Abstract

White mold or stem rot disease are ubiquitously distributed throughout the world and the causal organism of this disease Sclerotinia sclerotiorum (Lib.) de Bary, is known to infect over 400 plant species. Sclerotinia stem rot is one of the most devastating fungal diseases and poses a serious threat to the worldwide cultivation of oilseed Brassica including India. S. sclerotiorum pathogen usually infects the stem but in severe cases leaves and pods also affected at different developmental stages that deteriorate not only the oil quality but also causing the seed and oil yield losses up to 90% depending on the severity of the disease infestation. This study investigated the morphological and molecular characterization of pathogenic S. sclerotiorum (Lib) de Bary geographical isolates from oilseed Brassica including Brassica juncea (Indian mustard). The aim of this study was to compare isolates of S. sclerotiorum originated from different agro-climatic conditions and to analyse similarity or differences between them as well as to examine the virulence of this pathogen specifically in Brassica for the first time. The collection of S. sclerotiorum isolates from symptomatic Brassica plants was done and analyzed for morphological features, and molecular characterization. The virulence evaluation test of 65 isolates on four Brassica cultivars has shown 5 of them were highly virulent, 46 were virulent and 14 were moderately virulent. Phylogenetic analysis encompassing all the morphological features, SSR polymorphism, and ITS sequencing has shown the existence of high genetic diversity among the isolates that categorized all the isolates in three evolutionary lineages in the derived dendrogram. Further, genetic variability analysis based on sequences variation in ITS region of all the isolates has shown the existence of either insertions or deletions of the nucleotides in the ITS region has led to the interspecies variability and observed the variation were in a clade-specific manner. Together this analysis observed the existence of higher heterogeneity and genetic variability in S. sclerotiorum isolates collection and indicates the presence of clonal and sexual progenies of the pathogen in the mustard growing regions of India surveyed in this study. With a higher level of genetic variability and diversity among the S. sclerotiorum population needs robust screening approaches to identify the donor parent and utilize them in resistance breeding program for effectively counter the menace of stem rot disease in Brassica.

Highlights

  • India continues to be at a 3rd position after Canada and China in acreage (19.3%) and after China and Canada in production (11.1%) of rapeseed-mustard

  • The genetic diversity analysis by all the means followed in this study has clearly shown there were not many differences exists in terms of virulence and in groups that comprised from the isolates of different virulence nature belongs to different localities

  • The current investigation reports the studies on morphological features, pathogenicity variance over the Brassica cultivars, and genetic diversity based on molecular characteristics of the S. sclerotiorum isolates population collected from 65 locations of the 10 major Brassica growing states of India

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Summary

Introduction

India continues to be at a 3rd position after Canada and China in acreage (19.3%) and after China and Canada in production (11.1%) of rapeseed-mustard. A widely adopted monocropping practices and cultivation of rapeseed-mustard under irrigated condition has significantly increased the sclerotial population in the soil that has made SR very serious disease of oilseed Brassica crops in states including Rajasthan, Haryana, Punjab, Uttar Pradesh, Bihar, Assam, West Bengal and Madhya Pradesh (Sharma et al, 2015) This fungus has been long considered as prototypical necrotrophs as it begins highly pathogenic phase by releasing oxalic acids and cellulolytic enzymes immediately upon host cuticle penetration followed by mycelial proliferation inside the host cell followed by a saprophytic phase that supports the sclerotia formation (Hegedus and Rimmer, 2005). Polymorphism and genetic diversity of the S. sclerotiorum isolates at the morphological and DNA sequence level has not been comprehensively studied so far especially for the isolates from Brassica species of India

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