RESUMO
Though numerous bacteria have been used as probiotics by industries, at present, Saccharomyces boulardii and Saccharomyces cerevesiae are the only yeast probiotics which are industrially exploited. In view of this, yeast probiotics were isolated from traditional fermented foods and products collected from different parts of Karnataka, India. In this work, we have studied the probiotic attributes of ten yeast isolates isolated from different traditionally fermented foods and products. About 73 yeast isolates were initially isolated by serially diluting the samples and plating on the Potato Dextrose Agar (PDA) plates. The spot assay was performed to screen the yeast isolates against test pathogens. Ten isolates were selected based on their significant antimicrobial activity. These isolates were subjected to biochemical characterization and then assessed for probiotic properties. The ability of probiotics to endure at pH 2.0 and tolerate bile conditions (0.3%) are crucial attributes for the survival in the gastrointestinal tract (GIT). The yeast isolates were also assessed for cell surface hydrophobicity and autoaggregation capabilities. All the ten isolates showed endurance in GIT tract and > 40% of adhesion. The study further examined cholesterol assimilation, antioxidant and antagonistic properties of the yeasts. Subsequently, the molecular characterization was performed by isolating the DNA of yeast isolates by phenol-chloroform method and identified molecularly through sequencing of D1/D2 regions. The isolates tested negative for gelatinase and DNase and were non-haemolytic indicating they are safe for consumption. Among ten isolates, Meyerozyma guillermondii (MYSY23), Meyerozyma caribbica (MYSY22) and Meyerozyma guillermondii (MYSY19) showed significant results for all probiotic and functional characteristics with greater than 65% survivability in GIT tract and > 50% of antagonistic activity against test pathogens and also proved non-cytotoxic and safe. These findings suggest that yeasts with significant probiotic attributes could be recommended for various probiotic application.
RESUMO
Chia (Salvia hispanica L., Lamiaceae) is an important commercial and medicinal crop recently popularized in India and widely cultivated in Karnataka (Joy et al., 2022). During the field survey of chia crop diseases, characteristic virescence like symptoms were observed at Main Agricultural Research Station, UAS, Raichur as well as at Mysuru and HD Kote region. The incidence was ranged from 2 - 4 per cent in an area of 30 hectares. Typical symptoms associated with chia are malformed shoot and/or inflorescence axis with reduced floral parts with greenish florets. The stem axis become thick, flattened, leaves are reduced towards terminal region. A total of five phytoplasma suspected samples and five suspected healthy samples were used for identification purpose. The Plant Genomic DNA Miniprep Kit (Sigma Aldrich, USA) was used to extract the DNA from five symptomatic and five asymptomatic samples and the DNA was used as template to amplify the phytoplasma-specific 16S rDNA gene using P1/P7 primers (Deng and Hiruki, 1991; Schneider et al., 1995) followed by nested PCR using R16F2n/R16R2 primers (Gundersen and Lee 1996). The expected 1.25-kb amplicon was detected from the suspected symptomatic samples. Nested PCR products were purified and sequenced from both the directions using ABIX370 Genetic Analyzer (Applied Biosystems, Waltham, MA). The analysis revealed that all five sequences shared 100 per cent identity with Candidatus Phytoplasma aurantifolia (OM649850, ON975012) and Tomato big bud phytoplasma (EF193359). The in-silico RFLP pattern of F2n/R2 primed region of 16S rDNA gene analyzed by using iPhyClassifier (Zhao et al. 2009) revealed that the sequence shared 98.72 per cent nucleotide sequence similarity with coefficient value of 1.00 to the reference strain RFLP pattern of 16Sr group II, subgroup D (witches'-broom disease of lime; U15442). Based on 16SrDNA sequences and in-silico RFLP analysis, the phytoplasma associated with the chia virescence was identified as a member of 16SrII-D group. Further, SecA gene was also amplified from the samples using SecAfor1/SecArev3 primer pair (Hodgetts et al., 2008). All samples produced ~400 bp products and sequenced as detailed above. Sequence analysis by nBLAST revealed 100 per cent similarity to Ca. P. australasia (MW020545) and Ca. P. aurantifolia isolate Idukki Kerala 1 (MK726369) both representing 16SrII-D group phytoplasma. The representative sequence (16Sr: PP359693, PP359694; secA:PP386558, PP386559) were deposited in GenBank. Chia virescence phytoplasma belonging to Ca. phytoplasma australasia has not been reported anywhere. The phytopathological studies associated with chia crop are very limited. Joy et al. (2022) reported the occurrence of foot rot disease caused by Athelia rolfsii. Several hosts are recorded to be associated with 16SrII D phytoplasma which includes china aster, eggplant and crotalaria (Mahadevakumar et al., 2017, Yadav et al., 2016a, b). Now the wide occurrence of the phytoplasma in the area might have transmitted by vectors. The occurrence of virescence is of great importance as it affects the overall yield which reduces the market value. To our knowledge, this is the first report of a group 16SrII-D phytoplasma associated with chia virescence in India.
RESUMO
Alternaria alternata, a notorious phytopathogenic fungus, has been documented to infect several plant species, leading to the loss of agricultural commodities and resulting in significant economic losses. Lactic acid bacteria (LAB) hold immense promise as biocontrol candidates. However, the potential of LABs derived from fruits remains largely unexplored. In this study, several LABs were isolated from tropical fruit and assessed for their probiotic and antifungal properties. A total of fifty-five LABs were successfully isolated from seven distinct fruits. Among these, seven isolates showed inhibition to growth of A. alternata. Two strains, isolated from fruits: Ficus benghalensis, and Tinospora cordifolia exhibited promising antifungal properties against A. alternata. Molecular identification confirmed their identities as Lactiplantibacillus plantarum MYSVB1 and MYSVA7, respectively. Both strains showed adaptability to a wide temperature range (10-45°C), and salt concentrations (up to 7%), with optimal growth around 37 °C and high survival rates under simulated gastrointestinal conditions. Among these two strains, Lpb. plantarum MYSVB1 demonstrated significant inhibition (p < 0.01) of the growth of A. alternata. The inhibitory effects of cell-free supernatant (CFS) were strong, with 5% crude CFS sufficient to reduce fungal growth by >70% and complete inhibition by 10% CFS. Moreover, the CFS was inhibitory for both mycelial growth and conidial germination. CFS retained its activity even after long cold storage. The chromatographic analysis identified organic acids in CFS, with succinic acid as the predominant constituent, with lactic acid, and malic acid in descending order. LAB strains isolated from tropical fruits showed promising probiotic and antifungal properties, making them potential candidates for various applications in food and agriculture.
RESUMO
The primary objective of this study was to assess the probiotic attributes and antifungal activity of lactic acid bacteria (LAB) against the fungus, Trichophyton tonsurans. Among the 20 isolates screened for their antifungal attributes, isolate MYSN7 showed strong antifungal activity and was selected for further analysis. The isolate MYSN7 exhibited potential probiotic characteristics, having 75 and 70% survival percentages in pH3 and pH2, respectively, 68.73% tolerance to bile, a moderate cell surface hydrophobicity of 48.87%, and an auto-aggregation percentage of 80.62%. The cell-free supernatant (CFS) of MYSN7 also showed effective antibacterial activity against common pathogens. Furthermore, the isolate MYSN7 was identified as Lactiplantibacillus plantarum by 16S rRNA sequencing. Both L. plantarum MYSN7 and its CFS exhibited significant anti-Trichophyton activity in which the biomass of the fungal pathogen was negligible after 14 days of incubation with the active cells of probiotic culture (106 CFU/ml) and at 6% concentration of the CFS. In addition, the CFS inhibited the germination of conidia even after 72 h of incubation. The minimum inhibitory concentration of the lyophilized crude extract of the CFS was observed to be 8 mg/ml. Preliminary characterization of the CFS showed that the active component would be organic acids in nature responsible for antifungal activity. Organic acid profiling of the CFS using LC-MS revealed that it was a mixture of 11 different acids, and among these, succinic acid (9,793.60 µg/ml) and lactic acid (2,077.86 µg/ml) were predominant. Additionally, a scanning electron microscopic study revealed that CFS disrupted fungal hyphal structure significantly, which showed scanty branching and bulged terminus. The study indicates the potential of L. plantarum MYSN7 and its CFS to control the growth of T. tonsurans. Furthermore, in vivo studies need to be conducted to explore its possible applications on skin infections.
RESUMO
During past twenty years the opportunistic fungal infections have been emerging, causing morbidity and mortality. The fungi belonging to Aspergillus, Mucor, Rhizopus, Candida, Fusarium, Penicillium, Dermatophytes and others cause severe opportunistic fungal infections. Among these Aspergillus and Candida spp cause majority of the diseases. The continuum of fungal infections will prolong to progress in the surroundings of the growing inhabitants of immunocompromised individuals. Presently many chemical-based drugs were used as prophylactic and therapeutic agents. Prolonged usage of antibiotics may lead to some severe effect on the human health. Also, one of the major threats is that the fungal pathogens are becoming the drug resistant. There are many physical, chemical, and mechanical methods to prevent the contamination or to control the disease. Owing to the limitations that are observed in such methods, biological methods are gaining more interest because of the use of natural products which have comparatively less side effects and environment friendly. In recent years, research on the possible use of natural products such as probiotics for clinical use is gaining importance. Probiotics, one of the well studied biological products, are safe upon consumption and are explored to treat various fungal infections. The antifungal potency of major groups of probiotic cultures such as Lactobacillus spp, Leuconostoc spp, Saccharomyces etc. and their metabolic byproducts which act as postbiotics like organic acids, short chain fatty acids, bacteriocin like metabolites, Hydrogen peroxide, cyclic dipeptides etc. to inhibit these opportunistic fungal pathogens have been discussed here.
RESUMO
Moringa oleifera Lam. (Moringaceae) is one of the important leafy vegetable trees widely spread from India to Africa and widely used as food and medicine. During field investigation (July, 2021) of drumstick fields in Hiriyur (13°95'79" N; 76°64'45" E), Karnataka, fruits of drumstick plants showed a characteristic rot disease with an incidence ranging from 10-12% in an area of 5 hectares surveyed. Initially water soaked lesions turned to small necrotic lesions and later coalesced to form larger areas covered with white mycelial growth leading to softening and later mummification of fruits. Infected fruits were collected (n=5) and infected fruit parts (margins of healthy and infected tissues) were cut into small pieces, surface sterilized with sodium hypochlorite (2%, v/v) and blotter dried after three sterile water washes. An associated fungal species was isolated on PDA medium amended with Streptomycin (40 mg/L) and incubated at 28 ºC for 1 week. The fungal isolate grown on PDA had dense, white, aerial mycelium with light brown coloration on the reverse side of the agar medium. Morphological characteristics of conidia were determined for single-spore cultures grown on water agar media. Microconidia were single-celled, hyaline, non septate, ovoid, and 8.4 - 9.8 × 1.8 - 2.94 µm (n=20). Macroconidia were three- to five-septate, slightly curved, tapered at the apex, and 24.4 - 28 × 2 - 4 µm (n=20). Based on morphological characters' pathogen was identified as Fusarium sp. (Leslie and Summerell, 2006). Further, three representative isolate (SMG, MYS1 & MYS2) were subjected for molecular identification. The genomic DNA was extracted following CTAB method and ITS-rDNA was amplified using ITS1/ITS4 primers (White et al., 1990) and translation elongation factor (tef-1α) gene was amplified using EF1/EF2 primers (O'Donnell et al., 2009) respectively. ITS-rDNA sequence shared 100% sequence similarity (650bp / 650bp) with reference sequence F. incarnatum (ON226997) followed by 100% sequence identity to F. equiseti (KT277307 & MT953927). But, tef-1α gene sequence analysis in nBLAST showed that the sequence shared 100% (123/123bp) identity with F. incarnatum (F. incarnatum NEAU-TG1 MH920853; M2JP3 OP312673; WEH ON456146). Combined phylogenetic analysis revealed that the isolate shared a common clade with reference sequence of F. incarnatum in the Fusarium-incarnatum-equiseti species complex thus confirming the identity of the isolated pathogen as F. incarnatum (Rob. ex Desm.) Sacc. 1886. The sequences obtained in the present study are deposited in GenBank database (ITS: OP508729, OQ159019, OQ159020 and tef-1α: OP477394, OQ176254, OQ176255). To prove Koch's postulates, pathogenicity test was carried out by inoculating healthy drumstick fruits cv. Bhagya (n=10) with spore suspension (105 conidia/ml). Control fruits (n=5) were sprayed with sterile water. The experiments were conducted in triplicates and repeated twice. Inoculated and control fruits were kept in a moist chamber at 26 ± 2°C for 2 days and observed at regular intervals. Development of disease symptoms were recorded on 52 out of 60 inoculated fruits which were identical with symptoms seen in the field and all control fruits remained symptomless. Identity was confirmed after re-isolation by morphology and culture studies. To the best of our knowledge, this is the first report of F. incarnatum causing a fruit rot of drumstick in India. This disease affected the cost of drumstick production and contributed to the decline in production in India.
RESUMO
Biojarosite as a replacement for commercial iron catalyst in the oxidative degradation of the dye Rhodamine B was confirmed and established. Investigations on the oxidative degradation by Fenton's oxidation and UV-Fenton's oxidation with EDTA at neutral pH were conducted and degradation of target compound was evaluated. UV-Fenton's oxidation was shown to be efficient over Fenton's oxidation in the degradation of Rhodamine B with removal efficiency of 90.0%. Design of Experiments was performed with Box-Behnken design. Investigation was conducted for the predicted values separately for both Fenton's oxidation and UV-Fenton's oxidation and the Rhodamine B removal was taken as response. Variable parameters biojarosite, H2O2 dosage and EDTA were optimized in the range of 0.1-1 g/L, 2.94-29.4 mM and 10-100 mM, respectively. A quadratic regression model is fitted for both Fenton's and UV-Fenton's oxidation. Analysis of variance (ANOVA) is performed and model fit is discussed.
Assuntos
Peróxido de Hidrogênio , Ferro , Ferro/química , Peróxido de Hidrogênio/química , Ácido Edético , Oxirredução , Concentração de Íons de HidrogênioRESUMO
AIMS: In the study, seven Plant Growth Promoting Rhizobacteria (PGPR) Azotobacter species were screened against three strains of Fusarium verticillioides to test its antifungal activity. Azotobacter strains were tested for the degradation of fumonisin produced by F. verticillioides. Secondary metabolites were isolated and characterized from the Azotobacter strains for the first time. METHODS AND RESULTS: Potential seven Azotobacter species antifungal activity was tested following the dual culture assay against three strains of Fusarium verticillioides namely FVM-42, FVM-86 and MTCC156 estimating the substantial zone of inhibition. Azotobacter species AZT-31 and AZT-50 strains significantly inhibited the growth of F. verticillioides recording drastic growth enhancement of maize under in-vitro conditions by calculating the infection incidence, vigour index and germination percentage. As confirmation, dereplication studies were conducted for the reconfirmation of Azotobacter strains by isolating from rhizoplane. Azotobacter strains played a key role in the degradation of fumonisin produced by F. verticillioides reporting 98% degradation at 2 h of incubation with the pathogen. Furthermore, in the study first time, we have tried to isolate and characterize the secondary metabolites from the Azotobacter strains exhibiting six compounds from the species AZT-31 (2) and AZT-50 (4). Preliminary in-vitro experiments were carried out using the compounds extracted to check the reduction of infection incidence (90%) and increase in germination percentage upto 50 to 70% when compared to the test pathogen. CONCLUSION: Azotobacter strains referred as PGPR on influencing the growth of plant by producing certain substances that act as stimulators on inhibiting the growth of the pathogen. SIGNIFICANCE AND IMPACT OF THE STUDY: The future perspective would be the production of an active combination of carboxamide compound and Azotobacter species for preventively controlling the phytopathogenic fungi of plants and crops and also towards the treatment of seeds.
Assuntos
Azotobacter , Fumonisinas , Fusarium , Antifúngicos/farmacologia , Fumonisinas/metabolismo , Zea mays/microbiologiaRESUMO
Fermented foods are sources of functionally salient microbes. These microbes when ingested can regulate biomolecule metabolism which has a plethora of health benefits. Lactic acid bacteria species (LABs) isolated from fermented beetroot were biochemically characterized and validated using 16s rRNA sequence. Also, an in vitro assay was conducted to confirm the probiotic activity of the isolates. The cell-free supernatant (CS), cell-free extract (CE), and intact cell (IC) were evaluated for α-glucosidase and α-amylase inhibition. The six isolates RAMULAB01-06 were categorized to be Lactobacillus spp. by observing phenotypic and biochemical characters. Molecular validation using 16S rDNA sequencing, followed by homology search in NCBI database, suggested that the isolates are >95% similar to L. paracasei and L. casei. Also, isolates exhibited probiotic potential with a high survival rate (>96%) in the gastrointestinal condition, and adherence capability (>53%), colonization (>86%), antibacterial, and antibiotic activity. The safety assessments expressed that the isolates are safe. The α-glucosidase and α-amylase inhibition by CS, CE, and IC ranged from 3.97 ± 1.42% to 53.91 ± 3.11% and 5.1 ± 0.08% to 57.15 ± 0.56%, respectively. Hence, these species have exceptional antidiabetic potential which could be explicated to its use as a functional food and health-related food products.
RESUMO
Antifungal efficacy of Azotobacter salinestris against trichothecene-producing Fusarium spp. was investigated in maize, sorghum, and wheat. The three cereals were subjected to four treatments as control (T1), Fusarium alone (T2), combination of Fusarium and A. salinestris treatment (T3), and only A. salinestris (T4). All the treatments were evaluated for total mass of seedlings, root and shoot length, seed germination, and vigor index (VI), and extent of rhizoplane colonization by A. salinestris was investigated. Further, greenhouse studies were conducted to learn the efficacy of A. salinestris in vivo conditions. Antifungal efficacy was tested by the dual-culture method which resulted in significant reduction in Fusarium growth. Infection by Fusarium was reduced up to 50% in treated cereals such as maize, sorghum, and wheat, and there was also significant increase in seedling mass in the three hosts. Maize showed the highest VI (1859.715), followed by sorghum (1470.84), and wheat (2804.123) with A. salinestris treatment. In addition, seed germination was enhanced to 76% in maize, 69% in sorghum, and 68% in wheat, respectively. Efficacy of rhizoplane colonization showed successful isolation of A. salinestris with high CFU rate, and furthermore, significant colonization inhibition by Fusarium spp. was observed. In the greenhouse conditions, on the 45th day of the experimental set-up, the highest shoot length/root length recorded in maize was 155.70/70.0 cm, in sorghum 165.90/48.0 cm, and in wheat 77.85/56.0 cm, and the maximum root mass recorded was 17.53 g in maize, 4.52 g in sorghum, and 1.90 g in wheat. Our present study showed that seed treatment by A. salinestris, may be used as an alternate biocontrol method against Fusarium infection in maize, sorghum, and wheat.
RESUMO
Cluster bean (Cyamopsis tetragonoloba (L.) Taub.) is an important vegetable crop cultivated widely in India. During a field survey in November 2021, about 60% of plants exhibited characteristic powdery mildew disease symptoms and signs in a 15 ha field in Northern Karnataka (Raichur), India. Initially, the symptoms and signs appeared as tan lesions, which later became small, circular and chlorotic. The abaxial surface turned yellow and was covered with white mycelial growth. As the disease progressed, white mycelia grew on the adaxial leaf surface, stems and pods as well. In severe infections, drying and premature defoliation of infected leaves were observed. Infected leaf samples with mycelia were collected (n=8) and the fungus was subjected to morphological and molecular observations. Mycelia on leaves was characterized as epiphytic, amphigenous, producing dense, white patches on the upper and lower leaf surfaces, stem and young pods. Hyphae were hyaline, thin-walled, 1.8 to 4.2 µm wide with erect conidiophores consisting of a cylindrical foot-cell, straight flexuous at the base and measured 20 to 36 × 6 to 9 µm (n=30), followed by 1 to 2 shorter cells. Ellipsoid conidia were produced singly and measured 28 to 42 × 12 to 20 µm (n=30) without fibrosin bodies. Chasmothecia were not observed. A reference specimen was deposited at the Institution of Excellence, University of Mysore Herbarium (UOM-IOE 2022_1). The morphology and other characteristics of conidia were consistent with an Erysiphe species (Braun and Cook 2012). Genomic DNA was isolated from a conidial suspension harvested from the powdery mildew affected cluster bean samples. The ITS region was amplified from three samples using powdery mildew-specific primer pair PN23/PN34 and sequenced directly (Chen et al. 2008). nBLAST analysis revealed that the ITS sequence shared 100% similarity with the reference sequence (E. diffusa vouchers HMJAU02177 - KM260363, BRIP 71013 - MW009058) of Erysiphe diffusa (Cooke & Peck) U. Braun & S. Takam. In addition to 100% match to voucher specimens of E. diffusa, there were no vouchers from other species that also had 100% match. The representative sequences were deposited in GenBank with accession numbers OM669776 - OM669778. Koch's postulates were conducted on healthy cluster bean plants grown under greenhouse conditions. Conidia were harvested from infected leaves, suspended in water and sprayed on 40 to 50-day-old cluster bean plants (28 ± 2°C and >70% relative humidity). The development of powdery mildew symptoms was recorded on 22 plants after 10-14 days of post inoculation. Control plants inoculated with sterile water remained healthy without powdery mildew symptoms. Microscopic observation of spores from inoculated plants confirmed the pathogen as E. diffusa. The genus Erysiphe is known to infect many crop plants. E. diffusa has been reported to infect Vigna radiata, Glycine max and Phaseolus mungo in Australia (Kelly et al. 2021). No reports are available at USDA's host-fungus database for cluster bean and E. diffusa (Farr and Rossman 2022). To the best of our knowledge, this is the first report of E. diffusa associated with powdery mildew of cluster bean in India. Further comprehensive investigations will shed a light on the economic impact of powdery mildew disease on the cluster bean in India.
RESUMO
Probiotics are vital and beneficial organisms which offers the health benefits to the host organisms. The fungal probiotic field is one of the developing fields nowadays. Yeast has an enormous and diverse group of microorganisms that is attracting and expanding the attention from researchers and industries. Saccharomyces boulardii, the only patented strain belonging to yeast genera for the human use, has been broadly evaluated for its probiotic effect. Yeasts belonging to the genera Debaryomyces, Pichia, Yarrowia, Meyerozyma, Kluyveromyces etc.., have attained more interest because of their beneficial and probable probiotic features. These yeast probiotics produce VOCs (Volatile organic compounds), mycocins and antimicrobials which shows the antagonistic effect against pathogenic fungi and bacteria. Additionally, those yeasts have been recorded as good plant growth promoting microorganisms. Yeast has an important role in environmental applications such as bioremediation and removal of metals like chromium, mercury, lead etc., from waste water. Probiotic yeasts with their promising antimicrobial, antioxidant, anticancer properties, cholesterol assimilation and immunomodulatory effects can also be utilized as biotherapeutics. In this review article we have made an attempt to address important yeast probiotic attributes.
RESUMO
A total of 130 isolates were screened, twelve isolates were characterized for probiotic attributes and two isolates with best probiotic features were evaluated in the study. Isolates MYSRD108 and MYSRD71 survived gastric conditions and were susceptible to tested antibiotics. Isolates showed more vital cell surface traits such as autoaggregation of 89.2 and 88.5% and cell surface hydrophobicity of 61 and 64%. PCR amplification followed by 16sRNA sequencing results confirmed that the isolates as Lactobacillus casei (MYSRD 108) and Lactobacillus plantarum (MYSRD 71). During this study, the Cells and their Cell Free Supernatant (CFS) were examined for antimicrobial activity. Both the isolates inhibited different bacterial pathogens in which the growth of S. paratyphi was significantly reduced. Further, their CFS also showed inhibitory effects against S. paratyphi with agar well diffusion and Minimum Inhibitory Concentration using Broth micro dilution method. The antimicrobial compounds in the CFS was characterized to different constraints such as pH neutralization, heat treatment, Hydrogen peroxide test and storage stability at -20> °C and represented that the antagonistic acitivity against Salmonella is due to the presence of organic acids in the supernatants that lowered the pH. These strains were further examined for the inhibition of S. paratyphi biofilm. The results indicated that CFS reduced S. paratyphi biofilm by more than 75% and the number of Salmonella biofilm was effectively reduced using 15% concentration of CFS. These strains may be used to produce antimicrobial compounds which can be a substitute for chemical preservatives in food industry.
RESUMO
Linseed commonly called as flaxseed (Linum usitatissimum Linn.) is an important oilseed crop cultivated widely in Northern parts of Karnataka. During, 2019 (January-February), a characteristic disease was noticed with symptoms that resembled phytoplasma or like disease symptoms. The incidence was ranged from 6·5 to 16·5% in the experimental station of Raichur Agricultural University. The typical symptoms observed were virescence of floral parts, fasciation of the inflorescence axis, phyllody, stunted and flattened stem with reduced leaves. Symptomatic and healthy samples were collected and processed for molecular detection of phytoplasma. Total DNA was isolated from four infected plants and two healthy plants. The 16S rDNA region was amplified using P1/P7 followed by R16F2n/R16R2 primer pair which showed the amplification of expected amplicon size from all four infected samples. Furthermore, the SecA gene was amplified using SecA1/SecA3 primers. The PCR amplified products were subjected for direct sequencing from both directions and the consensus sequences were obtained and nBLAST search analysis revealed that the 16Sr RNA and SecA sequences were sharing maximum similarity (100%) with the reference sequence of Ca. P. cynodontis. The sequences were analysed phylogenetically by constructing a Phylogram independently by NJ method along with reference sequence of 16S rRNA region and SecA region retrieved from GenBank database showed that the phytoplasma sequence from linseed phyllody of the present study placed in a distinct clade along with reference sequence of "Ca. P. cynodontis" thus confirming the identity phylogenetically. Furthermore, iPhyClassifier and virtual RFLP proved that the phytoplasma belonged to 16SrXIV (subgroup A) phytoplasma. Previously linseed is known to be associated with 16SrII-D phytoplasma but the association of the 16SrXIV-A group of phytoplasma is not reported so far. Therefore, this is the new host record for Ca. P. cynodontis (16SrXIV-A) phytoplasma associated with linseed stem fasciation, phyllody from India.
Assuntos
Linho , Phytoplasma , DNA Bacteriano/genética , Humanos , Índia , Filogenia , Phytoplasma/genética , Doenças das Plantas , RNA Ribossômico 16S/genética , Análise de Sequência de DNARESUMO
The use of probiotics and antifungal capabilities of the lactic acid bacteria (LAB) isolated from different niches is a strategy to prepare functional cultures and biopreservatives for food/feed industries. In the present study, LAB strains isolated from an Indian traditional fermented food, Pozha, were evaluated for their probiotic properties and biocontrol potential. A total of 20 LAB isolates were selected from Pozha samples collected aseptically and screened for their antagonistic activity against Fusarium verticillioides. Among the bioactive isolates, Lacticaseibacillus brevis MYSN105 showed the highest antifungal activity in vitro, causing some morphological alterations such as damaged mycelia and deformed conidia. Cell-free supernatant (CFS) from L. brevis MYSN105 at 16% concentration effectively reduced the mycelial biomass to 0.369 g compared to 1.938 g in control. Likewise, the conidial germination was inhibited to 20.12%, and the seed treatment using CFS induced a reduction of spore count to 4.1 × 106 spores/ml compared to 1.1 × 109 spores/ml for untreated seeds. The internal transcribed spacer (ITS) copy number of F. verticillioides decreased to 5.73 × 107 and 9.026 × 107 by L. brevis MYSN105 and CFS treatment, respectively, compared to 8.94 × 1010 in control. The L. brevis MYSN105 showed high tolerance to in vitro gastrointestinal conditions and exhibited high adhesive abilities to intestinal epithelial cell lines. The comparative genome analysis demonstrated specific secondary metabolite region coding for bacteriocin and T3PKS (type III polyketide synthase) possibly related to survival and antimicrobial activity in the gut environment. Our results suggest that L. brevis MYSN105 has promising probiotic features and could be potentially used for developing biological control formulations to minimize F. verticillioides contamination and improve food safety measures.
RESUMO
Probiotic bacteria were isolated from different traditional fermented foods as there are several such foods that are not well explored for their probiotic activities. Hence, the present study was conducted to find the potential of lactic acid bacteria (LAB) as probiotics that were isolated from the sap extract of the coconut palm inflorescence - Neera, which is a naturally fermented drink consumed in various regions of India. A total of 75 isolates were selected from the Neera samples collected aseptically in the early morning (before sunrise). These isolates were initially screened for cultural, microscopic, and biochemical characteristics. The initial screening yielded 40 Gram-positive, catalase-negative isolates that were further subjected to acid - bile tolerance with resistance to phenol. Among 40 isolates, 16 survived screening using analysis of cell surface hydrophobicity, auto aggregation with adhesion to epithelial cells, and gastric-pancreatic digestion for gastrointestinal colonization. The isolates were also assessed for antimicrobial, antibiotic sensitivity, and anti-oxidative potential. The safety of these isolates was evaluated by their hemolytic and deoxyribonuclease (DNase) activities. Based on these results, seven isolates with the best probiotic attributes were selected and presented in this study. These LAB isolates, with 51.91-70.34% survival at low pH, proved their resistance to gastric conditions. The cell surface hydrophobicity of 50.32-77.8% and auto aggregation of 51.02-78.95% represented the adhesion properties of these isolates. All the seven isolates exhibited good antibacterial and antifungal activity, showing hydroxyl-scavenging activity of 32.86-77.87%. The results proved that LAB isolated from Neera exhibited promising probiotic properties and seem favorable for use in functional fermented foods as preservatives.
RESUMO
The investigation reports the application of biogenic jarosite, an iron hydroxy sulfate mineral in Fenton's Oxidation process. Ametryn, a herbicide detrimental to aquatic life and also to human is treated by Fenton's oxidation process using synthesized iron mineral, jarosite. The jarosite synthesis was carried out by using an isolated Acidithiobacillus ferrooxidans bacterial strain with ferrous as an iron supplement. The isolated strain was characterized by molecular techniques and biooxidation activity to ferrous to ferric iron was checked. On Fenton's treatment ametryn degradation upto 84.9% and COD removal to the extent of 56.1% was observed within 2â¯h of treatment and the reaction follows the pseudo first order kinetics with the curve best fit. The slight increase in kinetic rate constant on jarosite loading rate increase from 0.1â¯g/L to 0.5â¯g/L with H2O2 dosage of 100â¯mg/L confirms that jarosite has a catalytic role in the removal of ametryn. Mass spectroscopy analysis of treated synthetic ametryn solution at various intervals reveal the degradation follows dealkylation and hydroxylation pathway with the formation of three major intermediate compounds discussed here.