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1.
Pestic Biochem Physiol ; 170: 104681, 2020 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-32980063

RESUMO

Chitosan oligosaccharides (COS) can elicit plant immunity and defence responses in rice plants, but exactly how this promotes plant growth remains largely unknown. Herein, we explored the effects of 0.5 mg/L COS on plant growth promotion in rice seedlings by measuring root and stem length, investigating biochemical factors in whole plants via proteomic analysis, and confirming upregulated and downregulated genes by real-time quantitative PCR. Pathway enrichment results showed that COS promoted root and stem growth, and stimulated metabolic (biosynthetic and catabolic processes) and photosynthesis in rice plants during the seedling stage. Expression levels of genes related to chlorophyll a-b binding, RNA binding, catabolic processes and calcium ion binding were upregulated following COS treatment. Furthermore, comparative analysis indicated that numerous proteins involved in the biosynthesis, metabolic (catabolic) processes and photosynthesis pathways were upregulated. The findings indicate that COS may upregulate calcium ion binding, photosynthesis, RNA binding, and catabolism proteins associated with plant growth during the rice seedling stage.


Assuntos
Quitosana , Oryza/genética , Clorofila A , Regulação da Expressão Gênica de Plantas , Oligossacarídeos , Proteínas de Plantas/genética , Proteômica , Plântula/genética
2.
Nat Commun ; 11(1): 3439, 2020 07 10.
Artigo em Inglês | MEDLINE | ID: mdl-32651385

RESUMO

Various stress conditions induce the nuclear translocation of cytosolic glyceraldehyde-3-phosphate dehydrogenase (GAPC), but its nuclear function in plant stress responses remains elusive. Here we show that GAPC interacts with a transcription factor to promote the expression of heat-inducible genes and heat tolerance in Arabidopsis. GAPC accumulates in the nucleus under heat stress. Overexpression of GAPC enhances heat tolerance of seedlings and the expression of heat-inducible genes whereas knockout of GAPCs has opposite effects. Screening of Arabidopsis transcription factors identifies nuclear factor Y subunit C10 (NF-YC10) as a GAPC-binding protein. The effects of GAPC overexpression are abolished when NF-YC10 is deficient, the heat-induced nuclear accumulation of GAPC is suppressed, or the GAPC-NF-YC10 interaction is disrupted. GAPC overexpression also enhances the binding ability of NF-YC10 to its target promoter. The results reveal a cellular and molecular mechanism for the nuclear moonlighting of a glycolytic enzyme in plant response to environmental changes.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Arabidopsis/fisiologia , Gliceraldeído-3-Fosfato Desidrogenases/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Fator de Ligação a CCAAT/genética , Fator de Ligação a CCAAT/metabolismo , Núcleo Celular/genética , Núcleo Celular/metabolismo , Núcleo Celular/fisiologia , Citosol/metabolismo , Citosol/fisiologia , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Gliceraldeído-3-Fosfato Desidrogenases/genética , Resposta ao Choque Térmico/genética , Resposta ao Choque Térmico/fisiologia , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/metabolismo , Plantas Geneticamente Modificadas/fisiologia , Plântula/genética , Plântula/metabolismo , Plântula/fisiologia , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
3.
PLoS One ; 15(7): e0235896, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32730265

RESUMO

Mature sorghum herbage is known to contain several water-soluble secondary metabolites (allelochemicals). In this study, we investigated quantitative trait loci (QTLs) associated with allelochemical characteristics in sorghum using linkage mapping and linkage disequilibrium (LD)-based association mapping. A sorghum diversity research set (SDRS) of 107 accessions was used in LD mapping whereas, F2:3 lines derived from a cross between Japanese and African landraces were used in linkage mapping. The QTLs were further confirmed by positional (targeted) association mapping with Q+K model. The inhibitory effect of water-soluble extracts (WSE) was tested on germination and root length of lettuce seedlings in four concentrations (25%, 50%, 75% and 100%). A Significant range of variations was observed among genotypes in both types of mapping populations (P < 0.05). A total of 181 simple sequence repeats (SSRs) derived from antecedently reported map have been used for genotyping of SDRS. A genetic linkage map of 151 sorghum SSR markers was also developed on 134 F2 individuals. The total map length was 1359.3 cM, with an average distance of 8.2 cM between adjacent markers. LD mapping identified three QTLs for inhibition effect on germination and seven QTLs for root length of lettuce seedlings. Whereas, a total of six QTLs for inhibition of germination and ten QTLs for root length were detected in linkage mapping approach. The percent phenotypic variation explained by individual QTL ranged from 6.9% to 27.3% in SDRS and 9.9% to 35.6% in F2:3 lines. Regional association analysis identified four QTLs, three of them are common in other methods too. No QTL was identified in the region where major gene for sorgoleone (SOR1) has been cloned previously on chromosome 5.


Assuntos
Locos de Características Quantitativas , Sorghum/genética , Ligação Genética , Germinação/genética , Repetições de Microssatélites , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Plântula/genética , Plântula/crescimento & desenvolvimento , Sorghum/crescimento & desenvolvimento , Sorghum/fisiologia
4.
PLoS One ; 15(7): e0235565, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32614894

RESUMO

Powdery mildew is an important foliar disease of barley (Hordeum vulgare L.) caused by the biotrophic fungus Blumeria graminis f. sp. hordei (Bgh). The understanding of the resistance mechanism is essential for future resistance breeding. In particular, the identification of race-nonspecific resistance genes is important because of their regarded durability and broad-spectrum activity. We assessed the severity of powdery mildew infection on detached seedling leaves of 267 barley accessions using two poly-virulent isolates and performed a genome-wide association study exploiting 201 of these accessions. Two-hundred and fourteen markers, located on six barley chromosomes are associated with potential race-nonspecific Bgh resistance or susceptibility. Initial steps for the functional validation of four promising candidates were performed based on phenotype and transcription data. Specific candidate alleles were analyzed via transient gene silencing as well as transient overexpression. Microarray data of the four selected candidates indicate differential regulation of the transcription in response to Bgh infection. Based on our results, all four candidate genes seem to be involved in the responses to powdery mildew attack. In particular, the transient overexpression of specific alleles of two candidate genes, a potential arabinogalactan protein and the barley homolog of Arabidopsis thaliana's Light-Response Bric-a-Brac/-Tramtrack/-Broad Complex/-POxvirus and Zinc finger (AtLRB1) or AtLRB2, were top candidates of novel powdery mildew susceptibility genes.


Assuntos
Ascomicetos/genética , Hordeum/genética , Interações Hospedeiro-Patógeno/genética , Doenças das Plantas/microbiologia , Alelos , Ascomicetos/isolamento & purificação , Ascomicetos/patogenicidade , Análise por Conglomerados , Regulação da Expressão Gênica de Plantas , Predisposição Genética para Doença , Estudo de Associação Genômica Ampla , Genótipo , Desequilíbrio de Ligação , Mucoproteínas/genética , Mucoproteínas/metabolismo , Fenótipo , Doenças das Plantas/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plântula/genética , Virulência/genética
5.
Physiol Plant ; 170(2): 227-247, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32492180

RESUMO

Seedling pre-emergence is a critical phase of development for successful crop establishment because of its susceptibility to environmental conditions. In a context of reduced use of inorganic fertilizers, the genetic bases of the response of seedlings to nitrate supply received little attention. This issue is important even in legumes where nitrate absorption starts early after germination, before nodule development. Natural variation of traits characterizing seedling growth in the absence or presence of nitrate was investigated in a core collection of 192 accessions of Medicago truncatula. Plasticity indexes to the absence of nitrate were calculated. The genetic determinism of the traits was dissected by genome-wide association study (GWAS). The absence of nitrate affected seed biomass mobilization and root/shoot length ratio. However, the large range of genetic variability revealed different seedling performances within natural diversity. A principal component analysis (PCA) carried out with plasticity indexes highlighted four physiotypes of accessions differing in relationships between seedling elongation and seed biomass partitioning traits in response to the absence of nitrate. Finally, GWAS revealed 45 associations with single or combined traits corresponding to coordinates of accessions on PCA, as well as two clusters of genes encoding sugar transporters and glutathione transferases surrounding loci associated with seedling elongation traits.


Assuntos
Medicago truncatula/genética , Plântula/genética , Estudo de Associação Genômica Ampla , Germinação , Sementes
6.
PLoS One ; 15(6): e0234510, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32555619

RESUMO

Seeds stored in controlled conditions in gene banks, faster or slower lose their viability. The effects of seed moisture content levels (ca. 5, 8, 11%) combined with storage temperatures (-3°, -18°, -196°C) were investigated in terms of the description of seeds defined as orthodox under oxidative stress after seed storage, during germination, and initial seedling growth. Hydrogen peroxide (H2O2), thiobarbituric acid reactive substances (TBARS) and ascorbate (Asc) were analyzed in relation to seed germinability and seedlings emergence in three species: Malus sylvestris L., Prunus avium L. and Prunus padus L. The effect of seed storage conditions on H2O2 levels appeared in germinated seeds after the third year of storage in each species. The H2O2 levels were negatively correlated with the germination and seedling emergence of P. avium seeds after three years of storage under all examined combinations. The emergence of P. padus seedlings was not linked to any of the stress markers tested. The P. padus seed biochemical traits were least altered by storage conditions, and the seeds produced tolerant seedlings of relatively high levels of H2O2 and TBARS. To cope with different H2O2 levels, TBARS levels, and Asc levels in seeds of three species varying storage conditions different molecular responses, i.e. repairing mechanisms, were applied during stratification to compensate for the storage conditions and, as a result, seeds remained viable and seedlings were successfully established.


Assuntos
Malus/metabolismo , Prunus avium/metabolismo , Plântula/genética , Germinação/efeitos dos fármacos , Peróxido de Hidrogênio/metabolismo , Malus/crescimento & desenvolvimento , Oxirredução/efeitos dos fármacos , Prunus avium/crescimento & desenvolvimento , Plântula/crescimento & desenvolvimento , Sementes/genética , Sementes/crescimento & desenvolvimento , Sementes/metabolismo , Temperatura
7.
Plant Mol Biol ; 104(1-2): 39-53, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32564178

RESUMO

Plants are exposed to various environmental cues that lead to reactive oxygen species (ROS) accumulation. ROS production and detoxification are tightly regulated to maintain balance. Although studies of glucose (Glc) are always accompanied by ROS in animals, the role of Glc in respect of ROS in plants is unclear. We isolated gsm2 (Glc-hypersensitive mutant 2), a mutant with a notably chlorotic-cotyledon phenotype. The chloroplast-localized GSM2 was characterized as a transaldolase in the pentose phosphate pathway. With 3% Glc treatment, fewer or no thylakoids were observed in gsm2 cotyledon chloroplasts than in wild-type cotyledon chloroplasts, suggesting that GSM2 is required for chloroplast protection under stress. gsm2 also showed evaluated accumulation of ROS with 3% Glc treatment and was more sensitive to exogenous H2O2 than the wild type. Gene expression analysis of the antioxidant enzymes in gsm2 revealed that chloroplast damage to gsm2 cotyledons results from the accumulation of excessive ROS in response to Glc. Moreover, the addition of diphenyleneiodonium chloride or phenylalanine can rescue Glc-induced chlorosis in gsm2 cotyledons. This work suggests that GSM2 functions to maintain ROS balance in response to Glc during early seedling growth and sheds light on the relationship between Glc, the pentose phosphate pathway and ROS.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Homeostase , RNA Helicases/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Transaldolase/metabolismo , Ácido Abscísico/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Clorofila/metabolismo , Cloroplastos/genética , Cloroplastos/metabolismo , Cotilédone/metabolismo , Regulação da Expressão Gênica de Plantas , Germinação , Glucuronidase/metabolismo , Peróxido de Hidrogênio/metabolismo , Via de Pentose Fosfato/genética , Via de Pentose Fosfato/fisiologia , Fenótipo , RNA Helicases/genética , RNA de Plantas/genética , RNA de Plantas/isolamento & purificação , Plântula/genética , Plântula/metabolismo , Transaldolase/genética
8.
Plant Mol Biol ; 103(4-5): 545-560, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32504260

RESUMO

KEY MESSAGE: OsGTγ-2, a trihelix transcription factor, is a positive regulator of rice responses to salt stress by regulating the expression of ion transporters. Salinity stress seriously restricts rice growth and yield. Trihelix transcription factors (GT factors) specifically bind to GT elements and play a diverse role in plant morphological development and responses to abiotic stresses. In our previous study, we found that the GT-1 element (GAAAAA) is a key element in the salinity-induced OsRAV2 promoter. Here, we identified a rice OsGTγ family member, OsGTγ-2, which directly interacted with the GT-1 element in the OsRAV2 promoter. OsGTγ-2 specifically targeted the nucleus, was mainly expressed in roots, sheathes, stems and seeds, and was induced by salinity, osmotic and oxidative stresses and abscisic acid (ABA). The seed germination rate, seedling growth and survival rate under salinity stress was improved in OsGTγ-2 overexpressing lines (PZmUbi::OsGTγ-2). In contrast, CRISPR/Cas9-mediated OsGTγ-2 knockout lines (osgtγ-2) showed salt-hypersensitive phenotypes. In response to salt stress, different Na+ and K+ acclamation patterns were observed in PZmUbi::OsGTγ-2 lines and osgtγ-2 plants were observed. The molecular mechanism of OsGTγ-2 in rice salt adaptation was also investigated. Several major genes responsible for ion transporting, such as the OsHKT2; 1, OsHKT1; 3 and OsNHX1 were transcriptionally regulated by OsGTγ-2. A subsequent yeast one-hybrid assay and EMSA indicated that OsGTγ-2 directly interacted with the promoters of OsHKT2; 1, OsNHX1 and OsHKT1; 3. Taken together, these results suggest that OsGTγ-2 is an important positive regulator involved in rice responses to salt stress and suggest a potential role for OsGTγ-2 in regulating salinity adaptation in rice.


Assuntos
Aclimatação/fisiologia , Proteínas de Ligação a DNA/metabolismo , Oryza/fisiologia , Estresse Salino/fisiologia , Tolerância ao Sal/genética , Fatores de Transcrição/metabolismo , Ácido Abscísico/metabolismo , Aclimatação/genética , Adaptação Fisiológica , Sistemas CRISPR-Cas , Proteínas de Transporte de Cátions/metabolismo , Proteínas de Ligação a DNA/genética , Regulação da Expressão Gênica de Plantas , Oryza/genética , Oryza/crescimento & desenvolvimento , Desenvolvimento Vegetal , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/metabolismo , Salinidade , Plântula/genética , Sementes/metabolismo , Sódio/metabolismo , Trocadores de Sódio-Hidrogênio/metabolismo , Estresse Fisiológico/genética , Simportadores/metabolismo , Fatores de Transcrição/genética
9.
Gene ; 755: 144905, 2020 Sep 10.
Artigo em Inglês | MEDLINE | ID: mdl-32540372

RESUMO

DNA methylation, an important epigenetic modification, regulates a wide range of biological processes. Previous MSAP results showed that the occurrence of PaWB related to changes of DNA methylation level; however, the relationship between DNA methylation and gene expression remains obscure in paulownia. Therefore, in the present study, we applied WGBS and RNA-seq techniques to investigate the DNA methylation and gene expression changes between healthy Paulownia fortunei seedlings and the phytoplasma-infected ones. A map of methylated cytosines at the single base pair resolution of paulownia was constructed. Compared to the healthy seedlings, the DNA methylation level increased after phytoplasma infection, and the change of mCHH was the main methylation pattern. DMR analysis showed that 422,662 DMRs in the genome were identified, in which, 27,871 DMR-associated genes were differentially expressed. Finally, 436 genes with significant differences in their methylation levels and mRNA expression profiles were identified through integrated analysis of the DNA methylomic and transcriptomic. KEGG pathway analysis revealed that these genes are mainly involved in plant hormone signal transduction, carbon metabolism, and starch and sucrose metabolism pathways. Two of DMR-associated genes were verified by BS- PCR. Finally, we selected TRP 1 and R2R3-MYB protein were closely related to the occurrence of PaWB. Our findings provide valuable insight into the mechanism of PaWB at the epigenetic level.


Assuntos
Metilação de DNA/genética , Lamiales/genética , Phytoplasma/genética , China , Epigênese Genética/genética , Regulação da Expressão Gênica de Plantas/genética , Genoma de Planta/genética , Interações Hospedeiro-Patógeno/genética , Infecções/genética , Lamiales/microbiologia , Magnoliopsida/genética , MicroRNAs/genética , Phytoplasma/patogenicidade , Doenças das Plantas/genética , Plântula/genética , Transcriptoma/genética
10.
Gene ; 753: 144802, 2020 Aug 30.
Artigo em Inglês | MEDLINE | ID: mdl-32454178

RESUMO

Synchronous and timely regulation of multiple genes results in an effective defense response that decides the fate of the host when challenged with pathogens or unexpected changes in environmental conditions. One such gene, which is downregulated in response to multiple bacterial pathogens, is a putative nonspecific lipid transfer protein (nsLTP) of unknown function that we have named DISEASE RELATED NONSPECIFIC LIPID TRANSFER PROTEIN 1 (DRN1). We show that upon pathogen challenge, DRN1 is strongly downregulated, while a putative DRN1-targeting novel microRNA (miRNA) named DRN1 Regulating miRNA (DmiR) is reciprocally upregulated. Furthermore, we provide evidence that DRN1 is required for defense against bacterial and fungal pathogens as well as for normal seedling growth under salinity stress. Although nsLTP family members from different plant species are known to be a significant source of food allergens and are often associated with antimicrobial properties, our knowledge on the biological functions and regulation of this gene family is limited. Our current work not only sheds light on the mechanism of regulation but also helps in the functional characterization of DRN1, a putative nsLTP family member of hitherto unknown function.


Assuntos
Arabidopsis/genética , Proteínas de Transferência de Fosfolipídeos/genética , Estresse Salino/genética , Ácido Abscísico/metabolismo , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Resistência à Doença/genética , Secas , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Proteínas de Transferência de Fosfolipídeos/metabolismo , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Patologia Vegetal , Plantas Geneticamente Modificadas , Salinidade , Tolerância ao Sal/genética , Plântula/genética , Estresse Fisiológico/genética
11.
Ecotoxicol Environ Saf ; 200: 110779, 2020 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-32460045

RESUMO

Melon is of great value in food, medicine and industry. In recent years, the continuous cropping obstacles of melon is increasingly prominent, which seriously affects the cultivation. Autotoxicity is the key factor for the obstacles. Root is the first line against autotoxicity and main organs for autotoxins secretion. Some physiological responses and differentially expressed genes (DEGs) related to autotoxicity are only limited to root system. Considering the lack of relevant research, physiological researches combined with transcriptome sequencing of melon seedling after autotoxicity stress mediated by root exudates (RE) was performed to help characterize the response mechanism to autotoxicity in melon roots. The results showed that autotoxicity inhibited root morphogenesis of melon seedlings, induced the excessive accumulation of reactive oxygen species (ROS) and lipid peroxidation in roots, and activated most antioxidant enzymes. Compared with the control group, the osmoregulation substance content was always at a high level. DEGs response to autotoxicity in roots were distinguished from that in leaves. Functional annotation of these DEGs suggested that autotoxicity affected biological regulation in a negative manner. DEGs were mainly involved in the synthesis of antioxidants, DNA damage and metabolism, and stress response. These setbacks were associated with the deterioration of root morphogenesis, generation of dwarf and slender roots, and ultimately leading to plant death. The results may provide important information for revealing the response mechanism of root to autotoxicity, and provide theoretical basis for solving the continuous cropping obstacles in melon.


Assuntos
Produção Agrícola/métodos , Cucumis melo/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacos , Raízes de Plantas/efeitos dos fármacos , Poluentes do Solo/toxicidade , Transcriptoma/efeitos dos fármacos , Cucumis melo/genética , Cucumis melo/metabolismo , Perfilação da Expressão Gênica , Peroxidação de Lipídeos/efeitos dos fármacos , Osmorregulação/efeitos dos fármacos , Estresse Oxidativo/genética , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/genética , Folhas de Planta/metabolismo , Raízes de Plantas/genética , Raízes de Plantas/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Plântula/efeitos dos fármacos , Plântula/genética , Plântula/metabolismo
12.
PLoS One ; 15(5): e0233807, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32470009

RESUMO

Benzoxazinoids (BXs) are secondary metabolites with diverse functions, but are primarily involved in protecting plants, mainly from the family Poaceae, against insects and fungal pathogens. Rye is a cereal crop that is highly resistant to biotic stresses. However, its susceptibility to brown rust caused by Puccinia recondita f. sp. secalis (Prs) is still a major problem affecting its commercial production. Additionally, the genetic and metabolic factors related to this disease remain poorly characterized. In this study, we investigated whether and to what extent the brown rust infection and the inoculation procedure affect the contents of specific BXs (HBOA, GDIBOA, DIBOA, GDIMBOA, DIMBOA, and MBOA) and the expression of genes related to BX (ScBx1-5, ScIgl, and Scglu). We revealed that treatments with water and a urediniospore suspension usually downregulate gene expression levels. Moreover, HBOA and DIBOA contents decreased, whereas the contents of the remaining metabolites increased. Specifically, the MBOA content increased more after the mock treatment than after the Prs treatment, whereas the increase in GDIBOA and GDIMBOA levels was usually due to the Prs infection, especially at two of the most critical time-points, 17 and 24 h post-treatment. Therefore, GDIBOA and GDIMBOA are glucosides that are important components of rye defence responses to brown rust. Furthermore, along with MBOA, they protect rye against the stress associated with the inoculation procedure used in this study.


Assuntos
Basidiomycota/fisiologia , Benzoxazinas/metabolismo , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Doenças das Plantas/genética , Doenças das Plantas/microbiologia , Secale/genética , Interações Hospedeiro-Patógeno/genética , Plântula/genética , Plântula/microbiologia , Água
13.
PLoS One ; 15(5): e0228515, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32407318

RESUMO

BACKGROUND: Recently, it was found that 1% Phytagel plates used to conduct Arabidopsis thaliana seedling phenotypic analysis no longer reproduced previously published results. This Phytagel, which is produced in China (Phytagel C), has replace American-made Phytagel (Phytagel), which is no longer commercially available. In this study, we present the impact of Phytagel produced in the United States vs. China on seedling phenotypic analysis. As a part of this study, an alternative gelling agent has been identified that is capable of reproducing previously published seedling morphometrics. RESULTS: Phytagel and Phytagel C were investigated based on their ability to reproduce the subtle phenotype of the sob3-4 esc-8 double mutant. Fluence-rate-response analysis of seedlings grown on 1% Phytagel C plates failed to replicate the sob3-4 esc-8 subtle phenotype seen on 1% Phytagel. Furthermore, root penetrance analysis showed a significant difference between sob3-4 esc-8 seedlings grown on 1% Phytagel and 1% Phytagel C. It was also found that 1% Phytagel C was significantly harder than 1% Phytagel. As a replacement for Phytagel C, Gellan was tested. 1% Gellan was able to reproduce the subtle phenotype of sob3-4 esc-8. Furthermore, there was no significant difference in root penetration of the wild type or sob3-4 esc-8 seedlings between 1% Phytagel and 1% Gellan. This may be due to the significant reduction in hardness in 1% Gellan plates compared to 1% Phytagel plates. Finally, we tested additional concentrations of Gellan and found that seedlings on 0.6% Gellan looked more uniform while also being able to reproduce previously published results. CONCLUSIONS: Phytagel has been the standard gelling agent for several studies involving the characterization of subtle seedling phenotypes. After production was moved to China, Phytagel C was no longer capable of reproducing these previously published results. An alternative gelling agent, Gellan, was able to reproduce previously published seedling phenotypes at both 1% and 0.6% concentrations. The information provided in this manuscript is beneficial to the scientific community as whole, specifically phenomics labs, as it details key problematic differences between gelling agents that should be performing identically (Phytagel and Phytagel C).


Assuntos
Arabidopsis/crescimento & desenvolvimento , Géis/farmacologia , Reprodução/efeitos dos fármacos , Plântula/crescimento & desenvolvimento , Arabidopsis/efeitos dos fármacos , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Proteínas de Ligação a DNA/genética , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Fenômica , Fenótipo , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Reprodução/genética , Plântula/efeitos dos fármacos , Plântula/genética
14.
PLoS One ; 15(5): e0232756, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32407323

RESUMO

Mitogen-activated protein kinase (MAPK) is a form of serine/threonine protein kinase that activated by extracellular stimulation acting through the MAPK cascade (MAPKKK-MAPKK-MAPK). The MAPK cascade gene family, an important family of protein kinases, plays a vital role in responding to various stresses and hormone signal transduction processes in plants. In this study, we identified 14 CmMAPKs, 6 CmMAPKKs and 64 CmMAPKKKs in melon genome. Based on structural characteristics and a comparison of phylogenetic relationships of MAPK gene families from Arabidopsis, cucumber and watermelon, CmMAPKs and CmMAPKKs were categorized into 4 groups, and CmMAPKKKs were categorized into 3 groups. Furthermore, chromosome location revealed an unevenly distribution on chromosomes of MAPK cascade genes in melon, respectively. Eventually, qRT-PCR analysis showed that all 14 CmMAPKs had different expression patterns under drought, salt, salicylic acid (SA), methyl jasmonate (MeJA), red light (RL), and Podosphaera xanthii (P. xanthii) treatments. Overall, the expression levels of CmMAPK3 and CmMAPK7 under different treatments were higher than those in control. Our study provides an important basis for future functional verification of MAPK genes in regulating responses to stress and signal substance in melon.


Assuntos
Cucumis melo/enzimologia , Cucumis melo/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Estudo de Associação Genômica Ampla , Sistema de Sinalização das MAP Quinases/genética , Acetatos/farmacologia , Motivos de Aminoácidos , Sequência de Aminoácidos , Cromossomos de Plantas/genética , Cucumis melo/efeitos dos fármacos , Ciclopentanos/farmacologia , Secas , Éxons/genética , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Genes de Plantas , Íntrons/genética , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Proteínas Quinases Ativadas por Mitógeno/química , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Oxilipinas/farmacologia , Filogenia , Folhas de Planta/efeitos dos fármacos , Folhas de Planta/genética , Domínios Proteicos , Ácido Salicílico/farmacologia , Plântula/efeitos dos fármacos , Plântula/enzimologia , Plântula/genética , Cloreto de Sódio/farmacologia , Estresse Fisiológico/efeitos dos fármacos , Estresse Fisiológico/genética
15.
Gene ; 750: 144725, 2020 Aug 05.
Artigo em Inglês | MEDLINE | ID: mdl-32360839

RESUMO

The small auxin-up RNA (SAUR) family plays a vital role in the regulation of plant growth and development. We identified 80 MdSAUR genes in this study. Phylogenetic analysis indicated that the SAUR proteins from Arabidopsis, rice, and apple were divided into six groups. Of the 80 MdSAURs, 71 were randomly distributed along the 17 chromosomes, while the remaining genes were located along unassigned scafoolds. Among them, a comprehensive overview of SAUR gene family is presented, including gene structures, chromosome locations, duplication and selection pressure analyses, synteny and promoter analyses, and protein interaction. The expression profiles based on microarray data found that 80 genes showed increased expression levels in at least one tissue including seed, seedling, root, stem, leaf, flower, fruit 100daa, and harvested fruit. MdSAUR7 possibly regulate the development of flower organs, and MdSAUR15, MdSAUR24, and MdSAUR80 promote the growth of fruits by regulating cell division. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis indicated the expression levels of 79 MdSAUR genes in leaves under exogenous IAA treatment. MdSAUR4, MdSAUR22, MdSAUR37, MdSAUR38, MdSAUR49, and MdSAUR54 were up-regulated after IAA treatment compared with the control, indicating that they may play specific roles in the IAA signaling transduction pathway. This work provided a foundation for further investigations for the functional analyses of SAURs in apple.


Assuntos
Genes de Plantas/genética , Ácidos Indolacéticos/metabolismo , Malus/genética , Arabidopsis/genética , Frutas/metabolismo , Perfilação da Expressão Gênica/métodos , Regulação da Expressão Gênica de Plantas/genética , Família Multigênica/genética , Filogenia , Reguladores de Crescimento de Planta/metabolismo , Folhas de Planta/metabolismo , Proteínas de Plantas/genética , RNA/genética , Plântula/genética
16.
Plant Mol Biol ; 103(4-5): 561-580, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32405802

RESUMO

KEY MESSAGE: CmHKT1;1 selectively exports Na+ from plant cells. Upon NaCl stress, its expression increased in a salt-tolerant melon cultivar. Overexpression of CmHKT1;1 increased transgenic Arabidopsis salt tolerance through improved K+/Na+ balance. High-affinity K+ transporters (HKTs) are thought to be involved in reducing Na+ in plant shoots under salt stress and modulating salt tolerance, but their function in a moderately salt-tolerant species of melon (Cucumis melo L.) remains unclear. In this study, a Na+ transporter gene, CmHKT1;1 (GenBank accession number: MK986658), was isolated from melons based on genome data. The transcript of CmHKT1;1 was relatively more abundant in roots than in stems or leaves from melon seedlings. The tobacco transient expression system showed that CmHKT1;1 was plasma-membrane localized. Upon salt stress, CmHKT1;1 expression was more strongly upregulated in a salt-tolerant melon cultivar, 'Bingxuecui' (BXC) compared with a salt-sensitive cultivar, 'Yulu' (YL). Electrophysiological evidence demonstrated that CmHKT1;1 only transported Na+, rather than K+, when expressed in Xenopus laevis oocytes. Overexpression of CmHKT1;1 increased salt sensitivity in Saccharomyces cerevisiae and salt tolerance in Arabidopsis thaliana. Under NaCl treatments, transgenic Arabidopsis plants accumulated significantly lower concentrations of Na+ in shoots than wild type plants and showed a better K+/Na+ balance, leading to better Fv/Fm, root length, biomass, and enhanced plant growth. The CmHKT1;1 gene may serve as a useful candidate for improving crop salt tolerance.


Assuntos
Arabidopsis/metabolismo , Cucumis melo/metabolismo , Proteínas de Membrana Transportadoras/metabolismo , Plantas Geneticamente Modificadas/metabolismo , Potássio/metabolismo , Sódio/metabolismo , Arabidopsis/genética , Proteínas de Transporte de Cátions/genética , Proteínas de Transporte de Cátions/metabolismo , Clorofila/análise , Clonagem Molecular , Cucumis melo/genética , Regulação da Expressão Gênica de Plantas , Proteínas de Membrana Transportadoras/genética , Folhas de Planta/metabolismo , Proteínas de Plantas/genética , Raízes de Plantas/metabolismo , Brotos de Planta/genética , Saccharomyces cerevisiae/genética , Tolerância ao Sal , Plântula/genética , Plântula/metabolismo , Alinhamento de Sequência , Análise de Sequência de Proteína , Cloreto de Sódio/metabolismo , Estresse Fisiológico/genética , Estresse Fisiológico/fisiologia , Simportadores/genética , Simportadores/metabolismo , Tabaco/genética , Tabaco/metabolismo
17.
Mol Genet Genomics ; 295(5): 1163-1172, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32472284

RESUMO

With global warming as a major environment concern over the coming years, heat tolerance is an important trait for forest tree survival during the predicted future warmer weather conditions. Cryptomeria japonica is a coniferous species widely distributed throughout Japan, and thus, can adapt to a wide range of air temperatures. To elucidate genes involved in heat response in Cryptomeria japonica, transcriptome analysis was conducted for seedlings under heat shock conditions. To test whether heat acclimation affects levels of gene expression, half of the seedlings were pretreated with moderately high temperatures prior to heat shock. De novo assembly of the transcriptome generated 107,924 unigenes and the analysis of differentially expressed genes was conducted using these unigenes. A total of 5217 differentially expressed genes were identified. Most genes upregulated by heat shock, regardless of pre-heat treatment, were conserved to heat response genes of angiosperm species, such as heat shock factors (Hsf) and heat shock proteins (Hsp). Pre-heating of seedlings affected expression levels of several Hsfs and their induction was lower in pre-heated seedlings than in seedlings without pre-heat treatment. This suggests a conserved role of Hsfs in heat response and heat acclimation in seed plants. On the other hand, many unknown genes were upregulated in only seedlings without pre-heat treatment after heat exposure. Notably, expression of gypsy/Ty3 type retrotransposons was dramatically induced. These findings provide valuable information to develop a better understanding of the molecular mechanisms of heat response and acclimation in C. japonica.


Assuntos
Cryptomeria/fisiologia , Perfilação da Expressão Gênica/métodos , Proteínas de Plantas/genética , Cryptomeria/genética , Regulação da Expressão Gênica de Plantas , Aquecimento Global , Fatores de Transcrição de Choque Térmico/genética , Proteínas de Choque Térmico/genética , Resposta ao Choque Térmico , Plântula/genética , Plântula/fisiologia , Análise de Sequência de RNA
18.
PLoS One ; 15(5): e0233616, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32470066

RESUMO

Salt stress is a common abiotic stress that limits the growth, development and yield of maize (Zea mays L.). To better understand the response of maize to salt stress and the mechanism by which exogenous glycine betaine (GB) alleviates the damaging effects of salt stress, the morphology, physiological and biochemical indexes, and root transcriptome expression profiles of seedlings of salt-sensitive inbred line P138 and salt-tolerant inbred line 8723 were compared under salt stress and GB-alleviated salt stress conditions. The results showed that under salt stress the growth of P138 was significantly inhibited and the vivo ion balance was disrupted, whereas 8723 could prevent salt injury by maintaining a high ratio of K+ to Na+. The addition of a suitable concentration of GB could effectively alleviate the damage caused by salt stress, and the mitigating effect on salt-sensitive inbred line P138 was more obvious than that on 8723. Transcriptome analysis revealed that 219 differentially expressed genes (DEGs) were up-regulated and 153 DEGs were down-regulated in both P138 and 8723 under NaCl treatment, and that 487 DEGs were up-regulated and 942 DEGs were down-regulated in both P138 and 8723 under salt plus exogenous GB treatment. In 8723 the response to salt stress is mainly achieved through stabilizing ion homeostasis, strong signal transduction activation, increasing reactive oxygen scavenging. GB alleviates salt stress in maize mainly by inducing gene expression changes to enhance the ion balance, secondary metabolic level, reactive oxygen scavenging mechanism, signal transduction activation. In addition, the transcription factors involved in the regulation of salt stress response and exogenous GB mitigation mainly belong to the MYB, MYB-related, AP2-EREBP, bHLH, and NAC families. We verified 10 selected up-regulated DEGs by quantitative real-time polymerase chain reaction (qRT-PCR), and the expression results were basically consistent with the transcriptome expression profiles. Our results from this study may provide the theoretical basis for determining maize salt tolerance mechanisms and the mechanism by which GB regulates salt tolerance.


Assuntos
Betaína/metabolismo , Tolerância ao Sal , Plântula/genética , Transcriptoma , Zea mays/genética , Regulação da Expressão Gênica de Plantas , Potássio/metabolismo , Plântula/fisiologia , Sódio/metabolismo , Zea mays/fisiologia
19.
PLoS One ; 15(4): e0230958, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32294092

RESUMO

Soil salinization is a serious problem for cultivation of rice, as among cereals rice is the most salt sensitive crop, and more than 40% of the total agricultural land amounting to approximately 80 million ha the world over is salt affected. Salinity affects a plant in a varieties of ways, including ion toxicity, osmotic stress and oxidative damage. Since miRNAs occupy the top place in biochemical events determining a trait, understanding their role in salt tolerance is highly desirable, which may allow introduction of the trait in the rice cultivars of choice through biotechnological interventions. High throughput sequencing of sRNAs in the root and shoot tissues of the seedlings of the control and NaCl treated Pokkali, a salt-tolerant rice variety, identified 75 conserved miRNAs and mapped 200 sRNAs to the rice genome as novel miRNAs. Expression of nine novel miRNAs and two conserved miRNAs were confirmed by Northern blotting. Several of both conserved and novel miRNAs that expressed differentially in root and/or shoot tissues targeted transcription factors like AP2/EREBP domain protein, ARF, NAC, MYB, NF-YA, HD-Zip III, TCP and SBP reported to be involved in salt tolerance or in abiotic stress tolerance in general. Most of the novel miRNAs expressed in the salt tolerant wild rice Oryza coarctata, suggesting conservation of miRNAs in taxonomically related species. One of the novel miRNAs, osa-miR12477, also targeted L-ascorbate oxidase (LAO), indicating build-up of oxidative stress in the plant upon salt treatment, which was confirmed by DAB staining. Thus, salt tolerance might involve miRNA-mediated regulation of 1) cellular abundance of the hormone signaling components like EREBP and ARF, 2) synthesis of abiotic stress related transcription factors, and 3) antioxidative component like LAO for mitigation of oxidative damage. The study clearly indicated importance of osa-miR12477 regulated expression of LAO in salt tolerance in the plant.


Assuntos
MicroRNAs/genética , Oryza/genética , Tolerância ao Sal/genética , Northern Blotting/métodos , Regulação da Expressão Gênica de Plantas/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Estresse Oxidativo/genética , Salinidade , Plântula/genética , Estresse Fisiológico/genética , Fatores de Transcrição/genética
20.
PLoS One ; 15(4): e0230820, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32298285

RESUMO

The high toxicity of cadmium (Cd) and its ready uptake by plants has become a major agricultural problem. To investigate the genetic architecture and genetic regulation of Cd tolerance in barley, we conducted quantitative trait loci (QTL) analysis in the phenotypically polymorphic Oregon Wolfe Barley (OWB) mapping population, derived from a cross between Rec and Dom parental genotypes. Through evaluating the Cd tolerance of 87 available doubled haploid lines of the OWB mapping population at the seedling stage, one minor and one major QTL were detected on chromosomes 2H and 6H, respectively. For chlorosis and necrosis traits, the major QTL explained 47.24% and 38.59% of the phenotypic variance, respectively. RNA-Seq analysis of the parental seedlings under Cd treatment revealed 542 differentially expressed genes between Cd-tolerant Rec and Cd-susceptible Dom genotypes. By analyzing sequence variations in transcribed sequences of the parental genotypes, 155,654 SNPs and 1,525 InDels were identified between the two contrasting genotypes and may contribute to Cd tolerance. Finally, by integrating the data from the identified QTLs and RNA-Seq analysis, 16 Cd tolerance-related candidate genes were detected, nine of which were metal ion transporters. These results provide promising candidate genes for further gene cloning and improving Cd tolerance in barley.


Assuntos
Cádmio/toxicidade , Tolerância a Medicamentos/genética , Hordeum/efeitos dos fármacos , Hordeum/genética , Locos de Características Quantitativas/genética , Mapeamento Cromossômico/métodos , Cromossomos de Plantas/efeitos dos fármacos , Cromossomos de Plantas/genética , Haploidia , Oregon , Fenótipo , Polimorfismo de Nucleotídeo Único/genética , RNA-Seq/métodos , Plântula/efeitos dos fármacos , Plântula/genética , Estresse Fisiológico/genética , Sequenciamento Completo do Exoma/métodos
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