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1.
Life (Basel) ; 12(11)2022 Nov 07.
Artigo em Inglês | MEDLINE | ID: mdl-36362967

RESUMO

The worldwide climate changes every year due to global warming, waterlogging, drought, salinity, pests, and pathogens, impeding crop productivity. Brassica napus is one of the most important oil crops in the world, and rapeseed oil is considered one of the most health-beneficial edible vegetable oils. Recently, miRNAs have been found and confirmed to control the expression of targets under disruptive environmental conditions. The mechanism is through the formation of the silencing complex that mediates post-transcriptional gene silencing, which pairs the target mRNA and target cleavage and/or translation inhibition. However, the functional role of miRNAs and targets in B. napus is still not clarified. This review focuses on the current knowledge of miRNAs concerning development regulation and biotic and abiotic stress responses in B. napus. Moreover, more strategies for miRNA manipulation in plants are discussed, along with future perspectives, and the enormous amount of transcriptome data available provides cues for miRNA functions in B. napus. Finally, the construction of the miRNA regulatory network can lead to the significant development of climate change-tolerant B. napus through miRNA manipulation.

2.
Int J Mol Sci ; 23(19)2022 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-36232853

RESUMO

Stress-associated protein (SAP) genes-encoding A20/AN1 zinc-finger domain-containing proteins-play pivotal roles in regulating stress responses, growth, and development in plants. They are considered suitable candidates to improve abiotic stress tolerance in plants. However, the SAP gene family in sweetpotato (Ipomoea batatas) and its relatives is yet to be investigated. In this study, 20 SAPs in sweetpotato, and 23 and 26 SAPs in its wild diploid relatives Ipomoea triloba and Ipomoea trifida were identified. The chromosome locations, gene structures, protein physiological properties, conserved domains, and phylogenetic relationships of these SAPs were analyzed systematically. Binding motif analysis of IbSAPs indicated that hormone and stress responsive cis-acting elements were distributed in their promoters. RT-qPCR or RNA-seq data revealed that the expression patterns of IbSAP, ItbSAP, and ItfSAP genes varied in different organs and responded to salinity, drought, or ABA (abscisic acid) treatments differently. Moreover, we found that IbSAP16 driven by the 35 S promoter conferred salinity tolerance in transgenic Arabidopsis. These results provided a genome-wide characterization of SAP genes in sweetpotato and its two relatives and suggested that IbSAP16 is involved in salinity stress responses. Our research laid the groundwork for studying SAP-mediated stress response mechanisms in sweetpotato.


Assuntos
Arabidopsis , Ipomoea batatas , Ipomoea , Ácido Abscísico/metabolismo , Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Proteínas de Choque Térmico/metabolismo , Hormônios/metabolismo , Ipomoea/genética , Ipomoea batatas/genética , Ipomoea batatas/metabolismo , Filogenia , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Tolerância ao Sal/genética , Estresse Fisiológico/genética , Zinco/metabolismo , Dedos de Zinco/genética
3.
Virulence ; 13(1): 1358-1378, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-35943142

RESUMO

Aspergillus flavus infects various crops with aflatoxins, and leads to aspergillosis opportunistically. Though H3K36 methylation plays an important role in fungal toxin metabolism and virulence, no data about the biological function of H3K36 methylation in A. flavus virulence has been reported. Our study showed that the Set2 histone methyltransferase family, AshA and SetB, involves in morphogenesis and mycotoxin anabolism by regulating related transcriptional factors, and they are important for fungal virulence to crops and animals. Western-blotting and double deletion analysis revealed that AshA mainly regulates H3K36me2, whereas SetB is mainly responsible for H3K36me3 in the nucleus. By construction of domain deletion A. flavus strain and point mutation strains by homologous recombination, the study revealed that SET domain is indispensable in mycotoxin anabolism and virulence of A. flavus, and N455 and V457 in it are the key amino acid residues. ChIP analysis inferred that the methyltransferase family controls fungal reproduction and regulates the production of AFB1 by directly regulating the production of the transcriptional factor genes, including wetA, steA, aflR and amylase, through H3K36 trimethylation in their chromatin fragments, based on which this study proposed that, by H3K36 trimethylation, this methyltransferase family controls AFB1 anabolism through transcriptional level and substrate utilization level. This study illuminates the epigenetic mechanism of the Set2 family in regulating fungal virulence and mycotoxin production, and provides new targets for controlling the virulence of the fungus A. flavus.AUTHOR SUMMARYThe methylation of H3K36 plays an important role in the fungal secondary metabolism and virulence, but no data about the regulatory mechanism of H3K36 methylation in the virulence of A. flavus have been reported. Our study revealed that, in the histone methyltransferase Set2 family, AshA mainly catalyzes H3K36me2, and involves in the methylation of H3K36me1, and SetB mainly catalyzes H3K36me3 and H3K36me1. Through domain deletion and point mutation analysis, this study also revealed that the SET domain was critical for the normal biological function of the Set2 family and that N455 and V457 in the domain were critical for AshA. By ChIP-seq and ChIP-qPCR analysis, H3K36 was found to be trimethylation modified in the promotors and ORF positions of wetA, steA, aflR and the amylase gene (AFLA_084340), and further qRT-PCR results showed that these methylation modifications regulate the expression levels of these genes. According to the results of ChIP-seq analysis, we proposed that, by H3K36 trimethylation, this methyltransferase family controls the metabolism of mycotoxin through transcriptional level and substrate utilization level. All the results from this study showed that Set2 family is essential for fungal secondary metabolism and virulence, which lays a theoretical groundwork in the early prevention and treatment of A. flavus pollution, and also provides an effective strategy to fight against other pathogenic fungi.


Assuntos
Aspergillus flavus , Micotoxinas , Amilases/metabolismo , Animais , Aspergillus flavus/genética , Histona Metiltransferases/metabolismo , Histonas/genética , Histonas/metabolismo , Metilação , Metiltransferases/genética , Metiltransferases/metabolismo , Micotoxinas/genética , Micotoxinas/metabolismo , Metabolismo Secundário , Virulência
4.
Sci Rep ; 12(1): 11549, 2022 07 07.
Artigo em Inglês | MEDLINE | ID: mdl-35798807

RESUMO

Accurately obtaining the spatial distribution information of fruit tree planting is of great significance to the development of fruit tree growth monitoring, disease and pest control, and yield estimation. In this study, the Sentenel-2 multispectral remote sensing imageries of different months during the growth period of the fruit trees were used as the data source, and single month vegetation indices, accumulated monthly vegetation indices (∑VIs), and difference vegetation indices between adjacent months (∆VIs) were constructed as input variables. Four conventional vegetation indices of NDVI, PSRI, GNDVI, and RVI and four improved vegetation indices of NDVIre1, NDVIre2, NDVIre3, and NDVIre4 based on the red-edge band were selected to construct a decision tree classification model combined with machine learning technology. Through the analysis of vegetation indices under different treatments and different months, combined with the attribute of Feature_importances_, the vegetation indices of different periods with high contribution were selected as input features, and the Max_depth values of the decision tree model were determined by the hyperparameter learning curve. The results have shown that when the Max_depth value of the decision tree model of the vegetation indices under the three treatments was 6, 8, and 8, the model classification was the best. The accuracy of the three vegetation index processing models on the training set were 0.8936, 0.9153, and 0.8887, and the accuracy on the test set were 0.8355, 0.7611, and 0.7940, respectively. This method could be applied to remote sensing classification of fruit trees in a large area, and could provide effective technical means for monitoring fruit tree planting areas with medium and high resolution remote sensing imageries.


Assuntos
Frutas , Tecnologia de Sensoriamento Remoto , Tecnologia de Sensoriamento Remoto/métodos
5.
Front Plant Sci ; 13: 841969, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35498654

RESUMO

The present study was undertaken to determine the scope of sweetpotato cultivation in arid regions of China. For this purpose, we investigated yield, anthocyanin compositions and physicochemical properties of starch in purple-fleshed sweetpotato (PFSP) "Xuzishu8" under humid (zi8-X) and arid (zi8-D) environments of China. The experiment was conducted in three replications in both environments during 2019 and 2020. The yield and anthocyanidins contents of PFSP were significantly higher in the arid conditions as compared to humid. Zi8-X and zi8-D both revealed the presence of three anthocyanidins, namely, cyanidin (Cy), peonidin (Pn), and pelargonidin (Pg). Cy and Pn accounted for 36.40 and 63.54% of the total anthocyanidins in zi8-X, while in zi8-D, they were found as 26.13 and 73.80%, respectively. The quantitative analysis of these anthocyanins was performed using HPLC-ESI-MS/MS which revealed eighteen anthocyanins such as nine Cy, eight Pn and one Pg. Out of which, eleven anthocyanins showed a significant difference under both conditions. Starch and amylopectin contents were found to be increased by 15.39 and 4.71%, respectively, while the amylose concentration was reduced by 15.54% under the arid environment. The diameter of the starch granule and the peak viscosity were significantly higher under arid as compared to humid conditions. On the basis of results of this study, it seems quite practicable to develop PFSP cultivation in desert regions.

6.
Plant Physiol Biochem ; 168: 143-154, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34628175

RESUMO

GIGANTEA (GI) is known to play significant roles in various molecular pathways. Nevertheless, the underlying mechanism of the transcriptional regulation of GI remains obscure in sweetpotato. In the present study, a 1518-bp promoter sequence was obtained from the Ipomoea batatas GIGANTEA (IbGI) gene, and several potential cis-elements responsive to light, phytohormones and abiotic stresses were identified by in silico analysis. In order to functionally validate the IbGI promoter, the 5' deletion analysis of the promoter was performed by cloning the full-length promoter (D0) and its four deletion fragments, D1 (1235 bp), D2 (896 bp), D3 (549 bp) and D4 (286 bp), upstream of the ß-glucuronidase (GUS) reporter gene. Then, these were stably transformed in Arabidopsis plants. All transgenic seedlings exhibited stable GUS activity in the condition of control, but with decreased activity in the condition of most treatments. Interestingly, merely D1 seedlings that contained an abscisic acid responsive cis-element (ABRE-element) had an extremely powerful GUS activity under the treatment of ABA, which implies that fragment spanning nucleotides of -1235 to -896 bp might be a crucial component for the responses of ABA. Eight different types of potential transcriptional regulators of IbGI were isolated by Y1H, including TGA2.2, SPLT1 and GADPH, suggesting the complex interaction mode of protein-DNA on the IbGI promoter. Taken together, these present results help to better understand the transcriptional regulation mechanism of the IbGI gene, and provides an insight into the IbGI promoter, which can be considered as an alternation for breeding transgenic plants.


Assuntos
Arabidopsis , Ipomoea batatas , Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Ipomoea batatas/genética , Melhoramento Vegetal , Reguladores de Crescimento de Plantas/farmacologia , Plantas Geneticamente Modificadas/genética , Estresse Fisiológico/genética
7.
Int J Mol Sci ; 22(13)2021 Jun 22.
Artigo em Inglês | MEDLINE | ID: mdl-34206151

RESUMO

The saccharification of sweetpotato storage roots is a common phenomenon in the cooking process, which determines the edible quality of table use sweetpotato. In the present study, two high saccharified sweetpotato cultivars (Y25, Z13) and one low saccharified cultivar (X27) in two growth periods (S1, S2) were selected as materials to reveal the molecular mechanism of sweetpotato saccharification treated at high temperature by transcriptome sequencing and non-targeted metabolome determination. The results showed that the comprehensive taste score, sweetness, maltose content and starch change of X27 after steaming were significantly lower than those of Y25 and Z13. Through transcriptome sequencing analysis, 1918 and 1520 differentially expressed genes were obtained in the two periods of S1 and S2, respectively. Some saccharification-related transcription factors including MYB families, WRKY families, bHLH families and inhibitors were screened. Metabolic analysis showed that 162 differentially abundant metabolites related to carbohydrate metabolism were significantly enriched in starch and sucrose capitalization pathways. The correlation analysis between transcriptome and metabolome confirmed that the starch and sucrose metabolic pathways were significantly co-annotated, indicating that it is a vitally important metabolic pathway in the process of sweetpotato saccharification. The data obtained in this study can provide valuable resources for follow-up research on sweetpotato saccharification and will provide new insights and theoretical basis for table use sweetpotato breeding in the future.


Assuntos
Metabolismo dos Carboidratos , Temperatura Alta , Ipomoea batatas/metabolismo , Raízes de Plantas/metabolismo , Transcriptoma , Manipulação de Alimentos , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Metabolômica , Amido/metabolismo , Sacarose/metabolismo
8.
J Immunother Cancer ; 8(2)2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-33323464

RESUMO

BACKGROUND: In patients with hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC), virus-specific cytotoxic T lymphocytes (CTLs) fail to eliminate HCC cells expressing HBV antigens. As the expression of viral antigen in HBV-associated HCC may decrease to allow tumor to escape immune attacks, we hypothesized that an HBV surface antigen (HBsAg)-specific affinity-improved-T-cell receptor (TCR) will enable T cells to target HCC more effectively than corresponding wild-type-TCR. We also postulated that TCR promiscuity can be exploited to efficiently capture HBV variants that can hinder CTL-based therapeutics. METHODS: We applied flexi-panning to isolate affinity-improved TCRs binding to a variant antigen, the human leukocyte antigen (HLA)-A*02:01-restricted nonapeptide HBs371-379-ILSPFLPLL, from libraries constructed with a TCR cloned using the decapeptide HBs370-379-SIVSPFIPLL. The potency and safety of the affinity-improved-TCR engineered T-cells (Ai-TCR-T) were verified with potentially cross-reactive human and HBV-variant peptides, tumor and normal cells, and xenograft mouse models. RESULTS: Ai-TCR-T cells retained cognate HBV antigen specificity and recognized a wide range of HBV genotypic variants with improved sensitivity and cytotoxicity. Cell infusions produced complete elimination of HCC without recurrence in the xenograft mouse models. Elevated accumulation of CD8+ Ai-TCR-T cells in tumors correlated with tumor shrinkage. CONCLUSION: The in vitro and in vivo studies demonstrated that HBsAg-specific Ai-TCR-T cells had safety profiles similar to those of their wild-type counterparts and significantly enhanced potency. This study presents an approach to develop new therapeutic strategies for HBV-related HCC.


Assuntos
Carcinoma Hepatocelular/virologia , Vírus da Hepatite B/patogenicidade , Neoplasias Hepáticas/virologia , Linfócitos T/metabolismo , Engenharia Tecidual/métodos , Animais , Carcinoma Hepatocelular/imunologia , Carcinoma Hepatocelular/patologia , Humanos , Neoplasias Hepáticas/patologia , Masculino , Camundongos , Ensaios Antitumorais Modelo de Xenoenxerto
9.
Sci Rep ; 10(1): 12573, 2020 07 28.
Artigo em Inglês | MEDLINE | ID: mdl-32724138

RESUMO

Purple-fleshed sweet potato (PFSP) is an important food crop, as it is a rich source of nutrients and anthocyanin pigments. Drought has become a major threat to sustainable sweetpotato production, resulting in huge yield losses. Therefore, the present study was conducted to identify drought stress-responsive genes using next-generation (NGS) and third-generation sequencing (TGS) techniques. Five cDNA libraries were constructed from seedling leaf segments treated with a 30% solution of polyethylene glycol (PEG-6000) for 0, 1, 6, 12, and 48 h for second-generation sequencing. Leaf samples taken from upper third of sweet potato seedlings after 1, 6, 12, and 48 h of drought stress were used for the construction of cDNA libraries for third-generation sequencing; however, leaf samples from untreated plants were collected as controls. A total of 184,259,679 clean reads were obtained using second and third-generation sequencing and then assembled into 17,508 unigenes with an average length of 1,783 base pairs. Out of 17,508 unigenes, 642 (3.6%) unigenes failed to hit any homologs in any databases, which might be considered novel genes. A total of 2, 920, 1578, and 2,418 up-regulated unigenes and 3,834, 2,131, and 3,337 down-regulated unigenes from 1 h, 6 h, 12 h, and 48 h library were identified, respectively in drought stress versus control. In addition, after 6, 12, and 48 h of drought stress, 540 up-regulated unigenes, 486 down-regulated unigenes and 414 significantly differentially expressed unigenes were detected. It was found that several gene families including Basic Helix-loop-helix (bHLH), basic leucine zipper (bZIP), Cystein2/Histidine2 (C2H2), C3H, Ethylene-responsive transcription factor (ERF), Homo domain-leucine zipper (HD-ZIP), MYB, NAC (NAM, ATAF1/2, and CUC2), Thiol specific antioxidant and WRKY showed responses to drought stress. In total, 17,472 simple sequence repeats and 510,617 single nucleotide polymorphisms were identified based on transcriptome sequencing of the PFSP. About 96.55% of the obtained sequences are not available online in sweet potato genomics resources. Therefore, it will enrich annotated sweet potato gene sequences and enhance understanding of the mechanisms of drought tolerance through genetic manipulation. Moreover, it represents a sequence resource for genetic and genomic studies of sweet potato.


Assuntos
Ipomoea batatas/fisiologia , Proteínas de Plantas/genética , Poliploidia , RNA de Plantas/genética , Secas , Regulação da Expressão Gênica de Plantas , Ipomoea batatas/genética , Proteínas de Plantas/metabolismo , RNA de Plantas/metabolismo , Análise de Sequência de RNA , Estresse Fisiológico , Transcriptoma
10.
Front Microbiol ; 11: 234, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32132990

RESUMO

The filament fungal pathogen, Aspergillus flavus, spreads worldwide and contaminates several important crops. Histone posttranslational modifications are deeply involved in fungal development and virulence, but the biological function of the histone methyltransferase AflSet1 in A. flavus is still unknown. In the study, Aflset1 deletion strain was constructed through homologous recombination, and it was found that AflSet1 up-regulates hyphae growth, and promotes conidiation by sporulation regulation genes: abaA and brlA. It was also found that AflSet1 involves in sclerotia formation and AFB1 biosynthesis via sclerotia related transcriptional factors and orthodox AFB1 synthesis pathway, respectively. Crop models revealed that AflSet1 plays critical roles in colonization and AFB1 production on crop kernels. Lipase activity analysis suggested that AflSet1 affects fungal virulence to crops via digestive enzymes. Stresses tests revealed that AflSet1 is deeply involved in fungal resistance against osmotic, oxidative and cell membrane stress. The preparation of N_SET, SET domain deletion mutants and H988K mutant revealed that both domains play critical roles in fungal development and AFB1 production, and that H988 is very important in executing biological functions on morphogenesis and AFB1 synthesis. Subcellular location analysis revealed that AflSet1 is stably accumulated in nuclei in both spore germination and hyphae growth stages, even under the stress of SDS. Through immunoblot analysis, it was found that AflSet1 methylates H3K4me2 and me3 as well as H3K9me2. This study provides a solid evidence to discover the biological functions of histone methyltransferase in pathogenic fungi.

11.
BMC Genomics ; 21(1): 197, 2020 Mar 04.
Artigo em Inglês | MEDLINE | ID: mdl-32131729

RESUMO

BACKGROUND: Purple-fleshed sweetpotato (PFSP) is one of the most important crops in the word which helps to bridge the food gap and contribute to solve the malnutrition problem especially in developing countries. Salt stress is seriously limiting its production and distribution. Due to lacking of reference genome, transcriptome sequencing is offering a rapid approach for crop improvement with promising agronomic traits and stress adaptability. RESULTS: Five cDNA libraries were prepared from the third true leaf of hexaploid sweetpotato at seedlings stage (Xuzi-8 cultivar) treated with 200 mM NaCl for 0, 1, 6, 12, 48 h. Using second and third generation technology, Illumina sequencing generated 170,344,392 clean high-quality long reads that were assembled into 15,998 unigenes with an average length 2178 base pair and 96.55% of these unigenes were functionally annotated in the NR protein database. A number of 537 unigenes failed to hit any homologs which may be considered as novel genes. The current results indicated that sweetpotato plants behavior during the first hour of salt stress was different than the other three time points. Furthermore, expression profiling analysis identified 4, 479, 281, 508 significantly expressed unigenes in salt stress treated samples at the different time points including 1, 6, 12, 48 h, respectively as compared to control. In addition, there were 4, 1202, 764 and 2195 transcription factors differentially regulated DEGs by salt stress at different time points including 1, 6, 12, 48 h of salt stress. Validation experiment was done using 6 randomly selected unigenes and the results was in agree with the DEG results. Protein kinases include many genes which were found to play a vital role in phosphorylation process and act as a signal transductor/ receptor proteins in membranes. These findings suggest that salt stress tolerance in hexaploid sweetpotato plants may be mainly affected by TFs, PKs, Protein Detox and hormones related genes which contribute to enhance salt tolerance. CONCLUSION: These transcriptome sequencing data of hexaploid sweetpotato under salt stress conditions can provide a valuable resource for sweetpotato breeding research and focus on novel insights into hexaploid sweetpotato responses to salt stress. In addition, it offers new candidate genes or markers that can be used as a guide to the future studies attempting to breed salt tolerance sweetpotato cultivars.


Assuntos
Sequenciamento do Exoma/métodos , Perfilação da Expressão Gênica/métodos , Ipomoea batatas/fisiologia , Proteínas de Plantas/genética , Regulação da Expressão Gênica de Plantas , Biblioteca Gênica , Sequenciamento de Nucleotídeos em Larga Escala , Ipomoea batatas/genética , Anotação de Sequência Molecular , Poliploidia , Estresse Salino
12.
J Exp Bot ; 71(9): 2723-2739, 2020 05 09.
Artigo em Inglês | MEDLINE | ID: mdl-31974553

RESUMO

Due to its subtropical origins, rice (Oryza sativa) is sensitive to low-temperature stress. In this study, we identify LOC_Os04g24110, annotated to encode the UDP-glycosyltransferase enzyme UGT90A1, as a gene associated with the low-temperature seedling survivability (LTSS) quantitative trait locus qLTSS4-1. Differences between haplotypes in the control region of OsUGT90A1 correlate with chilling tolerance phenotypes, and reflect differential expression between tolerant and sensitive accessions rather than differences in protein sequences. Expression of OsUGT90A1 is initially enhanced by low temperature, and its overexpression helps to maintain membrane integrity during cold stress and promotes leaf growth during stress recovery, which are correlated with reduced levels of reactive oxygen species due to increased activities of antioxidant enzymes. In addition, overexpression of OsUGT90A1 in Arabidopsis improves freezing survival and tolerance to salt stress, again correlated with enhanced activities of antioxidant enzymes. Overexpression of OsUGT90A1 in rice decreases root lengths in 3-week-old seedlings while gene-knockout increases the length, indicating that its differential expression may affect phytohormone activities. We conclude that higher OsUGT90A1 expression in chilling-tolerant accessions helps to maintain cell membrane integrity as an abiotic stress-tolerance mechanism that prepares plants for the resumption of growth and development during subsequent stress recovery.


Assuntos
Oryza , Membrana Celular , Temperatura Baixa , Resposta ao Choque Frio , Regulação da Expressão Gênica de Plantas , Glicosiltransferases/genética , Oryza/genética , Plântula/genética
13.
Front Plant Sci ; 10: 1086, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31552077

RESUMO

Lipid remodeling plays an important role in the adaptation of plants to environmental factors, but the mechanism by which lipid remodeling mediates salt stress response remains unclear. In this study, we compared the root and leaf lipidome profiles of salt-tolerant and salt-sensitive sweet potato cultivars (Xu 22 and Xu 32, respectively) under salinity stress. After salt treatment, the leaf lipidome showed more significant remodeling than the root lipidome in both cultivars. Compared with Xu 32 leaves, Xu 22 leaves generally maintained higher abundance of phospholipids, glycolipids, sphingolipids, sterol derivatives, and diacylglycerol under salinity conditions. Interestingly, salinity stress significantly increased phosphatidylserine (PS) abundance in Xu 22 leaves by predominantly triggering the increase of PS (20:5/22:6). Furthermore, Xu 32 leaves accumulated higher triacylglycerol (TG) level than Xu 22 leaves under salinity conditions. The exogenous application of PS delayed salt-induced leaf senescence in Xu 32 by reducing salt-induced K+ efflux and upregulating plasma membrane H+-ATPase activity. However, the inhibition of TG mobilization in salinized-Xu 22 leaves disturbed energy and K+/Na+ homeostasis, as well as plasma membrane H+-ATPase activity. These results demonstrate alterations in the leaf lipidome of sweet potato under salinity condition, underscoring the importance of PS and TG in mediating salt-defensive responses in sweet potato leaves.

14.
PLoS One ; 14(5): e0216470, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31063496

RESUMO

BACKGROUND: Human B-cell lymphoma 6 (BCL6) gene, usually coding protein of 706 amino acids, is closely associated with large B cell lymphoma. Researches showed that protein mutation or change of expression levels usually happened in the mounting non-hodgkin lymphoma (NHL). Thus BCL6 is considered to be involved in germinal center (GC)-derived lymphoma. RESULTS: The BCL61-350 gene codons were optimized for prokaryotic system. After expression of BCL61-350 in E. coli, the BCL61-350 protein was purified with Ni column. Then the BCL61-350 protein, mixing with QuickAntibody-Mouse5W adjuvant, was injected into Balb/c mice. After immunization and cell fusion, a stable cell line named 1E6A4, which can secrete anti-BCL6 antibody, was obtained. The isotype of 1E6A4 mAb was determined as IgG2a, and the affinity constant reached 5.12×1010 L/mol. Furthermore, the specificity of the mAb was determined with ELISA, western blot and immunohistochemistry. Results indicated that the 1E6A4 mAb was able to detect BCL6 specifically and sensitively. CONCLUSIONS: BCL61-350 antigen has been successfully generated with an effective and feasible method, and a highly specific antibody named 1E6A4 against BCL6 has been screened and characterized in this study, which was valuable in clinical diagnosis.


Assuntos
Anticorpos Monoclonais Murinos , Imunoglobulina G , Linfoma de Células B/diagnóstico , Linfoma de Células B/imunologia , Proteínas Proto-Oncogênicas c-bcl-6/imunologia , Animais , Anticorpos Monoclonais Murinos/química , Anticorpos Monoclonais Murinos/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Centro Germinativo/imunologia , Centro Germinativo/metabolismo , Centro Germinativo/patologia , Humanos , Imunoglobulina G/química , Imunoglobulina G/imunologia , Imuno-Histoquímica , Linfoma de Células B/metabolismo , Linfoma de Células B/patologia , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Proto-Oncogênicas c-bcl-6/biossíntese
16.
Plant Physiol Biochem ; 135: 395-403, 2019 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-30634134

RESUMO

Anthocyanins are synthesized by multi-enzyme complexes localized at the cytoplasmic surface of the endoplasmic reticulum (synthesis site), and transported to the destination site, the vacuole. Three mechanisms for the vacuolar accumulation of anthocyanin in plant species have been proposed. Previous studies have indicated that glutathione S-transferase (GST) genes from model and ornamental plants are involved in anthocyanin transportation. In the present study, an anthocyanin-related GST, IbGSTF4, was identified and characterized based on transcriptome results. Phylogenetic analysis revealed that IbGSTF4 was most closely correlated to PhAN9 and CkmGST3, the anthocyanin-related GST of Petunia hybrida and Cyclamen. Furthermore, the expression analysis revealed that IbGSTF4 is strongly expressed in pigmented tissues, when compared to green organs, which is in agreement to the ability to correlate with anthocyanin accumulation. A GST activity assay uncovered that the IbGST4 protein owned similar activities with the GST family. Furthermore, the molecular functional complementation of Arabidopsis thaliana mutant tt19 demonstrated that IbGSTF4 might play a vital role in the vacuole sequestration of anthocyanin in sweetpotato. Moreover, the dual luciferase assay revealed that the LUC driven by the promoter of IbGSTF4 could not be directly activated by IbMYB1, suggesting that the regulatory mechanism of anthocyanin accumulation and sequestration in sweetpotato was intricate.


Assuntos
Antocianinas/metabolismo , Glutationa Transferase/genética , Ipomoea batatas/enzimologia , Proteínas de Plantas/genética , Arabidopsis/genética , DNA de Plantas/genética , Genes de Plantas/genética , Genes de Plantas/fisiologia , Glutationa Transferase/metabolismo , Ipomoea batatas/genética , Ipomoea batatas/metabolismo , Filogenia , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Reação em Cadeia da Polimerase em Tempo Real , Análise de Sequência de DNA , Transcriptoma
17.
Genes Genomics ; 41(1): 107-111, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30264213

RESUMO

Lipoic acid synthase (LipA) plays a role in lipoic acid synthesis and potentially affects the levels of acetyl-CoA, the critical precursor of tricarboxylic acid (TCA) cycle. Considering the potential effect of LipA on TCA cycle, whether the enzyme is involved in the growth and aflatoxin B1 (AFB1) biosynthesis, the significant events in Aspergillus flavus is yet known. The study was designed to explore the role of lipA gene in A. flavus, including growth rate, conidiation, sclerotia formation, and biosynthesis of AFB1. LipA coding lipoic acid synthetase was knocked out using homologous recombination. The role of lipA gene in A. flavus morphogenesis (including colony size, conidiation, and sclerotia formation) was explored on various media, and the bio-function of lipA gene in the biosynthesis of AFB1 was analyzed by thin layer chromatography analysis. The growth was suppressed in △lipA. The formation of conidia and sclerotia was also reduced when lipA gene was deleted. Moreover, AFB1 was down-regulated in ΔlipA compared with WT controls. LipA plays a role in the development of A. flavus and AFB1 biosynthesis, contributing to the full understanding of the lipA bio-function in A. flavus.


Assuntos
Aspergillus/genética , Proteínas Fúngicas/genética , Sulfurtransferases/genética , Aspergillus/enzimologia , Aspergillus/crescimento & desenvolvimento , Proteínas Fúngicas/metabolismo , Mutação , Esporos Fúngicos/crescimento & desenvolvimento , Esporos Fúngicos/metabolismo , Sulfurtransferases/metabolismo
18.
Data Brief ; 21: 88-91, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30338280

RESUMO

Increasing demands for bio-products such as starch and anthocyanin stimulate researches towards purple-fleshed sweetpotato. Therefore, the aim of this complementary work is to identify the relationship between leaf area index, T/R value, starch and anthocyanin content in purple sweetpotato cultivar (cv "Xz3" from China) during the different growth stages. The traits were investigated every 15 days starting from 60 to 135 days after transplanting. The current data considered as a complementary for the main work "Quantifying cultivation technique and growth dynamics of purple-fleshed sweetpotato (Ipomoea batatas L.) in China" (Tang et al., 2018) [1].

19.
Toxins (Basel) ; 10(7)2018 07 20.
Artigo em Inglês | MEDLINE | ID: mdl-30036940

RESUMO

Aspergillus flavus produces mycotoxins especially aflatoxin B1 and infects crops worldwide. As a PHD transcription factor, there is no report on the role of Rum1 in the virulence of Aspergillus spp. yet. This study explored the biological function of Rum1 in A. flavus through the construction of rum1 deletion mutants and rum1 complementation strains with the method of homologous recombination. It was found, in the study, that Rum1 negatively regulates conidiation through abaA and brlA, positively regulates sclerotia formation through nsdC, nsdD, and sclR, triggers aflatoxin biological synthesis, and enhances the activity of amylase. Our findings suggested that Rum1 plays a major role in the growth of mycelia, conidia, and sclerotia production along with aflatoxin biosynthesis in A. flavus.


Assuntos
Aflatoxinas/biossíntese , Aspergillus flavus/fisiologia , Proteínas Fúngicas/fisiologia , Fatores de Transcrição/fisiologia , Morfogênese , Mutação , Micélio/crescimento & desenvolvimento , Esporos Fúngicos/crescimento & desenvolvimento
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