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1.
Gene ; 766: 145141, 2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-32911031

RESUMO

Jatropha curcasseeds are abundant in biodiesel, and low seed yields are linked to poor quality female flowers, which creates a bottleneck for Jatropha seed utilization. Therefore, identifying the genes associated with flowering is crucial for the genetic enrichment of seed yields. Here, we identified an AGAMOUS homologue gene (JcAG) from J. curcas. We found that reproductive organs had higher JcAG expression than vegetative organs, particularly the carpel. Rosette leaves were small and misshapen in 35S:JcAG transgenic lines in comparison with those in wild-type plants. JcAG overexpression caused an extremely early flowering, delayed perianth and stamen filament development, small flowers, and significantly shorter Arabidopsis plants with little fruit. In the JcAG-overexpressing line, the homeotic transformation of sepals into pistillate organs was observed, and floral meristem and organ identity genes were regulated. This study provides insights into the JcAG's function and benefits to our knowledge of the underlying the genetic mechanisms related to floral sex differentiation in Jatropha.


Assuntos
Expressão Ectópica do Gene/genética , Flores/genética , Regulação da Expressão Gênica de Plantas/genética , Genes de Plantas/genética , Jatropha/genética , Proteínas de Plantas/genética , Arabidopsis/genética , Meristema/genética , Fenótipo , Plantas Geneticamente Modificadas/genética , Sementes/genética
2.
PLoS One ; 15(12): e0244021, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-33362265

RESUMO

Random regression models (RRM) are a powerful tool to evaluate genotypic plasticity over time. However, to date, RRM remains unexplored for the analysis of repeated measures in Jatropha curcas breeding. Thus, the present work aimed to apply the random regression technique and study its possibilities for the analysis of repeated measures in Jatropha curcas breeding. To this end, the grain yield (GY) trait of 730 individuals of 73 half-sib families was evaluated over six years. Variance components were estimated by restricted maximum likelihood, genetic values were predicted by best linear unbiased prediction and RRM were fitted through Legendre polynomials. The best RRM was selected by Bayesian information criterion. According to the likelihood ratio test, there was genetic variability among the Jatropha curcas progenies; also, the plot and permanent environmental effects were statistically significant. The variance components and heritability estimates increased over time. Non-uniform trajectories were estimated for each progeny throughout the measures, and the area under the trajectories distinguished the progenies with higher performance. High accuracies were found for GY in all harvests, which indicates the high reliability of the results. Moderate to strong genetic correlation was observed across pairs of harvests. The genetic trajectories indicated the existence of genotype × measurement interaction, once the trajectories crossed, which implies a different ranking in each year. Our results suggest that RRM can be efficiently applied for genetic selection in Jatropha curcas breeding programs.


Assuntos
Jatropha/genética , Modelos Genéticos , Melhoramento Vegetal , Variação Biológica da População , Variação Genética
3.
Gene ; 742: 144588, 2020 Jun 05.
Artigo em Inglês | MEDLINE | ID: mdl-32179173

RESUMO

Jatropha curcas is an important bioenergy oil plant, and often planted on barren land to save the area of arable land. It is significant to improve the adaptability of J. curcas to various abiotic stresses. In the present study, we transferred a J. curcas gene, encoding a CBF2 transcription factor, into Nicotiana benthamiana. Under drought treatment, the JcCBF2 transgenic lines showed improved survival rate, leaf water retention and active oxygen scavenging capacity, but reduced photosynthesis and transpiration rate, suggesting that JcCBF2 played an important role in improving plant drought tolerance. Overexpressing JcCBF2 decreased leaf area and increased leaf thickness. To explore the possible mechanisms for the change of leaf anatomical structure, the leaves of wild-type and overexpression lines under drought stress were RNA sequenced. Genes involved in the plant hormones signal transduction were found to be enriched. Cytokinin and indole-3-acetic acid were the major plant hormones whose abundance increased. Quantitative RT-PCR analysis showed expression of NbMYB21, NbMYB86 and NbMYB44 and both abscisic acid (ABA) and jasmonic acid (JA) related genes in the overexpression lines were increased under drought stress. These results indicated that JcCBF2 was able to positively regulate plant drought response by changing the leaf anatomical structure and possibly through JA and ABA signalling pathways. Our work may help us to understand the drought tolerant mechanism.


Assuntos
Jatropha/genética , Proteínas de Plantas/genética , Estresse Fisiológico/genética , Tabaco/genética , Transativadores/genética , Secas , Regulação da Expressão Gênica de Plantas , Jatropha/anatomia & histologia , Folhas de Planta/anatomia & histologia , Folhas de Planta/genética , Folhas de Planta/metabolismo , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Tabaco/anatomia & histologia , Tabaco/crescimento & desenvolvimento , Transativadores/metabolismo
4.
Int J Mol Sci ; 21(2)2020 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-31963715

RESUMO

Jatropha curcas L. is monoecious with a low female-to-male ratio, which is one of the factors restricting its seed yield. Because the phytohormone cytokinins play an essential role in flower development, particularly pistil development, in this study, we elevated the cytokinin levels in J. curcas flowers through transgenic expression of a cytokinin biosynthetic gene (AtIPT4) from Arabidopsis under the control of a J. curcas orthologue of TOMATO MADS BOX GENE 6 (JcTM6) promoter that is predominantly active in flowers. As expected, the levels of six cytokinin species in the inflorescences were elevated, and flower development was modified without any alterations in vegetative growth. In the transgenic J. curcas plants, the flower number per inflorescence was significantly increased, and most flowers were pistil-predominantly bisexual, i.e., the flowers had a huge pistil surrounded with small stamens. Unfortunately, both the male and the bisexual flowers of transgenic J. curcas were infertile, which might have resulted from the continuously high expression of the transgene during flower development. However, the number and position of floral organs in the transgenic flowers were well defined, which suggested that the determinacy of the floral meristem was not affected. These results suggest that fine-tuning the endogenous cytokinins can increase the flower number and the female-to-male ratio in J. curcas.


Assuntos
Proteínas de Arabidopsis/genética , Arabidopsis/metabolismo , Citocininas/metabolismo , Jatropha/genética , Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Vias Biossintéticas , Citocininas/genética , Flores/genética , Flores/fisiologia , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Inflorescência , Jatropha/fisiologia , Plantas Geneticamente Modificadas/fisiologia , Regiões Promotoras Genéticas , Reprodução Assexuada
5.
Sci Rep ; 9(1): 19617, 2019 12 23.
Artigo em Inglês | MEDLINE | ID: mdl-31871315

RESUMO

Salinity is one of the major factors negatively affecting crop productivity. WRKY transcription factors (TFs) are involved in salicylic acid (SA) mediated cellular reactive oxygen species homeostasis in response to different stresses, including salinity. Therefore, the effect of NaCl, NaCl + SA and SA treatments on different photosynthesis-related parameters and wax metabolites were studied in the Jatropha curcas WRKY (JcWRKY) overexpressing tobacco lines. JcWRKY transgenics showed improved photosynthesis rate, stomatal conductance, intercellular CO2 concentration/ambient CO2 concentration ratio (Ci/Ca ratio), electron transport rate (ETR), photosynthesis efficiency (Fv/Fm), photochemical quenching (qP), non-photochemical quenching (NPQ) and quantum yield of PSII electron transport (ΦPSII) in response to salinity stress, while exogenous SA application had subtle effect on these parameters. Alkane, the major constituent of wax showed maximum accumulation in transgenics exposed to NaCl. Other wax components like fatty alcohol, carboxylic acid and fatty acid were also higher in transgenics with NaCl + SA and SA treatments. Interestingly, the transgenics showed a higher number of open stomata in treated plants as compared to wild type (WT), indicating less perception of stress by the transgenics. Improved salinity tolerance in JcWRKY overexpressing tobacco transgenics is associated with photosynthetic efficiency and wax accumulation, mediated by efficient SA signalling. The transgenics showed differential regulation of genes related to photosynthesis (NtCab40, NtLhcb5 and NtRca1), wax accumulation (NtWIN1) and stomatal regulation (NtMUTE, NtMYB-like, NtNCED3-2 and NtPIF3). The present study indicates that JcWRKY is a potential TF facilitating improved photosynthesis with the wax metabolic co-ordination in transgenics during stress.


Assuntos
Fotossíntese , Proteínas de Plantas , Plantas Geneticamente Modificadas , Estresse Salino , Tabaco , Fatores de Transcrição , Ceras/metabolismo , Jatropha/genética , Proteínas de Plantas/biossíntese , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Tabaco/genética , Tabaco/metabolismo , Fatores de Transcrição/biossíntese , Fatores de Transcrição/genética
6.
BMC Genomics ; 20(Suppl 9): 958, 2019 Dec 24.
Artigo em Inglês | MEDLINE | ID: mdl-31874631

RESUMO

BACKGROUND: Jatropha curcas is an oil-bearing plant, and has seeds with high oil content (~ 40%). Several advantages, such as easy genetic transformation and short generation duration, have led to the emergence of J. curcas as a model for woody energy plants. With the development of high-throughput sequencing, the genome of Jatropha curcas has been sequenced by different groups and a mass of transcriptome data was released. How to integrate and analyze these omics data is crucial for functional genomics research on J. curcas. RESULTS: By establishing pipelines for processing novel gene identification, gene function annotation, and gene network construction, we systematically integrated and analyzed a series of J. curcas transcriptome data. Based on these data, we constructed a J. curcas database (JCDB), which not only includes general gene information, gene functional annotation, gene interaction networks, and gene expression matrices but also provides tools for browsing, searching, and downloading data, as well as online BLAST, the JBrowse genome browser, ID conversion, heatmaps, and gene network analysis tools. CONCLUSIONS: JCDB is the most comprehensive and well annotated knowledge base for J. curcas. We believe it will make a valuable contribution to the functional genomics study of J. curcas. The database is accessible at http://jcdb.xtbg.ac.cn.


Assuntos
Jatropha/genética , Bases de Conhecimento , Perfilação da Expressão Gênica , Redes Reguladoras de Genes , Genes de Plantas , Genômica , Jatropha/metabolismo , Modelos Biológicos , Proteínas de Plantas/genética , Mapeamento de Interação de Proteínas , Software
7.
BMC Plant Biol ; 19(1): 468, 2019 Nov 04.
Artigo em Inglês | MEDLINE | ID: mdl-31684864

RESUMO

BACKGROUND: In higher plants, inflorescence architecture is an important agronomic trait directly determining seed yield. However, little information is available on the regulatory mechanism of inflorescence development in perennial woody plants. Based on two inflorescence branching mutants, we investigated the transcriptome differences in inflorescence buds between two mutants and wild-type (WT) plants by RNA-Seq to identify the genes and regulatory networks controlling inflorescence architecture in Jatropha curcas L., a perennial woody plant belonging to Euphorbiaceae. RESULTS: Two inflorescence branching mutants were identified in germplasm collection of Jatropha. The duo xiao hua (dxh) mutant has a seven-order branch inflorescence, and the gynoecy (g) mutant has a three-order branch inflorescence, while WT Jatropha has predominantly four-order branch inflorescence, occasionally the three- or five-order branch inflorescences in fields. Using weighted gene correlation network analysis (WGCNA), we identified several hub genes involved in the cytokinin metabolic pathway from modules highly associated with inflorescence phenotypes. Among them, Jatropha ADENOSINE KINASE 2 (JcADK2), ADENINE PHOSPHORIBOSYL TRANSFERASE 1 (JcAPT1), CYTOKININ OXIDASE 3 (JcCKX3), ISOPENTENYLTRANSFERASE 5 (JcIPT5), LONELY GUY 3 (JcLOG3) and JcLOG5 may participate in cytokinin metabolic pathway in Jatropha. Consistently, exogenous application of cytokinin (6-benzyladenine, 6-BA) on inflorescence buds induced high-branch inflorescence phenotype in both low-branch inflorescence mutant (g) and WT plants. These results suggested that cytokinin is an important regulator in controlling inflorescence branching in Jatropha. In addition, comparative transcriptome analysis showed that Arabidopsis homologous genes Jatropha AGAMOUS-LIKE 6 (JcAGL6), JcAGL24, FRUITFUL (JcFUL), LEAFY (JcLFY), SEPALLATAs (JcSEPs), TERMINAL FLOWER 1 (JcTFL1), and WUSCHEL-RELATED HOMEOBOX 3 (JcWOX3), were differentially expressed in inflorescence buds between dxh and g mutants and WT plants, indicating that they may participate in inflorescence development in Jatropha. The expression of JcTFL1 was downregulated, while the expression of JcLFY and JcAP1 were upregulated in inflorescences in low-branch g mutant. CONCLUSIONS: Cytokinin is an important regulator in controlling inflorescence branching in Jatropha. The regulation of inflorescence architecture by the genes involved in floral development, including TFL1, LFY and AP1, may be conservative in Jatropha and Arabidopsis. Our results provide helpful information for elucidating the regulatory mechanism of inflorescence architecture in Jatropha.


Assuntos
Citocininas/metabolismo , Redes Reguladoras de Genes , Genes de Plantas , Inflorescência/crescimento & desenvolvimento , Jatropha/genética , Reguladores de Crescimento de Planta/metabolismo , Transcriptoma , Perfilação da Expressão Gênica , Inflorescência/genética , Jatropha/crescimento & desenvolvimento , Mutação , Proteínas de Plantas/genética
8.
Sci Rep ; 9(1): 15973, 2019 11 04.
Artigo em Inglês | MEDLINE | ID: mdl-31685957

RESUMO

Jatropha curcas, an economically important biofuel feedstock with oil-rich seeds, has attracted considerable attention among researchers in recent years. Nevertheless, valuable information on the yield component of this plant, particularly regarding ovule development, remains scarce. In this study, transcriptome profiles of anther and ovule development were established to investigate the ovule development mechanism of J. curcas. In total, 64,325 unigenes with annotation were obtained, and 1723 differentially expressed genes (DEGs) were identified between different stages. The DEG analysis showed the participation of five transcription factor families (bHLH, WRKY, MYB, NAC and ERF), five hormone signaling pathways (auxin, gibberellic acid (GA), cytokinin, brassinosteroids (BR) and jasmonic acid (JA)), five MADS-box genes (AGAMOUS-2, AGAMOUS-1, AGL1, AGL11, and AGL14), SUP and SLK3 in ovule development. The role of GA and JA in ovule development was evident with increases in flower buds during ovule development: GA was increased approximately twofold, and JA was increased approximately sevenfold. In addition, the expression pattern analysis using qRT-PCR revealed that CRABS CLAW and AGAMOUS-2 were also involved in ovule development. The upregulation of BR signaling genes during ovule development might have been regulated by other phytohormone signaling pathways through crosstalk. This study provides a valuable framework for investigating the regulatory networks of ovule development in J. curcas.


Assuntos
Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Jatropha/genética , Óvulo Vegetal/genética , Desenvolvimento Vegetal/genética , Transcriptoma , Sequência de Aminoácidos , Biologia Computacional/métodos , Citocininas/metabolismo , Flores/genética , Flores/metabolismo , Genes de Plantas , Anotação de Sequência Molecular , Óvulo Vegetal/ultraestrutura , Reguladores de Crescimento de Planta/metabolismo
9.
DNA Cell Biol ; 38(11): 1278-1291, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31584843

RESUMO

The plants being sessile cannot escape from the adverse environmental stresses, hence get negatively affected in terms of their growth and yield. Transcriptional control simultaneously regulate different cellular processes, minimizing the deleterious effects of these stresses. The salicylic acid (SA)-inducible WRKY family of transcription factors auto or crossregulate the stress signaling in response to abiotic and biotic stresses, facilitating enhanced stress tolerance. In this study, we characterized the group III WRKY gene, JcWRKY2 from ecological and economical valued shrub Jatropha curcas. The JcWRKY2 tobacco transgenics showed improved physiological growth parameters, elevated chlorophyll content, improved antioxidative activities, and increased endogenous SA with both salt and SA stress. Interestingly, the pretreatment with SA and hydrogen peroxide facilitated improved germination of transgenic seeds with salinity stress. The transgenics showed differential regulation of antioxidative enzymes, calcium/calmodulin, dehydrins, and phospholipase genes with salt and SA stress. The increased SA content in transgenics on stress treatments, enhanced the antioxidant capacity leading to reduced susceptibility to stresses. Thus, JcWRKY2 transgenics participate in SA-mediated, improved antioxidative status during salinity stress with reduced reactive oxygen species damage.


Assuntos
Jatropha/genética , Estresse Salino/genética , Tolerância ao Sal/genética , Tabaco/genética , Fatores de Transcrição/fisiologia , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Germinação/efeitos dos fármacos , Germinação/genética , Estresse Oxidativo/efeitos dos fármacos , Estresse Oxidativo/genética , Plantas Geneticamente Modificadas , Salinidade , Cloreto de Sódio/farmacologia , Estresse Fisiológico/genética , Tabaco/efeitos dos fármacos , Tabaco/crescimento & desenvolvimento , Fatores de Transcrição/genética
10.
BMC Plant Biol ; 19(1): 298, 2019 Jul 08.
Artigo em Inglês | MEDLINE | ID: mdl-31286900

RESUMO

BACKGROUND: Homeodomain-leucine zipper (HD-ZIP) transcription factors play important roles in the growth, development and stress responses of plants, including (presumably) physic nut (Jatropha curcas), which has high drought and salinity tolerance. However, although physic nut's genome has been released, there is little knowledge of the functions, expression profiles and evolutionary histories of the species' HD-ZIP genes. RESULTS: In this study, 32 HD-ZIP genes were identified in the physic nut genome (JcHDZs) and divided into four groups (I-IV) based on phylogenetic analysis with homologs from rice, maize and Arabidopsis. The analysis also showed that most of the JcHDZ genes were closer to members from Arabidopsis than to members from rice and maize. Of the 32 JcHDZ genes, most showed differential expression patterns among four tissues (root, stem cortex, leaf, and seed). Expression profile analysis based on RNA-seq data indicated that 15 of the JcHDZ genes respond to at least one abiotic stressor (drought and/or salinity) in leaves at least at one time point. Transient expression of a JcHDZ16-YFP fusion protein in Arabidopsis protoplasts cells showed that JcHDZ16 is localized in the nucleus. In addition, rice seedlings transgenically expressing JcHDZ16 had lower proline contents and activities of antioxidant enzymes (catalase and superoxide dismutase) together with higher relative electrolyte leakage and malondialdehyde contents under salt stress conditions (indicating higher sensitivity) than wild-type plants. The transgenic seedlings also showed increased sensitivity to exogenous ABA, and increases in the transcriptional abundance of several salt stress-responsive genes were impaired in their responses to salt stress. Further data on JcHDZ16-overexpressing plants subjected to salt stress treatment verified the putative role of JcHDZ genes in salt stress responses. CONCLUSION: Our results may provide foundations for further investigation of functions of JcHDZ genes in responses to abiotic stress, and promote application of JcHDZ genes in physic nut breeding.


Assuntos
Jatropha/genética , Oryza/genética , Proteínas de Plantas/genética , Fatores de Transcrição/genética , Estudo de Associação Genômica Ampla , Jatropha/metabolismo , Oryza/metabolismo , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Fatores de Transcrição/metabolismo , Transcriptoma
11.
Sci Rep ; 9(1): 10220, 2019 07 15.
Artigo em Inglês | MEDLINE | ID: mdl-31308439

RESUMO

Edible/non-toxic varieties of Jatropha curcas L. are gaining increasing attention, providing both oil as biofuel feedstock or even as edible oil and the seed kernel meal as animal feed ingredient. They are a viable alternative to the limitation posed by the presence of phorbol esters in toxic varieties. Accurate genotyping of toxic/non-toxic accessions is critical to breeding management. The aim of this study was to identify SNP markers linked to seed toxicity in J. curcas. For SNP discovery, NGS technology was used to sequence the whole genomes of a toxic and non-toxic parent along with a bulk of 51 toxic and 30 non-toxic F2 plants. To ascertain the association between SNP markers and seed toxicity trait, candidate SNPs were genotyped on 672 individuals segregating for seed toxicity and two collections of J. curcas composed of 96 individuals each. In silico SNP discovery approaches led to the identification of 64 candidate SNPs discriminating non-toxic and toxic samples. These SNPs were mapped on Chromosome 8 within the Linkage Group 8 previously identified as a genomic region important for phorbol ester biosynthesis. The association study identified two new SNPs, SNP_J22 and SNP_J24 significantly linked to low toxicity with R2 values of 0.75 and 0.54, respectively. Our study released two valuable SNP markers for high-throughput, marker-assisted breeding of seed toxicity in J. curcas.


Assuntos
Jatropha/genética , Jatropha/toxicidade , Sementes/toxicidade , Biocombustíveis/toxicidade , Biomarcadores , Ligação Genética/genética , Genótipo , Óleos Vegetais/metabolismo , Polimorfismo de Nucleotídeo Único/genética , Sementes/genética
12.
Int J Mol Sci ; 20(9)2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-31052421

RESUMO

Trehalose-6-phosphate (T6P) phosphatase (TPP), a dephosphorylating enzyme, catalyzes the dephosphorylation of T6P, generating trehalose. In Jatropha, we found six members of the TPP family. Five of them JcTPPA, JcTPPC, JcTPPD, JcTPPG, and JcTPPJ are highly expressed in female flowers or male flowers, or both, suggesting that members of the JcTPP family may participate in flower development in Jatropha. The wide expression of JcTPPJ gene in various organs implied its versatile roles and thus was chosen for unraveling its biological functions during developmental process. We constructed an overexpression vector of JcTPPJ cDNA driven by the cauliflower mosaic virus (CaMV) 35S promoter for genetic transformation. Compared with control Arabidopsis plants, 35S:JcTPPJ transgenic Arabidopsis plants presented greater sucrose contents in their inflorescences and displayed late-flowering and heterostylous phenotypes. Exogenous application of sucrose to the inflorescence buds of wild-type Arabidopsis repressed the development of the perianth and filaments, with a phenocopy of the 35S:JcTPPJ transgenic Arabidopsis. These results suggested that the significantly increased sucrose level in the inflorescence caused (or induced) by JcTTPJ overexpression, was responsible for the formation of heterostylous flower phenotype. However, 35S:JcTPPJ transgenic Jatropha displayed no obvious phenotypic changes, implying that JcTPPJ alone may not be sufficient for regulating flower development in Jatropha. Our results are helpful for understanding the function of TPPs, which may regulate flower organ development by manipulating the sucrose status in plants.


Assuntos
Arabidopsis/genética , Expressão Ectópica do Gene , Flores/genética , Jatropha/genética , Fenótipo , Monoéster Fosfórico Hidrolases/genética , Proteínas de Plantas/genética , Arabidopsis/crescimento & desenvolvimento , Jatropha/crescimento & desenvolvimento , Monoéster Fosfórico Hidrolases/metabolismo , Desenvolvimento Vegetal/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Sacarose/metabolismo
13.
Sci Rep ; 9(1): 4510, 2019 03 14.
Artigo em Inglês | MEDLINE | ID: mdl-30872797

RESUMO

MicroRNAs (miRNAs) are endogenously short noncoding regulatory RNAs implicated in plant development and physiology. Nine small RNA (sRNA) libraries from three typical seed developmental stages (young, intermediate, and mature) were generated by deep sequencing to identify the miRNAs of J. curcas, a potential oilseed crop for the production of renewable oil. Strict criteria were adopted to identify 93 high confidence miRNAs including 48 conserved miRNAs and 45 novel miRNAs. Target genes of these miRNAs were involved in a broad range of physiological functions, including gene expression regulation, primary & secondary metabolism, growth & development, signal transduction, and stress response. About one third (29 out of 93) miRNAs showed significant changes in expression levels during the seed developmental process, indicating that the miRNAs might regulate its targets by their changes of transcription levels in seed development. However, most miRNAs were found differentially expressed in the late stage of seed development, suggesting that miRNAs play more important roles in the stage when seed accumulating organic matters and suffering dehydration stress. This study presents the first large scale identification of high confidence miRNAs in the developing seeds of J. curcas.


Assuntos
Perfilação da Expressão Gênica/métodos , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Jatropha/crescimento & desenvolvimento , MicroRNAs/genética , Regulação da Expressão Gênica no Desenvolvimento , Regulação da Expressão Gênica de Plantas , Biblioteca Gênica , Redes Reguladoras de Genes , Jatropha/genética , RNA de Plantas/genética , Sementes/genética , Sementes/crescimento & desenvolvimento , Análise de Sequência de RNA
14.
Mol Biol Rep ; 46(2): 1649-1660, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30756333

RESUMO

Hexokinase, the key rate-limiting enzyme of plant respiration and glycolysis metabolism, has been found to play a vital role in plant sugar sensing and sugar signal transduction. Using Jatropha curcas genome database and bioinformatics method, J. curcas HXK gene family (JcHXK) was identified and its phylogenetic evolution, functional domain, signal peptide at the N-terminal, and expression analysis were conducted. The results showed that a total of 4 HXK genes (JcHXK1, JcHXK2, JcHXK3, and JcHKL1) with 9 exons were systematically identified from J. curcas. JcHXK1, JcHXK3, and JcHKL1 with putative transmembrane domain at the N-terminal belonged to the type of secretory pathway protein, and JcHXK2 contained putative chloroplast targeting peptide. Quantitative real-time PCR (qRT-PCR) analysis revealed that all the four JcHXKs were expressed in different tissues of the leaves, roots, and seeds; however, JcHXK1 and JcHKL1 expression were higher in the roots, whereas JcHXK2 and JcHXK3 showed over-expression in the leaves and seeds, respectively. Furthermore, all the four JcHXKs were up-regulated in the leaves after cold stress at 12 °C; however, only JcHXK3 remarkably demonstrated cold-induced expression in the roots, which reached the highest expression level at 12 h (2.28-fold). According to the cis-acting element analysis results, JcHXK2 contained the most low temperature responsive elements, which was closely related to the cold resistance in J. curcas. A pET-28a-JcHXK2 prokaryotic recombinant expression vector was successfully constructed and a 57.0 kDa protein was obtained, JcHXK2 revealed catalytic activity towards glucose and fructose, with a higher affinity for glucose than fructose. The subcellular localization assays revealed that JcHXK2 was localized in the chloroplast. The results of this study might provide theoretical foundation for further studies on gene cloning and functional verification of HXK family in J. curcas.


Assuntos
Resposta ao Choque Frio/genética , Hexoquinase/genética , Jatropha/genética , Clonagem Molecular/métodos , Temperatura Baixa , Biologia Computacional/métodos , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Estudo de Associação Genômica Ampla/métodos , Jatropha/metabolismo , Filogenia , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/genética
15.
Genes (Basel) ; 10(1)2019 01 21.
Artigo em Inglês | MEDLINE | ID: mdl-30669588

RESUMO

Jatropha curcas is an important perennial, drought tolerant plant that has been identified as a potential biodiesel crop. We report here the hybrid de novo genome assembly of J. curcas generated using Illumina and PacBio sequencing technologies, and identification of quantitative loci for Jatropha Mosaic Virus (JMV) resistance. In this study, we generated scaffolds of 265.7 Mbp in length, which correspond to 84.8% of the gene space, using Benchmarking Universal Single-Copy Orthologs (BUSCO) analysis. Additionally, 96.4% of predicted protein-coding genes were captured in RNA sequencing data, which reconfirms the accuracy of the assembled genome. The genome was utilized to identify 12,103 dinucleotide simple sequence repeat (SSR) markers, which were exploited in genetic diversity analysis to identify genetically distinct lines. A total of 207 polymorphic SSR markers were employed to construct a genetic linkage map for JMV resistance, using an interspecific F2 mapping population involving susceptible J. curcas and resistant Jatropha integerrima as parents. Quantitative trait locus (QTL) analysis led to the identification of three minor QTLs for JMV resistance, and the same has been validated in an alternate F2 mapping population. These validated QTLs were utilized in marker-assisted breeding for JMV resistance. Comparative genomics of oil-producing genes across selected oil producing species revealed 27 conserved genes and 2986 orthologous protein clusters in Jatropha. This reference genome assembly gives an insight into the understanding of the complex genetic structure of Jatropha, and serves as source for the development of agronomically improved virus-resistant and oil-producing lines.


Assuntos
Resistência à Doença , Jatropha/genética , Locos de Características Quantitativas , Geminiviridae , Jatropha/imunologia , Jatropha/virologia , Repetições de Microssatélites
16.
Int J Biol Macromol ; 123: 648-656, 2019 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-30414420

RESUMO

Purple acid phosphatase (PAP) family genes play a crucial role in the phosphorus (P) foraging and recycling. There are 25 putative Jatropha curcas PAP genes (JcrPAP) were identified and classified into three groups based on their molecular weights. Subcellular localization prediction indicated that most of the JcrPAPs were localized to secretory pathway. Several PAPs possess signal peptide motifs and varied numbers of N-glycosylation and transmembrane helix motifs. JcrPAP proteins have 3-5 active pocket sites comprising 1 to 11 binding residues which interact with different ligands such as iron (Fe), N-acetyl l-d-Glucosamine (NAG), zinc (Zn) and manganese (Mn). The core structure of the predicted JcrPap28 was similar to the Ipomoea batatas Pap protein. Most of the JcrPAP genes showed higher expression in the root tissues compared to stem and leaf tissues. Several JcrPAP genes were upregulated under low phosphorus conditions. JcrPAP genes such as JcrPap26b, JcrPap27b, and JcrPap28 have shown multifold induction in low phosphorus treated plants which suggest that these genes might be involved in phosphorus metabolism. The present study provided the structural variations and expression regulation of JcrPAP genes in the economically viable biodiesel crop and it would be helpful for the crop improvement under phosphorus limiting conditions.


Assuntos
Fosfatase Ácida/química , Genoma de Planta/genética , Glicoproteínas/química , Jatropha/química , Sinais Direcionadores de Proteínas , Fosfatase Ácida/genética , Domínio Catalítico , Regulação da Expressão Gênica de Plantas , Glicoproteínas/genética , Glicosilação , Jatropha/genética , Família Multigênica/genética , Fósforo/química , Fósforo/metabolismo , Folhas de Planta/química , Folhas de Planta/genética
17.
Plant Biotechnol J ; 17(2): 517-530, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30059608

RESUMO

Jatropha curcas (physic nut), a non-edible oilseed crop, represents one of the most promising alternative energy sources due to its high seed oil content, rapid growth and adaptability to various environments. We report ~339 Mbp draft whole genome sequence of J. curcas var. Chai Nat using both the PacBio and Illumina sequencing platforms. We identified and categorized differentially expressed genes related to biosynthesis of lipid and toxic compound among four stages of seed development. Triacylglycerol (TAG), the major component of seed storage oil, is mainly synthesized by phospholipid:diacylglycerol acyltransferase in Jatropha, and continuous high expression of homologs of oleosin over seed development contributes to accumulation of high level of oil in kernels by preventing the breakdown of TAG. A physical cluster of genes for diterpenoid biosynthetic enzymes, including casbene synthases highly responsible for a toxic compound, phorbol ester, in seed cake, was syntenically highly conserved between Jatropha and castor bean. Transcriptomic analysis of female and male flowers revealed the up-regulation of a dozen family of TFs in female flower. Additionally, we constructed a robust species tree enabling estimation of divergence times among nine Jatropha species and five commercial crops in Malpighiales order. Our results will help researchers and breeders increase energy efficiency of this important oil seed crop by improving yield and oil content, and eliminating toxic compound in seed cake for animal feed.


Assuntos
Euphorbiaceae/enzimologia , Jatropha/enzimologia , Família Multigênica , Fósforo-Oxigênio Liases/metabolismo , Biocombustíveis , Mapeamento Cromossômico , Euphorbiaceae/genética , Euphorbiaceae/crescimento & desenvolvimento , Perfilação da Expressão Gênica , Jatropha/genética , Jatropha/crescimento & desenvolvimento , Lipídeos/biossíntese , Anotação de Sequência Molecular , Ésteres de Forbol/metabolismo , Fósforo-Oxigênio Liases/genética , Filogenia , Melhoramento Vegetal , Óleos Vegetais/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Sementes/enzimologia , Sementes/genética , Sementes/crescimento & desenvolvimento
18.
Plant Cell Physiol ; 60(2): 462-475, 2019 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-30476253

RESUMO

The lipid biosynthesis pathway in plants has been studied in detail; however, the factors that regulate the pathway at the transcription level are largely unknown. LEAFY COTYLEDON1 (LEC1), WRINKLED1 (WRI1) and FUSCA3 (FUS3) are considered master regulators to control seed oil content in Arabidopsis. Beside these master regulators, several other transcription factors that may regulate the pathway in plants are poorly studied. In the present work, we have shown the involvement of an uncharacterized Jatropha curcas R2R3MYB gene (JcMYB1) in seed oil biosynthesis. Seed oil analysis and expression profiling of fatty acid (FA) and triacylglycerol (TAG) biosynthetic genes in transgenic Arabidopsis and tobacco plants revealed that JcMYB1 enhances seed oil accumulation and alters FA composition by regulating the expression of endogenous pathway genes in transgenics. Using virus-induced gene silencing (VIGS) in Jatropha, we demonstrated that the suppression of JcMYB1 reduced lipid content with altered FA composition. Agro-infiltration and yeast one-hybrid assay results showed that JcMYB1 protein directly binds to the diacylglycerol acyltransferase1 (DGAT1) promoter, a rate-limiting enzyme of TAG biosynthesis, and activates its expression. These results suggested that JcMYB1 may augment the lipid content by regulating lipid biosynthetic genes. Additionally, manipulation of JcMYB1 in oil crop plants may be used for the potential improvement of oil production and quality.


Assuntos
Genes de Plantas/fisiologia , Jatropha/genética , Lipídeos/biossíntese , Proteínas de Plantas/genética , Fatores de Transcrição/genética , Arabidopsis , Ácidos Graxos/biossíntese , Genes de Plantas/genética , Jatropha/fisiologia , Redes e Vias Metabólicas , Proteínas de Plantas/fisiologia , Plantas Geneticamente Modificadas , Tabaco , Fatores de Transcrição/fisiologia , Triglicerídeos/biossíntese , Técnicas do Sistema de Duplo-Híbrido
19.
PLoS One ; 13(12): e0208549, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30521604

RESUMO

Jatropha (Jatropha curcas) is an oil-bearing plant used for biodiesel production. Construction of its standard karyotype and identification of the euchromatin/heterochromatin distribution associated with gene expression and meiotic recombination are essential to fully characterize its genome. Here, we developed a J. curcas karyotype based on meiotic pachytene chromosomes. In addition, a karyotype of J. integerrima, a useful species for jatropha breeding, was also constructed. Five out of eleven J. curcas chromosomes were metacentric, but only two were metacentric in J. integerrima. Almost all of the heterochromatin was distributed around the pericentric regions. The interstitial and distal regions were euchromatic without heterochromatic knobs, except for small heterochromatin regions associated with the subtelomeric repeat sequence JcSat1. These pericentric heterochromatin distribution patterns, together with chromosome structure data and the results of FISH probing with rDNA and JcSat1, allowed us to classify all chromosomes of both species. The two species had two 35S rDNA loci and one 5S rDNA locus; one 35S rDNA locus in J. integerrima was located on the interstitial region of the short arms. In addition, JcSat1 was found at only the heterochromatic ends of the J. curcas chromosome, not the J. integerrima chromosome. Despite the same chromosome number, the two pachytene chromosome-based karyotypes suggest variation in chromosome structure and distribution of repetitive DNAs in these two species.


Assuntos
Cromossomos de Plantas , Jatropha/genética , Cariótipo , Variação Genética , Hibridização in Situ Fluorescente , Estágio Paquíteno , Sequências Repetitivas de Ácido Nucleico
20.
Plant Cell Physiol ; 59(12): 2549-2563, 2018 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-30541045

RESUMO

Jatropha curcas is a promising feedstock for biofuel production because its oil is highly suitable for processing bio-jet fuels and biodiesel. However, Jatropha exhibits a long juvenile stage in subtropical areas. miR172, a conserved small non-protein-coding RNA molecule with 21 nucleotides, regulates a wide range of developmental processes. To date, however, no studies have examined the function of miR172 in Jatropha. There are five miR172 precursors encoding two mature miR172s in Jatropha, which are expressed in all tissues, with the highest expression level in leaves, and the levels are up-regulated with age. Overexpression of JcmiR172a resulted in early flowering, abnormal flowers, and altered leaf morphology in transgenic Arabidopsis and Jatropha. The expression levels of miR172 target genes were down-regulated, and the flower identity genes were up-regulated in the JcmiR172a-overexpressing transgenic plants. Interestingly, we showed that JcmiR172 might be involved in regulation of stem vascular development through manipulating the expression of cellulose and lignin biosynthesis genes. Overexpression of JcmiR172a enhanced xylem development and reduced phloem and pith development. This study helped elucidate the functions of miR172 in perennial plants, a known age-related miRNA involved in the regulation of perennial plant phase change and organ development.


Assuntos
Jatropha/crescimento & desenvolvimento , Jatropha/genética , MicroRNAs/metabolismo , Reprodução/genética , Madeira/crescimento & desenvolvimento , Madeira/genética , Arabidopsis/genética , Sequência de Bases , Tamanho Celular , Flores/genética , Regulação da Expressão Gênica de Plantas , Genes de Plantas , MicroRNAs/genética , Fenótipo , Fotoperíodo , Folhas de Planta/anatomia & histologia , Caules de Planta/anatomia & histologia , Plantas Geneticamente Modificadas , Sementes/genética , Sementes/crescimento & desenvolvimento , Xilema/crescimento & desenvolvimento
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