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.

Emergence of yellowing disease in cucurbitaceous vegetables caused by Crinivirus and Polerovirus in India.

Totally 102 symptomatic samples of cucurbitaceous vegetables showing yellowing were collected from fields of Uttar Pradesh and screened by RT-PCR assay for the presence of Crinivirus and Polerovirus. Among them, Crinivirus (16%) and Polerovirus (23%) were tested positive with the universal primer pairs. Based on the sequence analysis of amplified product, two Crinivirus (Cucurbit chlorotic yellows virus - CCYV and Cucurbit yellow stunting disorder virus - CYSDV) and two Polerovirus (Cucurbit aphid-borne yellows virus - CABYV and Luffa aphid-borne yellows virus - LABYV) species were characterized. Phylogenetic analysis revealed less genetic distance among the Indian isolates of CCYV, CYSDV and LABYV whereas CABYV closely related to Chinese isolates. To the best of our knowledge, this study documents infection of CCYV on cucumber, round melon and muskmelon; CYSDV on satputia and sponge gourd; CABYV on ivy gourd; and LABYV on ridge gourd, satputia and muskmelon for the first time in India.

Exploring the Potentiality of Native Actinobacteria to Combat the Chilli Fruit Rot Pathogens under Post-Harvest Pathosystem.

Chilli is an universal spice cum solanaceous vegetable crop rich in vitamin A, vitamin C, capsaicin and capsanthin. Its cultivation is highly threatened by fruit rot disease which cause yield loss as high as 80-100% under congenial environment conditions. Currently actinobacteria are considered as eco-friendly alternatives to synthetic fungicides at pre and post-harvest pathosystems. Hence, this research work focuses on the exploitation of rhizospheric, phyllospheric and endophytic actinobacteria associated with chilli plants for their antagonistic activity against fruit rot pathogens viz., Colletotrichum scovillei, Colletotrichum truncatum and Fusarium oxysporum. In vitro bioassays revealed that the actinobacterial isolate AR26 was found to be the most potent antagonist with multifarious biocontrol mechanisms such as production of volatile, non-volatile, thermostable compounds, siderophores, extracellular lytic enzymes. 16S rRNA gene sequence confirmed that the isolate AR26 belongs to Streptomyces tuirus. The results of detached fruit assay revealed that application of liquid bio-formulation of Stretomyces tuirus @ 10 mL/L concentration completely inhibited the development of fruit rot symptoms in pepper fruits compared to methanol extracts. Hence, the present research work have a great scope for evaluating the biocontrol potential of native S. tuirus AR26 against chilli fruit rot disease under field condition as well against a broad spectrum of post-harvest plant pathogens.

Characterization of chaetoglobosin producing Chaetomium globosum for the management of Fusarium-Meloidogyne wilt complex in tomato.

AIM: Simultaneous management of FOL and RKN causing wilt complex in tomato by chaetoglobosin-producing Chaetomium globosum. METHODS AND RESULTS: Random survey was carried out to isolate Fusarium and Chaetomium. Twelve Fusarium isolates were characterized, and FOL4 (virulent) was molecularly identified. Wilt complex by FOL, RKN was assessed individually and in combination under greenhouse. RKN (1000 juveniles ml-1) inoculation followed by FOL4 (5 × 105 spores ml-1) accounted for 90% incidence. The chaetoglobosin-producing Chaetomium was isolated, characterized morphologically and molecularly. Among 55 isolates, nine showed >50% inhibition against FOL, and crude culture filtrate showed a significant reduction in RKN egg hatching (15.66%) and juvenile mortality (100%). Chaetomium Cg 40 was confirmed as C. globosum using SCAR marker (OK032373). Among 40 volatile compounds, hexadecanoic acid and 1,2-epoxy-5,9-cyclododecadiene exhibited antifungal and nematicidal properties in GC-MS. High-performance liquid chromatography revealed chaetoglobosin A (0.767 µg µl-1), and the presence of bioactive molecules chaetoglobosin (528.25 m/z), chaetomin (710 m/z), chaetocin (692.8 m/z), chaetoviridin (432.85 m/z), and chaetomugilin (390 m/z) was confirmed by LC/MS/MS. Cg 40 and Cg 6 were able to synthesize the pks1a, b gene responsible for chaetoglobosin, sporulation, and melanin biosynthesis was confirmed by PCR. The application of an aqueous formulation as seed treatment, seedling dip, and soil drenching (application) recorded lowest wilt incidence (11.11%) and gall index (1) with the maximum growth parameter (plant height 51.9 cm), fruit yield (287.5 g), and lycopene content (11.46 mg/100 g). CONCLUSIONS: Cg 40 and Cg 6, containing polyketides, secondary metabolites, antibiotics, chaetoglobosin, and plant growth-promoting ability, showed antifungal and nematicidal properties against the FOL-RKN wilt complex in tomato in vitro and pot culture experiments.

Detection of QTLs associated with mungbean yellow mosaic virus (MYMV) resistance using the interspecific cross of Vigna radiata × Vigna umbellata.

Mungbean (Vigna radiata) and ricebean (V. umbellata) were utilized to obtain an inter-specific recombinant inbred line (RIL) population with the objective of detecting quantitative trait loci (QTL) associated with mungbean yellow mosaic virus (MYMV) resistance. To precisely map QTLs, accurate genetic linkage maps are essential. In the present study, genotyping-by-sequencing (GBS) platform was utilized to develop the genetic linkage map. The map contained 538 single nucleotide polymorphism (SNP) markers, consisted of 11 linkage groups and spanned for 1291.7 cM with an average marker distance of 2.40 cM. The individual linkage group ranged from 90.2 to 149.1 cM in length, and the SNP markers were evenly distributed in the genetic linkage map, with 30-79 SNP markers per chromosome. The QTL analysis using the genetic map and 2 years (2015 and 2016) of phenotyping data identified five QTLs with phenotypic variation explained (PVE) from 10.11 to 20.04%. Of these, a QTL on chromosome 4, designated as qMYMV4-1, was major and stably detected in the same marker interval in both years. This QTL region harbours possible candidate genes for controlling MYMV resistance. The linkage map and QTL/gene (s) for MYMV resistance identified in this study should be useful for QTL fine mapping and cloning for further studies.

A new seed-transmissible begomovirus in bitter gourd (Momordica charantia L.).

A begomovirus isolate collected from bitter gourd plants showing yellowing, puckering and stunting symptoms from Coimbatore district, Tamil Nadu, India was characterized. The full-length genome of the virus isolate was amplified by rolling circle amplification using phi29 DNA polymerase. The virus isolate exhibited 98% identity in the nucleotide sequence of DNA-A component with the Coccinia mosaic Virudhunagar virus (GenBank accession no. KY860899). The DNA-B component was very distinct and shared only 60% identity with the begomovirus, Coccinia mosaic Tamil Nadu virus (GenBank accession no. KM244719). The virus renamed as new species Bitter gourd yellow mosaic virus (BgYMV) was detected in seeds from infected plants and in the grow-out test seedlings by ELISA and virus-specific PCR. The seed infectivity was 79.16% and transmission rate to seedling was 32.05%. The virus titre as indicated by A405 absorption value was high (0.854-0.280) in different seed parts. Results clearly indicated seed transmission of the begomovirus, BgYMV.

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.

Grapevine leafroll-associated virus 1 occurs as genetically diverse populations.

The genetic diversity of 34 isolates of Grapevine leafroll-associated virus 1 (GLRaV-1) from different wine, table, and ornamental grape cultivars in California, New York, and Washington States in the United States was investigated. Segments of the heat-shock protein 70 homolog (HSP70h) gene, coat protein (CP) gene, coat protein duplicate 2 (CPd2) gene, and open reading frame 9 (p24) were amplified by reverse-transcription polymerase chain reaction, cloned, and sequenced. A pairwise comparison of nucleotide sequences revealed intra- and interisolate sequence diversity, with CPd2 and HSP70h being the most and the least divergent, respectively, among the four genomic regions studied. The normalized values for the ratio of nonsynonymous substitutions per nonsynonymous site to synonymous substitutions per synonymous site indicated different purifying selection pressures acting on each of the four genomic regions, with the CP and CPd2 being subjected to the strongest and weakest functional constraints, respectively. A global phylogenetic analysis of sequences from the four genomic regions revealed segregation of GLRaV-1 isolates into three major clades and a lack of clearly defined clustering by geographical origin. In contrast, only two lineages were apparent when the CP and CPd2 gene sequences were used in phylogenetic analyses. Putative recombination events were revealed among the HSP70h, CP, and p24 sequences. The genetic landscape of GLRaV-1 populations presented in this study provides a foundation for better understanding of the epidemiology of grapevine leafroll disease across grape-growing regions in the United States. In addition, this study will benefit grape clean plant programs across the country in improving the sanitary status of planting materials provided to nurseries and grape growers.