Hepatitis B virus (HBV) infection remains a global health concern, with persistent covalently closed circular DNA (cccDNA) hindering curative treatments. Coumarin is one of these natural compounds that could be utilized as a pharmaceutical agent due to its stability, solubility, availability and low toxicity. Previous studies demonstrate their efficacy, but the challenges of specificity and resistance remain unexplored. Further research is needed to unlock coumarins full potential as antiviral agents for chronic HBV infection and removal of cccDNA from infected hepatocytes.

The present paper deals with frequency of abnormalities at anaphase I of meiosis in control and gamma ray treated plant of "S-22" cultivator of Lycopersicon esculentum(Tomato). Tomatoes are rich source of various vitamins and have good amount of Lycopene. The fruits are used in salad, cooked vegetable and ketchup. Lycopersicon esculentum belongs to family Solanaceae. Tomatoes plants are cultivated for its edible fruits. The plants of tomatoes are generally 90 cm tall and much branched. Leave are hairy and pinnately compound and 10-25 cm long. Leaves are strong odorous. The five petaled flowers are yellow and fruit are berries. In present investigation different doses of Gamma rays treatment such as 10 Kr, 15 Kr, 20 Kr, 25 Kr and 30 Kr given to "S-22" Cultivars of Lycopersicon esculentum, which is commonly cultivated in Gaya, Bihar. Some interesting abnormalities at anaphase I of meiosis in R1 and R2 generation were found out. Frequency of abnormalities at anaphase I in control and gamma ray treated were noted out. However the frequencies of abnormalities work less in R2 than R1 generation.

A pot experiment was conducted during 2014-15 in the Department of Agriculture Botany, Janta Mahavidiyalaya Ajitmal, Auraiya (C.S.J.M. University, Kanpur). Eight genotypes of wheat viz. KRL1-4, K8434, K88, K9644, K9465, K9006, HD2733 and HD2329 were tested to study their response to different levels of salt i.e. 3, 6, 9 and 12 dsm1 in addition to control. Lower levels of salinity did not affect the growth and physiology of wheat. Higher salinity caused a deleterious effect on growth parameters such as plant height, number of tillers per plant and dry weight. Increased salt concentration caused a great reduction in relative water content, chlorophyll content and rate of photosynthesis. Genotypes K9006, K8434, KRL1-4, K88 and HD2733 showed better performance in all the regard indicating their hardness towards salt. However, genotype K9644 showed poor performance under salt stress.

During November 2007, brinjal little leaf (BLL) symptoms (Fig. 1) were observed in approximately 20% of the brinjal (Solanum melongena) plants growing in the fields of Bihar, India, leading to the suspicion of a phytoplasma infection. To test for the presence of phytoplasma, genomic DNA was isolated from the leaf midribs of ten plants with and four plants without symptoms, and the phytoplasma DNA amplified by nested PCR with the universal primers P1/P7 (Deng & Hiruki, 1991) followed by R16mF2/R16mR1 (Gundersen & Lee, 1996), as previously described (Khan et al., 2004). The nested PCR amplicons of 1.4 kb corresponding to the phytoplasma 16S ribosomal DNA were cloned into pDRIVE vector (Qiagen GmbH, Germany). No PCR amplicons were observed for the symptomless plants. Twelve positive clones containing 16S ribosomal DNA of phytoplasma were sequenced and found sharing a 99.93% of sequence identity. Sequences of two clones were deposited in GenBank (Accession Nos. JQ518317 and JQ518318). In silico RFLP patterns were generated from the phytoplasma 16S ribosomal sequences using the gel plotting program pDRAW32 (http://www.acaclone.com/) and a phylogenetic tree was constructed using the neighbour-joining method of MEGA 4 (Tamura et al., 2007). BLAST analysis revealed that the Bihar phytoplasma detected in brinjal showed 98% 16S rDNA sequence identity with those of phytoplasmas from group 16SrI ('Candidatus Phytoplasma asteris'). The Bihar phytoplasma also showed only 84%, 74% and 72% 16S rDNA sequence identity respectively with those of the previously reported BLL phytoplasmas in India (EF186820, EU375486) and Bangladesh (AF228052) belonging to the 16SrVI group ('Ca. Phytoplasma trifolii'). Phylogenetic analysis (Fig. 2) evidenced that the phytoplasma associated with little leaf in brinjal in Bihar separated as a new phylogenetic branch within the 16SrI group cluster. In silico restriction fragment length polymorphism (RFLP) patterns were generated (Wei et al., 2007) for the Bihar BLL phytoplasma and the 16SrVI BLL phytoplasma reported earlier in India (EF186820) as well as the 16SrI Indian phytoplasmas identified in sandal spike (EF198362) and withania (DQ151998) (Fig. 3) with 13 restriction enzymes (AluI, BamHI, BfaI, DraI, EcoRI, HaeIII, HhaI, HinfI, HpaI, HpaII, KpnI, I, and TaqI). All the RFLP profiles of the Bihar BLL phytoplasma were similar to those of the 16SrI phytoplasmas, except for the AluI and KpnI RFLP patterns that differed from those exhibited by the 16SrVI BLL phytoplasma (EF186820). RFLP and the sequence results confirmed that the Bihar BLL phytoplasma is closely related to the phytoplasma group 16SrI and may represent a new subgroup within this group. This is the first report of a 16SrI phytoplasma affecting brinjal in India. The fact that two different phytoplasma groups (16SrVI and 16SrI) have been associated with little leaf diseases in brinjal may have further significant impact on disease epidemiology and control in India. The authors are grateful to the Department of Biotechnology, Government of India for funding.

New Disease ReportsVolume 25, Issue 1 p. 9-9 ArticleOpen Access First report of Radish leaf curl virus infecting okra in India J. Kumar, Corresponding Author J. Kumar jitsingh27281@gmail.com National Agri-Food Biotechnology Institute, Mohali, 160071 Punjab, IndiaSearch for more papers by this authorA. Kumar, A. Kumar School of Biochemistry, Devi Ahilya Vishwavidyalaya, Indore, IndiaSearch for more papers by this authorS.P. Singh, S.P. Singh National Agri-Food Biotechnology Institute, Mohali, 160071 Punjab, IndiaSearch for more papers by this authorJ.K. Roy, J.K. Roy National Agri-Food Biotechnology Institute, Mohali, 160071 Punjab, IndiaSearch for more papers by this authorA. Lalit, A. Lalit Department of Biochemistry and Genetics, Barkatullah University, Bhopal, IndiaSearch for more papers by this authorD. Parmar, D. Parmar Department of Biochemistry and Genetics, Barkatullah University, Bhopal, IndiaSearch for more papers by this authorN.C. Sharma, N.C. Sharma Department of Biochemistry and Genetics, Barkatullah University, Bhopal, IndiaSearch for more papers by this authorR. Tuli, R. Tuli National Agri-Food Biotechnology Institute, Mohali, 160071 Punjab, IndiaSearch for more papers by this author J. Kumar, Corresponding Author J. Kumar jitsingh27281@gmail.com National Agri-Food Biotechnology Institute, Mohali, 160071 Punjab, IndiaSearch for more papers by this authorA. Kumar, A. Kumar School of Biochemistry, Devi Ahilya Vishwavidyalaya, Indore, IndiaSearch for more papers by this authorS.P. Singh, S.P. Singh National Agri-Food Biotechnology Institute, Mohali, 160071 Punjab, IndiaSearch for more papers by this authorJ.K. Roy, J.K. Roy National Agri-Food Biotechnology Institute, Mohali, 160071 Punjab, IndiaSearch for more papers by this authorA. Lalit, A. Lalit Department of Biochemistry and Genetics, Barkatullah University, Bhopal, IndiaSearch for more papers by this authorD. Parmar, D. Parmar Department of Biochemistry and Genetics, Barkatullah University, Bhopal, IndiaSearch for more papers by this authorN.C. Sharma, N.C. Sharma Department of Biochemistry and Genetics, Barkatullah University, Bhopal, IndiaSearch for more papers by this authorR. Tuli, R. Tuli National Agri-Food Biotechnology Institute, Mohali, 160071 Punjab, IndiaSearch for more papers by this author First published: 01 March 2012 https://doi.org/10.5197/j.2044-0588.2012.025.009Citations: 9AboutSectionsPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Okra (Abelmoschus esculentus) is one of the important vegetable crops of India cultivated in 452 hectares and yielding 4803 tonnes (Anonymous, 2010). In Bihar State, India, leaf curl disease on okra was observed in the field causing crop losses of about 30% in 2009 and 35% in 2010-11. Characteristic symptoms of this disease were leaf curling and overall stunting of plants that bore no fruit (Fig. 1A). To test for a begomovirus-infection, total DNA was extracted from symptom-bearing leaves of six infected plants (2 plants ' 3 fields). A PCR approach was used to amplify viral genomes (primers F1For/Rev and F2For/Rev; Kumar et al., 2011) or alpha- and betasatellites (primers 'nanofor'/'nanorev' and 01/04; Kumar et al., 2010). All samples yielded PCR-products for a begomovirus and associated satellites, which were cloned and sequenced. The sequence deposited in GenBank for the monopartite begomovirus (Accession No. HQ257375) showed 97% and 92% nucleotide identity to Radish leaf curl virus (RaLCV) depositions GU732203 and EF175733, respectively. The alphasatellite (Accession No. HQ728354) possessed 98% and 96% nucleotide identity to Cotton leaf curl Burewala alphasatellite (CLCuBwA) sequences HM004548 and FN658728, respectively. The betasatellite (Accession No. HQ257376) exhibited 96% and 94% nucleotide identity to Tomato leaf curl Bangladesh betasatellite (ToLCBDB) sequences GU732208 and EF190215, respectively. Phylogenetic analysis of the begomovirus genome revealed a close relationship with RaLCV,GU732203 and EF175733 but a distant relationship with other okra infecting begomoviruses in India, AF241479 and FJ176236 (Fig. 2A). Phylogenetic analysis of the alphasatellite showed close relationship with CLCuBwA (Fig. 2B) and the betasatellite with ToLCBDB (Fig. 2C). Figure 1Open in figure viewerPowerPoint Figure 2Open in figure viewerPowerPoint Rolling circle amplification (RCA) was performed (TempliPhi amplification kit; GE Healthcare, USA) to construct infectious clones. RCA products were partially digested with BamHI to obtain monomer and head-to-tail tandem repeat dimers of full-length begomoviral DNA. Monomers and dimers were cloned into the pCAMBIA1301 vector. Infectious head-to-tail tandem repeat clones of both satellites were prepared analogously. Sequencing of 30 begomoviral monomer clones confirmed the presence of identical viral DNAs. Healthy whiteflies (~25) were used for virus and satellite transmission from field-collected infected plants to healthy tobacco and okra. Infectivity testing was performed by inoculation of tobacco and okra (10 plants each) with a mixture of begomovirus and alpha- and betasatellite infectious clones. Plants of both assays yielded typical symptoms of leaf curling and stunting identical to those observed previously in the field. Thus RaLCV and its associated satellites were confirmed as the causal agent of okra leaf curl disease. However, the functional role of both satellites for symptom development remains to be determined. This is the first report providing the evidence for RaLCVinfecting okra in India. Acknowledgments Authors are grateful to the Executive Director, NABI for providing facilities. Thanks to Dr. A K Pandey for critical reading of the manuscript. References Anonymous, 2010. Okra production status by Indian Horticulture database, National Horticulture Board. [http://nhb.gov.in/area-pro/ch2.pdf]. Kumar J, Kumar A, Roy JK, Tuli R, Khan JA, 2010. Identification and molecular characterization of begomovirus and associated satellite DNA molecules infecting Cyamopsis tetragonoloba. Virus Genes 41, 118– 125. [doi:10.1007/s11262-010-0482-7] Kumar A, Snehi SK, Raj SK, Kumar J, Khan JA, 2011. Association of Cotton leaf curl Burewala virus and its satellite molecules with leaf distortion symptoms of cotton in India. New Disease Reports 24, 18. [doi:10.5197/j.2044-0588.2011.024.018] Citing Literature Volume25, Issue1January 2012-June 2012Pages 9-9 FiguresReferencesRelatedInformation

Diseased cotton plants showing typical leaf curl symptoms were collected from experimental plot of Agriculture Research Station-Sriganganagar, Rajasthan. Complete DNA-A component from samples taken from two areas were amplified through rolling circle amplification (RCA) using templiphi kit (GE Healthcare) and characterized. DNA-A of one isolate consists of 2751 nucleotides and second isolate of 2759 nucleotide. Both sequences comprised six ORF's. Genome organization of DNA-A of one isolate shows high sequence similarity with other characterized local begomovirus isolates of Rajasthan, while other isolate shows high sequence similarity with CLCuV reported from Pakistan. The maximum similarity of first isolate, CLCuV-SG01, shows highest sequence identity with Cotton leaf curl Abohar (Rajasthan) virus, and second isolate, CLCuV-SG02, shows highest sequence identity with cotton leaf curl virus from Pakistan. Both isolates showed 85% similarities with each other. The sequence data revealed probable infiltration of some strains of Cotton leaf curl virus from Pakistan to India, or co-existence of different isolates under similar geographical conditions. While CLCuV-SG01 shows highest nt sequence similarity with CLCuV Rajasthan (Abohar), nt identity of V1 ORF (encoding coat protein) of SG01 shows the highest nt identity (100%) with CLCuV Multan (Bhatinda) and Abohar virus while AC1 region also showed difference. Complete nucleotide sequence of SG01 shows only 86% similarity with CLCuV Multan virus. Similarity search revealed significant difference in AV1 and AC1 regions with respect to DNA-A suggesting an evolutionary history of recombination. Computer based analysis, recombination detection Program (RDP) supports the recombination hypothesis, indicated that recombination with other begomoviruses had taken place within V1 ORF and AC1 ORF of CLCuV-SG01 and AC1 ORF of CLCuV-SG02 and also in noncoding intergenic region (IR).

Leaves of sunn hemp (Crotalaria juncea) showing geminiviral symptoms were collected from Lucknow, India during rainy season in 2008. DNA template isolated from the symptomatic leaf tissues were subjected to polymerase chain reaction (PCR) using specific primers to amplify coat protein (CP) gene of DNA-A as well as betasatellite DNA associated with the leaf curl disease. CP gene showed 97% sequence identity with that of Cotton leaf curl Burewala virus (CLCuBwV). Further, the betasatellite DNA molecule revealed sequence similarity with previously characterized betasatellite DNA of begomoviruses affecting malvaceous crops from different regions of India and Pakistan. Maximum similarity (>90%) of betasatellite DNA under study was observed with Cotton leaf curl Multan betasatellite (CLCuMB-[Pak: Mul17:08) and other betasatellite DNA from Pakistan thus confirming possible infection of C. juncea with begomovirus. A complementary sense open reading frame (ORF) βC1 is present at nucleotide position 194–550. Sequence comparison of this ORF with other members of begomoviruses further confirmed association of a begomovirus with C. juncea. The betasatellite DNA when expressed under the control of CaMV35S promoter Nicotiana tabacum, showed leaf deformities. Our results demonstrated that a malvaceous betasatellite is adapted by a nonmalvaceous host and causes similar disease symptoms.

Leaves of sunn hemp () showing geminiviral symptoms were collected from Lucknow, India during rainy season in 2008. DNA template isolated from the symptomatic leaf tissues were subjected to polymerase chain reaction (PCR) using specific primers to amplify coat protein (CP) gene of DNA-A as well as betasatellite DNA associated with the leaf curl disease. CP gene showed 97% sequence identity with that of (CLCuBwV). Further, the betasatellite DNA molecule revealed sequence similarity with previously characterized betasatellite DNA of begomoviruses affecting malvaceous crops from different regions of India and Pakistan. Maximum similarity (>90%) of betasatellite DNA under study was observed with Cotton leaf curl Multan betasatellite (CLCuMB-[Pak: Mul17:08) and other betasatellite DNA from Pakistan thus confirming possible infection of with begomovirus. A complementary sense open reading frame (ORF) βC1 is present at nucleotide position 194–550. Sequence comparison of this ORF with other members of begomoviruses further confirmed association of a begomovirus with . The betasatellite DNA when expressed under the control of CaMV35S promoter , showed leaf deformities. Our results demonstrated that a malvaceous betasatellite is adapted by a nonmalvaceous host and causes similar disease symptoms.

Monopartite begomoviruses comprise DNA-A as the main genome and associated satellite DNAs. Viral DNA extracted from guar (Cyamopsis tetragonoloba) showing leaf curl symptoms exhibited positive amplification of coat protein (CP) gene of DNA-A component, suggesting the presence of begomovirus. Full length DNA-A was amplified by primer pair re-designed from CP gene nucleotide sequence. The associated alphasatellite and betasatellite DNA molecules were amplified and sequenced, confirming the presence of monopartite begomovirus. Sequence comparisons showed 89% identity with other begomoviruses. The Neighbor-Joining tree based on full length DNA-A nucleotide sequence showed that the guar infecting begomovirus clustered separately from other known begomoviruses. The betasatellite shared a high (96%) nucleotide identity to Cotton leaf curl Multan betasatellites. The alphasatellite showed 91% nucleotide identity to alphasatellite associated with begomovirus infecting Okra. Recombination analyses showed three recombinant fragments in DNA-A, two in betasatellite, and four in alphasatellite. The results suggest that the begomovirus identified in this study was a new recombinant virus. Its name was proposed as Cyamopsis tetragonoloba leaf curl virus (CyTLCuV).

Variances due to general and specific combining abilities were significant for most of the characters. The ratio of 62 sca/2 gca was greater than unity. Parents, PSPL, LC2-1 PBOG-61 were good general combiners for days to early female flowering; PBOG-40, Pusa Naveen and LC2-1 for early harvest; NDBG-56 and PBOG-40 for fruit length; LC2-1 for fruit girth; PBOG-40, PBOG-61 and PSPL for weight per fruit; ARBGH-7 for number of fruits; PSPL and NDBG-56 for vine length PBOG-40, ARBGH-7 and PBOG-61 for yield per vine and hectare. Cross ARBGH-7 x LC2-1 for fruit weight and days to first female flowering; ARBGH-7 x NDBG-56 for fruit number; NDBG-56 x Pusa Naveen for vine length and PSPL x LC2-1 for first harvest.