Host specificity and genetic diversity of race 4 strains of Ralstonia solanacearum

Ralstonia solanacearum race 4 isolates were obtained from Zingiberaceae plants in India during bacterial wilt outbreaks. Polyphasic phenotypic and genotypic analysis revealed intraracial diversity and dominance of biovar 3 over biovar 4. Biovar 3 strains were isolated from very severely wilted Zingiberaceae plants in the field and found to be present across diverse geographical, host and seasonal boundaries. It was hypothesized that these isolates belong to a single, ‘fast wilting’, lineage. Using one ‘fast wilting’ isolate in controlled inoculations, rapid wilt was observed in ginger within 5–7 days. Wilting was also observed in several other closely and distantly related hosts such as turmeric (Curcuma longa), aromatic turmeric (Curcuma aromatica), black turmeric (Curcuma caesia), sand ginger (Kaempferia galanga), white turmeric (Curcuma zeodaria), awapuhi (Zingiber zerumbet), greater galangal (Alpinia galanga), globba (Globba sp.), small cardamom (Elettaria cardamomum) and large cardamom (Ammomum subulatum) of the Zingiberaceae family, and in tomato (Solanum lycopersicum). Molecular analysis, including multiplex PCR‐based phylotyping, sequence analysis of 16S rDNA, 16–23S intergenic spacer and the recN gene, and multilocus sequence typing, revealed minimal differences between fast wilting isolates, confirming that almost all belong to the same lineage. Biovar 4 was isolated from plants showing slow wilt progression and self‐limiting wilting in restricted geographical locations instead, and was identified to be genetically distinct from the fast wilting biovar 3 isolates. To the authors' knowledge, this is the first report of host range and genetic analysis of R. solanacearum race 4 in India.     

Identification of mungbean lines with tolerance or resistance to yellow mosaic in fields in India where different begomovirus species and different <Emphasis Type="Italic">Bemisia tabaci</Emphasis> cryptic species predominate

The segmentation of symptoms during image analysis of diseased plant leaves is an essential process for detection and classification of diseases. However, there are challenges involved in the task, many of them related to the variability of image and host/symptom characteristics and conditions. As a result of those challenges, the methods proposed in the literature so far focus on a specific problem and are usually bounded by tight constraints regarding image capture conditions. This research explores a new automatic method for segmenting disease symptoms on plant leaves that was designed to be applicable in a wide range of situations. The proposed technique employs only color channel manipulations and Boolean operations applied on binary masks, thus being simpler and more robust compared to many previously described automatic methods. Its effectiveness is demonstrated by tests performed over a large database containing images of 77 different diseases of 11 plant species. A comparison with manual segmentation is also presented, further reinforcing the advantages of the proposed approach.     

Toxicity and dissipation of soil insecticides applied in the management of arecanut white grub, <Emphasis Type="Italic">Leucopholis burmeisteri</Emphasis> Brenk. (Coleoptera: Scarabaeidae)

White grubs, Leucopholis spp. are subterranean pests of arecanut grown in South India. Grub infestation leads to yellowing, stem tapering, and crown size and yield reduction. Use of chemical insecticide to mage the white grubs gives varying degree of success. Hence an attempt was made to screen newer and safer insecticides. Imidacloprid (LC₅₀ at 120 h = 16.849 ppm on III instar larvae), chlorpyriphos (LC₅₀ = 14.242) and bifenthrin (LC₅₀ = 12.797 ppm) were identified as effective insecticides. Evaluation of these insecticide in the field over two year period indicated the following efficacy in reducing larval population: chlorpyriphos @ 4 kg a.i/ha (83.31%) > bifenthrin @ 4 kg a.i./ha (82.83%) > imidacloprid @ 0.24 kg a.i./ha (75.84%) > bifenthrin @ 2 kg a.i./ ha (74.26%) > chlorpyriphos @ 2 kg a.i./ha (69.15%) > chlorpyriphos @ 1 kg a.i./ha (61.79%) > imidacloprid @ 0.12 kg a.i./ha (56.54%) > bifenthrin @ 1 kg a.i./ha (54.34%) > imidacloprid @ 0.06 kg a.i./ha (41.47%). Bifenthrin in soil persisted for a longer period than chlorpyriphos. On the day of application, 59.46 ppm bifenthrin residue was recovered from soil. On 10th day, it was 7.29 ppm which decreased to 2.59 ppm on 30th day and was beyond detection limit on 65th day. Chlorpyriphos exhibited a rapid degradation in the initial stage; 27.46 ppm residue on the day of application, which further reduced to 0.964 ppm on 10th day, and was below the detection limit on the 30th day. Growth of Trichoderma harzianum was not affected by bifenthrin even up to 40 ppm concentration. However, chlorpyriphos affected the growth of Trichoderma at higher than 5 ppm dose. Similarly, imidacloprid inhibited the colony growth from 2 ppm onwards. Having high lipophylic property and contact toxicity, bifenthrin would be an ideal alternative insecticide to chlorpyriphos for the management of white grubs in palm garden, which is safe and long persisting.     

Morphological and biochemical basis of resistance in okra to cotton jassid, <Emphasis Type="Italic">Amrasca biguttula biguttula</Emphasis> (Ishida)

Fifteen okra germplasm entries viz. accessions: IC0506027, IC0506118 and EC0306728; Abelmoschus spp.: Abelmoschus tuberculatus, Abelmoschus moschatus, Abelmoschus angulosus, Abelmoschus tetraphyllus, Abelmoschus manihot and Abelmoschus caillei; genotypes: POL-6 and POL-7; and four cultivated varieties: Punjab 8, Punjab Padmini, Punjab 7 and Pusa Sawani were screened against jassid, Amrasca biguttula biguttula (Ishida) in field at Vegetable Research Farm, Department of Vegetable Science, Punjab Agricultural University, Ludhiana, India during Kharif 2015. Different morphological and biochemical parameters of leaves of the selected entries were also studied. The correlation between jassid nymphal population and mid vein hair density, total phenols and tannins was negative and significant (r = ?0.67, ?0.83, ?0.75, respectively); negative and non-significant for hair length, angle of insertion of hair, total sugars and silica (r = ?0.40, ?0.49, ?0.63 and ?0.59, respectively) and positive and highly significant for lacination index, reducing sugars and lignins (r = 0.82, 0.95 and 0.90, respectively). Abelmoschus spp. Abelmoschus tetraphyllus, Abelmoschus angulosus and Abelmoschus moschatus were found to be field resistant on the basis of significantly lower pooled jassid nymphal population (1.56–1.99), jassid injury index (1.16–1.27) and susceptibility index (2.70–2.92). High degree of resistance in Abelmoschus tetraphyllus, Abelmoschus angulosus and Abelmoschus moschatus was found to be associated with high hair density (4.75–7.50), longer hair (1285.00–1513.20 μm), more erect hair (83.40–95.20°), broad leaves, high total sugars (15.21–18.36 mg/g), total phenols (1.52–1.58 mg/g), tannins (26.12–31.48 mg/g) and silica (32.66–33.17 mg/g) and low levels of reducing sugars (2.50–3.39 mg/g). Abelmoschus tuberculatus, A. manihot, IC0506027 and EC0306728 were found moderately field resistant with variable levels of morphological and biochemical parameters. High hair density, broad leaves, moderate levels of total sugars, reducing sugars, total phenols, tannins and silica seems to be associated with moderate levels of resistance in these entries. The variable levels of above mentioned parameters in moderately resistant entries also indicate that a single factor is not responsible for resistance but combination of different factors may be conferring resistance to jassid.     

Low pathotype diversity in a recombinant Puccinia striiformis population through convergent selection at the eastern Himalayan centre of diversity (Nepal)

Worldwide Puccinia striiformis f. sp. tritici (Pst) epidemics have been reported to be driven by few genetic lineages, while a high diversity is evident at the Pst Himalayan centre of diversity. This study investigated the relationship between pathotype diversity and genetic structure in Nepal, the eastern Himalayan region, which has been largely unexplored. Despite the high genetic diversity and recombinant structure detected through microsatellite genotyping, characterization of virulence phenotypes for 62 isolates identified only eight pathotypes, with two pathotypes predominant over all the populations. This is in contrast to the Pakistani and Chinese recombinant populations, where high pathotype diversity is associated with genetic diversity. The most prevalent Nepali pathotype was not a unique clonal lineage, but was represented by seven multilocus genotypes from four distinct genetic subgroups, suggesting strong directional selection on virulence genes, resulting in convergent pathotypes in distinct genetic groups. This convergent selection is discussed in comparison with clonal French and recombinant Pakistani populations. Additionally, the Nepali Pst population carried virulence to 17 out of 24 tested yellow rust resistance genes (Yr), with the absence of virulence to Victo and Early Premium and resistance genes Yr5, Yr10, Yr15, Yr24 and Yr26. Virulence to Yr2, Yr7, Yr27 and YrSu were fixed in all isolates, in line with the deployment of these resistance genes in Nepal. The results reflect the influence of resistance gene deployment on selection of virulence and pathotypes in a recombinant pathogen population, which must be considered in the context of durable resistance gene deployment.     

Potential biopesticides for crucifer flea beetle, <Emphasis Type="Italic">Phyllotreta cruciferae</Emphasis> (Coleoptera: Chrysomelidae) management under dryland canola production in Montana

The crucifer flea beetle, Phyllotreta cruciferae (Goeze), is an economically important and dominant pest of canola (Brassica napus L) in the Northern Great Plains of the USA. The current flea beetle management strategy is based on using synthetic chemical treated seeds and if necessary, foliar spray of chemicals at canola seedlings in early spring for targeting adult population. However, there is an increasing demand for development of alternative management strategies for P. cruciferae pertaining to concerns over the development of resistance to synthetic insecticides and non-target effects on pollinators and other beneficial insects. Replicated field trials were conducted to test the efficacy of several commercially available biopesticides including Entrust® (spinosad), entomopathogenic nematode Steinernema feltiae?+?Barricade® (polymer gel 1%), Aza-Direct® (azadirachtin), Pyganic 1.4® EC (pyrethrin), Grandevo® SC (Chromobacterium subtsugae), Venerate® XC (Heat killed Burkholderia sp. strain A396 as seed treatment and foliar application) and Gaucho® (imidacloprid) (chemical check) for the P. cruciferae management at two locations (Conrad and Sweetgrass) of Montana in 2016. Biopesticide products were evaluated based on canola leaf area injury ratings and seed yield levels. Although, there was no clear trend of canola yield increase, selected biopesticide treatments were effective in maintaining low leaf area injury ratings as compared to untreated control. Entrust was able to maintain low leaf area injury ratings (8.5–14.5%) when compared to untreated control (16.0–21.4%) at both the locations. Entomopathogenic nematodes, Steinernema feltiae?+?Barricade® and Venerate® applied as foliar treatments maintained significantly lower feeding injury pressure at Sweetgrass (11.8%) and Conrad (13.4%) locations respectively, when compared to the untreated control. Our study results suggest that these biopesticide treatment results were comparable in efficacy to the chemical seed treatment Gaucho®. Other two biopesticide products- Aza-Direct® and Pyganic 1.4® EC treatments did not provide effective control of P. cruciferae at both the locations.     

Effect of host plants on developmental and population parameters of invasive leafminer, <Emphasis Type="Italic">Tuta absoluta</Emphasis> (Meyrick) (Lepidoptera: Gelechiidae)

Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is one of the most serious pests of tomato, capable of causing 100% crop losses under favourable conditions. We studied the effect of different host plants on developmental and population parameters of this pest at 25?±?0.5 °C temperature, 70?±?5% relative humidity and 12 L: 12D photoperiod. Host plant had significant effect on the developmental biology and the population growth parameters of the pest. The leafminer developed fastest on tomato leaves and slowest on potato tubers. Population growth parameters like intrinsic rate of increase, net reproductive rate, finite rate of increase, doubling time and weekly multiplication rate of T. absoluta were highest on tomato leaves and lowest on potato tubers. Mean generation time was minimum on tomato leaves and maximum on potato tubers. Females developed on tomato leaves were more fecund than other hosts. Though, tomato was found to be the most suitable host plant of T. absoluta, yet, the pest developed and grew successfully on other alternate hosts like potato (Solanum tuberosum L.), brinjal (Solanum melongena L.) and pepino (Solanum muricatum Aiton). These hosts can, therefore, play an important role in the survival, population build up and overwintering of the miner. Further, under favourable conditions the miner can become a serious pest on these crops and need to be monitored on these crops as well.     

Influence of abiotic factors on behaviour and adult emergence pattern of coconut white grub, <Emphasis Type="Italic">Leucopholis coneophora</Emphasis> Burmeister (Coleoptera: Scarabaeidae)

Leucopholis coneophora Burmeister is a subterranean pest associated with coconut based cropping systems in south India. Feeding damage causes yellowing of fronds and yield reduction. To develop appropriate IPM strategy a basic knowledge on insect behaviour is essential. Four years studies indicated that, adult emergence of L. coneophora was commenced with summer shower in April in Kerala. Delay in summer shower delayed the emergence. After a pause in May, the emergence resumed with the setting of south west monsoon in June. The beetles did not emerge during dry spells in between the rainy days, when the soil temperature (at 10 cm depth) was ≥34.5 °C. Emergence of the beetles started at an illuminance of 124.37?±?75.5 l in evening and remained active till 2?±?0.4 l with a maximum swarming at 32.6?±?15.1 l. Female emergence and mating occurred at 12.04?±?8.1 l. Female based sex pheromone mediated communication is evident. Strong competition among the males for mating with emerging female, which was evident by a wider operational sex ratio in the initial period (1:10.11) that narrowed down to 1:4.33 in later days. The beetles neither congregate on any host plant nor exhibit phototaxis. Number of beetles entrapped in light traps varied from 1.5–16.5% and hand picking is highly significant over light trapping. Hence hand picking of beetles daily in the evening for 2 weeks commencing from the onset of south west monsoon in Kerala, in Indian subcontinent is suggested as a tool in IPM.     

Effect of two-component cultivar mixtures on development of wheat yellow rust disease in the field and greenhouse in the Nepal Himalayas

Wheat yellow rust (WYR), caused by Puccinia striiformis f. sp. tritici (PST), is a major disease of wheat, and deployment of a single cultivar often leads to disease epidemics. Effect of inoculum level, foliar fungicide spray, and wheat cultivar mixtures were evaluated on disease development in the field and greenhouse in Nepal. Treatments were arranged in a split–split plot design with three replications in both experiments. Two inoculum levels of PST (low and high) were main plot factors; nontreated control and foliar spray of fungicides (Mancozeb and Bayleton) were subplot factors; and two-component cultivar mixtures, composed of different ratios of a susceptible (S) and a resistant (R) cultivars (90:10, 80:20, and 50:50, 100:0, and 0:100) were sub–subplot factors. WYR severity was assessed at different time intervals, and disease development was calculated as area under the disease progress curve (AUDPC). Inoculum level did not cause significant differences in AUDPC in the field but did in the greenhouse. Foliar spray of fungicides reduced the AUDPC in the greenhouse and field. In both experiments, AUDPC values were low in cultivar mixtures compared with a pure stand of a susceptible cultivar. As the proportion of resistant cultivar increased compared with the susceptible cultivar in the S:R mixture component, disease severity decreased with a consequent increase in grain yield. The greater yield obtained with cultivar mixtures compared with only the susceptible cultivar, independent of inoculum level and fungicide spray in the field, revealed a promising strategy to manage WYR in Nepal.