Bacillus subtilis NBRI-W9 simultaneously activates SAR and ISR against Fusarium chlamydosporum NBRI-FOL7 to increase wilt resistance in tomato.

AIMS: The study aimed to determine the pathogenicity of Fusarium species currently prevalent in tomato fields having history of chemical fungicide applications and determine the bio-efficacy of Bacillus subtilis NBRI-W9 as a potent biological control agent. METHODS AND RESULTS: Fusarium was isolated from surface-sterilized infected tomato plants collected from fields. Pathogenicity of 30 Fusarium isolates was determined by in vitro and in vivo assays. Following Koch's postulates, F. chlamydosporum (FOL7) was identified as a virulent pathogen. The biological control of FOL 7 by B. subtilis NBRI-W9 (W9) and the colonization potential of W9 were established using spontaneous rifampicin-resistant mutants. W9 showed 82% inhibition of FOL7 on a dual-culture plate and colonization levels in tomato plants of ∼5.5, ∼3.3, and ∼2.2 log10 CFU/g in root, stem, and leaf tissue, respectively. Antagonistic activity was shown by scanning electron microscopy (SEM) and cell-wall-degradative enzymes. W9 reduced FOL7 infection in net-house and field experiments by 60% and 41%, respectively. Biochemical investigation, defence enzymes, defence gene expression analysis, SEM, and field studies provide evidence of hyperparasitism and induced resistance as the mode of biological control. The study also demonstrates that the potent biocontrol agent W9, isolated from Piper, can colonize tomato plants, control fungal disease by inducing induced systemic resistance (ISR) and systemic acquired resistance (SAR) simultaneously, and increase crop yield by 21.58% under field conditions. CONCLUSIONS: This study concludes that F. chlamydosporum (NBRI-FOL7) is a potent, fungicide-resistant pathogen causing wilt in tomatoes. NBRI-W9 controlled FOL7 through mycoparasitism and simultaneously activated ISR and SAR in plants, providing an attractive tool for disease control that acts at multiple levels.

Studies on the gametophytic development, mating behaviour and reproductive biology of Macrothelypteris torresiana (Gaudich.) Ching (Thelypteridaceae).

Macrothelypteris torresiana is a terrestrial fern belongs to the family Thelypteridaceae. It is very imperative to know the germination and growth pattern of this fern to make strategy for ex- situ conservation. In the present study germination of spore, developmental pattern of prothallium with particular emphasis on emergence of antheridia as well as archegonia, their fertilization and development of sporophytes were studied. The spores were spheroidal or reniform with broadly winged fimbriate perispore having average size of 48 ± 3.6 × 39 ± 4.2 µm. The spores started germinating just after 6 days of plating and percentage of germination was 94%. The germination pattern of spore was Vittaria type and development of gametophyte was Aspidium type. After 27 days of plating, a very distinct cordate-shaped adult gametophyte with deep apical notch and unicellular papillate hairs throughout the gametophyte was developed. The rhizoids appeared away from the apical notch of the gametophyte. The gametophyte first developed archegonia on 32nd day, whereas antheridia were developed on 42nd day. In isolate culture the gametophytes did not develop any sporophyte whereas in composite culture juvenile sporophytes were emerged on 77th day. This suggests the plant has ability to reproduce only through inter-gametophytic selfing and crossing, hence decreasing the plant's capacity for colonization in a particular locality.