples. Quantitative PCR experiments showed that SsERF52 expression was up-regulated under salt stress, but suppressed under drought stress. SsSoloist4 had the most considerable upregulation in response to treatment with the exogenous hormones ABA and GA. Within 3 hours of ABA or PEG6000 treatment, SsSoloist4 expression was up-regulated, indicating that this gene could play a role in ABA and GA-associated drought stress response mechanisms. Analysis of AP2/ERF gene expression patterns under different treatments indicated that SsAP2/ERF genes play an important role in drought and salt stress responses of S. spontaneum. Conclusions: In this study, a total of 218 members of the AP2 / ERF superfamily were identified in sugarcane, and their genetic structure, evolution characteristics, and expression patterns were studied and analyzed. The results of this study provide a foundation for future analyses to elucidate the importance of AP2/ERF transcription factors in the function and molecular breeding of sugarcane.

The plant specific LATERAL ORGAN BOUNDARIES (LOB)-domain (LBD) proteins belong to a family of transcription factors that play important roles in plant growth and development, as well as in responses to various stresses. However, a comprehensive study of LBDs in Brassica napus has not yet been reported. In the present study, 126 BnLBD genes were identified in B. napus genome using bioinformatics analyses. The 126 BnLBDs were phylogenetically classified into two groups and nine subgroups. Evolutionary analysis indicated that whole genome duplication (WGD) and segmental duplication played important roles in the expansion of the BnLBD gene family. On the basis of the RNA-seq analyses, we identified BnLBD genes with tissue or developmental specific expression patterns. Through cis-acting element analysis and hormone treatment, we identified 19 BnLBD genes with putative functions in plant response to abscisic acid (ABA) treatment. This study provides a comprehensive understanding on the origin and evolutionary history of LBDs in B. napus, and will be helpful in further functional characterisation of BnLBDs.

WRINKLED1 (WRI1), an AP2/ERE transcription factor, is one of the most important regulators of oil accumulation. It has been extensively studied in angiosperms, but its evolution and overview features in plants remain unknown. In this study, WRI1s, as well as WRI1-likes in non-WRI1 species, were investigated in 64 genome-sequenced plants. Their origin, distribution, duplication, evolution, functional domains, motifs, properties, and cis-elements were analyzed. Results suggest that WRI1 and WRI1-like may originate from Chlorophyta, and WRI1-likes in angiosperms resemble phylogenetically and structurally WRI1s from Chlorophyta and non-vascular plants. WRI1 or WRI1-like may be essential to vascular plants but not to non-vascular plants. Two YRG elements and two RAYD elements, as well as their phosphorylation sites and the 14-3-3 binding motif, are relatively conserved from Chlorophyta to angiosperm. The predicted DNA-binding domains are slightly shorter than the combination of one YRG element and one RAYD element. WRI1 gradually evolves from alkalinity to acidity. More motifs were developed in N-terminuses and C-terminuses in vascular plants. A short acidic amino-acid-enriched domain in the C-terminal region is predicted to be the putative transactivation domain. The VYL exon appears randomly in different WRI1 transcripts and it is not important for the function of WRI1. In addition, more cis-elements developed during WRI1 evolution may suggest its more complicated regulation and physiological functions. These results will assist future function studies of WRI1 and evolution studies of fatty acid biosynthesis regulation in plants.

Odorant‐binding proteins (OBPs) and chemosensory proteins (CSPs) of insects are thought to play roles in olfactory recognition affecting host choice, copulation, reproduction and other behaviors. Previous descriptions of OBPs and CSPs in the whitefly Bemisia tabaci often provided no or incomplete genetic information. In this study, we present a genome‐wide and transcriptome‐wide investigation of the OBPs and CSPs in B. tabaci MEAM1 (Middle East‐Asia Minor1 species). Eight OBP and 19 CSP genes were identified that covered all previous sequences. Phylogenetic analyses showed that the CSP genes had a lineage‐specific expansion (BtabBCSP1, BtabBCSP3, BtabBCSP13, BtabBCSP17, BtabBCSP18 and BtabBCSP19). Expression profiling of OBPs and CSPs by transcriptome sequencing and quantitative real‐time polymerase chain reaction (qPCR) revealed that expression patterns differed among developmental stages of B. tabaci MEAM1. Five OBP genes and 11 CSP genes significantly differed between males and females; four of the 19 CSP genes were highly expressed in adults, while two were highly expressed in nymphs. The expression profiles of the OBP and CSP genes in different tissues of B. tabaci MEAM1 adults were analyzed by qPCR. Four OBP genes found in B. tabaci MEAM1 were highly expressed in the head. Conversely, only two CSPs were enriched in the head, while the other six CSPs were specifically expressed in other tissues. Our results provide a foundation for future research on OBPs and CSPs in B. tabaci.

DNA methylation plays an important role in regulating gene expression during tissue development and differentiation in eukaryotes. In contrast to domestic animals, epigenetic studies have been seldom conducted in wild animals. In the present study, we conducted the genome-wide profiling of DNA methylation for five tissues of sika deer using the fluorescence-labeled methylation-sensitive amplified polymorphism (F-MSAP) technique. Overall, a total of 104,131 fragments were amplified including 41,951 methylated fragments using 32 pairs of selected primers. The average incidence of DNA methylation was approximately 38.18% in muscle, 40.32% in heart, 41.86% in liver, 41.20% in lung, and 41.68% in kidney, respectively. Also, the significant differences of the DNA methylation levels were found between the different tissue types (P

In the present study, we have identified and characterized two 14-3-3 isoforms, namely isoform A and C from chickpea (Cicer arietinum L.), that might play a crucial role during disease resistance. Further, in silico analysis of these 14-3-3 proteins was accomplished, including motif identification and structure prediction from deduced amino acid sequences. Expression profiling of the two representative 14-3-3 isoforms in the roots of wilt resistant and susceptible chickpea varieties upon Fusarium oxysporum f. sp ciceri race 1 (FOC1) challenge, revealed time dependent isoform specific differential expression in induced chickpea roots upon FOC1 colonization

The sea cucumber Apostichopus japonicus is a deposit-feeder and vital for marine benthic ecosystems. Hypoxia can influence the behaviour and even lead to massive mortality in A. japonicus in the wild. It is important to understand the molecular responses of A. japonicus when exposed to acute changes in dissolved oxygen (DO) concentration. In this study, RNA-seq provided a general overview of the gene expression profiles of the respiratory tree of A. japonicus exposed to DO of 8 mg l−1 (DO8), 4 mg l−1 (DO4) and 2 mg l−1 (DO2) conditions. Differentially expressed genes (DEGs) screening with the NOISeq method identified 51, 59 and 61 DEGs according to the criteria of fold change ≥2 and divergence probability ≥0.8 in the comparisons of DO2 vs DO4, DO2 vs DO8 and DO4 vs DO8, respectively. Gene ontology analysis showed that ‘cellular process’ and ‘binding’ had the most enriched DEGs in the categories of ‘biological process’ and ‘molecular function’, respectively (catalytic activity also had the most enriched DEGs in the category of ‘molecular function’ based on the comparison of DO2 vs DO8), while ‘cell’ and ‘cell part’ had the most enriched DEGs in the category of ‘cellular component’. The DEGs were mapped to 79, 81 and 104 pathways in the KEGG database, and 8, 29 and 16 pathways were significantly enriched, respectively. The DO-specific DEGs identified in this study of the respiratory tree are important targets for further research into the biochemical mechanisms involved in the response of the sea cucumber to changes in the DO concentration.

DNA methylation is an important epigenetic modification that has been implicated in many biological processes. Methylation changes during stress responses and developmental processes have been well studied. Here, we present a novel comparison of methylation patterns in two poplar species. We detected the global methylation patterns in leaves of Populus trichocarpa and Populus euphratica by methylated DNA immune precipitation sequencing (MeDIPseq). A total of eleven significant modules were detected in the two species. Our results indicate that the methylation of CG/CHH/CHG contexts was much more significant in P. trichocarpa than in P. euphratica, but methylation changes in response to saline stress were consistent in both species.

Melon (Cucumis melo L.) is a very important crop throughout the world and has great economic importance, in part due to its nutritional properties. It prefers well-drained soil with low acidity and has a strong demand for water during fruit set. Therefore, a correct water balance—involving aquaporins—is necessary to maintain the plants in optimal condition. This manuscript describes the identification and comparative analysis of the complete set of aquaporins in melon. 31 aquaporin genes were identified, classified and analysed according to the evolutionary relationship of melon with related plant species. The individual role of each aquaporin in the transport of water, ions and small molecules was discussed. Finally, qPCR revealed that almost all melon aquaporins in roots and leaves were constitutively expressed. However, the high variations in expression among them point to different roles in water and solute transport, providing important features as that CmPIP1;1 is the predominant isoform and CmTIP1;1 is revealed as the most important osmoregulator in the tonoplast under optimal conditions. The results of this work pointing to the physiological importance of each individual aquaporin of melon opening a field of knowledge that deserves to be investigated., This study was funded by the Spanish Ministerio de Ciencia, Innovación y Universidades (RTC-2017-6119-2), was co-financed by SAKATA SEED IBERICA S.L.U. and was developed under the auspices of the Spanish Higher Council for Scientific Research (CSIC)

Background: OVATE family proteins (OFPs) are plant-specific proteins with the conserved OVATE domain that regulating plant growth and development. Although these OFPs have been studied in several species, the biological functions of this OFP gene family remain largely unknown in cucumber (Cucumis sativus L.). Results: In this study, we identified 19 CsOFPs in cucumber. This CsOFPs are distributed on seven chromosomes and can be divided into four subgroups. Most CsOFP genes are expressed in reproductive organs although have different expression patterns. Cis-elements analysis showed that there are six kinds of hormone response elements in CsOFPs and exogenous gibberellin treatment leads to a ‘first increase then decrease’ expression pattern of CsOFP7, CsOFP11 and CsOFP12. Ectopic expression of CsOFP11 in Arabidopsis resulted in shorter and blunt siliques. Conclusions: Together, these results indicated that CsOFPs may play important roles in cucumber fruit development.

The whitefly (Bemisia tabaci), an important invasive pest that causes severe damage to crops worldwide, has developed resistance to a variety of insecticides. Carboxylesterases (COEs) are important multifunctional enzymes involved in the growth, development, and xenobiotic metabolism of insects. However, systematic studies on the COEs of B. tabaci are scarce. Here, 42 putative COEs in different functional categories were identified in the Mediterranean species of B. tabaci (B. tabaci MED) based on a genome database and neighbor-joining phylogeny. The expression patterns of the COEs were affected by the development of B. tabaci. The expression levels of six COEs were positively correlated with the concentration of imidacloprid to which B. tabaci adults were exposed. The mortality of B. tabaci MED adults fed dsBTbe5 (67.5%) and dsBTjhe2 (58.4%) was significantly higher than the adults fed dsEGFP (41.1%) when treated with imidacloprid. Our results provide a basis for functional research on COEs in B. tabaci and provide new insight into the imidacloprid resistance of B. tabaci.

Background Diversification on the basis of utilization is a hallmark of Beta vulgaris (beet), as well as other crop species. Often, crop improvement and management activities are segregated by crop type, thus preserving unique genome diversity and organization. Full interfertility is typically retained in crosses between these groups and more traits may be accessible if the genetic basis of crop type lineage were known, along with available genetic markers to effect efficient transfer (e.g., via backcrossing). Beta vulgaris L. (2n =18) is a species complex composed of diverged lineages (e.g., crop types), including the familiar table, leaf (chard), fodder, and sugar beet crop types. Using population genetic and statistical methods with whole genome sequence data from pooled samples of 23 beet cultivars and breeding lines, relationships were determined between accessions based on identity-by-state metrics and shared genetic variation among lineages. Results Distribution of genetic variation within and between crop types showed extensive shared (e.g. non-unique) genetic variation. Lineage specific variation (e.g. apomorphy) within crop types supported a shared demographic history within each crop type, while principal components analysis revealed strong crop type differentiation. Relative contributions of specific chromosomes to genome wide differentiation were ascertained, with each chromosome revealing a different pattern of differentiation with respect to crop type. Inferred population size history for each crop type helped integrate selection history for each lineage, and highlighted potential genetic bottlenecks in the development of cultivated beet lineages. Conclusions A complex evolutionary history of cultigroups in Beta vulgaris was demonstrated, involving lineage divergence as a result of selection and reproductive isolation. Clear delineation of crop types was obfuscated by historical gene flow and common ancestry (e.g. admixture and introgression, and sorting of ancestral polymorphism) which served to share genome variation between crop types and, likely, important phenotypic characters. Table beet was well differentiated as a crop type, and shared more genetic variation within than among crop types. The sugar beet group was not quite as well differentiated as the table beet group. Fodder and chard groups were intermediate between table and sugar groups, perhaps the result of less intensive selection for end use.

Although the regulatory function of miRNAs and their targets have been characterized in model plants, a possible underlying role in the cotton response to herbivore infestation has not been determined. To investigate this, we performed small RNA and degradome sequencing between resistant and susceptible cotton cultivar following infestation with the generalist herbivore whitefly. In total, 260 miRNA families and 241 targets were identified. Quantitative-PCR analysis revealed that several miRNAs and their corresponding targets exhibited dynamic spatio-temporal expression patterns. Moreover, 17 miRNA precursors were generated from 29 long intergenic non-coding RNA (lincRNA) transcripts. Genome-wide analysis also led to the identification of 85 phased small interfering RNA (phasiRNA) loci. Among these, nine PHAS genes were triggered by miR167, miR390, miR482a, and two novel miRNAs, including those encoding a leucine-rich repeat (LRR) disease resistance protein, an auxin response factor (ARF) and MYB transcription factors. Through combined modeling and experimental data, we explored and expanded the miR390-tasiARF cascade during the cotton response to whitefly. Virus-induced gene silencing (VIGS) of ARF8 in whitefly-resistant cotton plants increased auxin and jasmonic acid (JA) accumulation, resulting in an increased tolerance to whitefly infestation. These results highlight the provides a useful transcriptomic resource for plant-herbivore interaction.

Growth-regulating factors (GRFs) are plant-specific transcription factors that perform important functions in plant growth and development. Herein, we identified and characterised 24 AhGRF genes in peanut (Arachis hypogaea). AhGRF family genes were divided into six classes with OLQ and WRC domains. Transcriptome expression profile showed that more AhGRF genes, such as AhGRF5a gene, were at higher expression during pod development in Arachis monticola than cultivated species, especially at the pod rapid-expansion stage. AhGRF5a and AhGRF5b genes expressed at higher levels in pods than roots, leaves and stems tissues, existing in the difference between Arachis monticola and H8107. Exogenous GA3 application can activate AhGRF5a and AhGRF5b genes and H8107 line showed more positive response than Arachis monticola species. These results imply that these two AhGRF genes may be active during the peanut pod development.