Harnessing Nanoencapsulated Bacillus spp. Consortia To Combat Groundnut Bud Necrosis Orthotospovirus in Tomato.

Tomato (Solanum lycopersicum L.), a globally significant vegetable crop, faces a substantial threat from viral diseases, specifically Groundnut bud necrosis orthotospovirus (GBNV). Traditional approaches such as removal of infected plants, use of barrier crops, and insecticides have been employed but they have not proven to be consistently effective. Consequently, an alternative approach involving the stimulation of host resistance through the Plant Growth Promoting Rhizobacteria (PGPR) was adopted. From the previous study, B. subtilis (BST8), B. subtilis (Bbv57), and B. amyloliquefaciens (Ka1) were found to be effective against GBNV in cowpea. To enhance the shelf life of Bacillus spp. and improve the water retention capacity of tomato leaf surfaces, these bacteria were encapsulated within nanosilica, an identified host defense inducer. An effective inverse Pickering emulsion with a 2.5% (w/v) silica concentration was developed and characterized using diverse techniques, viz., phase contrast, scanning electron microscopy, confocal microscopy, contact angle goniometry, and variable angle ellipsometry. The prepared emulsion was then tested for its antiviral efficacy against GBNV in cowpea and tomatoes. Nanoencapsulated Bacillus consortia significantly reduced GBNV lesions in cowpea to 0.63 per leaf compared to the control (6.63). DAC-ELISA revealed a virus titer of 0.75 (3.33 times lower than the control), indicating antiviral efficacy. In tomato (var. PKM1), the consortia achieved an impressive 77.91% disease reduction (19% DSI) at 14 days post-inoculation (DPI), surpassing both nanoemulsion and consortia alone (DSIs: 67 and 30%, respectively). Nanoencapsulated Bacillus consortia demonstrated the lowest GBNV titer in tomatoes (0.86 vs control-3.32) through DAC-ELISA. This study introduces a promising strategy for the effective management of GBNV in cowpea and tomatoes using nanoencapsulated Bacillus consortia, underscoring its potential as an effective solution in crop protection.

Identification of salivary proteins of the cowpea aphid Aphis craccivora by transcriptome and LC-MS/MS analyses.

Aphid salivary proteins mediate the interaction between aphids and their host plants. Moreover, these proteins facilitate digestion, detoxification of secondary metabolites, as well as activation and suppression of plant defenses. The cowpea aphid, Aphis craccivora, is an important sucking pest of leguminous crops worldwide. Although aphid saliva plays an important role in aphid plant interactions, knowledge of the cowpea aphid salivary proteins is limited. In this study, we performed transcriptomic and LC-MS/MS analyses to identify the proteins present in the salivary glands and saliva of A. craccivora. A total of 1,08,275 assembled transcripts were identified in the salivary glands of aphids. Of all these assembled transcripts, 53,714 (49.11%) and 53,577 (49.48%) transcripts showed high similarity to known proteins in the Nr and UniProt databases, respectively. A total of 2159 proteins were predicted as secretory proteins from the salivary gland transcriptome dataset, which contain digestive enzymes, detoxification enzymes, previously known effectors and elicitors, and potential proteins whose functions have yet to be determined. The proteomic analysis of aphid saliva resulted in the identification of 171 proteins. Tissue-specific expression of selected genes using RT-PCR showed that three genes were expressed only in the salivary glands. Overall, our results provide a comprehensive repertoire of cowpea aphid salivary proteins from the salivary gland and saliva, which will be a good resource for future effector functional studies and might also be useful for sustainable aphid management.

Analysis of genetic diversity and population structure of Magnaporthe grisea, the causal agent of foxtail millet blast using microsatellites.

Foxtail millet blast caused by Magnaporthe grisea is becoming a severe problem in foxtail millet growing regions of India. The genetic diversity and population structure of foxtail millet infecting M. grisea is crucial for developing effective management strategies, such as breeding blast-resistant cultivars. We analyzed thirty-two M. grisea isolates from ten foxtail millet-growing districts in Tamil Nadu, India for genetic diversity using twenty-nine microsatellite or simple sequence repeat (SSR) markers. A total of 103 alleles were identified with a mean of 3.55 alleles/locus. Gene diversity ranged from 0.170 to 0.717, while major allelic frequencies ranged from 0.344 to 0.906. The polymorphism information content (PIC) ranged from 0.155 to 0.680, with a mean value of 0.465. Population structure analysis of the genomic data sets revealed two major populations (SP1 and SP2) with different levels of ancestral admixture among the 32 blast isolates. Phylogenetic analysis classified the isolates into three major clusters. Analysis of molecular variance (AMOVA) showed high genetic variation among individuals and less among populations. Principal Coordinate Analysis (PCoA) revealed 27.16% genetic variation among populations. The present study provides the first report on the genetic diversity and population structure of the foxtail millet-infecting M. grisea population in Tamil Nadu, which could be useful for the development of blast-resistant foxtail millet cultivars.

Biomolecules from Chaetomium globosum Possessing Antimicrobial Compounds Potentially Inhibits Fusarium Wilt of Tomato.

Wilt disease caused by Fusarium solani, a soil-borne plant pathogenic fungus, is a serious disease in tomato causing economic losses. In the present study, among the nine isolates of Chaetomium globosum Kunze (Chg1-Chg9), screened against F. solani No. Fs-1, Chg2 exhibited the maximum inhibition (49.2 %), followed by the isolates Chg6 (47.4%) and Chg1 (46.3%) in dual culture. Further, the crude secondary metabolites from these three isolates showed maximum reduction of the mycelial growth of Fs-1 compared to control. Gas Chromatography-Mass Spectrometry (GC-MS) analysis of the metabolites from Chg2 revealed the presence of major compounds, viz., benzothiazole, 2-(2-hydroxyethylthio) (7.51%); 9,12,15- octadecatrienoic acid, 2,3-bis[(trimethylsilyl)oxy] propyl ester (3.13%); and hexadecanoic acid, 1- (hydroxymethyl)-1,2-ethanediyl ester (2.69%). Fourier transform infrared (FTIR) spectroscopy revealed the presence of alcohol, secondary amine, aliphatic primary amine, carboxylic acid, allene, conjugated ketone, imine/oxime, sulphate, and halo compound groups with a weak to strong range of intensity. Pure compound of benzothiazole @ 5000 ppm exhibited higher antagonistic activity of Fs-1which decreased at lower concentration. In pot culture experiment, dipping tomato seedlings cv. PKM1 in 2% ethyl acetate-fractionated biomolecules of Chg2 recorded a minimum disease incidence of 20.0%, whereas seedlings dipped in the culture filtrate of Chg2 showed a disease incidence of 26.6% compared to control (86.6%). Besides, these seedlings treated with the culture filtrate showed the highest plant height of 37.68 cm and the maximum number of leaves and flowers (39.6 and 5.48, respectively), followed by those treated with the biomolecules of Chg2 (C. globosum) and the culture filtrate of T. asperellum. The results revealed that the antifungal efficacy of C. globosum Chg2 is due to the presence of antimicrobial metabolites including benzothiazole. Exploring the use of this novel antifungal compound in the management of plant diseases is highly warranted.

Bio-Prospecting of Endospore-Based Formulation of Bacillus sp. BST18 Possessing Antimicrobial Genes for the Management of Soil-Borne Diseases of Tomato.

Tomato is affected by various diseases which cause economical loss to the farming community. In the present study, twenty isolates of Bacillus sp. were isolated from the rhizosphere of tomato and screened against soil-borne pathogens viz., Pythium aphanidermatum No.5, Fusarium oxysporum f. sp. lycopersici No. FOL-8, and Sclerotium rolfsii No. S-MK in tomato. The results revealed that three Bacillus sp. viz., BST8, BST18, and BST19 were promising in reducing the mycelial growth of the pathogens (up to 48% reduction) when compared to control under in vitro. The isolates possessed antimicrobial peptide genes which were detected through PCR. Novel compounds and secondary metabolites responsible for antifungal action were identified through GC-MS and FTIR analysis. Endospores have been isolated from the Bacillus sp. BST18 and standardized for the development of formulation. Pot culture experiment revealed sequential application of endospore-based bioformulation as seed treatment, (10 ml/kg), seedling dip (10 ml/lit), soil application on 30 days (100 ml/pot) along with foliar spray (0.2%) on 60 days recorded the lowest disease incidence of wilt (9.5%), and collar rot (11.5%) as against 65.5% and 75%, respectively, in the inoculated control. Field experiments revealed sequential application of endospore-based formulation of Bacillus sp. BST 18 as seed treatment, (10 ml/kg), seedling dip (10 ml/lit), soil application (400 ml/acre) at 30 days after transplantation (DAT) along with foliar spray (0.2%) on 60 DAT recorded the lowest wilt disease incidence of 15.97 &17.07 percent as against 49.77 & 51.10 percent in the control.

Comparative Proteomic Analysis of Different Isolates of Fusarium oxysporum f.sp. lycopersici to Exploit the Differentially Expressed Proteins Responsible for Virulence on Tomato Plants.

The vascular wilt of tomato caused by Fusarium oxysporum f.sp. lycopersici is an important soil borne pathogen causes severe yield loss. The molecular characterization and their interaction with its host is necessary to develop a protection strategy. 20 isolates of F. oxysporum f.sp. lycopersici (FOL) were isolated from wilt infected tomato plants across Tamil Nadu. They were subjected to cultural, morphological, molecular and virulence studies. The results revealed that all the isolates produced both micro and macro conidia with different size, number of cells. The colors of the culture and growth pattern were also varied. In addition, chlamydospores were observed terminally and intercalary. The PCR analysis with F. oxysporum species-specific primer significantly amplified an amplicon of 600 bp fragment in all the isolates. Based on the above characters and pathogenicity, isolate FOL-8 was considered as virulent and FOL-20 was considered as least virulent. Proteomics strategy was adopted to determine the virulence factors between the isolates of FOL-8 and FOL-20. The 2D analyses have showed the differential expression of 17 different proteins. Among them, three proteins were down regulated and 14 proteins were significantly up regulated in FOL-8 than FOL-20 isolate. Among the 17 proteins, 10 distinct spots were analyzed by MALDI-TOF. The functions of the analyzed proteins, suggested that they were involved in pathogenicity, symptom expression and disease development, sporulation, growth, and higher penetration rate on tomato root tissue. Overall, these experiments proves the role of proteome in pathogenicity of F. oxysporum f.sp. lycopersici in tomato and unravels the mechanism behinds the virulence of the pathogen in causing wilt disease.