R. P. Thakur

Downy mildews of India

Abstract The downy mildew diseases are caused by different fungus species in several genera in the class Oomycetes. They comprise a major group of diseases affecting a large number of crops. In India, downy mildews have been reported on several economically important crops, including maize, sorghum, pearl millet, onion, soybean, cucurbits, and grapes, causing severe economic losses in some regions and seasons. Pathogens reported to cause downy mildews in India include species in the genera, Perenosclerospora, Perenospora, Pseudoperonospora, Plasmopara, Sclerophthora and Sclerospora. In view of the economic importance of the crops, and prevalence and severity of the disease, the downy mildews have been classified into high, moderate and low research priority problems. In this article we present a brief review of the work done in India related to pathogen biology, epidemiology and management methods, and provide an outlook for future research.

Phylogenetic relationships of graminicolous downy mildews based on cox2 sequence data

Graminicolous downy mildews (GDM) are an understudied, yet economically important, group of plant pathogens, which are one of the major constraints to poaceous crops in the tropics and subtropics. Here we present a first molecular phylogeny based on cox2 sequences comprising all genera of the GDM currently accepted, with both lasting (Graminivora, Poakatesthia, and Viennotia) and evanescent (Peronosclerospora, Sclerophthora, and Sclerospora) sporangiophores. In addition, all other downy mildew genera currently accepted, as well as a representative sample of other oomycete taxa, have been included. It was shown that all genera of the GDM have had a long, independent evolutionary history, and that the delineation between Peronosclerospora and Sclerospora is correct. Sclerophthora was found to be a particularly divergent taxon nested within a paraphyletic Phytophthora, but without support. The results confirm that the placement of Peronosclerospora and Sclerospora in the Saprolegniomycetidae is incorrect. Sclerophthora is not closely related to Pachymetra of the family Verrucalvaceae, and also does not belong to the Saprolegniomycetidae, but shows close affinities to the Peronosporaceae. In addition, all GDM are interspersed throughout the Peronosporaceae s lat., suggesting that a separate family for the Sclerosporaceae might not be justified.

Resistance to Grain Mold and Downy Mildew in a Mini-Core Collection of Sorghum Germplasm

Grain mold and downy mildew are important biotic constraints to grain sorghum (Sorghum bicolor) production worldwide and are best managed through host plant resistance. A sorghum mini-core collection composed of 242 germplasm accessions developed from a core collection of 2,246 landrace accessions from 58 countries was evaluated to identify sources of grain mold and downy mildew resistance. Of the 242 accessions, 140 that flowered during the rainy season (the other 102 accessions were photoperiod sensitive) were screened for grain mold resistance in a grain mold nursery under field epiphytotic conditions during 2007 and 2008. All 242 accessions were screened for downy mildew in the greenhouse using a sandwich inoculation technique. Fifty accessions were resistant to grain mold (≤10% mean severity). These resistant accessions represented four basic and six intermediate races of sorghum that originated from 21 countries and exhibited considerable diversity for agronomic and morphological traits. Downy mildew resistance (mean incidence ≤10%) was observed only in six (IS 28747, IS 31714, IS 23992, IS 27697, IS 28449, and IS 30400) of the 242 accessions. One accession, IS 23992, exhibited resistance to both the diseases. The morphologically and agronomically diverse accessions that are resistant to grain mold or downy mildew should be useful to sorghum disease resistance breeding programs.

Identification and Characterization of Toxigenic Fusaria Associated with Sorghum Grain Mold Complex in India

Fusarium species are dominant within the sorghum grain mold complex. Some species of Fusarium involved in grain mold complex produce mycotoxins, such as fumonisins. An attempt was made to identify Fusarium spp. associated with grain mold complex in major sorghum-growing areas in India through AFLP-based grouping of the isolates and to further confirm the species by sequencing part of α-Elongation factor gene and comparing the sequences with that available in the NCBI database. The dendrogram generated from the AFLP data clustered the isolates into 5 groups. Five species of Fusarium—F. proliferatum, F. thapsinum, F. equiseti, F. andiyazi and F. sacchari were identified based on sequence similarity of α-Elongation factor gene of the test isolates with those in the NCBI database. Fusarium thapsinum was identified as predominant species in Fusarium—grain mold complex in India and F. proliferatum as highly toxigenic for fumonisins production. Analysis of molecular variance (AMOVA) revealed 54% of the variation in the AFLP patterns of 63 isolates was due to the differences between Fusarium species, and 46% was due to differences between the strains within a species.

Characterisation of pathogenic and molecular diversity in Sclerospora graminicola, the causal agent of pearl millet downy mildew

Genotypic diversity among 46 isolates of Sclerospora graminicola collected from seven states in India during 1992–2005 was determined through pathotyping and AFLP analysis. A high level of variation was observed among the isolates for downy mildew incidence, latent period and virulence index. Based on the reaction on a set of nine pearl millet lines, 46 isolates were classified in 21 pathotypes. Quantitative differences in virulence levels of the test isolates were assessed by calculating the virulence index (disease incidence × latent period − 1). A dendrogram generated by the average linkage cluster analysis of virulence index clustered the 46 isolates into eight groups. Region-specific grouping of five isolates from Gujarat and six from Rajasthan was observed within two distinct groups. Temporal variation was also observed among the isolates collected from the same location and same host over the years. A total of 297 bands were scored following selective amplification with three primer combinations E-TT/M-CAG, E-AT/M-CAG and E-TG/M-CAT and all of them were polymorphic. Cluster analysis of AFLP data clustered the test isolates into seven groups. Analysis of molecular variance indicated that variation in the S. graminicola populations was largely due to differences among the isolates within the states.

Ergot reaction of pearl millet hybrids affected by fertility restoration and genetic resistance of parental lines

SummaryPearl millet (Pennisetum glaucum (L.) R. Br.) hybrids based on the A1 cytoplasmic-nuclear male-sterile (CMS) lines are more susceptible to smut (Tolyposporium penicillariae Bref.) than open-pollinated varieties. Seventy eight pairs of hybrids, made onto male-sterile (A) lines and their counterpart maintainer (B) lines, were evaluated to examine the effects of male sterility and genetic resistance of parental lines on the smut severity of hybrids. The A-line hybrids had higher smut severity and lower selfed seedset than the counterpart B-line hybrids, indicating that it is the CMS-mediated male sterility rather than the A1 cytoplasm per se that caused greater smut severity of A-line hybrids. However, with the use of resistant parental lines even male-sterile hybrids of A-lines, in several cases, were as resistant as some of the highly resistant male-fertile hybrids of B-lines. It would be possible to produce smut resistant hybrids (< 10% severity) on A-lines, albeit in low frequency, even if only one parent of a hybrid were resistant. However, the probability of producing such hybrids would be higher when both parents were resistant to smut. Thus, improvement in smut resistance of parental lines and fertility restoration ability of pollinators would provide the most effective genetic approach to smut disease management in hybrids.

SSR allelic diversity in relation to morphological traits and resistance to grain mould in sorghum

Allelic variation at 46 simple sequence repeat (SSR) marker loci well distributed across the sorghum genome was used to assess genetic diversity among 92 sorghum lines, 74 resistant and 18 susceptible to grain mould. Of the 46 SSR markers, 44 were polymorphic, with the number of alleles ranging from 2 to 20 with an average of 7.55 alleles per locus. Genetic diversity among the sorghum lines was high as indicated by polymorphic information content (PIC) and gene diversity values. PIC values of polymorphic SSR markers ranged from 0.16 to 0.90, with an average of 0.54. Gene diversity among the sorghum lines varied from 0.16 to 0.91, with an average score of 0.58 per SSR marker. AMOVA indicated that 12% of the total variation observed among the sorghum lines was accounted for between grain mould resistant and susceptible types. Diversity based on six morphological traits and grain mould scores indicated major roles of panicle type and glumes coverage, followed by grain colour, in clustering of the lines. Seven grain mould resistant/ susceptible pairs with dissimilarity indices >0.50, but with similar flowering time, plant height, and panicle type/ inflorescence within each pair, were selected for use in developing recombinant inbred line mapping populations to identify genomic regions (and quantitative trait loci) associated with sorghum grain mould resistance.

GENETIC AND PATHOGENIC VARIABILITY AMONG ISOLATES OF SCLEROSPORA GRAMINICOLA, THE DOWNY MILDEW PATHOGEN OF PEARL MILLET

Downy mildew, caused by the oomycete pathogen Sclerospora graminicola (Sacc.) Schroet, is economically the most important disease of pearl millet (Pennisetum glaucum (L.) R. Br.) in Asia and Africa. The pathogen induces systemic infection in pearl millet plants that manifests itself through foliar chlorosis and panicle malformation (also called “green-ear” or “crazy top”). The fungus is heterothallic (Michelmore et al., 1982) and reproduces both by sexual and asexual means, and is therefore highly variable (Ball and Pike, 1984; Thakur et al., 1999). The commercial cultivation of a number of genetically homogeneous single-cross F1 hybrids of pearl millet in India has led to increased virulence in S. graminicola populations, thus shortening the useful life of the hybrid cultivars (Thakur et al., 1999). Monitoring virulence changes in the pathogen population, identifying resistance to specific and multiple pathotypes, and directing pearl millet breeding programmes towards strategic utilization and deployment of resistance genes form the basis of a long-term downy mildew management research at ICRISAT (Thakur, 1999).

Virulence Diversity in North Indian Isolates of Sclerospora Graminicola, the Pearl Millet Downy Mildew Pathogen

On-farm surveys were conducted in the Uttar Pradesh (India) during the two rainy seasons 2007 and 2008 to monitor pearl millet (Pennisetum glaucum) downy mildew incidence. Twenty-one isolates of Sclerospora graminicola, the pearl millet downy mildew pathogen, were collected from different hybrid cultivars. These isolates were established on seedlings of the highly susceptible line 7042S grown in the greenhouse and were characterized for their virulence diversity using a set of seven host differential lines. Quantitative differences in virulence among pathogen isolates were determined by calculating virulence index (percent disease incidence × latent period-1). Results were submitted to cluster analysis using the Average Linkage method to determine similarity among pathogen isolates. The two highly virulent isolates, Sg 492 from Aligarh and Sg 510 from Badaun, representing geographically diverse locations were selected for use in greenhouse screening of pearl millet breeding lines.

Biochemical characterisation of grain mould resistant and susceptible genotypes and PGPR-induced resistance in the host to Curvularia lunata and Fusarium proliferatum

Resistance to biotic stresses in plants is either due to the presence of preformed biochemical compounds or induced in response to external stimulus. In this study, 13 grain mould resistant and seven susceptible lines of sorghum were analysed for biochemical defence mechanism. The levels of total phenols and phenylalanine ammonia lyase were almost same in the resistant and susceptible genotypes. However, two additional isoforms of peroxidase were found in the three of the 13 resistant genotypes. The isoform peroxidase corresponding to the R f value of 0.25 was found in the resistant genotypes IS 13969, ICSB 377 and IS 8219-1, and two genotypes IS 13969 and ICSB 377 had an additional isoform corresponding to the R f value of 0.32. The results indicated the genotype specific association of peroxidases with grain mould resistance in sorghum. Nine bacterial strains (Bacillus pumilus SB 21, Bacillus megaterium HiB 9, Bacillus subtilis BCB 19, Pseudomonas plecoglossicida SRI 156, Brevibacterium antiquum SRI 158, B. pumilus INR 7, P. fluorescens UOM SAR 80, P. fluorescens UOM SAR 14, B. pumilus SE 34) were tested to induce systemic resistance in sorghum cultivars 296B and Bulk Y against the highly pathogenic grain mould pathogens Curvularia lunata and Fusarium proliferatum, respectively. The bacterial isolates were effective in inducing resistance in sorghum. Among the strains tested, SRI 158 was found highly effective in reducing grain mould severity in both the genotypes.