Manas Kumar Bag

Evaluation of 19,460 Wheat Accessions Conserved in the Indian National Genebank to Identify New Sources of Resistance to Rust and Spot Blotch Diseases

A comprehensive germplasm evaluation study of wheat accessions conserved in the Indian National Genebank was conducted to identify sources of rust and spot blotch resistance. Genebank accessions comprising three species of wheat–Triticum aestivum, T. durum and T. dicoccum were screened sequentially at multiple disease hotspots, during the 2011–14 crop seasons, carrying only resistant accessions to the next step of evaluation. Wheat accessions which were found to be resistant in the field were then assayed for seedling resistance and profiled using molecular markers. In the primary evaluation, 19,460 accessions were screened at Wellington (Tamil Nadu), a hotspot for wheat rusts. We identified 4925 accessions to be resistant and these were further evaluated at Gurdaspur (Punjab), a hotspot for stripe rust and at Cooch Behar (West Bengal), a hotspot for spot blotch. The second round evaluation identified 498 accessions potentially resistant to multiple rusts and 868 accessions potentially resistant to spot blotch. Evaluation of rust resistant accessions for seedling resistance against seven virulent pathotypes of three rusts under artificial epiphytotic conditions identified 137 accessions potentially resistant to multiple rusts. Molecular analysis to identify different combinations of genetic loci imparting resistance to leaf rust, stem rust, stripe rust and spot blotch using linked molecular markers, identified 45 wheat accessions containing known resistance genes against all three rusts as well as a QTL for spot blotch resistance. The resistant germplasm accessions, particularly against stripe rust, identified in this study can be excellent potential candidates to be employed for breeding resistance into the background of high yielding wheat cultivars through conventional or molecular breeding approaches, and are expected to contribute toward food security at national and global levels.

Correction: Use of molecular markers in identification and characterization of resistance to rice blast in India

Rice blast disease caused by Magnaporthe oryzae is one of the most destructive disease causing huge losses to rice yield in different parts of the world. Therefore, an attempt has been made to find out the resistance by screening and studying the genetic diversity of eighty released rice varieties by National Rice Research Institute, Cuttack (NRVs) using molecular markers linked to twelve major blast resistance (R) genes viz Pib, Piz, Piz-t, Pik, Pik-p, Pikm Pik-h, Pita/Pita-2, Pi2, Pi9, Pi1 and Pi5. Out of which, nineteen varieties (23.75%) showed resistance, twenty one were moderately resistant (26.25%) while remaining forty varieties (50%) showed susceptible in uniform blast nursery. Rice varieties possessing blast resistance genes varied from four to twelve and the frequencies of the resistance genes ranged from 0 to 100%. The cluster analysis grouped the eighty NRVs into two major clusters at 63% level of genetic similarity coefficient. The PIC value for seventeen markers varied from 0 to 0.37 at an average of 0.20. Out of seventeen markers, only five markers, 195R-1, Pi9-i, Pita3, YL155/YL87 and 40N23r corresponded to three broad spectrum R genes viz. Pi9, Pita/Pita2 and Pi5 were found to be significantly associated with the blast disease with explaining phenotypic variance from 3.5% to 7.7%. The population structure analysis and PCoA divided the entire 80 NRVs into two sub-groups. The outcome of this study would help to formulate strategies for improving rice blast resistance through genetic studies, plant-pathogen interaction, identification of novel R genes, development of new resistant varieties through marker-assisted breeding for improving rice blast resistance in India and worldwide.

Natural infection of croton yellow vein mosaic virus and its cognate betasatellite in germplasm of different Crambe spp in India

Crambe is an important crop grown worldwide for industrial oil and seed meal. Besides the fungal and bacterial diseases, the crop is reported to be infected by tobacco mosaic virus, beet western yellows virus and turnip mosaic virus under experimental condition. Till now, there was no report of natural infection of any begomovirus in this crop. In the present study, a leaf curl disease was observed in germplasm accessions of three species of Crambe (C. abyssinica, C. glabrata and C. hispanica). Based on the symptoms and presence of whitefly population in the field, begomovirus infection was suspected. Molecular characterization through RCA approach, indicated presence of croton yellow vein mosaic virus (CYVMV, KJ747958) and croton yellow vein mosaic betasatellite (CroYVMB, KM229762). Co-agroinoculation of partial dimeric construct of CYVMV with complete dimeric construct of CroYVMB, produced typical leaf curl symptoms in C. abyssinica, whereas, agroinoculation of partial dimeric construct of CYVMV alone could not produce symptoms in the same plant. In contrast, the CYVMV construct alone could produce symptom in Nicotiana benthamiana, a model host for plant virus studies. In N. benthamiana co-inoculation of CroYVMV with CYVMV construct develop more severe symptoms. However, neither the CYVMV construct alone nor the co-inoculation with CroYVMB produce any symptom in Arabidopsis thaliana even with different methods of inoculation. Inoculated Arabidopsis thaliana also did not yield any amplification of the virus as assessed through PCR and rolling circle amplification (RCA). Thus it confirmed that for successful infection in crambe, CYVMV requires betasatellite, while in N. benthamiana, it does not require betasatellite for symptom induction and in Arabidopsis thaliana CYVMV alone or in presence of betasatellite did not replicate and produce any symptom. This study constitutes the first confirmed record of natural infection of a begomovirus in crambe and further confirmed that cognate betasatellite of CYVMV has differential role in infectivity in different hosts.

Assessment of phenotypic diversity in pearl millet [Pennisetum glaucum (L.) R. Br.] germplasm of Indian origin and identification of trait-specific germplasm

Abstract. Evaluation of genetic diversity within germplasm collections and identification of trait-specific germplasm is a basic requirement for plant breeders. A total of 221 Indian pearl millet collections from the National Genebank were characterised and evaluated for 27 agro-morphological descriptors. Considerable variation was observed for all characters. Frequency distribution analysis showed predominance of cylindrical and compact spike, grey seeds, earliness (less than 40 days to spike emergence). Hierarchical clustering method was used for classifying 221 pearl millet accessions based on agronomic and disease resistance traits, which resulted into three clusters. Clusters 1, 2 and 3 comprised 91, 54 and 76 accessions respectively. There was high correspondence between the geographic collection sites of accessions and their inclusion in particular clusters. In addition, principal component analysis was used for data reduction and generating biplot. First four principal components explained 66.43% of total variability. Among the traits analysed, plant height, nodes/plant, days to spike emergence, number of tillers, leaf width and leaf length are major contributor towards phenotypic diversity. Further the trait-specific germplasm were identified for agronomic traits, disease resistance, popping and antioxidants activity, namely for earliness (IC343664, IC343689, IC343661, IC309064), spike girth (IC283693, IC283842, IC367638), dual purpose with high grain and fodder yield (IC283705, IC283745, IC283885 and IC335901 and so on). Four accessions of pearl millet germplasm viz., IC309064, IC393365, IC306465 and IC283866, were observed as multiple disease resistant. This study suggested that application of appropriate techniques and their interpretations provide more efficient way to identify potential accessions and improve the utilisation of germplasm collections in plant breeding.

Novel Trichoderma strains isolated from tree barks as potential biocontrol agents and biofertilizers for direct seeded rice

This study is the first time report of utilization of Trichoderma spp. isolated from different tree bark from Odisha state of India for rice crop health management and higher productivity. Six isolates of Trichoderma spp. were identified based on the morphological characteristics and species determination was performed by molecular assays. One of the isolated strains determined as Trichoderma erinaceum outperformed others. Trichoderma erinaceum controlled three soil borne plant pathogens i.e. Rhizoctonia solani, Sclerotium rolfsii and Sclerotium oryzae effectively under controlled condition and R. solani and Helminthosporium oryzae under filed condition. Seed treatments with the formulated isolates improved the germination rate of rice and enhanced vigour. These parameters along with higher chlorophyll content could be related to higher yield observed in two rice varieties; Karuna and Sahabhagidhan. Among the six isolates tested, Trichoderma erinaceum treatment recorded highest yield. Significantly higher expression of some stress related enzymes was observed in Trichoderma treated plants which helped in better crop growth both under biotic and abiotic stresses. These isolates helped both the varieties to accumulate more nutrients. This study proves that Trichoderma erinaceum obtained from tree bark may be incorporated in integrated rice crop management both as biocontrol agent and biofertilizer.

Diversity analysis and evaluation of wild Abelmoschus species for agro-morphological traits and major biotic stresses under the north western agro-climatic condition of India

The wild species in general is considered to be the reservoir of genes especially for biotic and abiotic stresses. In okra, the predominant biotic stresses are yellow vein mosaic disease (YVMD), shoot and fruit borer and leaf hopper. Sixty eight (68) accessions belonging to four wild Abelmoschus species [Abelmoschus caillei (A. Chev.) Stevels, Abelmoschus manihot (L.) Medik., Abelmoschus moschatus (L.) Medik. and Abelmoschus tuberculatus Pal et Singh] and eight okra varieties were characterized and evaluated for phenological characters including biotic stresses under natural epiphytotic condition. The wild species examined consisted of 18 accessions (16 exotic and 2 indigenous) of A. caillei, 29 accessions of A. manihot, 16 accessions of A. moschatus and 5 accessions of A. tuberculatus. All the wild Abelmoschus species exhibited high diversity (as measured by Shannon Diversity Index) for 3 qualitative characters viz. intensity of stem colour, leaf shape, epicalyx shape, 13 quantitative characters and 3 biotic stress parameters. Among the wild species, A. caillei and A. tuberculatus showed maximum and minimum diversity for qualitative characters, respectively. There was significant variation for 19 out of 24 quantitative characters studied. Inter-species diversity pattern as estimated through Ward’s Minimum Variance Dendrogram and Principal Component Analysis revealed clear differentiation among the species with minimum overlapping indicating close association between geographical origins and clustering pattern. Intra-species diversity indicated role of specific adaptation in sub-clustering. Resistance to YVMD was found in accessions belonging to three wild species viz. A. caillei, A. manihot and A. moschatus while resistance to shoot and fruit borer and leaf hopper was found in accessions of all the four wild species. The resistant accessions can further be used for introgressing biotic stress resistance through pre-breeding into cultivated okra species.