Abstract

Using gamma-ray-induced mutagenesis, we have developed a mutant (named G2) of Trichoderma virens that produced two- to three-fold excesses of secondary metabolites, including viridin, viridiol, and some yet-to-be identified compounds. Consequently, this mutant had improved antibiosis against the oomycete test pathogen Pythium aphanidermatum. A transcriptome analysis of the mutant vis-à-vis the wild-type strain showed upregulation of several secondary-metabolism-related genes. In addition, many genes predicted to be involved in mycoparasitism and plant interactions were also upregulated. We used tamarind seeds as a mass multiplication medium in solid-state fermentation and, using talcum powder as a carrier, developed a novel seed dressing formulation. A comparative evaluation of the wild type and the mutant in greenhouse under high disease pressure (using the test pathogen Sclerotium rolfsii) revealed superiority of the mutant over wild type in protecting chickpea (Cicer arietinum) seeds and seedlings from infection. We then undertook extensive field evaluation (replicated micro-plot trials, on-farm demonstration trials, and large-scale trials in farmers’ fields) of our mutant-based formulation (named TrichoBARC) for management of collar rot (S. rolfsii) in chickpea and lentil (Lens culinaris) over multiple locations in India. In certain experiments, other available formulations were included for comparison. This formulation consistently, over multiple locations and years, improved seed germination, reduced seedling mortality, and improved plant growth and yield. We also noticed growth promotion, improved pod bearing, and early flowering (7–10 days) in TrichoBARC-treated chickpea and lentil plants under field conditions. In toxicological studies in animal models, this formulation exhibited no toxicity to mammals, birds, or fish.

Highlights

  • Trichoderma spp. are among the most widely used bioagents in today’s agriculture throughout the world (Mukherjee et al, 2013)

  • We have combined genetic enhancement with novel formulation strategy, and through extensive evaluation over many years in several locations, we demonstrate the commercial potential of such a novel formulation

  • In developing countries, including India, food grains like sorghum are extensively used for growing Trichoderma at industry scale, while tamarind seeds are by-products of tamarind pulp that are very popular as a culinary ingredient throughout India

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Summary

Introduction

Trichoderma spp. are among the most widely used bioagents in today’s agriculture throughout the world (Mukherjee et al, 2013). The popularity of these fungi stems from their ability to kill other fungi (mycoparasitism), produce several hundred secondary metabolites (some are antimicrobial), induce local and systemic resistance in plants against invading pests and pathogens, improve nutrient (especially nitrogen) use efficiency, promote plant growth, and impart tolerance to abiotic stresses (Lorito et al, 2010). We report here improvement of a strain of Trichoderma virens that has been widely studied for biocontrol properties (Mukhopadhyay et al, 1992; Mukherjee et al, 2013; Sherkhane et al, 2017), using gamma-ray-induced mutagenesis. In addition to strain improvement, we report here a novel mass multiplication protocol and a formulation strategy for Trichoderma, and report on the non-toxic nature of the T. virens mutant formulation in mammals, birds, and fish

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