In silico identification of papaya genome-encoded microRNAs to target begomovirus genes in papaya leaf curl disease.

Papaya leaf curl disease (PaLCuD) is widespread and classified in the genus begomovirus (Geminiviridae), disseminated by the vector whitefly Bemisia tabaci. RNA interference (RNAi)-based antiviral innate immunity stands as a pivotal defense mechanism and biological process in limiting viral genomes to manage plant diseases. The current study aims to identify and analyze Carica Papaya locus-derived capa-microRNAs with predicted potential for targeting divergent begomovirus species-encoded mRNAs using a 'four integrative in silico algorithms' approach. This research aims to experimentally activate the RNAi catalytic pathway using in silico-predicted endogenous capa-miRNAs and create papaya varieties capable of assessing potential resistance against begomovirus species and monitoring antiviral capabilities. This study identified 48 predicted papaya locus-derived candidates from 23 miRNA families, which were further investigated for targeting begomovirus genes. Premised all the four algorithms combined, capa-miR5021 was the most anticipated miRNA followed by capa-miR482, capa-miR5658, capa-miR530b, capa-miR3441.2, and capa-miR414 'effective' papaya locus-derived candidate capa-miRNA and respected putative binding sites for targets at the consensus nucleotide position. It was predicted to bind and target mostly to AC1 gene of the complementary strand and the AV1 gene of the virion strand of different begomovirus isolates, which were associated with replication-associated protein and encapsidation, respectively, during PaLCuD. These miRNAs were also found targeting betaC1 gene of betasatellite which were associated with retardation in leaf growth and developmental abnormalities with severe symptoms during begomovirus infection. To validate target prediction accuracy, we created an integrated Circos plot for comprehensive visualization of host-virus interaction. In silico-predicted papaya genome-wide miRNA-mediated begomovirus target gene regulatory network corroborated interactions that permit in vivo analysis, which could provide biological material and valuable evidence, leading to the development of begomovirus-resistant papaya plants. The integrative nature of our research positions it at the forefront of efforts to ensure the sustainable cultivation of papaya, particularly in the face of evolving pathogenic threats. As we move forward, the knowledge gained from this study provides a solid foundation for continued exploration and innovation in the field of papaya virology, and to the best of our knowledge, this study represents a groundbreaking endeavor, undertaken for the first time in the context of PaLCuD research.

Multifarious Responses of Forest Soil Microbial Community Toward Climate Change.

Forest soils are a pressing subject of worldwide research owing to the several roles of forests such as carbon sinks. Currently, the living soil ecosystem has become dreadful as a consequence of several anthropogenic activities including climate change. Climate change continues to transform the living soil ecosystem as well as the soil microbiome of planet Earth. The majority of studies have aimed to decipher the role of forest soil bacteria and fungi to understand and predict the impact of climate change on soil microbiome community structure and their ecosystem in the environment. In forest soils, microorganisms live in diverse habitats with specific behavior, comprising bulk soil, rhizosphere, litter, and deadwood habitats, where their communities are influenced by biotic interactions and nutrient accessibility. Soil microbiome also drives multiple crucial steps in the nutrient biogeochemical cycles (carbon, nitrogen, phosphorous, and sulfur cycles). Soil microbes help in the nitrogen cycle through nitrogen fixation during the nitrogen cycle and maintain the concentration of nitrogen in the atmosphere. Soil microorganisms in forest soils respond to various effects of climate change, for instance, global warming, elevated level of CO2, drought, anthropogenic nitrogen deposition, increased precipitation, and flood. As the major burning issue of the globe, researchers are facing the major challenges to study soil microbiome. This review sheds light on the current scenario of knowledge about the effect of climate change on living soil ecosystems in various climate-sensitive soil ecosystems and the consequences for vegetation-soil-climate feedbacks.

Endophytic Nanotechnology: An Approach to Study Scope and Potential Applications.

Nanotechnology has become a very advanced and popular form of technology with huge potentials. Nanotechnology has been very well explored in the fields of electronics, automobiles, construction, medicine, and cosmetics, but the exploration of nanotecnology's use in agriculture is still limited. Due to climate change, each year around 40% of crops face abiotic and biotic stress; with the global demand for food increasing, nanotechnology is seen as the best method to mitigate challenges in disease management in crops by reducing the use of chemical inputs such as herbicides, pesticides, and fungicides. The use of these toxic chemicals is potentially harmful to humans and the environment. Therefore, using NPs as fungicides/ bactericides or as nanofertilizers, due to their small size and high surface area with high reactivity, reduces the problems in plant disease management. There are several methods that have been used to synthesize NPs, such as physical and chemical methods. Specially, we need ecofriendly and nontoxic methods for the synthesis of NPs. Some biological organisms like plants, algae, yeast, bacteria, actinomycetes, and fungi have emerged as superlative candidates for the biological synthesis of NPs (also considered as green synthesis). Among these biological methods, endophytic microorganisms have been widely used to synthesize NPs with low metallic ions, which opens a new possibility on the edge of biological nanotechnology. In this review, we will have discussed the different methods of synthesis of NPs, such as top-down, bottom-up, and green synthesis (specially including endophytic microorganisms) methods, their mechanisms, different forms of NPs, such as magnesium oxide nanoparticles (MgO-NPs), copper nanoparticles (Cu-NPs), chitosan nanoparticles (CS-NPs), ß-d-glucan nanoparticles (GNPs), and engineered nanoparticles (quantum dots, metalloids, nonmetals, carbon nanomaterials, dendrimers, and liposomes), and their molecular approaches in various aspects. At the molecular level, nanoparticles, such as mesoporous silica nanoparticles (MSN) and RNA-interference molecules, can also be used as molecular tools to carry genetic material during genetic engineering of plants. In plant disease management, NPs can be used as biosensors to diagnose the disease.

Development and screening of byproduct for its secondary metabolites, antioxidant and anti-diabetic potential from anthracnose-infected fruits of pomegranate: a sustainable approach.

The main focus of the present study was to analyze the antioxidant and anti-diabetic potential of fermentative byproduct, developed from anthracnose-infected pomegranate fruits. The analysis of fermented juice showed a reduction in total phenolic content, total flavanoid content, anthocyanins, and antioxidant potential over the time in 6 months as compared to fresh juice, while total protein and alcohol percent (11%) were increased. Measurements of antioxidant activity by DPPH, ABTS, superoxide radical scavenging activity, and reducing power assays were highly correlated to total phenolic content, with corresponding R 2 values as r DPPH = 0.88, r ABTS = 0.90, r SRS = 0.67, r RPA = 0.80. High-performance liquid chromatography clearly revealed that the increment of antioxidant activity is associated with the release of gallic acid, vanillin, and ferulic acid. LC-MS analysis identified 1263 metabolites in fresh juice, 1580 metabolites after 1 month of fermentation, and 1063 metabolites after 6 months of fermentation. Most of the detected metabolites are linked with antioxidant, anti-diabetic, phenolics, flavanoids, cardiac glycosides, anticancer, and anti-vomiting activity. Mainly, naphthofluorescein, CAY10599, CAY10506, aminofluoropropionic acid, and 8-azaadenosine anti-diabetic compounds were found in fresh juice and fermented juice. Administration of fresh juice and fermented juice for 1 month helped in the reduction of blood plasma glucose level from 112.6 to 94.73 mg/dL before food and 142.43 to 133.20 mg/dL after food as compared to prescribed medicine. The sensory attributes of fermented juice were well appreciated for taste, after taste, and flavor. Further research is necessary to improve the quality and stability of metabolites during storage. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-020-02629-z.

Biology and Interaction of the Natural Occurrence of Distinct Monopartite Begomoviruses Associated With Satellites in Capsicum annum From India.

Chili (Capsicum annuum L.) is an important vegetable and spice crop of tropical and sub-tropical regions. Chili plants showing upward leaf curling, leaf crinkling, and leaf yellowing symptoms, collected from Sikar district of Rajasthan, India, were found to be associated with begomovirus and satellite molecules. The presence of virus was confirmed by PCR using virus-specific primer. The full-length genomic DNA-A of three begomovirus (MM-1, CS-1 and RV-1) and two satellites (MM-2 and MM-3) were cloned which was identified from single symptomatic chili plant. The genome organization of isolated three viruses is similar to those of other Old World monopartite begomoviruses. The comparison of the sequences and closest phylogenetic relationships for the begomoviruses, betasatellite and alphasatellite DNAs revealed that MM-1 was designated as DNA-A of Chili leaf curl virus (ChiLCV), CS-1 is considered to be a new distinct species of Tomato leaf curl Gujrat virus (ToLCGV) whereas RV-1 as a new strain of Cotton leaf curl Multan virus (CLCuMuV). The DNA-A component of ChiLCV showed 8.6%, ToLCGV of 16.6% and CLCuMuV of 7.7% average evolutionary divergence, concomitantly, the betasatellite and alphasatellite molecule had 9.9% and 5.9% overall sequence divergence, respectively. Interestingly, most of the begomoviruses were found to be intra-species recombinants. The dN/dS ratio and Tajima D value of all viral DNA-A component and their associated betasatellite showed their selective control on evolutionary relationships. The nucleotide substitution rates were determined for the DNA-A genomes of ChiLCV (7.22 × 10-4 substitutions site-1 year-1), CLCuMuV (1.49 × 10-4 substitutions site-1 year-1), ToLCGV (7.47 × 10-4 substitutions site-1 year-1), the genome of associated ChiLCB (4.20 × 10-4 substitutions site-1 year-1) and CLCuMuA (1.49 × 10-4 substitutions site-1 year-1). Agro-inoculation studies indicate that the presence of DNA betasatellite induce severe symptoms in N. benthamiana and chili, suggesting prerequisite association for typical disease development.

Host Plant Strategies to Combat Against Viruses Effector Proteins.

Viruses are obligate parasites that exist in an inactive state until they enter the host body. Upon entry, viruses become active and start replicating by using the host cell machinery. All plant viruses can augment their transmission, thus powering their detrimental effects on the host plant. To diminish infection and diseases caused by viruses, the plant has a defence mechanism known as pathogenesis-related biochemicals, which are metabolites and proteins. Proteins that ultimately prevent pathogenic diseases are called R proteins. Several plant R genes (that confirm resistance) and avirulence protein (Avr) (pathogen Avr gene-encoded proteins [effector/elicitor proteins involved in pathogenicity]) molecules have been identified. The recognition of such a factor results in the plant defence mechanism. During plant viral infection, the replication and expression of a viral molecule lead to a series of a hypersensitive response (HR) and affect the host plant's immunity (pathogen-associated molecular pattern-triggered immunity and effector-triggered immunity). Avr protein renders the host RNA silencing mechanism and its innate immunity, chiefly known as silencing suppressors towards the plant defensive machinery. This is a strong reply to the plant defensive machinery by harmful plant viruses. In this review, we describe the plant pathogen resistance protein and how these proteins regulate host immunity during plant-virus interactions. Furthermore, we have discussed regarding ribosome-inactivating proteins, ubiquitin proteasome system, translation repression (nuclear shuttle protein interacting kinase 1), DNA methylation, dominant resistance genes, and autophagy-mediated protein degradation, which are crucial in antiviral defences.

PGPR-mediated induction of systemic resistance and physiochemical alterations in plants against the pathogens: Current perspectives.

Plant growth-promoting rhizobacteria (PGPR) are diverse groups of plant-associated microorganisms, which can reduce the severity or incidence of disease during antagonism among bacteria and soil-borne pathogens, as well as by influencing a systemic resistance to elicit defense response in host plants. An amalgamation of various strains of PGPR has improved the efficacy by enhancing the systemic resistance opposed to various pathogens affecting the crop. Many PGPR used with seed treatment causes structural improvement of the cell wall and physiological/biochemical changes leading to the synthesis of proteins, peptides, and chemicals occupied in plant defense mechanisms. The major determinants of PGPR-mediated induced systemic resistance (ISR) are lipopolysaccharides, lipopeptides, siderophores, pyocyanin, antibiotics 2,4-diacetylphoroglucinol, the volatile 2,3-butanediol, N-alkylated benzylamine, and iron-regulated compounds. Many PGPR inoculants have been commercialized and these inoculants consequently aid in the improvement of crop growth yield and provide effective reinforcement to the crop from disease, whereas other inoculants are used as biofertilizers for native as well as crops growing at diverse extreme habitat and exhibit multifunctional plant growth-promoting attributes. A number of applications of PGPR formulation are needed to maintain the resistance levels in crop plants. Several microarray-based studies have been done to identify the genes, which are associated with PGPR-induced systemic resistance. Identification of these genes associated with ISR-mediating disease suppression and biochemical changes in the crop plant is one of the essential steps in understanding the disease resistance mechanisms in crops. Therefore, in this review, we discuss the PGPR-mediated innovative methods, focusing on the mode of action of compounds authorized that may be significant in the development contributing to enhance plant growth, disease resistance, and serve as an efficient bioinoculants for sustainable agriculture. The review also highlights current research progress in this field with a special emphasis on challenges, limitations, and their environmental and economic advantages.

Identification, genetic diversity and recombination analysis of Watermelon Mosaic Virus isolates.

Watermelon mosaic virus (WMV) is an important virus causing adverse effects on cucurbits throughout the world. In this study, we recorded WMV infection in the watermelon (Citrullus lanatus)-growing area of Alwar and Sikar in districts of Rajasthan, India. The RT-PCR-based detection was performed to confirm the presence of WMV, by using potyvirus-degenerated coat protein primers. Further, the complete genome sequences of two WMV isolates were compared with previously reported genome sequences. The complete genome of each isolate was 10,030 nt long, excluding the poly-A tails. Phylogeny relationships of the WMV isolates in the present study revealed the presence of uneven evolutionary pressure among the different WMV viral genomic segments. The analysis revealed that all the WMV isolates were divided into three clusters and the Indian WMV isolates cluster together with the French isolate. Recombination analysis of WMV exhibited significant recombination hotspots in the P1, NIa-Pro and Nib-CP regions. Our finding highlights the importance of genetic variability and recombination analysis to provide a better understanding of WMV molecular diversity.

Genome of a known but distinct begomovirus associated with a novel satellite molecule infecting a new host bitter gourd (Momordica charantia).

Coccinia mosaic Virdhunagar virus (KY860899), Tomato leaf curl New Delhi virus (KY860898) and Tomato leaf curl Virdhunagar alphasatellite (KY848691) were found to be associated with leaf curl disease in Momordica charantia (bitter gourd). The complete nucleotide sequence of Coccinia mosaic Virdhunagar virus showed 82% identity with Coccinia mosaic Tamil Nadu virus (KM244719), whereas Tomato leaf curl New Delhi virus was 96% identical to Tomato leaf curl New Delhi virus (KP868764) and Tomato leaf curl Virdhunagar alphasatellite illustrated 81% similarity with Tomato leaf curl New Delhi alphasatellite (JQ041697). Phylogenetic and RDP analysis revealed the proximity of these begomoviruses with other monopartite begomoviruses and alphasatellites already reported from India. As per the threshold criteria laid down by International Committee on Taxonomy of Viruses for species demarcation in begomoviruses and satellite molecules, the identified virus isolates, Coccinia mosaic Virdhunagar virus and Tomato leaf curl Virdhunagar alphasatellite are proposed as new species. To the best of our knowledge, this is the first ever account of mixed infection of begomoviruses in Momordica charantia, a vegetable crop commonly cultivated throughout India.

Complete nucleotide sequence of a new geminivirus isolated from Vitis vinifera in India: a symptomless host of Grapevine red blotch virus.

Symptomless grape plants (Vitis vinifera) cultivated in Jind, Punjab, have been found to carry a Grapevine red blotch virus (GRBV). Evaluation of full length DNA sequence (3204 bp) of the virus (KU522121) has revealed similarity with mastrevirus, begomovirus, and other Grapevine red blotch viruses reported in the US and Canada. Similar to naturally growing plants, agroinfiltrated model plants with infectious clone of GRBV do not show any visible disease warning sign. To the best of our knowledge, this is the first report of a symptomless host Vitis vinifera from Indian vineyards harbouring a Grapevine geminivirus.

RNAi mediated gene silencing against betasatellite associated with Croton yellow vein mosaic begomovirus.

Plant viruses encode suppressors of posttranscriptional gene silencing, an adaptive antiviral defense responses that confines virus infection. Previously, we identified single-stranded DNA satellite (also known as DNA-ß) of ~1,350 nucleotides in length associated with Croton yellow vein mosaic begomovirus (CYVMV) in croton plants. The expression of genes from DNA-ß requires the begomovirus for packaged, replication, insect transmission and movement in plants. The present study demonstrates the effect of the ßC1 gene on the silencing pathway as analysed by using both transgenic systems and transient Agrobacterium tumefaciens based delivery. Plants that carry an intron-hairpin construct covering the ßC1 gene accumulated cognate small-interfering RNAs and remained symptom-free after exposure to CYVMV and its satellite. These results suggest that ßC1 interferes with silencing mechanism.

Complete nucleotide sequence of a begomovirus associated with satellites molecules infecting a new host Tagetes patula in India.

In the year 2012 leaf curl disease was observed on Marigold (Tagetes patula) in Lakshmangrh, Sikar province of India. Affected plants were severely stunted with apical leaf curl and crinkled leaves, symptoms typical of begomovirus infection. This is the first report of complete nucleotide sequence of a begomovirus associated with satellites molecules infecting a new host Tagetes patula in India.

Geminivirus database (GVDB): first database of family Geminiviridae and its genera Begomovirus.

Geminivirus Database (GVDB) is an online interactive database of Geminiviridae family. GVDB comprises of partial and complete nucleotide sequences along with duly annotated expressed genes of isolated Begomovirus species. The in silico homology modeling, docking and recombination results obtained for different begomoviral sequences are also mentioned. This database is endowed with comprehensive information about Geminivirus members which grounds infection in various plants species in India assorting from crops, ornamentals plants and common weeds. The home page of this database offers various links associated with current research projects and also the publications related to molecular and in silico study of Begomovirus infection. The main feature of GVDB includes flexible database designs based on platform of PHP allows easy retrieval of the information. The database is made available at www.wikigeminivirus.org.