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1.
Pestic Biochem Physiol ; 158: 149-155, 2019 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-31378351

RESUMO

Induced resistance is an effective measure for controlling plant diseases by utilizing the natural defense of the host and meets the strategic needs of pesticide application and safety for agricultural products worldwide. Ganoderma lucidum polysaccharide (GLP), which is the main active molecule of G. lucidum, has been widely used in functional food and clinical medicine. However, there are few reports of the use of GLP for the prevention and control of plant diseases. The purpose of this study is to explore the effect of GLP and its mechanism of inducing plant resistance. In this study, we found that GLP spray and irrigation root treatments can promote growth in cotton. After soaking in GLP, theseedling height and cotton fusarium wilt resistance both increased to some extent, effects that were dose dependent. After treatment of cotton with GLP, the activities of peroxidase (POD), superoxide dismutase (SOD) and polyphenol oxidase (PPO) in leaves increased significantly, whereas the content of malondialdehyde (MDA) decreased. In addition, QRT-PCR results showed significantly increased relative expression of genes related to the jasmonic acid pathway in cotton. Therefore, we speculate that GLP can induce plant resistance by stimulating the jasmonate pathway.


Assuntos
Fusarium/efeitos dos fármacos , Gossypium/microbiologia , Polissacarídeos/farmacologia , Reishi/química , Antioxidantes/metabolismo , Gossypium/metabolismo , Peroxidação de Lipídeos/efeitos dos fármacos , Malondialdeído/metabolismo , Doenças das Plantas/microbiologia , Polissacarídeos/química , Superóxido Dismutase/metabolismo
2.
BMC Plant Biol ; 19(1): 350, 2019 Aug 13.
Artigo em Inglês | MEDLINE | ID: mdl-31409298

RESUMO

BACKGROUND: The pentatricopeptide repeat (PPR) gene family, which contains multiple 35-amino acid repeats, constitutes one of the largest gene families in plants. PPR proteins function in organelles to target specific transcripts and are involved in plant development and growth. However, the function of PPR proteins in cotton is still unknown. RESULTS: In this study, we characterized a PPR gene YELLOW-GREEN LEAF (GhYGL1d) that is required for cotton plastid development. The GhYGL1d gene has a DYW domain in C-terminal and is highly express in leaves, localized to the chloroplast fractions. GhYGL1d share high amino acid-sequence homology with AtECB2. In atecb2 mutant, overexpression of GhYGL1d rescued the seedling lethal phenotype and restored the editing of accD and ndhF transcripts. Silencing of GhYGL1d led to the reduction of chlorophyll and phenotypically yellow-green leaves in cotton. Compared with wild type, GhYGL1d-silenced cotton showed significant deformations of thylakoid structures. Furthermore, the transcription levels of plastid-encoded polymerase (PEP) and nuclear-encoded polymerase (NEP) dependent genes were decreased in GhYGL1d-silenced cotton. CONCLUSIONS: Our data indicate that GhYGL1d not only contributes to the editing of accD and ndhF genes, but also affects the expression of NEP- and PEP-dependent genes to regulate the development of thylakoids, and therefore regulates leaf variegation in cotton.


Assuntos
Cloroplastos/genética , Gossypium/genética , Proteínas de Plantas/fisiologia , Cloroplastos/metabolismo , Cloroplastos/fisiologia , Gossypium/anatomia & histologia , Gossypium/metabolismo , Folhas de Planta/anatomia & histologia , Folhas de Planta/genética , Folhas de Planta/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo
3.
Plant Sci ; 286: 108-117, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31300136

RESUMO

It has been reported that drought stress (DS) reduces cotton yield by negatively affecting reproductive activities. Some studies have investigated the effects of DS on pollen physiology and biochemistry, but studies exploring the impact of drought on pistil biochemistry and its relationship with pollen tube growth rates in vivo are scarce. In order to investigate these objectives, a greenhouse study was conducted with a drought sensitive cotton cultivar, Yuzaomian 9110. Two water treatments were imposed at flowering stage, 1. control, where plants were irrigated with optimum quantity of water and 2. DS treatment, where plants were irrigated with 50% of the optimum quantity of water. Results indicated that stored starch content at the early stage of pollen tube growth (12:00 h) was 31.6% lower in drought-stressed pistils than control pistils, and it was highly correlated with pollen tube growth rate. The decline in starch accumulation of drought-stressed pistils could be attributed to the impeded transport of photosynthetic carbon assimilates. Moreover, decreased ADP-glucose pyrophosphorylase and soluble starch synthase activities also resulted in curtailing starch accumulation in drought-stressed pistils. Furthermore, pistil sucrose concentration was significantly higher in droughted plants relative to control plants at 12:00 and 18:00 h (during the rapid growth period), which was due to lower activities of sucrose synthase and acid invertase, and the down-regulated expressions of sucrose synthase genes, GhSusA, GhSusB and GhSusD, and acid invertase genes, GhINV1 and GhINV2, in drought-stressed pistils, limiting as a result the hydrolysis of sucrose into hexose. Drought-stressed pistils sampled at 18:00 h had lower α-amylase activity compared to control pistils, resulting in decreased starch decomposition, which, in conjunction with the decreased hydrolysis of sucrose, led to lower glucose and fructose contents in drought-stressed pistils at 18:00 h. Finally, lower pyruvate level in drought-stressed pistils could not produce enough acetyl-CoA in the tricarboxylic acid cycle to yield sufficient energy (ATP) for pollen tube growth. We conclude that DS disrupts the carbohydrate balance of pistil, reducing as a consequence carbon and energy supply for pollen tube elongation in the style, which will ultimately result in reproductive failure.


Assuntos
Metabolismo dos Carboidratos , Secas , Gossypium/genética , Gossypium/metabolismo , Flores/metabolismo , Tubo Polínico/crescimento & desenvolvimento , Tubo Polínico/metabolismo
4.
Plant Sci ; 286: 7-16, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31300144

RESUMO

Cotton fibers are developed epidermal cells of the seed coat and contain large amounts of cellulose and minor lignin-like components. Lignin in the cell walls of cotton fibers effectively provides mechanical strength and is also presumed to restrict fiber elongation and secondary cell wall synthesis. To analyze the effect of lignin and lignin-like phenolics on fiber quality and the transcriptional regulation of lignin synthesis in cotton fibers, we characterized the function of a bHLH transcription factor, GhbHLH18, during fiber elongation stage. GhbHLH18 knock-down plants have longer and stronger fibers, and accumulate less lignin-like phenolics in mature cotton fibers than control plants. By mining public transcriptomic data for developing fibers, we discovered that GhbHLH18 is coexpressed with most lignin synthesis pathway genes. Furthermore, we showed that GhbHLH18 strongly binds to the E-box in the promoter region of GhPER8 and activates its expression. Transient over expression of GhPER8 protein in tobacco leaves significantly decreased the content of coniferyl alcohol and sinapic alcohol-the substrate respectively for G-lignin and S-lignin biosynthesis. These results suggest that GhbHLH18 is negatively associated with fiber quality by activating peroxidase-mediated lignin metabolism, thus the paper represents an alternative strategy to improve fiber quality.


Assuntos
Fibra de Algodão/análise , Gossypium/genética , Lignina/biossíntese , Proteínas de Plantas/genética , Fatores de Transcrição/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica , Gossypium/metabolismo , Proteínas de Plantas/metabolismo , Fatores de Transcrição/metabolismo
5.
Plant Sci ; 286: 89-97, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31300146

RESUMO

Cottonseed oil is one of the most important renewable resources for edible oil and biodiesel. To detect QTLs associated with cottonseed oil content (OC) and identify candidate genes that regulate oil biosynthesis, a panel of upland cotton germplasm lines was selected among those previously used to perform GWASs in China. In the present study, 13 QTLs associated with 53 common SNPs on 13 chromosomes were identified in multiple environments based on 15,369 polymorphic SNPs using the Cotton63 KSNP array. Of these, the OC QTL qOC-Dt5-1 delineated by nine SNPs occurred in a confidence interval of 4 SSRs with previously reported OC QTLs. A combined transcriptome and qRT-PCR analysis revealed that a peroxidase gene (GhPRXR1) was predominantly expressed during the middle-late stage (20-35 days post anthesis) of ovule development. The overexpression of GhPRXR1 in yeast significantly increased the OC by 20.01-37.25 %. Suppression of GhPRXR1 gene expression in the virus-induced gene-silenced cotton reduced the OC by 18.11%. Our results contribute to identifying more OC QTLs and verifying a candidate gene that influences cottonseed oil biosynthesis.


Assuntos
Estudo de Associação Genômica Ampla , Gossypium/genética , Fosfoenolpiruvato Carboxilase/genética , Óleos Vegetais/química , Proteínas de Plantas/genética , China , Gossypium/química , Gossypium/enzimologia , Gossypium/metabolismo , Fosfoenolpiruvato Carboxilase/metabolismo , Proteínas de Plantas/metabolismo , Locos de Características Quantitativas
6.
Plant Physiol Biochem ; 141: 231-239, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31195253

RESUMO

Anther/pollen development is a highly programmed process in flowering plants. However, the molecular mechanism of regulating anther/pollen development is still largely unclear so far. Here, we report a cotton WRKY transcription factor (GhWRKY22) that functions in anther/pollen development. Quantitative RT-PCR and GUS activity analyses revealed that GhWRKY22 is predominantly expressed in the late developing anther/pollen of cotton. The transgenic Arabidopsis plants expressing GhWRKY22 displayed the male fertility defect with the fewer viable pollen grains. Expression of the genes involved in jasmonate (JA) biosynthesis was up-regulated, whereas expression of the JA-repressors (JAZ1 and JAZ8) was down-regulated in the transgenic Arabidopsis plants expressing GhWRKY22, compared with those in wild type. Yeast one-hybrid and ChIP-qPCR assays demonstrated that GhWRKY22 modulated the expression of JAZ genes by directly binding to their promoters for regulating anther/pollen development. Yeast two-hybrid assay indicated that GhMYB24 could interact with GhJAZ8-A and GhJAZ13-A. Furthermore, expression of AtMYB24, AtPAL2 and AtANS2 was enhanced in the transgenic Arabidopsis plants, owing to GhWRKY22 overexpression. Taking the data together, our results suggest that GhWRKY22 acts as a transcriptional repressor to regulate anther/pollen development possibly by modulating the expression of the JAZ genes.


Assuntos
Gossypium/metabolismo , Pólen/fisiologia , Fatores de Transcrição/metabolismo , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Núcleo Celular/metabolismo , Ciclopentanos/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Hipocótilo/metabolismo , Oxilipinas/metabolismo , Fenótipo , Infertilidade das Plantas/genética , Proteínas de Plantas/metabolismo , Caules de Planta/metabolismo , Plantas Geneticamente Modificadas , Regiões Promotoras Genéticas , Sementes/metabolismo , Ativação Transcricional , Transgenes , Técnicas do Sistema de Duplo-Híbrido
7.
Plant Physiol Biochem ; 141: 370-379, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31212241

RESUMO

Proteins will provide a new perspective and deeper understanding for the research of crop alkali tolerance. The aims of this study were to determine and identify the differentially abundant proteins and adaptive mechanisms to alkali tolerance between young and old leaves of cotton. The 4704 proteins were identified, in which 1490 were significantly changed in young leaves and 563 were changed in old leaves in response to alkali stress. The differentially abundant proteins were classified into 10 functional categories in the young leaves, and only 3 functional categories were involved in the old leaves. In the photoreaction system, the accumulations of differential proteins, especially Psb proteins, were higher in young leaves than in old leaves. Compared with old leaves, the carbon metabolism was enhanced significantly through an increased chlorophyll content and increased expression of key proteins for carbon metabolism in young leaves. Furthermore, alkali stress revealed more complex effects on the nitrogen metabolism in young leaves than that in old leaves. Our results demonstrated that during adaptation of cotton to alkali stress, young and old leaves have distinct mechanisms of molecular metabolism regulation. The metabolic flexibility was more remarkable in young leaves than in old leaves; therefore, the alkali tolerance of young leaves is more efficient. These data will increase our understanding of alkali-tolerant mechanisms in higher plants.


Assuntos
Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Gossypium/metabolismo , Folhas de Planta/metabolismo , Proteínas de Plantas/metabolismo , Adaptação Fisiológica , Álcalis/metabolismo , Apoptose , Clorofila/química , Regulação para Baixo , Concentração de Íons de Hidrogênio , Nitrogênio/metabolismo , Peptídeos/química , Sais/química , Tripsina/química
8.
Plant Biol (Stuttg) ; 21(5): 805-812, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31050863

RESUMO

Identifying the mechanisms of compensation to insect herbivory remains a major challenge in plant biology and evolutionary ecology. Most previous studies have addressed plant compensatory responses to one or two levels of insect herbivory, and the underlying traits mediating such responses remain elusive in many cases. We evaluated responses associated with compensation to multiple intensities of leaf damage (0% control, 10%, 25%, 50%, 75% of leaf area removed) by means of mechanical removal of foliar tissue and application of a caterpillar (Spodoptera exigua) oral secretions in 3-month-old wild cotton plants (Gossypium hirsutum). Four weeks post-treatment, we measured plant growth and multiple traits associated with compensation, namely: changes in above- and belowground, biomass and the concentration of nutrients (nitrogen and phosphorus) and non-structural carbon reserves (starch and soluble sugars) in roots, stems and leaves. We found that wild cotton fully compensated in terms of growth and biomass allocation when leaf damage was low (10%), whereas moderate (25%) to high leaf damage in some cases led to under-compensation. Nonetheless, high levels of leaf removal (50% and 75%) in most cases did not cause further reductions in height and allocation to leaf and stem biomass relative to low and moderate damage. There were significant positive effects of leaf damage on P concentration in leaves and stems, but not roots, as well as a negative effect on soluble sugars in roots. These results indicate that wild cotton fully compensated for a low level of leaf damage but under-compensated under moderate to high leaf damage, but can nonetheless sustain growth despite increasing losses to herbivory. Such responses were possibly mediated by a re-allocation of carbohydrate reserves from roots to shoots.


Assuntos
Gossypium/fisiologia , Herbivoria , Animais , Gossypium/metabolismo , Nitrogênio/metabolismo , Fósforo/metabolismo , Folhas de Planta/metabolismo , Raízes de Plantas/metabolismo , Caules de Planta/metabolismo , Spodoptera
9.
Plant Physiol Biochem ; 141: 114-121, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31146093

RESUMO

The spatial and temporal expression of insecticidal gene Cry1Ac in transgenic Bacillus thuringiensis (Bt) cotton varies with plant organs, plant age, as well as environmental conditions. The research was undertaken to study the impact of nitrogen deficit on Cry1Ac endotoxin concentration in seed in Yangtze River valley region in China in 2015 and 2016. To uncover the underlying mechanism, the nitrogen metabolism process including protein synthesis and degradation was characterized. Based on the results, nitrogen deficit decreased the boll weight, boll volume, boll number per plant, seed Cry1Ac endotoxin concentration, glutamate oxaloacetate transaminase (GOT) and glutamic pyruvic transaminase (GPT) activities, soluble protein concentration, but increased peptidase and protease activities, and free amino acid content in seed. Our results suggested that the decline of seed Cry1Ac endotoxin expression associated with decreased nitrogen was due to the enhanced protein decomposition and reduced protein synthesis, especially the altered activities of GOT and peptidase. There was a significant negative correlation between seed Cry1Ac endotoxin concentration and boll shell Cry1Ac endotoxin content under nitrogen deficiency. Therefore, seed Cry1Ac endotoxin concentration and boll shell Cry1Ac endotoxin concentration should be balanced to guarantee the insecticidal efficiency.


Assuntos
Proteínas de Bactérias/metabolismo , Endotoxinas/metabolismo , Gossypium/genética , Proteínas Hemolisinas/metabolismo , Nitrogênio/química , Bacillus thuringiensis , Proteínas de Bactérias/química , China , Gossypium/metabolismo , Peptídeo Hidrolases , Plantas Geneticamente Modificadas/metabolismo , Sementes/metabolismo
10.
BMC Genomics ; 20(1): 421, 2019 May 28.
Artigo em Inglês | MEDLINE | ID: mdl-31138116

RESUMO

BACKGROUND: Cotton is the most essential textile crop worldwide, and phytohormones are critical for cotton fiber development. One example is the role of auxin in fiber initiation, but we know little molecular basis. MicroRNAs (miRNAs) have a significant function in cotton development; nevertheless their role in fiber initiation remains unclear. Here, exogenous IAA was applied to cotton plant before anthesis. Utilizing small RNA sequencing, the mechanism underlying miRNA-mediated regulation of fiber initiation under exogenous IAA treatment was investigated. RESULTS: With exogenous IAA application, the endogenous IAA and GA contents of IAA treated (IT) ovules were higher than control (CK) ovules at the fiber initiation stage, while endogenous ABA content was lower in IT than CK. Using scanning electron microscopy, we found the fiber number and size were significantly promoted in IT at 0 DPA. Fiber quality analysis showed that fiber length, uniformity, strength, elongation, and micronaire of IT were higher than CK, though not statistically significant, while lint percent was significantly higher in IT. We generated six small RNA libraries using - 3, 0, and 3 DPA ovules of IT and CK, and identified 58 known miRNAs and 83 novel miRNAs together with the target genes. The differential expressed miRNAs number between IT and CK at - 3, 0, 3 DPA was 34, 16 and 24, respectively. Gene ontology and KEGG pathway enrichment analyses for the target genes of the miRNAs expressed in a differential manner showed that they were significantly enriched in 30 terms and 8 pathways. QRT-PCR for those identified miRNAs and the target genes related to phytohormones and fiber development was performed, and results suggested a potential role of these miRNAs in fiber initiation. CONCLUSIONS: The exogenous IAA application affected the relative phytohormone contents in ovule and promoted fiber initiation in cotton. Identification and profiling of miRNAs and their targets at the fiber initiation stage provided insights for miRNAs' regulation function of fiber initiation. These findings not only shed light on the regulatory network of fiber growth but also offer clues for cotton fiber amelioration strategies in cotton.


Assuntos
Gossypium/genética , Ácidos Indolacéticos/farmacologia , MicroRNAs/metabolismo , Reguladores de Crescimento de Planta/farmacologia , Perfilação da Expressão Gênica , Genes de Plantas , Gossypium/efeitos dos fármacos , Gossypium/crescimento & desenvolvimento , Gossypium/metabolismo , Sequenciamento de Nucleotídeos em Larga Escala , Óvulo Vegetal/efeitos dos fármacos , Óvulo Vegetal/genética , Óvulo Vegetal/crescimento & desenvolvimento , Óvulo Vegetal/ultraestrutura , Reguladores de Crescimento de Planta/metabolismo , Análise de Sequência de RNA
11.
Plant Physiol Biochem ; 139: 333-341, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30952085

RESUMO

Chronic elevated temperature and soil-waterlogging events often occur concomitantly in the Yangtze River Valley; however, a clear understanding of the effects of aforementioned co-occurring stresses on antioxidant defense in cotton has not been attained. To address this, two temperature conditions during the whole flowering and boll development periods, and three soil-waterlogging levels (0, 3, 6 d) starting on the day of anthesis were established. In the current study, no siginificant difference was observed on plant performance for 3 d soil-waterlogging, whereas 6 d soil-waterlogging event and elevated temperature in isolation negatively affected plant performance (i.e. leaf area declined by 33.3% and 14.7% in AW6 (soil waterlogging for 6 d under ambient temperature regime) and EC (soil well-watered (SRWC(75 ±â€¯5) %) under elevated temperature for 2-3 °C) relative to AC (soil well-watered (SRWC(75 ±â€¯5) %) under ambient temperature regime)) and induced ROS (reactive oxygen species) production and scavenging mechanisms in the subtending leaf of cotton. SOD (superoxide dismutase), CAT (catalase), and POX (peroxidase) activities were increased, and ASA (ascorbic acid) concentration was enhanced due to higher H2O2 (hydrogen peroxide) and O2- accumulations. Whereas, APX (ascorbate peroxidase), DHAR (dehydroascorbate reductase) and GR (glutathione reductase) activities were inhibited under elevated temperature regime or waterlogging event, especially in the treatment of EW6 (soil waterlogging for 6 d under elevated temperature for 2-3 °C), which resulted in increasing H2O2 concentration and higher O2- generation rate. However, plants acclimated to a short-term waterlogging stress (3 d) performed a cross tolerance to chronic elevated temperature regime (leaf number increased by 11.4%, whereas the abscission rate decreased by 4.6% in EW3 (soil waterlogging for 3 d under elevated temperature for 2-3 °C) compared with EC (soil well-watered (SRWC(75 ±â€¯5) %) under elevated temperature for 2-3 °C)). Moreover, plants undergone a brief soil-waterlogging (3 d) induced higher GR activity and increased ASA concentration, along with enhanced SOD, CAT, POX activities, limiting H2O2 and O2- accumulation and reducing oxidative damage to membrane lipids as evidenced by reduced MDA (malondialdehyde) concentration when cotton was subsequently exposed to chronic elevated temperature regime.


Assuntos
Gossypium/metabolismo , Folhas de Planta/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Antioxidantes/metabolismo , Ascorbato Peroxidases/metabolismo , Glutationa Redutase/metabolismo , Gossypium/enzimologia , Malondialdeído/metabolismo , Oxirredutases/metabolismo , Solo , Superóxido Dismutase/metabolismo , Temperatura Ambiente
12.
Int J Mol Sci ; 20(9)2019 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-31027387

RESUMO

Plant regeneration via somatic embryogenesis (SE) is a key step during genetic engineering. In the current study, integrated widely targeted metabolomics and RNA sequencing were performed to investigate the dynamic metabolic and transcriptional profiling of cotton SE. Our data revealed that a total of 581 metabolites were present in nonembryogenic staged calli (NEC), primary embryogenic calli (PEC), and initiation staged globular embryos (GE). Of the differentially accumulated metabolites (DAMs), nucleotides, and lipids were specifically accumulated during embryogenic differentiation, whereas flavones and hydroxycinnamoyl derivatives were accumulated during somatic embryo development. Additionally, metabolites related to purine metabolism were significantly enriched in PEC vs. NEC, whereas in GE vs. PEC, DAMs were remarkably associated with flavonoid biosynthesis. An association analysis of the metabolome and transcriptome data indicated that purine metabolism and flavonoid biosynthesis were co-mapped based on the Kyoto encyclopedia of genes and genomes (KEGG) database. Moreover, purine metabolism-related genes associated with signal recognition, transcription, stress, and lipid binding were significantly upregulated. Moreover, several classic somatic embryogenesis (SE) genes were highly correlated with their corresponding metabolites that were involved in purine metabolism and flavonoid biosynthesis. The current study identified a series of potential metabolites and corresponding genes responsible for SE transdifferentiation, which provides a valuable foundation for a deeper understanding of the regulatory mechanisms underlying cell totipotency at the molecular and biochemical levels.


Assuntos
Flavonoides/metabolismo , Gossypium/metabolismo , Purinas/metabolismo , Transdiferenciação Celular , Regulação da Expressão Gênica de Plantas/genética , Gossypium/genética , Metaboloma/genética , Metaboloma/fisiologia , Transcriptoma/genética
13.
Int J Mol Sci ; 20(7)2019 Apr 08.
Artigo em Inglês | MEDLINE | ID: mdl-30965557

RESUMO

The regulator of chromosome condensation 1 (RCC1) is the nucleotide exchange factor for a GTPase called the Ras-related nuclear protein, and it is important for nucleo-plasmic transport, mitosis, nuclear membrane assembly, and control of chromatin agglutination during the S phase of mitosis in animals. In plants, RCC1 molecules act mainly as regulating factors for a series of downstream genes during biological processes such as the ultraviolet-B radiation (UV-B) response and cold tolerance. In this study, 56 genes were identified in upland cotton by searching the associated reference genomes. The genes were found to be unevenly distributed on 26 chromosomes, except A06, A12, D03, and D12. Phylogenetic analysis by maximum-likelihood revealed that the genes were divided into five subgroups. The RCC1 genes within the same group shared similar exon/intron patterns and conserved motifs in their encoded proteins. Most genes of the RCC1 family are expressed differently under various hormone treatments and are negatively controlled by salt stress. Gh_A05G3028 and Gh_D10G2310, which encode two proteins located in the nucleus, were strongly induced under salt treatment, while mutants of their homoeologous gene (UVR8) in Arabidopsis and VIGS (virus induced gene silencing) lines of the two genes above in G. hirsutum exhibited a salt-sensitive phenotype indicating their potential role in salt resistance in cotton. These results provide valuable reference data for further study of RCC1 genes in cotton.


Assuntos
Gossypium/metabolismo , Fatores de Troca do Nucleotídeo Guanina/metabolismo , Proteínas de Plantas/metabolismo , Cromossomos de Plantas/genética , Cromossomos de Plantas/metabolismo , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Genoma de Planta/genética , Gossypium/genética , Fatores de Troca do Nucleotídeo Guanina/genética , Proteínas de Plantas/genética , Estresse Fisiológico/genética , Estresse Fisiológico/fisiologia
14.
BMC Genomics ; 20(1): 285, 2019 Apr 11.
Artigo em Inglês | MEDLINE | ID: mdl-30975097

RESUMO

BACKGROUND: Two-dimensional electrophoresis (2-DE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) are widely used in plant proteomics research. However, these two techniques cannot be simultaneously satisfied by traditional protein extraction methods when investigate cotton leaf proteome. RESULTS: Here, we evaluated the efficiency of three different protein extraction methods for 2-DE and LC-MS/MS analyses of total proteins obtained from cotton leaves. The protein yield of the borax/PVPP/phenol (BPP) method (0.14%) was significantly lower than the yields of the trichloroacetic acid/acetone (TCA) precipitation method (1.42%) and optimized TCA combined with BPP (TCA-B) method (0.47%). The BPP method was failed to get a clear 2-DE electrophoretogram. Fifty pairs of protein spots were randomly selected from the 2-DE gels of TCA- and TCA-B-extracted proteins for identification by MALDI TOF/TOF, and the results of 42 pairs were consistent. High-throughput proteomic analysis showed that 6339, 9282 and 9697 unique proteins were identified from the total cotton leaf proteins extracted by the TCA, BPP and TCA-B methods, respectively. Gene Ontology (GO) analysis revealed that the proteins specifically identified by TCA method were primarily distributed in the plasma membrane, while BPP and TCA-B methods specific proteins distributed in the cytosol, indicating the sub-cellular preference of different protein extraction methods. Further, ATP-dependent zinc metalloprotease FTSH 8 could be observed in the 2-DE gels of TCA and TCA-B methods, and could only be detected in the LC-MS/MS results of the BPP and TCA-B methods, showing that TCA-B method might be the optimized choice for both 2-DE and LC-MS/MS. CONCLUSION: Our data provided an improved TCA-B method for protein extraction that is compatible with 2-DE and LC-MS/MS for cotton leaves and similar plant tissues which is rich in polysaccharides and polyphenols.


Assuntos
Fracionamento Químico/métodos , Gossypium/metabolismo , Folhas de Planta/metabolismo , Proteínas de Plantas/isolamento & purificação , Proteínas de Plantas/metabolismo , Sequência de Aminoácidos , Cromatografia Líquida , Eletroforese em Gel Bidimensional , Proteínas de Plantas/química , Proteômica , Espectrometria de Massas em Tandem
15.
Int J Mol Sci ; 20(7)2019 Apr 04.
Artigo em Inglês | MEDLINE | ID: mdl-30987365

RESUMO

The somatic embryogenesis (SE) process of plants, as one of the typical responses to abiotic stresses with hormone, occurs through the dynamic expression of different proteins that constitute a complex regulatory network in biological activities and promotes plant totipotency. Plant SE includes two critical stages: primary embryogenic calli redifferentiation and somatic embryos development initiation, which leads to totipotency. The isobaric labels tandem mass tags (TMT) large-scale and quantitative proteomics technique was used to identify the dynamic protein expression changes in nonembryogenic calli (NEC), primary embryogenic calli (PEC) and globular embryos (GEs) of cotton. A total of 9369 proteins (6730 quantified) were identified; 805, 295 and 1242 differentially accumulated proteins (DAPs) were identified in PEC versus NEC, GEs versus PEC and GEs versus NEC, respectively. Eight hundred and five differentially abundant proteins were identified, 309 of which were upregulated and 496 down regulated in PEC compared with NEC. Of the 295 DAPs identified between GEs and PEC, 174 and 121 proteins were up- and down regulated, respectively. Of 1242 differentially abundant proteins, 584 and 658 proteins were up- and down regulated, respectively, in GEs versus NEC. We have also complemented the authenticity and accuracy of the proteomic analysis. Systematic analysis indicated that peroxidase, photosynthesis, environment stresses response processes, nitrogen metabolism, phytohormone response/signal transduction, transcription/posttranscription and modification were involved in somatic embryogenesis. The results generated in this study demonstrate a proteomic molecular basis and provide a valuable foundation for further investigation of the roles of DAPs in the process of SE transdifferentiation during cotton totipotency.


Assuntos
Transdiferenciação Celular/fisiologia , Gossypium/metabolismo , Proteínas de Plantas/metabolismo , Proteoma/metabolismo , Transdiferenciação Celular/genética , Desenvolvimento Embrionário/genética , Desenvolvimento Embrionário/fisiologia , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Gossypium/embriologia , Gossypium/genética , Proteínas de Plantas/genética , Proteômica
16.
Int J Mol Sci ; 20(6)2019 Mar 24.
Artigo em Inglês | MEDLINE | ID: mdl-30909634

RESUMO

High concentrations of heavy metals in the soil should be removed for environmental safety. Cadmium (Cd) is a heavy metal that pollutes the soil when its concentration exceeds 3.4 mg/kg. Although the potential use of cotton to remediate heavy Cd-polluted soils is known, little is understood about the molecular mechanisms of Cd tolerance. In this study, transcriptome analysis was used to identify Cd tolerance genes and their potential mechanisms in cotton. We exposed cotton plants to excess Cd and identified 4627 differentially expressed genes (DEGs) in the root, 3022 DEGs in the stem and 3854 DEGs in the leaves through RNA-Seq analysis. Among these genes were heavy metal transporter coding genes (ABC, CDF, HMA, etc.), annexin genes and heat shock genes (HSP), amongst others. Gene ontology (GO) analysis showed that the DEGs were mainly involved in the oxidation⁻reduction process and metal ion binding. The DEGs were mainly enriched in two pathways, the influenza A and pyruvate pathway. GhHMAD5, a protein containing a heavy-metal binding domain, was identified in the pathway to transport or to detoxify heavy metal ions. We constructed a GhHMAD5 overexpression system in Arabidopsis thaliana that showed longer roots compared to control plants. GhHMAD5-silenced cotton plants showed more sensitivity to Cd stress. The results indicate that GhHMAD5 is involved in Cd tolerance, which gives a preliminary understanding of the Cd tolerance mechanism in upland cotton. Overall, this study provides valuable information for the use of cotton to remediate soils polluted with Cd and potentially other heavy metals.


Assuntos
Adaptação Biológica/genética , Cádmio/metabolismo , Regulação da Expressão Gênica de Plantas , Gossypium/genética , Gossypium/metabolismo , Estresse Fisiológico , Arabidopsis/genética , Arabidopsis/metabolismo , Biologia Computacional/métodos , Perfilação da Expressão Gênica , Inativação Gênica , Gossypium/virologia , Fenótipo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Transcriptoma
17.
Plant Sci ; 281: 72-81, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30824063

RESUMO

Salinity is a major abiotic stress limiting plant growth and development that has caused severe damage to yield and quality of cotton fiber. Uncovering the mechanisms of response to salt stress is important in breeding salt-tolerant cotton varieties. Transcriptome analysis identified 2356 differentially expressed genes in cotton under salt stress, of which 9.4% were predicted transcription factors (TFs). Approximately 17.6% (39 out of 222) of the differentially expressed TFs belonged to the ethylene response factor (ERF) family. Expression pattern analysis showed significant changes in these ERFs during salt stress. Moreover, the number of down-regulated ERFs was more than that of the up-regulated ERFs. Two of the ERFs, GhERF4L and GhERF54L, showed increased (12-15 times) expression after 12 h of salt treatment. Silencing of GhERF4L and GhERF54L significantly reduced salt tolerance of cotton seedlings, indicating their role in regulating cotton response to salt stress. This study revealed the essential role of ERF transcription factors in the salt response mechanism of plants, and provided important genetic resources for breeding salt-tolerant cotton.


Assuntos
Gossypium/metabolismo , Fatores de Transcrição/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/genética , Regulação da Expressão Gênica de Plantas/fisiologia , Gossypium/genética , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Tolerância ao Sal/genética , Tolerância ao Sal/fisiologia , Fatores de Transcrição/genética
18.
Planta ; 249(5): 1551-1563, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30729290

RESUMO

MAIN CONCLUSION: A novel method for culturing ovules of Gossypium barbadense allowed in vitro comparisons with Gossypium hirsutum and revealed variable roles of microtubules in controlling cotton fiber cell expansion. Cotton fibers undergo extensive elongation and secondary wall thickening as they develop into our most important renewable textile material. These single cells elongate at the apex as well as elongating and expanding in diameter behind the apex. These multiple growth modes represent an interesting difference compared to classical tip-growing cells that needs to be explored further. In vitro ovule culture enables experimental analysis of the controls of cotton fiber development in commonly grown Gossypium hirsutum cotton, but, previously, there was no equivalent system for G. barbadense, which produces higher quality cotton fiber. Here, we describe: (a) how to culture the ovules of G. barbadense successfully, and (b) the results of an in vitro experiment comparing the role of microtubules in controlling cell expansion in different zones near the apex of three types of cotton fiber tips. Adding the common herbicide fluridone, 1-Methyl-3-phenyl-5-[3-(trifluoromethyl)phenyl]-4(1H)-pyridinone, to the medium supported G. barbadense ovule culture, with positive impacts on the number of useful ovules and fiber length. The effect is potentially mediated through inhibited synthesis of abscisic acid, which antagonized the positive effects of fluridone. Fiber development was perturbed by adding colchicine, a microtubule antagonist, to ovules of G. barbadense and G. hirsutum cultured 2 days after flowering. The results supported the zonal control of cell expansion in one type of G. hirsutum fiber tip and highlighted differences in the role of microtubules in modulating cell expansion between three types of cotton fiber tips.


Assuntos
Gossypium/citologia , Gossypium/metabolismo , Microtúbulos/metabolismo , Fibra de Algodão , Citoesqueleto/efeitos dos fármacos , Citoesqueleto/metabolismo , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Gossypium/efeitos dos fármacos , Microtúbulos/efeitos dos fármacos , Piridonas/farmacologia
19.
Planta ; 249(5): 1565-1581, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30737556

RESUMO

MAIN CONCLUSION: Evidence is presented that cotton fibre adhesion and middle lamella formation are preceded by cutin dilution and accompanied by rhamnogalacturonan-I metabolism. Cotton fibres are single cell structures that early in development adhere to one another via the cotton fibre middle lamella (CFML) to form a tissue-like structure. The CFML is disassembled around the time of initial secondary wall deposition, leading to fibre detachment. Observations of CFML in the light microscope have suggested that the development of the middle lamella is accompanied by substantial cell-wall metabolism, but it has remained an open question as to which processes mediate adherence and which lead to detachment. The mechanism of adherence and detachment were investigated here using glyco-microarrays probed with monoclonal antibodies, transcript profiling, and observations of fibre auto-digestion. The results suggest that adherence is brought about by cutin dilution, while the presence of relevant enzyme activities and the dynamics of rhamnogalacturonan-I side-chain accumulation and disappearance suggest that both attachment and detachment are accompanied by rhamnogalacturonan-I metabolism.


Assuntos
Gossypium/metabolismo , Polissacarídeos/metabolismo , Fibra de Algodão , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Glucanos/metabolismo , Xilanos/metabolismo
20.
BMC Genomics ; 20(1): 112, 2019 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-30727946

RESUMO

BACKGROUND: Improving cotton fiber length without reducing yield is one of the major goals of cotton breeding. However, genetic improvement of cotton fiber length by breeding has been a challenge due to the narrow genetic diversity of modern cotton cultivars and negative correlations between fiber quality and yield traits. A multi-parent advanced generation inter-cross (MAGIC) population developed through random mating provides an excellent genetic resource that allows quantitative trait loci (QTL) and causal genes to be identified. RESULTS: An Upland cotton MAGIC population, consisting of 550 recombinant inbred lines (RILs) derived from eleven different cultivars, was used to identify fiber length QTLs and potential genes that contribute to longer fibers. A genome wide association study (GWAS) identified a cluster of single nucleotide polymorphisms (SNPs) on chromosome (Chr.) D11 that is significantly associated with fiber length. Further evaluation of the Chr. D11 genomic region among lines of the MAGIC population detected that 90% of RILs have a D11 haplotype similar to the reference TM-1 genome (D11-ref), whereas 10% of RILs inherited an alternative haplotype from one of the parents (D11-alt). The average length of fibers of D11-alt RILs was significantly shorter compared to D11-ref RILs, suggesting that alleles in the D11-alt haplotype contributed to the inferior fiber quality. RNAseq analysis of the longest and shortest fiber length RILs from D11-ref and D11-alt populations identified 949 significantly differentially expressed genes (DEGs). Gene set enrichment analysis revealed that different functional categories of genes were over-represented during fiber elongation between the four selected RILs. We found 12 genes possessing non-synonymous SNPs (nsSNPs) significantly associated with the fiber length, and three that were highly significant and were clustered at D11:24-Mb, including D11G1928, D11G1929 and D11G1931. CONCLUSION: The results of this study provide insights into molecular aspects of genetic variation in fiber length and suggests candidate genes for genetic manipulation for cotton improvement.


Assuntos
Fibra de Algodão , Gossypium/genética , Polimorfismo de Nucleotídeo Único , Locos de Características Quantitativas , Transcriptoma , Alelos , Regulação da Expressão Gênica de Plantas , Genoma de Planta , Estudo de Associação Genômica Ampla , Gossypium/metabolismo , Melhoramento Vegetal , Proteínas de Plantas/genética
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