Cultural, Morphological and Pathogenic Variability in Alternaria cyamopsidis Causing Alternaria Blight of Clusterbean in Rajasthan

Background: Alternaria cyamopsidis (Rang. and Rao) causes Alternaria blight of clusterbean and it is one of the significant disease of clusterbean. Studies were conducted to compare the Cultural, morphological and pathogenic variability among ten isolates of Alternaria cyamopsidis from clusterbean, in five districts of Rajasthan viz., Bikaner, Barmer, Churu, Hanumangarh and Jaipur. Methods: During 2016-17 exhaustive survey was conducted in clusterbean growing areas of Rajasthan and collected diseased samples of clusterbean caused by Alternaria. All the samples were processed for isolation, purification and their pathogenicity was proved in cagehouse and laboratory and standard methods were adopted for cultural and morphological variability study. Result: All the isolates showed variation in their morphological characters, i.e., colony color and shape; conidial number, size, width, length, shape and septation on PDA. Out of ten isolates two isolate, viz., AlcyJp1 and AlcyJp2 showed maximum colony diameter 89.50 and 86.30 mm, respectively. All the isolates varied in their spore length and width and virulent on the tested variety of clusterbean for virulence. AlcyJp1 was the most virulent and produced maximum (65.50%) disease intensity, followed by AlcyJp2 isolate (61.22%).

Evaluation of the effects of pigments produced by environmental bacteria on cancer cells

Cancers are a collection of incapacitating diseases in which cells initiate to divide and spread to adjacent tissues in an uncontrolled manner. Many researches demonstrated the potential of bacterial pigments as promising anticancer agents Therefore, in this study, the cytotoxic effects of bacterial pigments were evaluated in breast and colon cancer cell lines. In this study, a total of 90 samples were collected from air, water, and soil from 28 different geographical areas of Iran. Forty isolates were selected based on differences in colony color and geographic regions. The MTT assay was used to evaluate the effect of pigment on MCF-7 and SW-48 cells. Bacteria whose pigment had the highest cytotoxic effect on cell lines were selected. Accurate identification was performed using PCR, and their relative purity was measured using TLC. Bacteria isolated from any three ecosystems are capable of producing pigments. Pigment-producing bacteria are more abundant in the soil than air and water. Among pigment-producing bacteria, 3 isolates had the highest cytotoxicity on MCF-7 cells, and 3 isolates had the greatest effect on SW-48 cells. The results of sequencing of isolates at the BLAST site showed that 6 isolates with cytotoxic effects were identified (Micrococcus xinjiangensis, Dietzia, Arthrobacter agilis, Exigubacterium mexicanum, Bacillus beijingensis). Chromatography shows that these pigmented bacteria have different pigment components.Pigment extraction from bacteria can be used as a complementary therapy or other therapies for breast and colon cancer in the future.

Phenotypic and Molecular Identification of Pathogenic Fusarium Species Isolated from Various Pulse Growing Geographic Areas of India

Fusarium phythopathogenic fungi is responsible for high economic loss of cereal food crop. The objective of this study was aimed at isolation, morphological and molecular identification of Fusarium species. 13 different Fusarium spp. i.e. F. solani, F. chlamydosporum, F. tabacinum, F. fujikuroi, F. oxysporum, F. verticillioides, F. brachygibbosum, Fusarium sp. and F. incarnatum were isolated and identified from diseased samples of chickpea, pigeonpea, rice, lentil and garden pea crop. Colony characteristics like colony color, colony growth diameters, mycelium type, sporulation, pigmentation, odour were obtained after culture purification. Shape, size and septation of microconidia and macroconidia, position, shape, occurrence and size of chlamydospores, conidiophore branching were examined microscopically. MS10, BI01 and KA14 isolates were slow growing, BI02 and UP07 were moderate growing and BI03, HA04, MS06, MS09, MS11 and KA(Gul)13 were fast to very fast growing on PDA after 7-10 days. Chlamydospores were found in most of the isolates. Colonies were abundant, loosely tufted, fluffy, pannose, vinaceous floccose, powdery and some were flat appressed, arachnoid. Pigmentation of most of the isolates was pinkish white to dark pink, carmine to violet in colour. Phylogenetic analysis was done by maximum likelihood method using the ITS-rDNA region of Fusarium isolates and multiple sequence alignment of ITS DNA sequences was done using Clustal_W program and all identified sequences were submitted in NCBI GenBank database.

Application of Crude Enzymatic Extract In Mushroom Processing To Recover High Value Products

In the present study, a fungal strain was isolated from mushroom waste dump-site and was described based on the morphological and molecular characteristics. The crude enzymatic extract was prepared by fermenting pineapple peels using the newly isolated fungal strain under solid-state condition. The enzymatic saccharification conditions of mushroom were optimized using the central composite design based on the response surface methodology. The isolate had black colony color, conidial head biseriate and small conidia which are synonymous with Aspergillus niger. The phylogenetic analysis using the rDNA ITS sequencing further revealed that the isolate was identical (≥99%) to A. niger. The crude extract displayed CMCase, Fpase and xylanase activities of 20.73U/mL, 34.57U/mL and 118.03U/mL respectively. The saccharification using the crude extract at optimal conditions of pH 6.5, temperature 50oC, enzyme loading of 5% (v/v) and time of 12h achieved maximum glucose yield of 1.639 mg mL-1 which is 1.1 folds higher than the predicted value. This study demonstrated the potential use of crude enzymatic extract from the newly isolated A. niger as a viable and efficient low-cost approach to mushroom processing using enzymes.

Image Analysis to Quantify Coral Bleaching Using Greyscale Model v1

This protocol outlines a method of quantitatively measuring the degree of bleaching of a coral colony non-destructively in the field using image analysis. Previous studies have shown that mean intensity grey (MIG), also known as percent whiteness, is highly correlated with chlorophyll a and Symbiodiniaceae density (Chow et al. 2016, Amid et al. 2018), and therefore can be used to quantify the bleaching intensity of a coral colony. Color analysis can be done using digital photographs of live coral colonies either in situ (e.g., Maguire et al. 2003) or ex-situ in the lab (Amid et al. 2018; this protocol). Photographs must be taken prior to any preservation or processing of tissue, such as freezing, use of preservatives or fixatives, airbrushing etc., to ensure no alteration of the original coral color occurs. In this protocol, corals are photographed in front of a white reference standard and the resulting color images are subsequently converted to 8-bit greyscale and analyzed. There are two steps to this protocol: 1) Photographing live coral fragments 2) Image analysis of mean grey value This protocol was written by Dr. Rowan McLachlan and was reviewed by Dr. Andréa Grottoli. Acknowledgments I would like to thank Dr. Eugene Katrukha for kindly taking the time to teach me this method, and providing me feedback on how to produce higher quality images for analysis.

Boeremia parva sp. nov., a novel species of the family Didymellaceae isolated from soil

Fungal strains, designated KNU-NL4 and KNU-OL2, belonging to the family Didymellaceae were isolated from a soil sample collected in Miryang, Korea. Phylogenetic analyses based on a concatenated dataset of DNA sequences of ITS regions and partial sequences of ACT, CAL, TEF1-α, and β-TUB genes showed that the isolates reside in a clade together with Boeremia species but occupy the distinct phylogenetic position. Morphologically, the novel strains produce bigger conidiomata (average size 169.8 μm) than the closely related B. rhapontica (126.59 μm) and smaller than the other close neighbor B. coffeae (187.5 μm). Both novel strains also differed from them by smaller colony size and colony color on OA and MEA. The detailed descriptions, illustrations, and discussions regarding the morphological and phylogenetic analyses of the closely related species are provided to support the novelty of the isolated species. The results of phylogenetic analysis and morphological observations indicate that strains KNU-NL4 and KNU-OL2 represent a novel species in the genus Boeremia, for which the name Boeremia parva sp. nov. is proposed.

Investigation on Cultural Characterization and Parasexual Recombination in Pyricularia Grisea, Causal Agent of Blast of Rice

Blast of rice caused by Pyricularia grisea is one of the most devastating diseases of rice. Because of importance of the disease and the fact that pyricularia grisea is considered to be notorious and model species. The variability in cultural characteristics of fifty isolates of P. grisea were taken from different regions of Jharkhand state. Out of fifty isolates of P. grisea, colony color of six isolates were found to be as greyish white color, three isolates were blackish grey, six isolates as white color, three isolates as whitish grey color, five isolates were whitish black and twenty seven isolates were recorded as blackish white in color. The growth pattern of 47 isolates of P. grisea showed circular growth pattern and three isolates have irregular growth pattern but elevation of the mycelium differs from flat to raised. Out of fifty isolates of P. grisea, sector formation was observed in seventeen isolates and no sector formation was observed in rest isolates. The radial growth of fifty isolates of P. grisea were ranged from 76.0 mm to 90.0 mm. Out of fifty isolates, group I included six isolates, group II and III included eleven and thirty three isolates, respectively. Parasexual recombination rarely causes genetic and phenotypic variation through hyphal anastomosis in India. Parasexual recombination is the principle cause in rice blast fungus and cause infection of resistant rice variety (IR-64) in India .

Ability of The Saccharomyces Cerevisiae Y904 to Tolerate and Adapt to High Concentrations of Selenium

The rational use of by-products is essential for the development of a sustainable society. Worldwide, the alcoholic fermentation industry generates a large surplus of yeasts, on the scale of millions of tons. So there is a need for beneficial applications to humanity of this surplus. Yeasts, in turn, have the ability to bioaccumulate minerals and enable their bioavailability after cell autolysis. Among these minerals, we highlight selenium (Se), which participates in the formation of antioxidant enzymes. The objectives of the work were to define the minimum and maximum concentration of Se that yeasts (Saccharomyces cerevisiae – Y904) support and the concentrations that they tolerate once adapted. To this end, a test of tolerance to Se was carried out, using treatments with different concentrations of Se. The adaptive process started at the maximum concentration obtained in the tolerance test of 60 µg mL− 1, with an increasing addition of 6 µg mL− 1, reaching up to 246 µg mL− 1 of Se. The macromorphological characteristics and number of colony forming units were evaluated. It was identified that yeasts without adaptation grew on substrate containing up to 60 µg mL− 1 of Se and those adapted, up to 246 µg mL− 1 of Se. In addition to the reduction in yeast growth speed, from the concentration of 84 µg mL− 1 of Se in the medium, morphological changes in colony color were observed. It is concluded that non-adapted yeasts support up to 60 µg mL− 1 of Se and, after the adaptive process, they support 246 µg mL− 1 of Se in the medium.

Phenotypic diversity of bacteria in root nodules of Dalbergia ecastaphyllum (L.) Taub. (Fabaceae)

Many species belonging to family Fabaceae are able to establish symbiotic relationships with nitrogen-fixing bacteria. Studies developed with Dalbergia ecastaphyllum (L.) Taub., for example, have demonstrated the symbiotic potential of this species. In this sense, this study aimed to analyze the phenotypic characteristics of rhizobia isolated from D. ecastaphyllum and to identify whether these bacterial isolates are capable of establishing symbiotic relationships with Vigna unguiculata (L.) Walp. D. ecastaphyllum seeds were sown in soil samples collected in Japaratinga, Alagoas, Brazil, in three zones located at different distances from the high tide line. At 60 days after the emergence of the plants, nodulation in the roots of D. ecastaphyllum was analyzed and the bacteria were isolated. Subsequently, the phenotypic characterization of the bacteria was carried out based on some criteria (growth time, type of pH, colony color, type of mucus and amount of mucus). For the nodulation test, the species V. unguiculata was used. In total, 17 phenotypic types of bacterial isolates were identified, of which 8 are fast-growing and 9 are intermediate-growth. Regarding the type of pH, 7 phenotypes are acidic pH, 8 are neutral pH and 2 are alkaline pH. As for the coloring of the bacterial colonies, 5 are yellow, 7 white and 5 pink. It was also observed the presence of consistent and aqueous mucus in colonies from the three different areas. Only 7 bacterial phenotypic types and the commercial inoculant were efficient in the nodulation of V. unguiculata.

Microscopic Characterization of Fusarium sp. Associated with Yellow Disease of Pepper (Piper nigrum L.) in South Bangka Regency

Pepper production has decreased recently, especially due to yellow diseases of Fusarium sp. Thus, this research aimed to isolate and characterize Fusarium sp. from soil and root of healthy and diseased pepper plants. The sampling technique used was purposive sampling. Soil and root pepper samples were taken from lands in Payung and Ranggung Village, Payung District, South Bangka Regency. There were 3 varieties of pepper plant used, including Petaling 1, Nyelungkup, and Merapin Daun Kecil. The characterization of Fusarium sp. isolate included macroscopic and microscopic observation. Macroscopic observation included colony color, colony base color, and growth rate/colony diameter size (cm), while microscopic observation included hyphae structure, and the shape and size of microconidia, macroconidia, chlamydospore, and conidiophore. The research found 66 isolates of Fusarium genus based on the colony color. Most of the isolates were white or purple and red. Colony color of Fusarium sp. showed white color, which then turned to orange color. All isolates showed septate hyphae. Isolates with macroconidia 3-4 septate and micronidia 0-1 septate showed the character of Fusarium oxysporum, while isolates with macroconidia 3-5 septate and microconidia 0-2 septate showed the character of Fusarium solani.