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1.
J Biotechnol ; 394: 92-102, 2024 Nov 10.
Artigo em Inglês | MEDLINE | ID: mdl-39181209

RESUMO

This study addresses the challenges posed by rainfall variability, leading to water deficits during critical stages of crop growth, resulting in a drastic reduction of cotton yield. In a comprehensive evaluation, thirty cotton genotypes, including five Gossypium arboreum (wild) and twenty-five Gossypium hirsutum (cultivated), were grown under rainfed and irrigated conditions. Drought tolerance indices (DTI) were evaluated, categorizing genotypes based on their resilience. Further, in-vitro screening at the seedling stage (20 days) under PEG-induced drought identified tolerant genotypes exhibiting elevated levels of free proline (19.07±5.31 mg.g-100fr.wt.), amino acids (34.59±6.51 mg.g-100fr.wt.), soluble proteins (13.73±2.65 mg.g-1fr.wt.), and glycine betaine (10.95±3.62 mg.g-100fr.wt.), in their leaves, positively correlating (p<0.001) with relative water content (87.70±3.45 %). Molecular analysis using drought-specific simple sequence repeats (SSR) markers revealed significant genetic variability in a cotton genotypes, with lower observed and higher expected heterozygosity. F statistics exposed a higher level of gene flow corresponding to low differentiation among populations. Among the genotypes group, wild GAM-67 and cultivated Deviraj emerged as the most potent, exhibiting the higher DTI and diverse gene pools. Study exhibited higher total gene diversity in drought-tolerant wild GAM-67 (0.8501) and greater expected heterozygosity (0.626) and gene flow (0.6731) in cultivated Deviraj, underlining their robust genetic adaptability to drought conditions. The integrated approach of field evaluations, in-vitro screening, and molecular analyses explained substantial genetic diversity, with the identification of promising genotypes displaying higher drought tolerance indices, elevated levels of stress-related biochemical osmolytes, and pronounced genetic adaptability, thereby contributing to the advancement of breeding initiatives for enhanced drought resilience in cotton.


Assuntos
Secas , Genótipo , Gossypium , Melhoramento Vegetal , Gossypium/genética , Gossypium/metabolismo , Repetições de Microssatélites/genética , Variação Genética , Estresse Fisiológico/genética , Resistência à Seca
2.
Funct Plant Biol ; 50(9): 736-751, 2023 09.
Artigo em Inglês | MEDLINE | ID: mdl-37536348

RESUMO

Nanotechnology provides tremendous potential in agriculture, mitigating climate change impact and improving abiotic stress management strategy. Chitosan nanoparticles (NCS) were synthesised using the ion gelation method and characterised for size (75.5nm in particle size analyser), shape (spherical under scanning electron microscopy) and stability (132.2mV zeta potential). Further, salicylic acid was incorporated into NCS to craft salicylic acid-functionalised chitosan nanoparticles (SA-NCS) and illustrated for size (517nm), shape (spherical) and stability (197.1mV). The influence of the exogenous application of SA-NCS (0.08%) was studied at the reproductive stage of three genotypes of cotton (Gossypium hirsutum ): (1) heat-tolerant Solar-651 BGII; (2) moderately heat-tolerant Solar-701 BGII; and (3) heat-susceptible Solar-805 BGII, exposed to different temperature regimes: (1) H1 (optimal), 32/20±2°C; (2) H2 (sub-optimal), 38/24±2°C; H3 (supra-optimal), 45/30±2°C. Heat stress significantly reduces carbon-fixing Rubisco, enzymes related to sucrose metabolism and pollen tube length. Considering three genotypes and reproductive stages (sepal and anther tissues), activities of Rubisco (sepals), invertase (sepals), sucrose phosphate synthase (anthers), sucrose content (sepals) and pollen tube length were elevated under high-temperature regimes, signifying better source to sink transposition of sucrose influenced by SA-NCS. The study provides new insights into SA-NCS to improve source-sink imbalance and restore sucrose metabolism for better growth of reproductive structure under heat stress in cotton.


Assuntos
Quitosana , Gossypium , Gossypium/genética , Gossypium/metabolismo , Quitosana/metabolismo , Ribulose-Bifosfato Carboxilase/metabolismo , Resposta ao Choque Térmico , Sacarose/metabolismo
3.
Arch Microbiol ; 205(6): 242, 2023 May 19.
Artigo em Inglês | MEDLINE | ID: mdl-37204527

RESUMO

Trichoderma isolates were inhibited variably in-vitro growth of soil-borne phytopathogen Macrophomina phaseolina (Maubl.) Ashby causes root rot in cotton. The growth inhibition of test-pathogen was found to be higher (90.36%) in T. viride NBAIITv23 followed by T. koningii MTCC796 (85.77%) under dual culture antagonism. The microscopic examination suggested that the antagonists Tv23 and MTCC796 adopted mycoparasitism as a strong mode of action to restrain pathogen growth. However, antagonists T. harzianum NBAIITh1 (77.89%) and T. virens NBAIITvs12 (61.74%) demonstrated strong antibiosis action for growth inhibition of the test pathogen. A significant positive correlation was established between the growth inhibition of M. phaseolina and the release of cell wall degrading enzymes- chitinase (p = 0.001), ß-1,3, glucanase (p = 0.01), and protease (p = 0.05) under the influence of pathogen cell wall. The chitinase and ß-1,3, glucanase activities were elevated 2.09 and 1.75 folds, respectively, in potent mycoparasitic Tv23 strain influenced by a pathogen cell wall compared to glucose as a carbon source. The three unique DNA-RAPD fragments OPA-07(1033), OPA-16(983), and OPO-15(239), amplified by potent mycoparasitic Tv23 strain, were subjected to DNA sequencing and derived functional 864 bp from OPA-16(983) and have sequence homology to ech42 gene with partial CDs of 262 amino acids (nucleotide accession No. KF723016.1 and protein accession No.AHF57046.1). Novel SCAR markers were developed from a functional sequence of OPA-16 fragments and validated across the genomic DNA of eleven Trichoderma antagonists. The novel SCAR markers evolved from the RAPD-SCAR interface to authenticate chitinolytic Trichoderma associated with mycoparasitic action for eco-friendly biocontrol activity.


Assuntos
Ascomicetos , Quitinases , Trichoderma , Trichoderma/genética , Técnica de Amplificação ao Acaso de DNA Polimórfico , Ascomicetos/fisiologia , Marcadores Genéticos , Quitinases/genética , Quitinases/metabolismo
4.
Pestic Biochem Physiol ; 191: 105368, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36963937

RESUMO

The study investigated potential microRNA-like small RNAs (milRNAs) from multi-stress-tolerant Tricho-fusants and parental strains (P1- Trichoderma virens NBAIITvs12 and P2- Trichoderma koningii MTCC796) for antagonistic activity during interaction with phytopathogen Sclerotium rolfsii. The Trichoderma was cultured in-vitro, with and without antagonism, against the pathogen and total RNA was extracted followed by small RNA library construction and sequencing. The milRNAs were identified by mapping high-quality unique reads against a reference genome. The milRNAs were recognized higher in antagonist Trichoderma during interaction with test pathogen compared to normal growth. The novel milRNAs candidates were found to vary during interaction with the pathogen and normal growth. The gene ontology and functional analysis illustrated that a total of 5828 potential targeted genes were recognized for 93 milRNAs of potent Fu21_IB and 3053 genes for 62 milRNAs of least fusant Fu28_IL. Functional annotation of milRNA-predicted genes integrating KEGG pathways indicates new insights into regulatory mechanisms, by interfering with milRNAs, associated with signal transduction, amino sugar metabolism, benzoate degradation, amino acid metabolism, and steroid and alkaloid metabolism for potential biocontrol of stress-tolerant Tricho-fusant FU21 during interaction with S. rolfsii. The present investigation is the first report of conserved and novel milRNAs from Tricho-fusants and parental strains interacting with S. rolfsii.


Assuntos
Basidiomycota , Hypocrea , MicroRNAs , Trichoderma , Trichoderma/genética , MicroRNAs/genética , Basidiomycota/genética , Hypocrea/genética
5.
Protein J ; 41(6): 638-658, 2022 12.
Artigo em Inglês | MEDLINE | ID: mdl-36251227

RESUMO

The resistant and susceptible genotypes of castor were utilized for leaf proteomic study during Fusarium wilt infection. The histopathological study was observed under SEM and it confirmed that the infection of Fusarium oxysporum f. sp. ricini was higher in the root of susceptible JI-35, while incompatible interaction is observed in resistant SKI-215 genotype. The acidic and neutral proteins were maximally up-expressed with 2 to 171 kDa in treated resistant and 2 to 150 kDa in treated susceptible interactions. In resistant genotype, the leaf proteins were recognized with 3.0- and 5.8-fold higher at infection stage and post-infection stage, respectively, as compared to susceptible genotype. The highly up expressions of leaf acidic (4.76 pI) and basic (8.77 pI) proteins were found with 224.94- and 61.68-fold change, respectively during the post-infection stage in treated resistance compared to its control. The protein spots at 4.76 pI and 8.77 pI were characterized with nanoLC-MS Triple TOF and were recognized as signalling molecules small GTP binding protein (23 kDa) and actin (8 kDa), respectively, on the basis of mass spectrometry and peptide sequences. However, basic and neutral proteins were up regulated as 30.11- and 20.30-fold changes in treated susceptible compared to its control. These proteins were identified as HSP90 (10 kDa) and LEA (27 kDa) proteins. The 148 kDa protein is recognized as histidine kinase in incompatible resistant interaction compared to compatible susceptible (serine threonine protein kinase, 65 kDa) as common acidic protein at 3.80 pI during infection stage. Some acidic proteins were maximally up-regulated in the leaf of resistant castor genotype and played a significant role in defense response.


Assuntos
Fusarium , Fusarium/metabolismo , Proteômica , Doenças das Plantas/genética , Ricinus , Genótipo , Folhas de Planta/genética , Espectrometria de Massas
6.
Arch Microbiol ; 204(6): 311, 2022 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-35538378

RESUMO

The Beauveria spp. were isolated from soil and insect cadavers and confirmed as Beauveria bassiana by molecular identification using a specific primer. The bioefficacy of 14 B. bassiana against whiteflies indicated the highest percent mortality in JAU2, followed by JAU1. The LC50 and LC90 values were found to be 0.043 × 105 and 0.05 × 1014 conidia.ml-1, respectively, in JAU2. Extracellular metabolites of B.bassiana are derived and used for the green synthesis of silver nanoparticles (AgNPs). The synthesized green AgNPs were characterized for size (24.8 nm), shape (scanning electron microscopy), stability (200 mV zeta), and purity (energy-dispersive X-ray spectroscopy, 3 keV). A total of 63 extracellular metabolites were identified using LC-MS/QTOF in potent JAU2 with recognition of alcohols, phenols, carboxylic acids, amines, alkynes, and amides as functional groups. The functional groups of green AgNPs were also confirmed in Fourier transforms infrared spectroscopy (FTIR) with the specific spectra in the electromagnetic spectrum. The relationship between identified metabolites of antagonist and the FTIR spectrum of the functional group indicated the involvement of extracellular novel compounds, viz., homoisocitrate, aconitine, phodexin A, capillone, solanocapsine, and anethole in the synthesis of green AgNPs. The efficacy of green AgNPs on whiteflies suggested that corrected percent mortality was observed at 60 µg Ag.ml-1 at 120 h, which corresponds to the LC50 value (66.42 µg Ag.ml-1). Results were interpreted to show that green AgNPs synthesized from extracellular metabolites of B.bassiana JAU2 gave better insecticidal activity at LC50 as compared to live antagonist JAU2.


Assuntos
Anopheles , Beauveria , Hemípteros , Nanopartículas Metálicas , Animais , Anopheles/metabolismo , Beauveria/metabolismo , Larva , Nanopartículas Metálicas/química , Extratos Vegetais/química , Folhas de Planta/química , Prata/química , Espectroscopia de Infravermelho com Transformada de Fourier
7.
Parasitol Res ; 121(7): 2019-2031, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35614146

RESUMO

The entomopathogenic Beauveria spp. were acquired from insect cadavers and soil rhizosphere of cotton, groundnut, and castor. Among Beauveria, five spp. derived from infected insects, eight Beauveria found from soil, and one strain of Beauveria bassiana collected from MTCC 9544. Beauveria were characterized for morphology and cuticle-degrading enzyme activity associated with virulence against Bemisia tabaci. The colony morphology, conidial arrangement, size, and shape confirmed all isolates as Beauveria. The chitinase (EC 3.2.1.14) and lipase (EC 3.1.1.3) activities were observed the highest in Beauveria JAU2, while higher protease (EC 3.4.21.4) activity found in JAU4 followed by JAU2 at 240 h. The bio-efficacy of Beauveria (1 × 107 conidia.ml-1) illustrated that potent JAU2 was examined with the highest % mortality and corrected mortality of B. tabaci at 144 h followed by JAU1. The LC90 and LC50were determined from potent (JAU1 and JAU2) and weak (JAU6), and it was found the lowest in JAU2. The most potent Beauveria JAU2, isolated from insect cadaver (Harmivora armigera), was illustrated higher virulence than other isolates. The Beauveria JAU2 were recognized as Beauveria bassiana based on the shape of conidia and size (2.00 to 2.09 µm dia) as examined in SEM. Study insight into recognition of potent Beauveria bassiana JAU2 was linked with cuticle-degrading enzyme activity for insecticidal action. The JAU2 isolate established the most positive correlation (P0.01: 0.864) between chitinase activity and corrected mortality of insect.


Assuntos
Beauveria , Quitinases , Animais , Insetos , Controle Biológico de Vetores , Solo , Esporos Fúngicos
8.
Biometals ; 35(3): 479-497, 2022 06.
Artigo em Inglês | MEDLINE | ID: mdl-35332436

RESUMO

The potent antagonist Bacillus isolated from the soil rhizosphere elucidated the highest antagonism against the phytopathogen Fusarium oxysporum f. sp. cumini and was identified as Bacillus subtilis strain JSD-RSCu-8D based on molecular recognition by 16S rRNA sequencing (NCBI Accession No. KT894724). Live Bacillus may not work as effectively against phytopathogen under unfavorable environmental conditions like temperature, humidity, or other abiotic stresses. The extracellular metabolites, obtained from culturing potent B. subtilis, were exploited for the creation of green nanosilver for proficient actions in a changing climate. The synthesized green nanosilver was illustrated for shape (spherical with 65.21 ± 3.71 nm under SEM), size (70.9 nm in PSA), purity (2.69 keV peak corresponded to the binding energy of silver under EDAX), and stability (44.2 mV as ZETA). The formation of green Ag-NPs from extracellular metabolites was confirmed by a comparative appraisal of the electromagnetic peak of the metabolite's functional groups, silver nitrate, and green nanoparticles in Fourier transform infrared spectroscopy. The novel mode of action of pathogen mycelium degradation was elucidated by the minimum inhibitory concentration (MIC) of green nanosilver as 40 µg Ag ml-1 to diminish F. oxysporum (SEM morphology). The green nanosilver at 2 DAI renowned the leakage of sugars from mycelia of the cell membrane and defeated the activity of respiratory chain dehydrogenases, followed by lipid peroxidation and the highest leakage of proteins at 3 DAI on MIC. The in-vivo study might allow for novel insight to utilize green nanosilver at MIC (40 µg Ag ml-1) as an eco-friendly and fungicide alternate way for antifungal action to demolish Fusarium wilt infection under harsh conditions.


Assuntos
Bacillus , Nanopartículas Metálicas , Nanoestruturas , Antifúngicos/química , Antifúngicos/farmacologia , Bacillus subtilis/genética , Bacillus subtilis/metabolismo , Nanopartículas Metálicas/química , Testes de Sensibilidade Microbiana , RNA Ribossômico 16S/genética , RNA Ribossômico 16S/metabolismo , Prata , Espectroscopia de Infravermelho com Transformada de Fourier
9.
Pestic Biochem Physiol ; 176: 104877, 2021 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-34119221

RESUMO

The Beauveria spp. were isolated from soil and insect cadavers of crop rhizosphere and characterized for parasitic enzyme activity and virulence against whiteflies (Bemisia tabaci). The colony morphology and molecular identification using ITS specific marker were carried out and confirmed entomopathogenic fungi as Beauveria bassiana. The bioefficacy of B. bassiana against whiteflies demonstrated highest corrected mortality and lowest LC50 in isolate B. bassiana JAU2 (SEM morphology) followed by JAU1 on 6th days. Parasitic enzymes chitinase and lipase were determined highest in JAU2 and protease activity examined higher in isolate JAU4 followed by JAU2 isolate on 6th days after inoculation. Comparative extracellular metabolomics carried out from potent (JAU1 and JAU2), moderate (JAU4 and JAU14) and weak (JAU6) B. bassiana isolates in normal suborder dextrose agar with yeast extrect (SDAY) and chitin induced media. Results illustrated that total 105 metabolites identified common for all five B. bassiana isolates differing in virulence. However, the color intensity of the metabolites changes in heat map showing differential concentration of that extracellular compound compared to other isolates. The volcano plot analysis illustrated 58 compounds significanlty diverse between potent JAU1 and JAU2 under two different culture conditions of which 34 compounds recognized up regulated in most potent JAU2 under chitin induced media. Out of 34 metabolites, ten compounds viz., fumaricine, resazurin, N-methyldioctylamine, penaresidun B, tetralin, squamocin B, oligomycin C, pubesenolide, epirbuterol and gentamicin C1a were recognized significantly upregulated in most potent JAU2 and reported for antimicrobial, nematicidal, larvicidalor insecticidal activities. The mass spectra and fragment structure were elucidated under LCMS-QTOF for some novel and unique compounds recognized in most potent B. bassiana JAU2, involved in parasitic activity against whiteflies.


Assuntos
Beauveria/enzimologia , Quitinases , Hemípteros , Controle Biológico de Vetores , Animais , Metabolômica
10.
J Cell Physiol ; 234(5): 7368-7383, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30370526

RESUMO

The current study aimed at developing diverse Trichoderma fusants for fungicides, drought, and salt tolerance with enhanced antagonistic activity against Sclerotium rolfsii Sacc. Trichoderma virens NBAII Tvs12 (mycoparasitic) and Trichoderma koningii MTCC796 (multistress tolerant) were used as parental strains for development of interspecific protoplast fusants. A total of 36 stable fusants were used for mycoparasitism, fungicides, and abiotic stresses (drought and salt) tolerance. The results revealed 20 homozygous progenies showing characteristics of either one parental strain and 14 heterozygous mutants depicting traits of both parental strains. A novel concept of inhibition coefficient was established using growth-related key parameters that represent the pathogen biology and the biocontrol-related biophysics of Trichoderma fusants. The results indicated a differential inhibition coefficient of the test pathogen and the highest (92.88%) inhibition coefficient of S. rolfsii was observed by interstable fusant Fu21. It also grew better under fungicides and abiotic stress (drought and salt) conditions. The molecular characterization and heterozygosity analysis evidenced the highest observed heterozygosity (0.5441) and gene flow (0.3872) in stable heterozygous Fu21. Principal coordinates analysis exhibited 62.7% of total variability. The ecofriendly heterozygous Trichoderma fusant (Fu21) might be useful for biocontrol of stem rot disease under adverse conditions or as a part of integrated disease management.


Assuntos
Basidiomycota/crescimento & desenvolvimento , Mutação , Controle Biológico de Vetores , Doenças das Plantas/prevenção & controle , Trichoderma/genética , Basidiomycota/patogenicidade , Desidratação/genética , Secas , Fungicidas Industriais/farmacologia , Heterozigoto , Homozigoto , Doenças das Plantas/microbiologia , Tolerância ao Sal/genética , Trichoderma/efeitos dos fármacos , Trichoderma/crescimento & desenvolvimento , Trichoderma/ultraestrutura
11.
Infect Genet Evol ; 66: 26-36, 2018 12.
Artigo em Inglês | MEDLINE | ID: mdl-30219319

RESUMO

Protoplast fusion is an imperative tool to develop Trichoderma inter-fusants having desire traits through genetic manipulation. Study designed to develop diverse Trichoderma fusants for fungicide tolerance (Mancozeb, Thiram, Tebuconazole, and Carbendazim) and enhanced mycoparasitic activity against Sclerotium rolfsii sacc. The mycoparasitic T. virens NBAII Tvs12 and fungicide tolerant T. koningii MTCC 796 were utilized for protoplast fusion. The derived inter-fusants were subjected to diploidization using d-camphor in minimal media followed by successive three sub culturing onto potato dextrose agar to obtain 36 stable fusants. The stable fusants were employed for conidial size, fungicide tolerance, mycoparasitism, gene specific SSR amplification and molecular heterozygosity analysis. The results explained that 22 homozygous mutants illustrated characteristic of either one parental strain and 14 heterozygous recombinants depicted traits of both parental strains. The antagonistic activity of fusants against S. rolfsii depicted highest growth inhibition (87.91%) by potent inter-fusant (Fu 21) with improved fungicide tolerance capacity. The molecular study revealed highest observed heterozygocity (0.544), coefficient of gene differentiation (0.526) and gene flow (0.387) by Fu 21 indicating better genetic exploitation of parental strains into that fusant with good genetic purity. Principal coordinate analysis of fusants and parental strains exhibited 65.07% total variation and confirmed the scattering pattern matched with UPGMA clustering pattern. The stable heterozygous Fu 21 derived from inter-fusion between Tvs 12 and MTCC 796 might be useful to practice eco-friendly bioformulation tolerance to fungicides for effective integrated stem rot disease management in groundnut.


Assuntos
Antibiose , Basidiomycota/fisiologia , Farmacorresistência Fúngica , Fungicidas Industriais/farmacologia , Variação Genética , Trichoderma/efeitos dos fármacos , Trichoderma/fisiologia , DNA Fúngico , Fluxo Gênico , Heterozigoto , Repetições de Microssatélites , Polimorfismo Genético
12.
J Food Sci Technol ; 55(2): 730-739, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29391638

RESUMO

The indigenous black jamun landraces (Syzygium Cumini L.), found in western Gujarat of Gir forest region (India), produced fruits with different size and shape. Fruit morphology like shape, volume, weight, length, girth were examined and black jamun categorized into six landraces viz., BJLR 1 (big fruit, > 11 g); BJLR 2 (medium to big fruit, 8-11 g); BJLR 3 (medium fruit, 6-8 g); BJLR 4 (medium to small fruit, 5-6 g); BJLR 5 (small fruit, 3-5 g) and BJLR 6 (very small fruit, < 3 g fruit weight). The landraces (BJLR 1 and 2) with larger size fruits were accumulated higher amount of moisture, total fat content, sugars, total protein, starch, free amino acid contents. Smaller fruits (BJLR 6) contained higher amount of ascorbic acid-137 and 132 mg%; anthocynin-47.7 and 2.35 mg%; crude fibre 3.05 and 10.5 g%; and total phenol-21.7 and 45.0 mg g-1 in their fruit pulp and seed part, respectively with better nutritional profile compared with big and moderate fruited landraces. Nutritional profile of six landraces indicated that fruit pulp accumulated higher amount of soluble sugars (6.51-17.6 mg g-1), anthocyanins (29.7-47.7 mg%) and free amino acids (7.54-18.9 mg%) while that of seeds exhibited higher amount of crude fibre (6017-10.5 g%), ascorbic acid (90-137 mg%), starch (22.8-29.4 g%), total protein (4.72-7.17 mg%), phenols (45-56.7 mg g-1). The black jamun landraces were subjected to ISSR based polymorphic finger prints and genetic diversity analysis. Total 144 bands were amplified across six landraces by 18 UBC primers, of which 94 were polymorphic with 64.2% average polymorphism. Cluster analysis demonstrates the BJLR 6 landraces distinguished from other landraces with 53% similarity.

13.
J Food Sci Technol ; 54(10): 3180-3191, 2017 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-28974803

RESUMO

ABSTRACT: Fruit phenolics are important dietary antioxidant and antidiabetic constituents. The fruit parts (pulp, seed, seed coat, kernel) of underutilized indigenous six black jamun landraces (Syzygium cumini L.), found in Gir forest region of India and differed in their fruit size, shape and weight, are evaluated and correlated with antidiabetic, DPPH radical scavenging and phenolic constituents. The α-amylase inhibitors propose an efficient antidiabetic strategy and the levels of postprandial hyperglycemia were lowered by restraining starch breakdown. The sequential solvent systems with ascending polarity-petroleum ether, ethyl acetate, methanol and water were performed for soxhlet extraction by hot percolation method and extractive yield was found maximum with methanolic fruit part extracts of six landraces. The methanolic extracts of fruit parts also evidenced higher antidiabetic activity and hence utilized for further characterization. Among the six landraces, pulp and kernel of BJLR-6 (very small, oblong fruits) evidenced maximum 53.8 and 98.2% inhibition of α-amylase activity, respectively. The seed attained inhibitory activity mostly contributed by the kernel fraction. The inhibition of DPPH radical scavenging activity was positively correlated with phenol constituents. An HPLC-PDA technique was used to quantify the seven individual phenolics. The seed and kernel of BJLR-6 exhibited higher individual phenolics-gallic, catechin, ellagic, ferulic acids and quercetin, whereas pulp evidenced higher with gallic acid and catechin as α-amylase inhibitors. The IC50 value indicates concentration of fruit extracts exhibiting ≥50% inhibition on porcine pancreatic α-amylase (PPA) activity. The kernel fraction of BJLR6 evidenced lowest (8.3 µg ml-1) IC50 value followed by seed (12.9 µg ml-1), seed coat (50.8 µg ml-1) and pulp (270 µg ml-1). The seed and kernel of BJLR-6 inhibited PPA at much lower concentrations than standard acarbose (24.7 µg ml-1) considering good candidates for antidiabetic herbal formulations.

14.
Infect Genet Evol ; 55: 75-92, 2017 11.
Artigo em Inglês | MEDLINE | ID: mdl-28864153

RESUMO

Trichoderma is one of the most exploited biocontrol agent for the management of plant diseases. Twenty strains of Trichoderma (six of T. harzianum, four of T. viride, three of T. virens, three of T. koningii, each one of T. hamatum, T. reesei, T. parceramosum and Trichoderma spp.) subjected to in vitro antagonism up to 12days after inoculation against Sclerotium rolfsii Sacc. causing stem rot in groundnut. A new concept was developed to determine inhibition coefficient representing pathogen biology and biocontrol related biophysical variables. Results explained differential inhibition coefficient of test pathogen by Trichoderma antagonists. The inhibition coefficient of test pathogen was examined highest (91.13%) by T. virens NBAII Tvs12 followed by T. virens MTCC 794 (89.33%) and T. koningii MTCC 796 (62.39%). Microscopic study confirmed biocontrol mechanism as mycoparasitism for Tvs12 and antibiosis for T. koningii MTCC 796. The sclerotial biogenesis of test pathogen was elevated during weak antagonism and diminished in interactions with strong antagonists. The inhibition coefficient of S. rolfsii was significantly negatively correlated with sclerotia formation and lipid peroxidation during the antagonism. Trichoderma strains were screened for fungicides (carbendazim and tebuconazole, thiram and mancozeb) and abiotic stress (drought and salt) tolerance. Results indicated that T. koningii MTCC 796 efficiently grew better than the other strains with maximum radial growth under adverse conditions. The genetic variability among the Trichoderma was determined using 34 gene specific markers which amplified 146 alleles. The SSR similarities explained substantial diversity (15 to 87%) across Trichoderma strains and pathogen S. rolfsii. Principal coordinates analysis (PCA) were comparable to the cluster analysis and first three most informative PC components explained 64.45% of the total variation. In PCA, potent antagonists appear to be distinct from other strains. Five SSR markers T1F/T1R(311), TvCTT56f/TvCTT56r(387), TvGAT18f/TvGAT18r(364), TvCA39f/TvCA39r(196) and TvAG29f/TvAG29r(418) found to be unique to distinguish best antagonist strain Tvs12. However, MTCC 796 was examined most stress tolerant strain with better inhibition coefficient which might be useful to control the disease under adverse conditions or as a part of integrated pest management.


Assuntos
Adaptação Biológica , Antibiose , Basidiomycota , Agentes de Controle Biológico , Variação Genética , Estresse Fisiológico , Trichoderma/classificação , Trichoderma/fisiologia , Basidiomycota/crescimento & desenvolvimento , Basidiomycota/ultraestrutura , Testes de Sensibilidade a Antimicrobianos por Disco-Difusão , Metabolismo dos Lipídeos , Peroxidação de Lipídeos , Repetições de Microssatélites , Trichoderma/ultraestrutura
15.
Infect Genet Evol ; 45: 383-392, 2016 11.
Artigo em Inglês | MEDLINE | ID: mdl-27720889

RESUMO

The biocontrol agent Trichoderma (T. harzianum, T. viride, T. virens, T. hamantum, T. koningii, T. pseudokoningii and Trichoderma species) inhibited variably (15.32 - 88.12%) the in vitro growth of Rhizoctonia solani causing root rot in cotton. The T. koningii MTCC 796 evidenced highest (88.12%) growth inhibition of test pathogen followed by T. viride NBAII Tv23 (85.34%). Scanning electron microscopic study confirmed mycoparasitism for MTCC 796 and Tv23 which were capable of completely overgrowing on R. solani by degrading mycelia, coiling around the hyphae with hook-like structures. The antagonists T. harzianum NBAII Th1 and, T. virens NBAII Tvs12 exhibited strong antibiosis and formed 2-4 mm zone of inhibition for 70.28% and 46.62%, respectively growth inhibition of test pathogen. Mycoparasitism is a strong mode of action for biocontrol activity compared with antibiosis. The antagonists Trichoderma strains were performed for start codon targeted (SCoT) polymorphism to acquire biocontrol genes from potent antagonist. The six unique SCoT fragments amplified by genomic DNA of best mycoparasitic antagonist MTCC 796 strain are subjected to DNA sequencing resulted to confirm two functional sequences for activity related to biocontrol genes. The phylogenetic and molecular evolution of functional 824 bp of SCoT-3(920) and 776 bp of SCoT-6(806) fragments signify sequence homology with biocontrol genes endochitinase (partial cds of 203 amino acids) and novel hmgR genes (partial cds of 239 amino acids), respectively and the same were annotated and deposited in NCBI GenBank database. The hmgR gene is liable to be express hmg - CoA reductase which is a key enzyme for regulation of terpene biosynthesis and mycoparasitic strains produced triterpenes during antagonism to inhibit growth of fungal pathogen as evidenced with GC-MS profile. The biocontrol genes are found in best antagonist T. koningii MTCC 796 for mycoparasitic activity to restrain the growth of test pathogen R. solani.


Assuntos
Antibiose/genética , Evolução Molecular , Polimorfismo Genético/genética , Rhizoctonia/fisiologia , Trichoderma/genética , Trichoderma/fisiologia , Controle Biológico de Vetores , Filogenia , Trichoderma/enzimologia
16.
Curr Genet ; 62(3): 619-41, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26803831

RESUMO

The study was performed to examine 11 isolates of Trichoderma for their bio-control potentials against Sclerotium rolfsii Sacc. causing stem rot in groundnut. The antagonists Trichoderma were subjected to sequence related amplified polymorphism (SRAP) based molecular diversity analysis and compared with their hardness to S. rolfsii with respect to secretary antifungal and antioxidant profile. T. virens NBAII Tvs 12 evident highest (87.91 %) growth inhibition of test pathogen followed by T. koningii MTCC 796 (67.03 %) at 7 days after inoculation (DAI). Microscopic study confirmed biocontrol mechanism as mycoparasitism for Tvs 12 and antibiosis for MTCC 796. The growth inhibition of test pathogen was significantly negatively correlated with sclerotia formation and lipid peroxidation during antagonism due to release of secretary bioactive antioxidants by antagonists to terminate oxidative burst generated by S. rolfsii and causing inhibition of sclerotium formation. The GC-MS profile identified antifungal and antioxidant constituents hexadecane, 1,2-benzenedicarboxylic acid, mono (2-ethylhexyl) ester, 1-hexadecanesulfonyl chloride, and octadecane in potent antagonists Tvs 12; and nonacosane and octadecane in MTCC 796 along with two novel compounds 1-pentadecene and 1-heneicosyl formate for biocontrol activity. Molecular diversity of Trichoderma isolates associated with antagonistic activity was assessed by SRAP markers. The 115 primer combinations generate total 1328 amplified products of which, 1095 are shared polymorphic and 199 are unique polymorphic. The 15 SRAP combinations produced 18 bands to diagnose best antagonist Tvs 12 and 13 SRAP combinations generated 19 unique bands for identification of MTCC 796. The mycoparasitic antagonist Tvs 12 would be the best antagonist and released unique antifungal and antioxidant constituents to combat pathogen infection. The SRAP based genetic diversity indicates Tvs12 strain clustered with T. viride NBAII Tv23 and shared only 52 % similarity with other isolates of Trichoderma. The SRAP similarities explained substantial diversity (19-68 %) across Trichoderma isolates.


Assuntos
Antibiose , Antifúngicos/metabolismo , Antioxidantes/metabolismo , Basidiomycota , Variação Genética , Trichoderma/genética , Trichoderma/metabolismo , Antifúngicos/química , Antioxidantes/química , Agentes de Controle Biológico , Contagem de Colônia Microbiana , Cromatografia Gasosa-Espectrometria de Massas , Marcadores Genéticos , Metabolismo dos Lipídeos , Peroxidação de Lipídeos , Fenótipo , Filogenia , Polimorfismo Genético , Trichoderma/classificação , Trichoderma/crescimento & desenvolvimento
17.
Microb Pathog ; 91: 26-34, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26620080

RESUMO

The study was conducted to examine the antioxidant enzymes induced by Trichoderma viride JAU60 as initial defense response during invasion of rot pathogen Aspergillus niger Van Tieghem in five groundnut varieties under pot culture. Seed treatment of T. viride JAU60 reduced 51-58% collar rot disease incidence in different groundnut varieties under pathogen infected soil culture. The activities of the antioxidant enzymes, viz., superoxide dismutase (SOD, EC 1.15.1.1), guaiacol peroxidase (GPX, EC 1.11.1.7) and ascorbate peroxidase (APX, EC 1.11.1.11), elevated in response to pathogen infection, in higher rate by tolerant varieties (J-11 and GG-2) compared with susceptible (GAUG-10, GG-13, GG-20) and further induced by T. viride treatment. Trichoderma treatment remarkably increased the 2.3 fold SOD, 5 fold GPX and 2.5 fold APX activities during disease development in tolerant varieties and the same was found about 1.2, 1.5 and 2.0 folds, respectively, in susceptible varieties. Overall, T. viride JAU60 treated seedlings (T3) witnessed higher activities of SOD (1.5 fold), GPX (3.25 fold) and APX (1.25 fold) than pathogen treatment (T2) possibly suggest the induction of antioxidant defense response by Trichoderma bio-controller to combat oxidative burst produced by invading pathogen.


Assuntos
Arachis/imunologia , Aspergillus niger/fisiologia , Doenças das Plantas/microbiologia , Trichoderma/fisiologia , Arachis/enzimologia , Arachis/genética , Arachis/microbiologia , Ascorbato Peroxidases/genética , Ascorbato Peroxidases/imunologia , Catalase/genética , Catalase/imunologia , Doenças das Plantas/genética , Doenças das Plantas/imunologia , Proteínas de Plantas/genética , Proteínas de Plantas/imunologia , Superóxido Dismutase/genética , Superóxido Dismutase/imunologia
18.
Infect Genet Evol ; 34: 314-25, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26160540

RESUMO

The study examine induction of defense enzymes involved in phenylpropanoid pathway and accumulation of pathogenesis related proteins in rot pathogen (Aspergillus niger Van Tieghem) challenged groundnut seedlings in response to Trichoderma viride JAU60. Seeds of five groundnut varieties differing in collar rot susceptibility were sown under non-infested, pathogen infested and pathogen+T. viride JAU60 seed treatment. Collar rot disease evident between 31.0% (J-11, GG-2) and 67.4% (GG-20) in different groundnut varieties under pathogen infested which was significantly reduced from 58.1% (J-11, GG-2) to 51.6% (GG-20) by Trichoderma treatment. The specific activities of polyphenol oxidase (EC 1.14.18.1) and ß-1,3 glucanase (EC 3.2.1.6) elevated 3.5 and 2.3-fold, respectively, at 3 days; phenylalanine ammonia lyase (EC 4.3.1.5) evident 1.6-fold higher at 6 days; and chitinase (EC 3.2.1.14) sustained 2.3-2.8 folds up to 9 days in Trichoderma treated+pathogen infested seedlings of tolerant varieties (J-11, GG-2) compared with moderate and susceptible (GAUG-10, GG-13, GG-20). T. viride JAU60 induces defense enzymes in a different way for tolerant and susceptible varieties to combat the disease. This study indicates the synergism activation of defense enzymes under the pathogenic conditions or induced resistance by T. viride JAU60 in a different groundnut varieties susceptible to collar rot disease.


Assuntos
Arachis/microbiologia , Doenças das Plantas/microbiologia , Trichoderma/fisiologia , Arachis/imunologia , Aspergillus/fisiologia , Resistência à Doença , Interações Microbianas , Doenças das Plantas/imunologia , Sementes/microbiologia
19.
Braz. j. microbiol ; 43(1): 43-52, Jan.-Mar. 2012. ilus, tab
Artigo em Inglês | LILACS | ID: lil-622789

RESUMO

Twelve isolates of Trichoderma (six of T. harzianum, five of T. viride, one of T. virens), which reduced variably the incidence of collar rot disease caused in peanut by Aspergillus niger Van Tieghem, were evaluated for their potential to produce lytic enzymes during in vitro antagonism. T. viride 60 inhibited highest (86.2%) growth of test fungus followed by T. harzianum 2J (80.4%) at 6 days after inoculation (DAI) on PDA media. The specific activities of chitinase, â-1,3-glucanase and protease were 11, 3.46 and 9 folds higher in T6 antagonist (T. viride 60 and A. niger interactions) followed by 8.72, 2.85 and 9 folds in T8 antagonist (T. harzianum 2J and A. niger interactions), respectively, compared to the activity produced by control petri plate T13 (A. niger alone) at 6 DAI. Activity of these lytic enzymes induced in antagonists' plates comprises the growth of Trichoderma isolates. However, cellulase and poly galacturonase were found least amount in these antagonists treatment. A significant positive correlation (p=0.01) between percentage growth inhibition of test fungus and lytic enzymes - (chitinase, â-1,3-glucanase and protease) in the culture medium of antagonist treatment established a relationship to inhibit growth of fungal pathogen by increasing the levels of these enzymes. Among the Trichoderma isolates, T. viride 60 was found best strain to be used in biological control of plant pathogen A. niger.


Assuntos
Arachis/enzimologia , Aspergillus niger/enzimologia , Aspergillus niger/isolamento & purificação , Glucanos/análise , Técnicas In Vitro , Trichoderma/enzimologia , Trichoderma/isolamento & purificação , Amostras de Alimentos
20.
Braz J Microbiol ; 43(1): 43-52, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-24031802

RESUMO

Twelve isolates of Trichoderma (six of T. harzianum, five of T. viride, one of T. virens), which reduced variably the incidence of collar rot disease caused in peanut by Aspergillus niger Van Tieghem, were evaluated for their potential to produce lytic enzymes during in vitro antagonism. T. viride 60 inhibited highest (86.2%) growth of test fungus followed by T. harzianum 2J (80.4%) at 6 days after inoculation (DAI) on PDA media. The specific activities of chitinase, ß-1,3-glucanase and protease were 11, 3.46 and 9 folds higher in T6 antagonist (T. viride 60 and A. niger interactions) followed by 8.72, 2.85 and 9 folds in T8antagonist (T. harzianum 2J and A. niger interactions), respectively, compared to the activity produced by control petri plate T13 (A. niger alone) at 6 DAI. Activity of these lytic enzymes induced in antagonists' plates comprises the growth of Trichoderma isolates. However, cellulase and poly galacturonase were found least amount in these antagonists treatment. A significant positive correlation (p=0.01) between percentage growth inhibition of test fungus and lytic enzymes - (chitinase, ß-1,3-glucanase and protease) in the culture medium of antagonist treatment established a relationship to inhibit growth of fungal pathogen by increasing the levels of these enzymes. Among the Trichoderma isolates, T. viride 60 was found best strain to be used in biological control of plant pathogen A. niger.

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