Development of PCR based SSR markers for Wilsonomyces carpophilus and a PCR based diagnosis protocol for the early detection of shot hole disease in stone fruit crops.

BACKGROUND: The conidial Ascomycota fungus Wilsonomyces carpophilus causing shot hole in stone fruits is a major constraint in the production of stone fruits worldwide. Shothole disease symptoms appear on leaves, fruits, and twigs. Successful isolation of the pathogen from different hosts on synthetic culture medium is a time consuming and tedious procedure for identification of the pathogen based on morpho-cultural characterization. METHODS AND RESULTS: The present research was carried out to develop a successful PCR based early detection protocol for the shot hole disease of stone fruits, viz., peach, plum, apricot, cherry, and almond using the pathogen specific SSR markers developed from the Wilsonomyces carpophilus genome using Genome-wide Microsatellite Analysing Tool package (GMATA) software. Diseased leaf samples of different stone fruits were collected from the SKUAST-K orchard and the pathogen was isolated on potato dextrose agar (PDA) medium and maintained on Asthana and Hawkers' medium with a total of 50 pathogen isolates comprised of 10 isolates each from peach, plum, apricot, cherry and almond. The DNA was extracted from both healthy and infected leaf samples of different stone fruits. The DNA was also extracted from the isolated pathogen cultures (50 isolates). Out of 2851 SSR markers developed, 30 SSRs were used for the successful amplification of DNA extracted from all the 50 pathogen isolates. These SSRs were used for the amplification DNA from shot hole infected leaf samples of different stone fruits, but the amplification was not observed in the control samples (DNA from healthy leaves), thus confirming the detection of this disease directly from the shot hole infected samples using PCR based SSR markers. To our knowledge, this forms the first report of SSR development for the Wilsonomyces carpophilus and their validation for the detection of shot hole disease directly from infected leaves. CONCLUSION: PCR based SSR makers were successfully developed and used for the detection of Wilsonomyces carpophilus causing shot hole disease in stone fruits including almond in nuts for the first time. These SSR markers could successfully detect the pathogen directly from the infected leaves of stone fruits namely peach, plum, apricot and cherry including almond from the nuts.

Effects of microplastics in freshwater fishes health and the implications for human health.

The presence of microplastics in aquatic environments has raised concerns about their abundance and potential hazards to aquatic organisms. This review provides insight into the problem that may be of alarm for freshwater fish. Plastic pollution is not confined to marine ecosystems; freshwater also comprises plastic bits, as the most of plastic fragments enter oceans via rivers. Microplastics (MPs) can be consumed by fish and accumulated due to their size and poor biodegradability. Furthermore, it has the potential to enter the food chain and cause health problems. Evidence of MPs s ingestion has been reported in >150 fish species from both freshwater and marine systems. However, microplastic quantification and toxicity in freshwater ecosystems have been underestimated, ignored, and not reported as much as compared to the marine ecosystem. However, their abundance, influence, and toxicity in freshwater biota are not less than in marine ecosystems. The interaction of MPs with freshwater fish, as well as the risk of human consumption, remains a mystery. Nevertheless, our knowledge of the impacts of MPs on freshwater fish is still very limited. This study detailed the status of the toxicity of MPs in freshwater fish. This review will add to our understanding of the ecotoxicology of microplastics on freshwater fish and give subsequent research directions.

Greenhouse Evaluation of Binucleate Rhizoctonia for Control of R. solani in Soybean.

Nine isolates of binucleate Rhizoctonia (BNR) from soybean were screened in the greenhouse for control of Rhizoctonia solani anastomosis groups AG-4 and AG-2-2. Eight of nine BNR isolates, when combined with AG-4 or AG-2-2, significantly increased emergence and survival of soybean (cv. Ozzie) and reduced disease severity compared with AG-4 or AG-2-2 alone. The interaction of soybean cultivar and BNR isolates in the presence of AG-4 and AG-2-2 was also studied using three isolates of BNR, BNR-4, BNR-8-2, and BNR-8-3, and seven soybean cultivars. There was no BNR × cultivar interaction. With AG-4, BNRs significantly increased emergence and survival of cultivars and reduced disease severity, whereas with AG-2-2, BNRs reduced disease severity. Control of R. solani by BNRs was achieved in both a potting soil mix and natural soil. In the initial screening experiments, two BNR isolates reduced emergence, but in all subsequent experiments using three BNR isolates alone, there were no negative effects on germination, survival, or height of soybean plants, and there was no evidence of pathogenicity. In several experiments, BNRs alone significantly increased height of plants compared with the noninoculated controls. BNRs were consistently isolated from hypocotyls and roots, indicating colonization of tissues was associated with control. These BNR isolates may have potential use in management of R. solani in soybean, but will require rigorous testing under field conditions and more extensive studies of their biology.

Zoonotic studies on influenza in pigs and birds, India, 1980-81.

Five hundred and twenty pig sera collected from Pune, Maharashtra State, India during 1980 were examined in Haemagglutination Inhibition (HI) tests to determine the antibody prevalence to nine human influenza virus strains covering the subtypes A(HON1), A(H1N1), A(H2N2), A(H3N2), type B and one swine influenza virus strain A(Hsw1N1). This study indicated considerable prevalence of antibodies to the four H3N2 strains isolated from 1973 onwards, particularly to the two recent H3N2 strains, limited prevalence of antibodies to H1N1 strain and absence of antibodies to the Hsw1N1 and HON1 influenza strains in the pig sera. Three hundred and eleven cloacal swab specimens collected from different species of domestic and wild birds from Kolar district, Karnataka State, India during 1980 and 1981 were investigated for influenza virus prevalence. No influenza virus was isolated from any of the specimen, but one strain of Newcastle disease virus was isolated from a chicken.