Molecular identification of blast resistance genes in rice landraces from northeastern India

Rice blast disease caused by the fungus Magnaporthe oryzae is one of the most devastating diseases causing huge losses worldwide. In the present study, major blast resistance genes were investigated in landraces originating from northeastern India. Based on phenotypic evaluation, 288 landraces were classified into three distinct groups: resistant (75), moderately resistant (127) and susceptible (86). The genetic frequencies of the 18 major blast resistance genes were between 6.2% and 27.4%, with only two genotypes possessing a maximum of nine blast resistance genes. The cluster and population structure analysis grouped the landraces into two groups. Through principal coordinate analysis, the scatter plots partitioned the resistant and moderately resistant landraces into different groups. Analysis of molecular variance showed maximum (96%) diversity within populations and least (4%) diversity between populations. Association analysis identified six markers, CRG4_2, RM72, tk59-2, pi21_79-3, RM1233 and RM6648, that are significantly associated with blast disease and explained a phenotypic variance of 1.1–6.5%. The associated genes could be used in marker-assisted rice breeding programmes for gene pyramiding to develop rice varietal resistance against blast disease. The present study represents a valuable blast resistance genetic resource that could be used for identification of new R genes, donors for blast resistance, and genomic studies.     

Temporal and spatial progress of the diseases caused by the crinivirus tomato chlorosis virus and the begomovirus tomato severe rugose virus in tomatoes in Brazil

Efficient management of whitefly-borne diseases remains a challenge due to the lack of a comprehensive understanding of their epidemiology, particularly of the diseases tomato golden mosaic and tomato yellowing. Here, by monitoring 16 plots in four commercial fields, the temporal and spatial distribution of these two diseases were studied in tomato fields in Brazil. In the experimental plots these diseases were caused by tomato severe rugose virus (ToSRV) and tomato chlorosis virus (ToCV), respectively. The incidence of each virus was similar in the plots within a field but varied greatly among fields. Plants with symptoms for both diseases were randomly distributed in three of four spatial analyses. The curves representing the progress of both diseases were similar and contained small fluctuations, indicating that the spread of both viruses was similar under field conditions. In transmission experiments of ToSRV and ToCV by Bemisia tabaci MEAM1 (former biotype B), these viruses had a similar transmission rate in single or mixed infections. It was then shown that primary and secondary spread of ToCV were not efficiently controlled by insecticide applications. Finally, in a typical monomolecular model of disease progress, simulation of the primary dissemination of ToSRV and ToCV showed that infected plants were predominantly randomly distributed. It is concluded that, although the manner of vector transmission differs between ToSRV (persistent) and ToCV (semipersistent), the main dispersal mechanisms are most probably similar for these two diseases: primary spread is the predominant mechanism, and epidemics of these diseases have been caused by several influxes of viruliferous whiteflies.     

Influence of temperature,humidity duration and growth stage on the infection and mycotoxin production by Fusarium langsethiae and Fusarium poae in oats

High occurrence of Fusarium poae (FP) and Fusarium langsethiae (FL) and their mycotoxins nivalenol (NIV) and T-2/HT-2 have been observed in Swiss oats. Early prediction of mycotoxin levels is important for farmers and the cereal industry to minimize the risk of contaminated food and feed. Therefore, climate chamber experiments were conducted to investigate the influence of different temperatures (10, 15, 20 °C) and durations (4, 8, 12 h) at 99% relative humidity (RH) on the infection of oats with FP and FL. In addition, to discover the most susceptible period of oats, artificial FL inoculations were conducted at different growth stages. Field experiments were performed to observe the dispersal of these fungal species within the field and to investigate the weather conditions that influence the dispersal. The climate chamber experiments revealed higher contamination with NIV and T-2/HT-2 in the 10 °C treatments and with a prolonged humidity duration of 12 h 99% RH. Inoculations of oat plants at early (DC 61) and mid (DC 65) anthesis, led to higher FL infection and T-2/HT-2 accumulation in the grains compared with treatments at earlier growth stages, which might be due to an increased susceptibility during anthesis. No indication for spore dispersal was observed in the field experiments. The results obtained, together with the cropping factors that influence infection and mycotoxin production, could be used as a first step in developing forecasting models to predict the contamination of oats with the mycotoxins NIV and T-2/HT-2.     

Temporal disease dynamics and relative importance of sexual and asexual reproduction of grape downy mildew (Plasmopara viticola) in an isolated vineyard in the North Georgia Mountains,USA

The North Georgia Mountains are the southernmost region along the United States East Coast where European wine grapes (Vitis vinifera) are grown commercially. Epidemics of downy mildew, caused by Plasmopara viticola, are frequent and severe, but little is known about the epidemiology and population biology of the pathogen in this region. Disease monitoring in an experimental vineyard from 2015 to 2017 indicated that times of disease onset and progress rates were highly variable across years and cultivars. Oospores were observed microscopically, and simulation with a process-based model indicated presence of conditions favourable for oospore germination in the spring and early summer each year. A total of 409 P. viticola isolates collected over three  years were genotyped with seven microsatellite markers, revealing very high genotypic diversity, which when combined with the observation of oospores is indicative of a sexually reproducing population. Among the 409 isolates, 225 multilocus genotypes (MLGs) were identified, of which 164 were detected only once and 61 were repeated (clonal). Eight MLGs (represented by 28 isolates) were detected across years, suggesting the possibility of asexual overwintering of P. viticola in this region. Across sampling dates, the percentage of isolates belonging to nonrepeated (unique) MLGs ranged from 27.3% to 63.2%. Even towards the end of the annual epidemic, the percentage of isolates in nonrepeated MLGs was still relatively high, around 30%. These MLGs may have originated from oospores germinating late during the growing season, although incomplete sampling at earlier dates and contribution by immigration cannot be fully excluded.     

Impact of weather variables and season on sporulation of Phytophthora pluvialis and Phytophthora kernoviae

Phytophthora pluvialis and Phytophthora kernoviae are the causal agents of important needle diseases on Pinus radiata in New Zealand. Little is known about the epidemiology of the diseases, making the development of control strategies challenging. To investigate the seasonality and climatic drivers of sporulation, inoculum traps, consisting of pine fascicles floating on water in plastic containers, were exchanged fortnightly at five sites in P. radiata plantations between February 2012 and December 2014. Sections of needle baits were plated onto selective media and growth of Phytophthora pluvialis and P. kernoviae recorded. To explore the generalizability of these data, they were compared to detection data for both pathogens from the New Zealand Forest Health Database (NZFHDB). Further, equivalent analyses on infection of Rhododendron ponticum by P. kernoviae in Cornwall, UK allowed the comparison of the epidemiology of P. kernoviae across different host systems and environments. In New Zealand, inoculum of P. pluvialis and P. kernoviae was detected between January–December and March–November, respectively. Inoculum of both species peaked in abundance in late winter. The probability of detecting P. pluvialis and P. kernoviae was greater at lower temperatures, while the probability of detecting P. pluvialis also increased during periods of wet weather. Similar patterns were observed in NZFHDB data. However, the seasonal pattern of infection by P. kernoviae in the UK was the opposite of that seen for sporulation in New Zealand. Phytophthora kernoviae was likely limited by warmer and drier summers in New Zealand, but by colder winter weather in the UK. These results emphasize the importance of considering both environmental drivers and thresholds in improving our understanding of pathogen epidemiology.     

Comparison of outcomes following treatment of septic calcaneal bursitis by needle or bursoscopic lavage: A retrospective study of 29 horses

Sepsis of the calcaneal bursae (CB) presents significant treatment challenges with limited clinical data available in the literature. The objective of this retrospective cohort study was to assess the clinical outcomes associated with CB lavage using either a through-and-through needle or bursoscopic technique. Clinical records of 29 horses treated for septic calcaneal bursitis using either technique between 2005 and 2019 were reviewed. Fisher’s exact test was used to assess statistical significance between first surgical technique and success at first surgery (i.e. not requiring >1 lavage), survival to discharge and return to work (RTW). Bursoscopy was performed in 13/29 (44.8%) cases, and needle lavage in 16/29 (55.2%). In the needle group, 12 (75%) horses were discharged following the first surgery. Four had repeat interventions; two (12.5%) had needle lavage and two (12.5%) had bursoscopy. Of the two horses to have repeat needle lavage, one was subjected to euthanasia and one discharged, and of the two that underwent bursoscopy, one was discharged and one received a third bursoscopy prior to discharge. In the bursoscopy group, seven (53.8%) were discharged and three (23.1%) were subjected to euthanasia following the first surgery. Three (23.1%) received a second bursoscopic lavage with one discharged, one subjected to euthanasia and one having a third bursoscopic lavage prior to discharge. Overall, 18/24 (75%) followed up cases RTW, 10 (55.5%) from the needle group, eight (44.4%) the bursoscopy group. No statistically significant differences between first surgical technique used and success at first surgery (no subsequent lavage(s) required), survival to discharge or return to work were detected. The main limitations of this study are that it is a retrospective study, has a small population with limited statistical power and potential selection bias. No statistically significant differences existed between the outcomes of the two techniques, contrary to the belief that bursoscopic lavage is superior. Larger, multicentred studies, with greater statistical power are required to further assess this relationship.     

Comparison of effects of dietary‐specific fatty acids on growth and lipid metabolism in Nile tilapia

The dominant fatty acids (FAs) in oils are often used to explain different nutritional effects of dietary oils in fish. However, the amounts of dominant FAs among oils are different, and the nutritional roles of these important FAs in fish have not been precisely compared at similar levels in feeding trials. In the present study, different amounts of palmitic acid were added to safflower oil (SO), olive oil (OO) and fish oil (FO) to obtain comparable amounts (about 550 g/kg of total FAs) of 18:2n‐6, 18:1n‐9 and 20:5n‐3 + 22:6n‐3 and subsequently fed to Nile tilapia (11.1 ± 0.01 g) for 8 weeks. The results showed similar growth among groups but FO group obtained lower fat deposition, serum ALT and AST activities, compared to OO. Lipogenesis‐related gene expressions were higher in OO group than FO group in liver, muscle and adipose tissue, but there were only few differences in these genes between SO and FO groups. Lipid catabolism genes in FO group were higher than OO and SO groups in adipose tissue, but not in muscle, and the significantly higher expressions of CPT1b and PPARα were only observed in liver. Overall, dietary 18:2n‐6, 20:5n‐3 and 22:6n‐3 were beneficial to normal growth and lipid metabolism, whereas high amount of 18:1n‐9 induced lipid deposition and liver damage in Nile tilapia.     

Crude garlic extract significantly inhibits replication of grapevine viruses

Efforts to control viral diseases of grapevine include the production of certified material and development of virus-resistant transgenic grapevines. However, effective antiviral agents, once the viruses have infected the plants, are still lacking. This study shows that a crude garlic extract has significant antiviral activity against grapevine viruses. Replication of grapevine leafroll-associated virus 2 (GLRaV-2) was obviously inhibited in grapevine cv. Cabernet Sauvignon calli treated with diluted (1:100) garlic extract. The relative RNA levels of GLRaV-2 and grapevine fleck virus (GFkV) in cv. Summer Black grapevine in in vitro-grown plantlets 10 days after treatment with diluted (1:100) garlic extract were about 22% and 20%, respectively, of that in controls. The viral RNA accumulation of GLRaV-2, GFkV, grapevine virus A (GVA), grapevine fanleaf virus (GFLV) and grapevine rupestris stem pitting-associated virus (GRSPaV) in field-grown grapevine cv. Centennial Seedless plants sprayed with diluted (1:100) garlic extract were about 31–40%, 26–38%, 18–31%, 17–42% and 15–18%, respectively, of that in controls. Moreover, the garlic extract treatment led to a significant decrease in viral RNA accumulation of GLRaV-3, GLRaV-2, GVA, GFkV, GFLV, GRSPaV and grapevine Pinot Gris virus in pot-grown grapevine cv. Shine Muscat plants, and viral disease symptoms in these plants were obviously attenuated. In addition, this extract significantly induced expression of pathogenesis-related protein genes and stimulated activity of antioxidant enzymes in grapevines. Taken together, these results indicate that the crude garlic extract acts as a significant inhibitor against a broad range of grapevine viruses.