SEVERITY AND INTENSITY OF FUNGAL DISEASES OF CUCURBIT CROPS OF HAMIRPUR REGION, HP, INDIA

Cucurbitaceae is the vegetable family enriched with numerous vitamins and minerals. In early rainy season cucurbits are the host of different fungal pathogens. In present study, four fungal diseases dominant on three members of cucurbitaceae family are observed in Bhoranj block of District Hamirpur, lie in lower foot hills of Himachal Pradesh. Four fungal diseases along with host plants are leaf spot of Lagenaria siceraria, downy mildews of Cucumis sativus, powdery mildews of Coccinia grandis and powdery mildews of Cucumis sativus. Among all these diseases, Disease Incidence and Disease Severity of Powdery mildews of Coccinia grandis are most dominant i.e. 40.3±1 and 51.2±1 respectively. Disease Incidence and Disease Severity of Leaf spot of Lagenaria siceraria is least i.e. 12.5±1 and 1.1±1, respectively.

Fungi Resistance to Multissite Fungicides

Multisite fungicides have been used for many years in fruit and vegetable crops worldwide. Cases of the fungi resistance development to these fungicides have been rare. From the 2002 season onwards, with the outbreak of Asian soybean rust in Brazil, caused by Phakopsora pachyrhizi, site-specific fungicides became the main weapon for its control. From 2002 to 2011, penetrant mobile site-specific fungicides were used and until today in double (DMI + QoI) or triple (DMI + QoI + SDHI) co-formulatoons in an area of more than 30 million hectares and with three sprays per area. This resulted, as expected, in the fungus sensitivity reduction, today with cross and multiple resistance to those site-specific fungicides. From the 2011 season in an attempt to recover control that for some chemicals and mixtures reached < 30%, research was started with site-specific + multi-site mixtures, taking as example Phytophthora infestans resistance development to metalaxyl in Europe showinig long-lasting solution found by the addition of multisite mancozeb. It is expected that the effective life of site-specific + multi-site mixtures may be as long in controlling soybean rust as it has been for potato, tomato and grape downy mildews. This review presents the concepts involved in the sensitivity reduction to fungicides. Some fungal species and fungicides involved are listed. Considering the P. pachyrhizi sporulation potential, the great soybean area sprayed and the number of sprays per area mainly with site-specific co-formulations and the reduced area sprayed with multisites, we discuss the need for annual monitoring of P. pachyrhizi sensitivity to the these chemicals.

Strobilurins: New group of fungicides

Strobilurin is a group of natural products and their synthetic analogs have been widely used to control and prevent fungal diseases. Strobilurins were firstly isolated in 1977 from the mycelium of Strobilurus tenacellus, a saprobic Basidiomycete fungus causing wood-rotting on forest trees. This group of pesticides was designed to manage fungal pathogens classes such as Ascomycetes, Basidiomycetes, and Oomycetes. Also, Strobilurin commercialized included derivatives such as are azoxystrobin, kresoxim-methyl, picoxystrobin, fluoxastrobin, oryzastrobin, dimoxystrobin, pyraclostrobin and trifloxystrobin. This group is a part of the larger group of QoI inhibitors, which act to inhibit the respiratory chain at the level of Complex III. Strobilurins group control an unusually wide array of fungal diseases, included water molds, downy mildews, powdery mildews, leaf spotting and rusts. This group are used on cereals, field crops, fruits, tree nuts, vegetables, turfgrasses and ornamentals. Also, Strobilurins found to enhance the plant growth in some cases.

“Jumping Jack”: Genomic Microsatellites Underscore the Distinctiveness of Closely Related Pseudoperonospora cubensis and Pseudoperonospora humuli and Provide New Insights Into Their Evolutionary Past

Downy mildews caused by obligate biotrophic oomycetes result in severe crop losses worldwide. Among these pathogens, Pseudoperonospora cubensis and P. humuli, two closely related oomycetes, adversely affect cucurbits and hop, respectively. Discordant hypotheses concerning their taxonomic relationships have been proposed based on host–pathogen interactions and specificity evidence and gene sequences of a few individuals, but population genetics evidence supporting these scenarios is missing. Furthermore, nuclear and mitochondrial regions of both pathogens have been analyzed using microsatellites and phylogenetically informative molecular markers, but extensive comparative population genetics research has not been done. Here, we genotyped 138 current and historical herbarium specimens of those two taxa using microsatellites (SSRs). Our goals were to assess genetic diversity and spatial distribution, to infer the evolutionary history of P. cubensis and P. humuli, and to visualize genome-scale organizational relationship between both pathogens. High genetic diversity, modest gene flow, and presence of population structure, particularly in P. cubensis, were observed. When tested for cross-amplification, 20 out of 27 P. cubensis-derived gSSRs cross-amplified DNA of P. humuli individuals, but few amplified DNA of downy mildew pathogens from related genera. Collectively, our analyses provided a definite argument for the hypothesis that both pathogens are distinct species, and suggested further speciation in the P. cubensis complex.

Application of target enrichment sequencing for population genetic analyses of the obligate plant pathogens Pseudoperonospora cubensis and P. humuli in Michigan

Technological advances in genome sequencing have improved our ability to catalog genomic variation and led to an expansion of the scope and scale of genetic studies. Yet, for agronomically important plant pathogens such as the downy-mildews the scale of genetic studies remains limited. This is, in part, due to the difficulties associated with maintaining obligate pathogens, and the logistical constraints involved in the genotyping of these species. To study the genetic variation of two Pseudoperonospora species (P. cubensis and P. humuli), we describe a targeted enrichment (TE) protocol able to genotype isolates using less than 50 ng of mixed pathogen and plant DNA for library preparation. We enriched 830 target genes across 128 samples and identified 2,514 high-quality SNP variants. We detected significant genetic differentiation (p=0.01) between P. cubensis subpopulations from Cucurbita moschata (clade I) and Cucumis sativus (clade II) in Michigan. No evidence of location-based differentiation was detected within the P. cubensis (clade II) subpopulation. A significant effect of location on the genetic variation of the P. humuli subpopulation was detected in the state (p=0.01). Mantel tests found evidence that the genetic distance among P. humuli samples was associated with the physical distance of the hop yards from which the samples were collected (p=0.005). The differences in the distribution of genetic variation of the P. humuli and P. cubensis subpopulations of Michigan suggest differences in the dispersal of these two species. Our TE protocol provides an additional tool for genotyping obligate pathogens and the execution of new genetic studies

Peronosclerospora philippinensis (Philippine downy mildew of maize).

Peronosclerospora philippinensis, a causal pathogen of maize downy mildews, is one of the major maize diseases reported in some maize-growing countries, especially in the Philippines. High disease incidence has been reported in many parts in the country specifically in northern Luzon and in many parts of Mindanao despite breakthroughs in controlling or mitigating the disease using cultural and chemical control (Pascual et al., 2005). P. philippinensis is considered the most virulent of the downy mildew pathogens affecting maize, causing substantial losses to crop production (Murray, 2009). Under normal conditions, a 40-60% yield reduction is observed; however, favourable conditions for disease development can amplify these losses to 80-100% (Exconde and Raymundo, 1974). As the pathogen is able to survive in seeds, is able to spread rapidly and occasionally forms resting spores that can survive for more than 1 year, the pathogen has the potential to become a threat to local maize production in warm temperate and tropical areas.

Peronosclerospora philippinensis (Philippine downy mildew of maize).

Peronosclerospora philippinensis, a causal pathogen of maize downy mildews, is one of the major maize diseases reported in some maize-growing countries, especially in the Philippines. High disease incidence has been reported in many parts in the country specifically in northern Luzon and in many parts of Mindanao despite breakthroughs in controlling or mitigating the disease using cultural and chemical control (Pascual et al., 2005). P. philippinensis is considered the most virulent of the downy mildew pathogens affecting maize, causing substantial losses to crop production (Murray, 2009). Under normal conditions, a 40-60% yield reduction is observed; however, favourable conditions for disease development can amplify these losses to 80-100% (Exconde and Raymundo, 1974). As the pathogen is able to survive in seeds, is able to spread rapidly and occasionally forms resting spores that can survive for more than 1 year, the pathogen has the potential to become a threat to local maize production in warm temperate and tropical areas.

Fantastic Downy Mildew Pathogens and How to Find Them: Advances in Detection and Diagnostics

Downy mildews affect important crops and cause severe losses in production worldwide. Accurate identification and monitoring of these plant pathogens, especially at early stages of the disease, is fundamental in achieving effective disease control. The rapid development of molecular methods for diagnosis has provided more specific, fast, reliable, sensitive, and portable alternatives for plant pathogen detection and quantification than traditional approaches. In this review, we provide information on the use of molecular markers, serological techniques, and nucleic acid amplification technologies for downy mildew diagnosis, highlighting the benefits and disadvantages of the technologies and target selection. We emphasize the importance of incorporating information on pathogen variability in virulence and fungicide resistance for disease management and how the development and application of diagnostic assays based on standard and promising technologies, including high-throughput sequencing and genomics, are revolutionizing the development of species-specific assays suitable for in-field diagnosis. Our review provides an overview of molecular detection technologies and a practical guide for selecting the best approaches for diagnosis.

Diploid chromosome-scale assembly of the Muscadinia rotundifolia genome supports chromosome fusion and disease resistance gene expansion during Vitis and Muscadinia divergence

Muscadinia rotundifolia, the muscadine grape, has been cultivated for centuries in the southeastern United States. M. rotundifolia is resistant to many of the pathogens that detrimentally affect Vitis vinifera, the grape species commonly used for winemaking. For this reason, M. rotundifolia is a valuable genetic resource for breeding. Single-molecule real-time reads were combined with optical maps to reconstruct the two haplotypes of each of the 20 M. rotundifolia cv. Trayshed chromosomes. The completeness and accuracy of the assembly were confirmed using a high-density linkage map of M. rotundifolia. Protein-coding genes were annotated using an integrated and comprehensive approach. This included using Full-length cDNA sequencing (Iso-Seq) to improve gene structure and hypothetical spliced variant predictions. Our data strongly support that Muscadinia chromosomes 7 and 20 are fused in Vitis and pinpoint the location of the fusion in Cabernet Sauvignon and PN40024 chromosome 7. Disease-related gene numbers in Trayshed and Cabernet Sauvignon were similar, but their clustering locations were different. A dramatic expansion of the Toll/Interleukin-1 Receptor-like Nucleotide-Binding Site Leucine-Rich Repeat (TIR-NBS-LRR) class was detected on Trayshed chromosome 12 at the Resistance to Uncinula necator 1 (RUN1)/ Resistance to Plasmopara viticola 1 (RPV1) locus, which confers strong dominant resistance to powdery and downy mildews. A genome browser for Trayshed, its annotation, and an associated Blast tool are available at .www.grapegenomics.com