Evaluation of actinomycete isolates obtained from herbal vermicompost for the biological control of Fusarium wilt of chickpea

A total of 137 actinomycetes cultures, isolated from 25 different herbal vermicomposts, were characterized for their antagonistic potential against Fusarium oxysporum f. sp. ciceri (FOC) by dual-culture assay. Of the isolates, five most promising FOC antagonistic isolates (CAI-24, CAI-121, CAI-127, KAI-32 and KAI-90) were characterized for the production of siderophore, cellulase, protease, hydrocyanic acid (HCN), indole acetic acid (IAA) and antagonistic potential against Rhizoctonia bataticola, which causes dry root rot in chickpea (three strains viz. RB-6, RB-24 and RB-115) and sorghum (one strain). All of the five FOC antagonistic isolates produced siderophore and HCN, four of them (except KAI-90) produced IAA, KAI-32 and KAI-90 produced cellulase and CAI-24 and CAI-127 produced protease. In the dual-culture assay, three of the isolates, CAI-24, KAI-32 and KAI-90, also inhibited all three strains of R. bataticola in chickpea, while two of them (KAI-32 and KAI-90) inhibited the tested strain in sorghum. When the FOC antagonistic isolates were evaluated further for their antagonistic potential in the greenhouse and wilt-sick field conditions on chickpea, 45-76% and 4-19% reduction of disease incidence were observed, respectively compared to the control. The sequences of 16S rDNA gene of the isolates CAI-24, CAI-121, CAI-127, KAI-32 and KAI-90 were matched with Streptomyces tsusimaensis, Streptomyces caviscabies, Streptomyces setonii, Streptomyces africanus and an identified species of Streptomyces, respectively using the BLAST searching. This study indicated that the selected actinomycete isolates have the potential for biological control of Fusarium wilt disease in chickpea.     

Development and characterization of tomato SSR markers from genomic sequences of anchored BAC clones on chromosome 6

Simple sequence repeat motifs are abundant in plant genomes and are commonly used molecular markers in plant breeding. In tomato, currently available genetic maps possess a limited number of simple sequence repeat (SSR) markers that are not evenly distributed in the genome. This situation warrants the need for more SSRs in genomic regions lacking adequate markers. The objective of the study was to develop SSR markers pertaining to chromosome 6 from bacterial artificial chromosome (BAC) sequences available at Solanaceae Genomics Network. A total of 54 SSR primer pairs from 17 BAC clones on chromosome 6 were designed and validated. Polymorphism of these loci was evaluated in a panel of 16 genotypes comprising of Solanum lycopersicum and its wild relatives. Genetic diversity analysis based on these markers could distinguish genotypes at species level. Twenty-one SSR markers derived from 13 BAC clones were polymorphic between two closely related tomato accessions, West Virginia 700 and Hawaii 7996 and were mapped using a recombinant inbred line population derived from a cross between these two accessions. The markers were distributed throughout the chromosome spanning a total length of 117.6 cM following the order of the original BAC clones. A major QTL associated with resistance to bacterial wilt was mapped on chromosome 6 at similar location of the reported Bwr-6 locus. These chromosome 6-specific SSR markers developed in this study are useful tools for cultivar identification, genetic diversity analysis and genetic mapping in tomato.     

Viral interference with MHC class I antigen presentation pathway: the battle continues

CD8+ cytotoxic T lymphocytes (CTLs) play a critical role in the defense against viral infections. In general, CD8+ CTLs recognize antigenic peptides in the context of the major histocompatibility complex (MHC) class I molecule. The MHC class I molecules are expressed on almost all the nucleated cells in the body. The trimolecular complex consisting of the class I heavy chain, beta2-microglobulin and the peptide are generated by the MHC class I antigen presentation pathway. This pathway is designed to sample the intracellular milieu and present the information to the CTLs trafficking the area. This rigorous sampling of intracellular environment enables the CTLs to quickly identify and eliminate the cells that synthesize non-self proteins as a result of a viral infection. Many viruses, including several viruses of veterinary importance, have evolved astounding strategies to interfere with the MHC class I antigen presentation pathway, as a means of evading the CTL response of the host. This review focuses on the diverse mechanisms of viral evasion of the MHC class I antigen presentation pathway with particular emphasis on viruses of veterinary importance.     

Antifungal and defense elicitor activities of pyrazines identified in endophytic Pseudomonas putida BP25 against fungal blast incited by Magnaporthe oryzae in rice

Journal of Plant Diseases and Protection - Black pepper-associated endophytic Pseudomonas putida BP25 displayed volatile-mediated antagonism against rice blast fungus Magnaporthe oryzae 1637. The...     

Immunoblot assays using recombinant antigens for the detection of Mycoplasma hyopneumoniae antibodies

The 36kDa L-lactate dehydrogenase (LDH) and a 29kDa partial fragment of an ABC transporter ATP-binding protein analogue/multidrug resistance protein homologue (PR2) of Mycoplasma hyopneumoniae were tested for their potential as diagnostic antigens. Recombinant LDH was genetically engineered to contain six histidine residues at its C-terminal end, expressed in Escherichia coli and purified to a high degree using Ni(2+)-chelate affinity chromatography. A partial 262 amino acid segment representing the C-terminal end of the PR2 protein was cloned as a glutathione S-transferase (GST) fusion protein, expressed in E. coli and purified by urea extraction. Purified recombinant LDH-6xHis and PR2-GST were then reacted with pig sera in immunoblot assays. Our immunoblots showed that both proteins detected anti-M. hyopneumoniae antibodies in field and experimentally infected pig sera but not in any of the SPF control sera. The two proteins were specific for M. hyopneumoniae as they did not react with sera of pigs infected with the closely related Mycoplasma flocculare and Mycoplasma hyorhinis which are frequently isolated in pigs but are not of particular concern.     

Measured values of the dry deposition velocities of atmospheric aerosols carrying natural and fallout radionuclides using artificial collectors

The dry deposition velocitiesVg of aerosols carrying (a) fallout beta activity from nuclear tests, (b) natural radioactivity due to thoron daughter²¹²Ph (Th-B) and (c) cosmic-ray produced⁷Be have been measured in Bombay, India, using artificial collectors consisting of trays with a thin layer of water to retain the deposited material. The location of Bombay is eminently suitable for such measurements in view of the existence of a long dry period of several months without any rainfall. The measuredVg values (cm s^?1)) are 0.063 ± 0.06 (1 S.D.) for fallout beta activity from 900 daily readings, 0.033 ± 0.03 (1 S.D.) for²¹²Pb from 80 daily measurements and 0.023 ± 0.014 (1 S.D.) for⁷Be from 23 bi-monthly measurements. A study of the associated meteorological parameters showed some correlation with wind velocity only in the case of radioactive fallout.     

A novel method for the identification and enumeration of microorganisms with potential for suppressing fungal plant pathogens

This paper describes a method that allowed counting of both the total culturable and antagonistic microorganisms in a given source such as compost. Fusarium solani, used as the test fungus, was spread-plated on quarter-strength (1/4) potato dextrose agar (PDA), its surface was exposed in a laminar flow for 4 h and then another layer (2–3 mm thick) of 1/4 PDA was poured over it, on which an appropriate dilution of a compost sample was spread-plated. Microorganisms in the compost samples appeared first, and were counted as total culturable organisms. Plates were further incubated until F. solani grew through the upper layer of PDA (generally in 4–8 days) and covered the whole plate including most of the microbial colonies, except for a few which had a halo around them. These were counted as antagonistic, and they were isolated and purified for further studies. The population of bacteria in the six specific compost samples (called Biodynamic or BD preparations by organic farmers) ranged from 3.45 log₁₀ (in BD502) to 8.59 log₁₀ (in BD504) per gram of materials. The population of antagonistic bacteria was counted for three of the six compost samples, and ranged from 3.24 log₁₀ (in BD502) to 6.90 log₁₀ (in BD500). Of the 67 bacterial isolates showing a halo that were assembled from different sources, 17 suppressed at least 1 of the 4 plant pathogenic fungi against which these were evaluated using the dual culture method.     

Characterization of Lasiodiplodia species causing leaf blight,stem rot and fruit rot of fig (Ficus carica) in Malaysia

Fig (Ficus carica) is an exotic deciduous plant that is grown worldwide. Fungal diseases pose a major threat to fig plants, affecting their fruit quality and production. This study was conducted to characterize the fungal isolates associated with leaf blight, stem rot and fruit rot of F. carica in Malaysia through morphological analysis, DNA sequencing, multigene phylogenetic analysis and pathogenicity tests. From September 2018 to March 2019, 30 blighted leaves and 30 rotted stems and fruits of F. carica were collected from several nurseries in Malaysia. Thirty fungal isolates that belonged to Lasiodiplodia theobromae (27 isolates) and L. brasiliensis (three isolates) were identified based on morphological characteristics, comparison of DNA sequences and phylogenetic analysis of the internal transcribed spacer (ITS), elongation translation factor 1-α (tef1-α), β-tubulin (tub2) and DNA-directed RNA polymerase II subunit (rpb2). Among the 27 isolates of L. theobromae, nine isolates were obtained from leaves, eight isolates from stems and 10 isolates from fruits, whereas the three isolates of L. brasiliensis were obtained from stems (two isolates) and a leaf (one isolate). The results of pathogenicity tests revealed that L. theobromae and L. brasiliensis isolates were responsible for leaf blight and stem rot of F. carica, whereas fruit rot was caused by L. theobromae isolates. The present study highlighted two different species, L. theobromae and L. brasiliensis, as the causal agents of leaf blight and stem rot of F. carica. Additionally, L. theobromae caused fruit rot of F. carica in Malaysia.     

Co-expression of ovine LPS receptor CD14 with Mannheimia haemolytica leukotoxin receptor LFA-1 or Mac-1 does not enhance leukotoxin-induced cytotoxicity

Leukotoxin (Lkt) and LPS are the major virulence determinants of Mannheimia haemolytica that contribute to the pathogenesis of bovine and ovine pneumonic pasteurellosis. We have previously identified bovine and ovine CD18 as the functional receptor for Lkt. LPS complexes with Lkt resulting in increased thermal stability and enhanced cytotoxic activity of Lkt. Cellular recognition of LPS involves several different molecules including CD14. We hypothesized that expression of ovine CD14 together with LFA-1 or Mac-1 would enhance Lkt-induced cytotoxicity. Ovine cDNA for CD14 was amplified by PCR and cloned into mammalian expression vectors. The 1122 bp cDNAs for bighorn sheep (BHS) and domestic sheep (DS) CD14 encode 373 amino acids which exhibit 99% identity with each other. Ovine CD14 plasmids were transfected either into HEK-293 cells, or previous HEK-293 transfectants stably expressing ovine LFA-1 or Mac-1. Flow cytometric analysis of transfectants confirmed the cell surface expression of CD14. The transfectants expressing LFA-1 or Mac-1 and the transfectants co-expressing CD14 with LFA-1 or Mac-1 did not show any significant difference in Lkt-induced cytotoxicity when incubated with LPS complexed Lkt. In contrast, incubation of the LFA-1 or Mac-1 and LFA-1/CD14 or Mac-1/CD14 transfectants with Lkt which lacks LPS, resulted in reduced cytotoxicity. None of the above transfectants showed any difference in [Ca2+](i) elevation when incubated with both types of Lkt preparations. Lkt did not induce any cytotoxicity or [Ca2+](i) elevation in ovine CD14 transfectants or parent HEK-293 cells. Based on these findings, we conclude that expression of CD14 together with LFA-1 or Mac-1 does not enhance Lkt-induced cytotoxicity, whereas LPS enhances cytotoxicity by complexing with Lkt.