A novel monopartite begomovirus and satellites associated with yellow mosaic disease of Sida spp. in India.

Begomoviruses (family Geminiviridae) cause severe diseases in many economically important crops and non-cultivated plants in the warmer regions of the world. Non-cultivated weeds have been reported to act as natural virus reservoirs. In January 2016, Sida plants with yellow mosaic symptoms were found at the edge of an agricultural field in Gujarat, India. Sequence analysis of the viral genomic components cloned from a diseased Sida plant indicated the presence of a distinct monopartite begomovirus (proposed as sida yellow mosaic Gujarat virus) along with a betasatellite (ludwigia leaf distortion betasatellite) and an alphasatellite (malvastrum yellow mosaic alphasatellite). Our results emphasize that this weed may harbor a begomovirus-alphasatellite-betasatellite complex. This host serves as a potential source of virus inoculum, which can be transmitted by whiteflies to other cultivated crops.

Bhendi Yellow Vein Mosaic Virus and Bhendi Yellow Vein Mosaic Betasatellite Cause Enation Leaf Curl Disease and Alter Host Phytochemical Contents in Okra.

Whitefly (Bemisia tabaci)-transmitted begomoviruses cause severe diseases in numerous economically important dicotyledonous plants. Okra enation leaf curl disease (OELCuD) has emerged as a serious threat to okra (Abelmoschus esculentus L. Moench) cultivation in the Indian subcontinent. This study reports the association of a monopartite begomovirus (bhendi yellow vein mosaic virus; BYVMV) and betasatellite (bhendi yellow vein mosaic betasatellite; BYVB) with OELCuD in the Mau region of Uttar Pradesh, India. The BYVMV alone inoculated Nicotiana benthamiana and A. esculentus cv. Pusa Sawani plants developed mild symptoms. Co-inoculation of BYVMV and BYVB resulted in a reduced incubation period, an increased symptom severity, and an enhanced BYVMV accumulation by Southern hybridization and quantitative real-time PCR. This is the first study that satisfies Koch's postulates for OELCuD in its natural host. Activities of various antioxidative enzymes were significantly increased in the virus-inoculated okra plants. Differential responses in various biochemical components (such as photosynthetic pigments, phenol, proline, and sugar) in diseased okra plants were observed. This change in phytochemical responses is significant in understanding its impact on virus pathogenesis and disease development.

Molecular genetic analysis and evolution of begomoviruses and betasatellites causing yellow mosaic disease of bhendi.

In India, Bhendi yellow vein mosaic disease (BYVMD) is one of the most economically important diseases of bhendi/okra and is caused by a complex of monopartite begomovirus (Bhendi yellow vein mosaic virus-BYVMV) and betasatellite (Bhendi yellow vein betasatellite-BYVB). In this study, we have analyzed the role of possible evolutionary factors involved in the evolution of BYVMV and BYVB isolates. Evidence of inter-species and inter-strain recombination events was detected among the viral isolates, and majority of these recombinant isolates possess microsatellites in their genome. Recombination analysis suggests that cotton-infecting and bhendi-infecting begomoviruses probably share a recent common ancestor. In addition to genetic differentiation and gene flow, high degree of genetic variability was detected among the viral population. A strong purifying selection seems to be acting on the viral coding regions. The nucleotide substitution rate of V1 gene (for BYVMV) and ßC1 gene (for BYVB) was estimated to be 7.55 × 10-4 and 2.25 × 10-3 nucleotide substitutions/site/year, respectively. The present study underlines that the evolution of BYVMD-associated viral components is driven by selection acting on the genetic variation generated by recombination and mutation.

An Accurate Methodology to detect Leaching of Nickel and Chromium Ions in the Initial Phase of Orthodontic Treatment: An in vivo Study.

AIM: The aim of this study was to evaluate the release of nickel and chromium ions in human saliva during fixed orthodontic therapy. MATERIALS AND METHODS: Ten patients with Angle's Class-I malocclusion with bimaxillary protrusion without any metal restorations or crowns and with all the permanent teeth were selected. Five male patients and five female patients in the age group range of 14 to 23 years were scheduled for orthodontic treatment with first premolar extraction. Saliva samples were collected in three stages: sample 1, before orthodontic treatment; sample 2, after 10 days of bonding sample; and sample 3, after 1 month of bonding. The samples were analyzed for the following metals nickel and chromium using inductively coupled plasma optical emission spectrometry (ICP-OES). RESULTS: The levels of nickel and chromium were statistically significant, while nickel showed a gradual increase in the first 10 days and a decline thereafter. Chromium showed a gradual increase and was statistically significant on the 30th day. CONCLUSION: There was greatest release of ions during the first 10 days and a gradual decline thereafter. Control group had traces of nickel and chromium. While comparing levels of nickel in saliva, there was a significant rise from baseline to 10th and 30th-day sample, which was statistically significant. While comparing 10th day to that of 30th day, there was no statistical significance. The levels of chromium ion in the saliva were more in 30th day, and when comparing 10th-day sample with 30th day, there was statistical significance. CLINICAL SIGNIFICANCE: Nickel and chromium levels were well within the permissible levels. However, some hypersensitive individuals may be allergic to this minimal permissible level.

Complexity of begomovirus and betasatellite populations associated with chilli leaf curl disease in India.

Chilli, which encompasses several species in the genus Capsicum, is widely consumed throughout the world. In the Indian subcontinent, production of chilli is constrained due to chilli leaf curl disease (ChiLCD) caused by begomoviruses. Despite the considerable economic consequences of ChiLCD on chilli cultivation in India, there have been scant studies of the genetic diversity and structure of the begomoviruses that cause this disease. Here we report on a comprehensive survey across major chilli-growing regions in India. Analysis of samples collected in the survey indicates that ChiLCD-infected plants are associated with a complex of begomoviruses (including one previously unreported species) with a diverse group of betasatellites found in crops and weeds. The associated betasatellites neither enhanced the accumulation of the begomovirus components nor reduced the incubation period in Nicotiana benthamiana. The ChiLCD-associated begomoviruses induced mild symptoms on Capsicum spp., but both the level of helper virus that accumulated and the severity of symptoms were increased in the presence of cognate betasatellites. Interestingly, most of the begomoviruses were found to be intra-species recombinants. The betasatellites possess high nucleotide variability, and recombination among them was also evident. The nucleotide substitution rates were determined for the AV1 gene of begomoviruses (2.60 × 10- 3 substitutions site- 1 year- 1) and the ßC1 gene of betasatellites [chilli leaf curl betasatellite (ChiLCB), 2.57 × 10- 4 substitution site- 1 year- 1; tomato leaf curl Bangladesh betasatellite (ToLCBDB), 5.22 × 10- 4 substitution site- 1 year- 1]. This study underscores the current understanding of Indian ChiLCD-associated begomoviruses and also demonstrates the crucial role of betasatellites in severe disease development in Capsicum spp.

Fabrication of zirconia composite membrane by in-situ hydrothermal technique and its application in separation of methyl orange.

The main objective of the work was preparation of zirconia membrane on a low cost ceramic support through an in-situ hydrothermal crystallization technique for the separation of methyl orange dye. To formulate the zirconia film on the ceramic support, hydrothermal reaction mixture was prepared using zirconium oxychloride as a zirconia source and ammonia as a precursor. The synthesized zirconia powder was characterized by X-ray diffractometer (XRD), N2 adsorption/desorption isotherms, Thermogravimetric analysis (TGA), Fourier transform infrared analysis (FTIR), Energy-dispersive X-ray (EDX) analysis and particle size distribution (PSD) to identify the phases and crystallinity, specific surface area, pore volume and pore size distribution, thermal behavior, chemical composition and size of the particles. The porosity, morphological structure and pure water permeability of the prepared zirconia membrane, as well as ceramic support were investigated using the Archimedes' method, Field emission scanning electron microscopy (FESEM) and permeability. The specific surface area, pore volume, pore size distribution of the zirconia powder was found to be 126.58m(2)/g, 3.54nm and 0.3-10µm, respectively. The porosity, average pore size and pure water permeability of the zirconia membrane was estimated to be 42%, 0.66µm and 1.44×10(-6)m(3)/m(2)skPa, respectively. Lastly, the potential of the membrane was investigated with separation of methyl orange by means of flux and rejection as a function of operating pressure and feed concentration. The rejection was found to decrease with increasing the operating pressure and increases with increasing feed concentrations. Moreover, it showed a high ability to reject methyl orange from aqueous solution with a rejection of 61% and a high permeation flux of 2.28×10(-5)m(3)/m(2)s at operating pressure of 68kPa.

Molecular characterization of Chilli leaf curl virus and satellite molecules associated with leaf curl disease of Amaranthus spp.

Amaranthus, collectively known as amaranth, is an annual or short-lived perennial plant used as leafy vegetables, cereals and for ornamental purposes in many countries including India. During 2011, leaf samples of Amaranthus plants displaying leaf curling, leaf distortion, leaf crinkling and yellow leaf margins were collected from Banswara district, Rajasthan in India. Full-length clones of a monopartite begomovirus, a betasatellite and an alphasatellite were characterized. The complete nucleotide sequence of the isolated begomovirus features as a typical 'Old World' begomovirus with the highest nucleotide per cent identity with Chilli leaf curl virus and hence, considered as an isolate of Chilli leaf curl virus. The complete nucleotide sequences of betasatellite and alphasatellite possess maximum nucleotide identity with Tomato yellow leaf curl Thailand betasatellite and Chilli leaf curl alphasatellite, respectively. This is the first report of the association of chilli-infecting begomovirus and satellite molecules infecting a new host, Amaranthus, causing leaf curl disease.

Host-specific adaptation of diverse betasatellites associated with distinct Indian tomato-infecting begomoviruses.

Tomato leaf curl viruses cause major crop loss hindering tomato cultivation worldwide. The 'Old World' begomoviruses are often associated with circular ssDNA satellite molecules called betasatellites. In the present study, replication compatibility of five different betasatellites with three distinct Indian tomato-infecting begomoviruses representing each of a monopartite, a mono-bipartite and a bipartite begomoviruses was studied. All the betasatellites could be trans-replicated by the begomoviruses in Nicotiana benthamiana plants, however, not uniformly in tomato. Tomato leaf curl Joydebpur betasatellite-Magrahat could not induce symptom with any of these begomoviruses in tomato, whereas only Tomato leaf curl Gujarat virus could trans-replicate Radish leaf curl betasatellite in this plant species. However, none of the betasatellites were found to complement the movement function of a bipartite begomovirus in tomato. Unlike tomato, the trans-replication/maintenance of betasatellites by these begomoviruses in N. benthamiana could be due to its compromised host defence machinery. Co-infection of betasatellites with these viruses did not enhance the helper virus accumulation, but the incubation period was reduced. The possible factors involved in this host-driven adaptability of betasatellites were also discussed.

Differential pathogenicity among Tomato leaf curl Gujarat virus isolates from India.

Tomato leaf curl Gujarat virus (ToLCGV) has been identified as one of the most destructive pathogens causing tomato leaf curl disease (ToLCD) in India. In the tomato growing regions of Dhanbad and Ramgarh, plants bearing severe symptoms of ToLCD such as leaf curling, leaf crinkling, yellowing and leaf rolling was observed in the farmer fields. The association of begomovirus in these samples was confirmed by PCR and the causal viruses were identified as the isolates of ToLCGV. However, association of cognate DNA B component could not be ascertained from these samples. Indeed, like other Old World begomoviruses, the present ToLCGV isolates were found to be associated with a particular betasatellite, Tomato yellow leaf curl Thailand betasatellite (TYLCTHB). Although DNA A of both ToLCGV isolates could alone infect tomato inducing systemic symptoms, the difference in virulence was observed. Co-inoculation of TYLCTHB reduced the incubation period without influencing the accumulation of helper virus DNA and hence, differential pathogenesis among ToLCGV isolates was governed by the helper component rather than betasatellite. ToLCGV infection with DNA B increases the accumulation of DNA A component of Dhanbad isolate but not of Ramgarh isolate. Results indicated that the begomovirus identified from Ramgarh sample was a mild strain of ToLCGV.

Diverse begomovirus-betasatellite complexes cause tomato leaf curl disease in the western India.

In the Indian sub-continent, tomato leaf curl disease (ToLCD) of tomato caused by begomoviruses has emerged as a major limiting factor for tomato cultivation. Despite the spread of this disease in the western India, a systematic study on the characterization of virus complexes with ToLCD is lacking. Here, we report the identification of a complex of begomoviruses including 19 DNA-A and 4 DNA-B as well as 15 betasatellites with ToLCD in the western part of the country. Additionally, a novel betasatellite and an alphasatellite were also identified. The recombination breakpoints were detected in the cloned begomoviruses and betasatellites. The cloned infectious DNA constructs cause disease on the tomato (a moderately virus-resistant cultivar) plants, thus fulfilling Koch's postulates for these virus complexes. Further, the role of non-cognate DNA B/betasatellite with ToLCD-associated begomoviruses on disease development was demonstrated. It also emphasizes the evolutionary potential of these virus complexes in breaking disease resistance and plausible expansion of its host range. This necessitates to investigate the mechanism of the interaction between resistance breaking virus complexes and the infected host.

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.

An improved plant regeneration and Agrobacterium - mediated transformation of red pepper (Capsicum annuum L.).

Capsicum annuum (red pepper) is an important spice cum vegetable crop in tropical and subtropical countries. Here, we report an effective and reproducible auxin free regeneration method for six different red pepper cultivars (ACA-10, Kashi Anmol, LCA-235, PBC-535, Pusa Jwala and Supper) using hypocotyl explants and an efficient Agrobacterium-mediated transformation protocol. The explants (hypocotyls, cotyledonary leaves and leaf discs) collected from axenic seedlings of six red pepper cultivars were cultured on either hormone free MS medium or MS medium supplemented with BAP alone or in combination with IAA. Inclusion of IAA in the regeneration medium resulted in callus formation at the cut ends of explants, formation of rosette leaves and ill defined shoot buds. Regeneration of shoot buds could be achieved from hypocotyls grown in MS medium supplemented with different concentrations of BAP unlike other explants which failed to respond. Incorporation of GA3 in shoot elongation medium at 0.5 mg/l concentration enhanced the elongation in two cultivars, LCA-235 and Supper, while other cultivars showed no significant response. Chilli cultivar, Pusa Jwala was transformed with ßC1 ORF of satellite DNA ß molecule associated with Chilli leaf curl Joydebpur virus through Agrobacterium tumefaciens. Transgene integration in putative transformants was confirmed by PCR and Southern hybridization analysis.

Two distinct monopartite begomovirus-betasatellite complexes in western India cause tomato leaf curl disease.

In India, begomovirus infection causing tomato leaf curl disease (ToLCD) is a major constraint for tomato productivity. Here, we have identified two distinct monopartite begomovirus and betasatellite complexes causing ToLCD in the western part of India. A new monopartite begomovirus (Tomato leaf curl Mumbai virus, ToLCMumV) and betasatellite (Tomato leaf curl Mumbai betasatellite, ToLCMumB) were isolated from the Mumbai sample. A distinct Tomato leaf curl Gandhinagar virus (ToLCGanV) and Tomato leaf curl Gandhinagar betasatellite (ToLCGanB) were identified from the Gandhinagar sample. Both of the cloned begomoviruses were recombinants. The demonstration of systemic infection caused by begomovirus (ToLCGanV or ToLCMumV) alone in N. benthamiana and tomato (a virus resistant variety) emphasizes that they were monopartite begomoviruses. Co-inoculation of cognate begomovirus and betasatellite reduces the incubation period and increases symptom severity. Thus, Koch's postulates were satisfied for these virus complexes. Further, an enhanced accumulation of ToLCGanV was detected in the presence of cognate ToLCGanB, however ToLCMumB did not influence the level of ToLCMumV in the agro-inoculated tomato plants. Our results indicate that the cloned viruses form potential virus resistance breaking disease complexes in India. This necessitates to investigate the spread of these disease complexes to major tomato growing regions in the country.

Plant Antiviral Immunity Against Geminiviruses and Viral Counter-Defense for Survival.

The family Geminiviridae includes plant-infecting viruses whose genomes are composed of one or two circular non-enveloped ssDNAs(+) of about 2.5-5.2 kb each in size. These insect-transmissible geminiviruses cause significant crop losses across continents and pose a serious threat to food security. Under the control of promoters generally located within the intergenic region, their genomes encode five to eight ORFs from overlapping viral transcripts. Most proteins encoded by geminiviruses perform multiple functions, such as suppressing defense responses, hijacking ubiquitin-proteasomal pathways, altering hormonal responses, manipulating cell cycle regulation, and exploiting protein-signaling cascades. Geminiviruses establish complex but coordinated interactions with several host elements to spread and facilitate successful infection cycles. Consequently, plants have evolved several multilayered defense strategies against geminivirus infection and distribution. Recent studies on the evasion of host-mediated resistance factors by various geminivirus proteins through novel mechanisms have provided new insights into the development of antiviral strategies against geminiviruses. This review summarizes the current knowledge concerning virus movement within and between cells, as well as the recent advances in our understanding of the biological roles of virus-encoded proteins in manipulating host-mediated responses and insect transmission. This review also highlights unexplored areas that may increase our understanding of the biology of geminiviruses and how to combat these important plant pathogens.

A novel integrated biodegradation-microfiltration system for sustainable wastewater treatment and energy recovery.

This work assessed the treatment of wastewater generated from three different industries viz., paper and pulp, biomass gasification and dairy by biodegradation followed by membrane filtration. Batch biodegradation was first carried out using wastewater as the potential substrate for oleaginous Rhodococcus opacus with lipid accumulation intracellular; subsequently, a microfiltration system was applied to recover the bacterial biomass grown as well as for residual chemical oxygen demand (COD) removal from the effluent. The combined process showed excellent results in terms of COD removal from the industrial wastewaters, with the values 56.8%, 46.1% and 68.9% for dairy, paper and pulp and biomass gasification wastewaters, respectively, by biodegradation. These values were further improved to 92.7%, 87.6% and 88.2%, respectively, following the microfiltration step performed by employing a low-cost ceramic membrane. In addition, lipids accumulated by the bacterium were extracted and characterized for biodiesel production potential. Lipid characterization using 1H NMR confirmed the presence of saturated fatty acids. Gas chromatography analysis of the transesterified lipids revealed the presence of methyl palmitate and methyl stearate. In addition, the estimated properties of the transesterified product affirmed its potential for biofuel application.

Optimization of ultrasound assisted extraction of pectin from custard apple peel: Potential and new source.

Pectin was extracted from the waste custard apple peel using ultrasound technique and optimized the extraction process by RSM. The various significant process parameters such as liquid-solid ratio, ultra-sonication time, temperature and pH of solution were studied in the range of 10-25 mL g-1, 10-30 min, 50-80 °C, and 1-3, respectively. The maximum yield of pectin (8.93%) was attained at the optimum condition of 23.52 mL g-1 of liquid-solid ratio, 18.04  min of ultra-sonication time, 63.22 °C of temperature and 2.3 pH of solution. The extracted and commercially available fresh pectin (for comparison purposes) were characterized by various analytical techniques namely, FTIR, DSC, XRD, SEM, and NMR to evaluate their functional groups, thermal properties, crystallinities, morphological and structural characteristics, respectively. The extracted pectin was a toxic free compound as determined by its anti nutritional property study and about 20 mg/mL of antioxidant presented in it.

Mucinous cystadenoma of pancreas with honeycombing appearance: Radiological-Pathological correlation.

Most mucinous cystadenomas of pancreas are solitary and multilocular with a few large compartments. Serous cystadenomas usually have a polycystic or microcystic (honeycomb) pattern consisting of collection of cysts (usually >6) that range from few millimetres up to 2 cm in size. Here we present a case of mucinous cystadenoma of pancreas showing an unusual appearance of honeycombing (which has not been described so far) using imaging studies such as endoscopic ultrasound and computed tomography with histopathological confirmation of the diagnosis.

Cross flow ultrafiltration of Cr (VI) using MCM-41, MCM-48 and Faujasite (FAU) zeolite-ceramic composite membranes.

This work describes the removal of Cr (VI) from aqueous solution in cross flow mode using MCM-41, MCM-48 and FAU zeolite membranes prepared on circular shaped porous ceramic support. Ceramic support was manufactured using locally available clay materials via a facile uni-axial compaction method followed by sintering process. A hydrothermal technique was employed for the deposition of zeolites on the ceramic support. The porosity of ceramic support (47%) is reduced by the formation of MCM-41 (23%), MCM-48 (22%) and FAU (33%) zeolite layers. The pore size of the MCM-41, MCM-48 and FAU membrane is found to be 0.173, 0.142, and 0.153 µm, respectively, which is lower than that of the support (1.0 µm). Cross flow ultrafiltration experiments of Cr (VI) were conducted at five different applied pressures (69-345 kPa) and three cross flow rates (1.11 × 10(-7) - 2.22 × 10(-7) m(3)/s). The filtration studies inferred that the performance of the fabricated zeolite composite membranes is optimum at the maximum applied pressure (345 kPa) and the highest rejection is obtained with the lowest cross flow rate (1.11 × 10(-7) m(3)/s) for all three zeolite membrane. The permeate flux of MCM-41, MCM-48 and FAU zeolite composite membranes are almost remained constant in the entire duration of the separation process. The highest removal of 82% is shown by FAU membrane, while MCM-41 and MCM-48 display 75% and 77% of Cr (VI) removal, respectively for the initial feed concentration of 1000 ppm with natural pH of the solution at an applied pressure of 345 kPa.

Potential linkage between compound microsatellites and recombination in geminiviruses: Evidence from comparative analysis.

The compound microsatellites consist of two or more individual microsatellites, originate from mutation or imperfection in simple repeat sequences. The reports on systematic analysis of the occurrence, size and density of compound microsatellite (cSSR) types are very rare. Our study indicates that cSSRs are clustered at specific regions in the begomovirus genomes. cSSRs were overrepresented in majority of begomovirus genomes indicating that they might have some functional significance. Further, non-random distribution pattern of cSSR in begomovirus genomes was significantly correlated with the recombination breakpoint positions in the genome. The analysis of cSSR regions in the viral genome indicates the presence of stem loop (hairpin) secondary structure. The significance of these findings in biology of geminiviruses is discussed based on our present understanding of recombination and repetitive DNA. To our knowledge, this is the first analysis suggesting the possible association between recombination and microsatellites in any viral genome.

Extrusion of impacted mandibular second molar using removable appliance.

The purpose of this article is to review the principles of case management of impacted mandibular molars and to illustrate their potential to respond well to treatment. Although the scope of treatment may be influenced by the patient's age, past dental history, severity of impaction, dentoalveolar development, and root form, the case reports demonstrate the inherent potential for good treatment outcome even in the most unfavorable circumstances.