Role of Lactic Acid Bacteria in Insecticide Residue Degradation.

Lactic acid bacteria are gaining global attention, especially due to their role as a probiotic. They are increasingly being used as a flavoring agent and food preservative. Besides their role in food processing, lactic acid bacteria also have a significant role in degrading insecticide residues in the environment. This review paper highlights the importance of lactic acid bacteria in degrading insecticide residues of various types, such as organochlorines, organophosphorus, synthetic pyrethroids, neonicotinoids, and diamides. The paper discusses the mechanisms employed by lactic acid bacteria to degrade these insecticides, as well as their potential applications in bioremediation. The key enzymes produced by lactic acid bacteria, such as phosphatase and esterase, play a vital role in breaking down insecticide molecules. Furthermore, the paper discusses the challenges and future directions in this field. However, more research is needed to optimize the utilization of lactic acid bacteria in insecticide residue degradation and to develop practical strategies for their implementation in real-world scenarios.

A distinct strain of tomato leaf curl New Delhi virus that causes mosaic disease in ash gourd and other cucurbitaceous crops.

Ash gourd (Benincasa hispida) is a cucurbitaceous crop cultivated as an edible vegetable rich in vitamins, minerals, dietary fibers and antioxidants. In a field survey conducted in the Udumalpet region of Tamil Nadu during 2019, the incidence of mosaic disease on ash gourd crop was observed to be 75%. The DNA-A and DNA-B components of begomovirus genome have been identified as associated with this disease. Both the cloned DNA-A and DNA-B genomic components shared highest pairwise sequence identities with the isolates of tomato leaf curl New Delhi virus (ToLCNDV), a bipartite begomovirus. Recombinant analysis showed that both the components are possibly evolved through intra-species recombination between ToLCNDV isolates. Tomato leaf curl Bangladesh betasatellite (ToLCBB) is not naturally associated with this sample. The results of infectivity studies on ash gourd and other cucurbitaceous crops demonstrates the Koch's postulates, when co-inoculation of DNA-A and DNA-B of ToLCNDV was undertaken. However, the inoculation of non-cognate ToLCBB along with DNA-A and DNA-B enhances the symptom expression and reduces the time taken for symptom development. Thus, Koch's postulates were proved for these virus complexes on cucurbitaceous crops. Furthermore, an enhanced accumulation of DNA-A component was detected in the cucurbits co-inoculated with ToLCNDV and ToLCBB. This report highlights the importance of investigating the spread of these disease complexes with other cucurbitaceous crops in India.

Exogenous delivery of dsRNA for management of mungbean yellow mosaic virus on blackgram.

MAIN CONCLUSION: Exogenous application of dsRNA molecules targeting MYMV genes offers a promising approach to effectively mitigate yellow mosaic disease in blackgram, demonstrating potential for sustainable plant viral disease management. The exogenous application of double-stranded RNA (dsRNA) molecules to control plant viral diseases is gaining traction due to its advantages over conventional methods, such as target specificity, non-polluting nature, and absence of residue formation. Furthermore, this approach does not involve genome modification. In this study, dsRNA molecules targeting the coat protein gene (dsCP) and replication initiator protein gene (dsRep) of mungbean yellow mosaic virus (MYMV) were synthesised using an in vitro transcription method. To evaluate the effectiveness of dsRNA treatment, blackgram plants exhibiting MYMV symptoms at the first trifoliate stage were subjected to exogenous application of dsRNA. Second, third, and fourth trifoliate leaves, which emerged at 7, 15, and 21 days after dsRNA application, respectively, were monitored for MYMV symptoms. Remarkably, a significant reduction in yellow mosaic disease (YMD) symptoms was observed in the newly emerged trifoliate leaves of MYMV-infected blackgram plants after treatment with dsRNA targeting both gene regions. This reduction was evident as a decrease in the intensity of yellow mosaic coverage on the leaf lamina compared to control. dsCP effectively reduced the MYMV titre in the treated plants for up to 15 days. However, dsRep demonstrated greater efficiency in conferring resistance to MYMV at 15 days post-application. These findings were supported by quantitative real-time PCR analysis, where the observed Ct values for DNA extracted from dsRep-treated plants were significantly higher compared to the Ct values of DNA from dsCP-treated plants at 15 days post-application. Similarly, higher viral copy numbers were observed in dsCP-treated plants 15 days after dsRNA treatment, in contrast to plants treated with dsRep.

Molecular studies on tobacco streak virus (TSV) infecting cotton in Tamil Nadu, India.

Tobacco streak virus (TSV), the causal agent of cotton necrosis, is of emerging importance in the recent years. Unfortunately, all the cotton varieties and hybrids are susceptible to this virus. Cotton plants cultivated in different districts of Tamil Nadu were surveyed during 2014-2016. Samples collected from different locations confirmed the presence of TSV in cotton. TSV infection was confirmed by direct antigen coating-enzyme linked immuno sorbent assay (DAC-ELISA), dot immuno binding assay (DIBA), and reverse transcriptase polymerase chain reaction (RT-PCR). The virus was morphologically confirmed by transmission electron microscope (TEM). TSV isolate collected from Coimbatore was sequenced to obtain the full-length genome. Full length analysis was done for RNA 1 and RNA 3, whereas there was problem in obtaining few nucleotides in 5' and 3' end in spite of using different primers. Critical domain search in the nucleotide sequences revealed the presence of characteristic viral methyl transferase domain and helicase domain (RNA 1), 'GDD' motif and 'DFSKFD' of viral replicase in RNA 2 and Zinc finger motif in RNA 3. Phylogenetic analysis reveals high nucleotide similarity with TSV isolates of India and USA. Supplementary Information: The online version contains supplementary material available at 10.1007/s13205-022-03437-3.

Molecular epidemiology on seasonal variation of yellow mosaic disease incidence in blackgram (Vigna mungo L. Hepper) with its vector Bemisia tabaci.

The yellow mosaic disease (YMD) of blackgram caused by Mungbean yellow mosaic virus has emerged as a serious threat to grain legume production, especially in Southeastern Asia. Seasonal incidence of YMD with its vector population was assessed in three different agroclimatic zones of Tamil Nadu in India for three consecutive cropping seasons namely, Rabi 2018 (October-December), Summer 2019 (March-May), and Kharif 2019 (June-August) at three different time intervals viz., 20, 40, and 60 days after sowing (DAS). For all three seasons, disease incidence and whitefly count were recorded for a resistant and susceptible variety of blackgram in fields without any vector control intervention. The highest disease incidence (87%) was observed in the Panpozhi location during the summer season followed by Vamban and Coimbatore locations. The whitefly count was made through both visual count and yellow sticky traps. The whitefly population was highest at 20 DAS and decreased with the increasing age of crop for all the three locations assessed. Molecular epidemiology was analyzed by determining latent infection of mungbean yellow mosaic virus (MYMV) using molecular diagnosis. Latent infection was found to be well pronounced in the Coimbatore location during the Kharif season, where the crop was asymptomatic in both the resistant and susceptible varieties for all the three time periods assessed. The latent infection of MYMV observed in Coimbatore and Vamban ranged from 16.6 to 83.3% in both resistant and susceptible varieties for all three seasons. In Panpozhi, the latent infection of MYMV ranged from 16.6 to 66.6% for the susceptible variety (CO-5) for all three seasons observed. However, in the Panpozhi location, the resistant variety (VBN-8) failed to record any latent infection.

Flagellin and elongation factor of Bacillus velezensis (VB7) reprogramme the immune response in tomato towards the management of GBNV infection.

The present study investigates the reprogramming of plant defense system, upon interaction with MAMP (Microbe Associated Molecular Pattern) gene products including, flagellin (Flg) and elongation factor (EF-Tu) of Bacillus velezensis (VB7) and groundnut bud necrosis virus (GBNV) in tomato (Shivam). The MAMP gene products induced the plant defense genes including, PAL, PPO, LOX, JAR, MYC2 and PDF 1.2. Secondary metabolites of Bacillus spp. at 1000 parts per million (ppm) concentration effectively reduced GBNV symptom expression in cowpea (CO7) up to 83.1 % compared to untreated, GBNV inoculated, control. The secondary metabolites from B. velezensis (VB7) and B. licheniformis (Soya 1) reduced GBNV symptoms in cowpea (CO7) up to 1.7 lesions/cm2 leaf area compared to 8.6 lesions/cm2 in virus inoculated control. Further, field study revealed that the combined application of B. velezensis (VB7) and B. licheniformis (Soya1) at 1% (10 mL/L) as, soil and foliar application reduced the percent disease incidence (PDI) up to 10.5 (PDI) compared to 28.4 (PDI) in untreated control plants. Besides, it also improved the plant growth and yield up to 23.5 tonnes/ha compared to 13.8 tonnes/ha in untreated control.

Mining the Genome of Bacillus velezensis VB7 (CP047587) for MAMP Genes and Non-Ribosomal Peptide Synthetase Gene Clusters Conferring Antiviral and Antifungal Activity.

Chemical pesticides have an immense role in curbing the infection of plant viruses and soil-borne pathogens of high valued crops. However, the usage of chemical pesticides also contributes to the development of resistance among pathogens. Hence, attempts were made in this study to identify a suitable bacterial antagonist for managing viral and fungal pathogens infecting crop plants. Based on our earlier investigations, we identified Bacillus amyloliquefaciens VB7 as a potential antagonist for managing Sclerotinia sclerotiorum infecting carnation, tobacco streak virus infecting cotton and groundnut bud necrosis infecting tomato. Considering the multifaceted action of B. amyloliquefaciens VB7, attempts were made for whole-genome sequencing to assess the antiviral activity against tomato spotted wilt virus infecting chrysanthemum and antifungal action against Fusarium oxysporum f. sp. cubense (Foc). Genome annotation of the isolate B. amyloliquefaciens VB7 was confirmed as B. velezensis VB7 with accession number CP047587. Genome analysis revealed the presence of 9,231,928 reads with an average read length of 149 bp. Assembled genome had 1 contig, with a total length of 3,021,183 bp and an average G+C content of 46.79%. The protein-coding sequences (CDS) in the genome was 3090, transfer RNA (tRNA) genes were 85 with 29 ribosomal RNA (rRNA) genes and 21 repeat regions. The genome of B. velezensis VB7 had 506 hypothetical proteins and 2584 proteins with functional assignments. VB7 genome had the presence of flagellin protein FlaA with 987 nucleotides and translation elongation factor TU (Ef-Tu) with 1191 nucleotides. The identified ORFs were 3911 with 47.22% GC content. Non ribosomal pepide synthetase cluster (NRPS) gene clusters in the genome of VB7, coded for the anti-microbial peptides surfactin, butirosin A/butirosin B, fengycin, difficidin, bacillibactin, bacilysin, and mersacidin the Ripp lanthipeptide. Antiviral action of VB7 was confirmed by suppression of local lesion formation of TSWV in the local lesion host cowpea (Co-7). Moreover, combined application of B. velezensis VB7 with phyto-antiviral principles M. Jalapa and H. cupanioides increased shoot length, shoot diameter, number of flower buds per plant, flower diameter, and fresh weight of chrysanthemum. Further, screening for antifungal action of VB7 expressed antifungal action against Foc in vitro by producing VOC/NVOC compounds, including hexadecanoic acid, linoelaidic acid, octadecanoic acid, clindamycin, formic acid, succinamide, furanone, 4H-pyran, nonanol and oleic acid, contributing to the total suppression of Foc apart from the presence of NRPS gene clusters. Thus, our study confirmed the scope for exploring B. velezensis VB7 on a commercial scale to manage tomato spotted wilt virus, groundnut bud necrosis virus, tobacco streak virus, S. sclerotiorum, and Foc causing panama wilt of banana.

Thrips diversity of cotton ecosystem and the role of parthenium pollen grains in the transmission of tobacco streak virus (TSV) infection in cotton.

Survey in the cotton fields of Coimbatore and Erode districts confirmed the presence of three different thrips species including, Scirtothrips dorsalis, Scirtothrips oligochaetus, and Frankliniella occidentalis. Their identity was confirmed through morphometric analysis and molecular characterization. Tobacco streak virus (TSV) was detected in the leaves and pollen grains of both parthenium and cotton plants collected from infected cotton fields. The presence of TSV was confirmed through immuno-detection by direct antigen coating enzyme-linked immuno sorbent assay (DAC-ELISA). Further confirmation was accomplished by reverse transcriptase polymerase chain reaction (RT-PCR) using TSV coat protein-specific primers. Other than parthenium, weed plants in the cotton field such as Trianthema portulacastrum, Boerhavia diffusa, and Amaranthus sp. were also confirmed for TSV infection through RT-PCR. Parthenium hysterophorus plants acts as a silent carrier of TSV and they occasionally produced symptoms. Among all the randomly collected thrips samples, two pooled samples were detected positive for TSV through RT-PCR. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-02967-6.

Molecular modelling of coat protein of the Groundnut bud necrosis tospovirus and its binding with Squalene as an antiviral agent: In vitro and in silico docking investigations.

Bud blight disease caused by groundnut bud necrosis virus (GBNV) is a serious constraint in the cultivation of agricultural crops such as legumes, tomato, chilies, potato, cotton etc. Owing to the significant damage caused by GBNV, an attempt was made to identify suitable organic antiviral agents through molecular modelling of the nucleocapsid Coat Protein of GBNV; molecular docking and molecular dynamics that disclosed the interaction of the ligands viz., Squalene and Ganoderic acid-A with coat protein of GBNV. Invitro inhibitory effect of Squalene and Ganoderic acid-A was examined in comparison with different concentrations, against GBNV in cowpea plants under glasshouse condition. The different concentrations of Squalene (50, 100, 150, 250 and 500 ppm) tested in vitro resulted in reduction of lesion numbers (1.69 cm2) as well as reduced virus titre in co-inoculation spray. The present study suggests the antiviral activity of Squalene by effectively fitting into binding site of coat protein of GBNV with favourable hydrophilic as well as strong hydrophobic interactions thereby challenging and blocking the binding of viral replication RNA with coat protein and propagation. The present organic antiviral molecules will be helpful in development of suitable eco-friendly formulations to mitigate GBNV infection disease in plants.

Association of cucurbit aphid-borne yellows virus with cucumber plants in India.

Symptoms like bright yellowing, puckering of the leaf, vein banding, and vein thickening were observed on different cucurbit hosts at the experimental farm of Indian Agricultural Research Institute, New Delhi during Kharif 2019. Leaf-dip electron microscopy of the symptomatic leaves revealed the association of isometric virus particles measuring ~ 25 nm with bitter gourd and cucumber samples. The RT-PCR assay using polerovirus generic primers covering the partial RdRp, intergenic region, and partial CP region was resulted the amplicons of ~ 1.1 kb. Subsequent cloning, sequencing, and sequence analysis revealed the association of cucurbit aphid-borne yellows virus (CABYV) with bitter gourd (Momordica charantia) and cucumber (Cucumis sativus) plants. These results constitute the first report of CABYV infection on cucumber plants from India. SUPPLEMENTARY INFORMATION: The online version of this article (10.1007/s13337-020-00645-4) contains supplementary material, which is available to authorized users.

Antiviral activity of basidiomycetous fungi against Groundnut bud necrosis virus in tomato.

Tomato is an important vegetable crop which is severely affected by Groundnut bud necrosis virus (GBNV). Until now effective antiviral agents have not been reported for the management of necrosis disease caused by GBNV. Therefore, a study was undertaken to manage the necrosis disease caused by GBNV using culture filtrate of basidiomycetous fungi viz., Coprinopsiscinerea, Ganoderma lucidum and Lentinula edodes. In vitro studies were conducted in the indicator host cowpea and primary host tomato in glasshouse under insect proof condition; co-inoculation spraying of culture filtrate of Ganoderma lucidum at 0.1% concentration reduced the lesion numbers and inhibited the virus population build-up when compared to inoculated control in the indicator host cowpea upto 77.83%. DAC-ELISA test was performed to quantify the virus titre, indicated reduced virus titre in co- inoculation spray of culture filtrate of G. lucidum treated cowpea with OD value 0.17 ± 0.01 at 405 nm and in tomato plants 0.14 ± 0.01 respectively. The viral copy numbers were quantified by qPCR. About 2.0 × 101 viral copy numbers were observed in tomato plants treated with G. lucidum (co-inoculation) which was lesser than untreated inoculated control plants (2.4 × 108). In order to identify the antiviral properties of G. lucidum, GCMS analysis was carried out and we found the triterpenoid compound Squalene. This is the first study to analyse and confirm the antiviral activity of G. lucidum against a plant virus.

Induction of in planta resistance by flagellin (Flg) and elongation factor-TU (EF-Tu) of Bacillus amyloliquefaciens (VB7) against groundnut bud necrosis virus in tomato.

Role of plant growth promoting rhizobacteria (PGPR) in growth promotion and induction of resistance against various plant pathogens have been extensively studied. However, MAMP (Microbe Associated Molecular Pattern) triggered immunity (MTI) against plant viruses are not well exploited. The present study enlightens the role of two MAMP genes including, flagellin (Flg) and elongation factor (EF-Tu) in the induction of plant defense against GBNV infecting tomato. Secondary metabolites of Bacillus amyloliquefaciens (VB7), effectively suppressed GBNV symptom expression up to 84% compared to untreated control in cowpea, the indicator host plant. Agrobacterium tumefaciens EHA105 clones expressing the MAMP genes were drenched in the root zone to assess the induction of resistance against GBNV in tomato. Treatment with A. tumefaciens EHA105 clones containing flagellin (Ag- Ba.Flg) and elongation factor-TU (Ag-Ba.EF-Tu) genes as soil drench and foliar spray, reduced virus titre,0.369 OD and 0.379 OD respectively as compared to control 1.249 OD. The disease severity was reduced up to 15% in Ag-Ba.Flg treated plants compared to 88.25% in inoculated control. Further there was an increased expression of defense associated genes including, MAPKK1, WRKY33BB, NPR1 and PR1.The present investigation clearly indicated the efficiency of MAMP genes in triggering defense mechanism in tomato against GBNV.

Evidence of seed transmission of dolichos yellow mosaic virus, a begomovirus infecting lablab-bean in India.

Yellow mosaic disease in field bean caused by begomoviruses belonging to the family Geminiviridae is a major threat to the cultivation of crop in South India. Appearance of bright yellow mosaic symptom in emerging seedlings in farmers field was suggestive of seed transmission of the begomovirus associated with the disease which was investigated in the present study. The begomovirus causing yellow mosaic disease was identified as dolichos yellow mosaic virus (DoYMV) and the presence of DoYMV in matured seeds was confirmed by DAS-ELISA with OD value up to 3.268. In PCR with DoYMV specific primer (DoYMV-CP) the virus was detected in different parts of the seeds viz., seed coat, endosperm and embryo. In embryo the virus was detectable up to 100 per cent followed by endosperm (69.23%). When the non symptomatic leaves of 30 days old grow-out test plants were subjected to DAS-ELISA, the virus was detected up to 46.6%. This is the first evidence of seed transmission of DoYMV.

A distinct seed-transmissible strain of tomato leaf curl New Delhi virus infecting Chayote in India.

Chayote (Sechium edule (Jacq. Sw.) is a single seeded cucurbitaceous vegetable crop mainly grown for its fruit. During 2015-2016, mosaic and leaf distortion type of symptoms were observed in chayote plants in hilly regions of Tamil Nadu. The disease incidence was 50. 3-100% and yield loss was about 69.9% in Dindigul district. The infected chayote plants showed yellow spots, yellow mosaic, leaf curling, puckering, and enations. The fruits of infected plants were malformed and were not marketable. The begomovirus causing the disease was identified as a variant of tomato leaf curl New Delhi virus. The chayote isolates of ToLCNDV share only 91 to 92% identity with other ToLCNDV isolates, deserving to be designated as distinct strain. The phylogenetic analysis on the basis of DNA A component nucleotides clearly indicated common origin of chayote, ridge gourd, ash gourd isolates of India along with Spanish isolates of ToLCNDV. This was contrasting to diverse origin of ToLCNDV isolates from other countries. The virus was sap transmissible to selected cucurbitaceous hosts. The whitefly population (Asia-I) reared in the glass house transmitted the virus to bottle gourd with 4 h of acquisition access period (AAP) and 24 h of inoculation feeding period (IFP). Heavy infestation of greenhouse whiteflies, Trialeurodes vaporariorum on infected chayote plant in Kodaikanal paved way to investigations on its role in vector transmission of ToLCNDV. The field population of T. vaporariorum was found to be viruliferous in PCR using virus specific primers. The greenhouse whitefly efficiently transmitted the virus with 4 h of AAP and 24 h IFP. Seed-borne nature of ToLCNDV was confirmed in PCR by using Roja's and ToLCNDV specific primers in different parts of the fruit viz., pericarp, mesocarp, seed coat, endosperm and embryo. This is the first report of seed transmissible nature of ToLCNDV, its implication in transboundary movement of the virus across several countries is discussed.

Biocontrol Potentials of Antimicrobial Peptide Producing Bacillus Species: Multifaceted Antagonists for the Management of Stem Rot of Carnation Caused by Sclerotinia sclerotiorum.

Bacillus species are widely exploited as biocontrol agents because of their efficiency in impeding various plant pathogens with multifaceted approach. In this study, Bacillus species were isolated from rhizosphere of various plants viz., carnations, cotton, turmeric, and bananas in Tamil Nadu state of India. Their potential to control the mycelial growth of Sclerotinia sclerotiorum was assessed in vitro by dual plate and partition plate techniques. B. amyloliquefaciens strain VB7 was much effective in inhibiting mycelial growth (45% inhibition of over control) and sclerotial production (100%). PCR detection of AMP genes revealed that B. amyloliquefaciens (VB7) had a maximum of 10 diverse antibiotic biosynthesis genes, namely, ituD, ipa14, bacA, bacD, bamC, sfP, spaC, spaS, alba, and albF, that resulted in production of the antibiotics iturin, bacilysin, bacillomycin, surfactin, subtilin, and subtilosin. Further, metabolites from B. amyloliquefaciens strains VB2 and VB7, associated with inhibition of S. sclerotiorum, were identified as phenols and fatty acids by gas chromatography mass spectrometry (GC-MS). Delivery of bacterial suspension of the effective strains of Bacillus spp. as root dip was found promising for the management of stem rot of cultivated carnations. Minimal percent disease incidence (4.6%) and maximum plant growth promotion was observed in the plants treated with B. amyloliquefaciens (VB7).