Cotton leaf curl disease (CLCuD) is a serious disease of cotton on the Indian subcontinent. In the present study, three cotton leaf curl viruses, cotton leaf curl Burewala virus (CLCuBuV), cotton leaf curl Kokhran virus (CLCuKoV) and cotton leaf curl Multan virus (CLCuMV), and their associated satellites, cotton leaf curl Multan betasatellite (CLCuMB) and cotton leaf curl Multan alphasatellite (CLCuMA), were detected. CLCuBuV with either intact (CLCuBuV-1) or mutant (CLCuBuV-2) transcriptional activator protein (TrAP) were detected in different plants. Agroinoculation with CLCuBuV-1 or CLCuBuV-2 together with CLCuMB and CLCuMA, resulted in typical leaf curling and stunting of tobacco plants. Inoculation with CLCuKoV or an isolate of CLCuMV (CLCuMV-2), together with CLCuMB and CLCuMA, induced severe leaf curling, while the other isolate of CLCuMV (CLCuMV-1), which was recombinant in origin, showed mild leaf curling in tobacco. To investigate the effect of intact or mutant TrAP and also the recombination events, CLCuBuV-1, CLCuBuV-2, CLCuMV-1 or CLCuMV-2 together with the satellites (CLCuMA and CLCuMB) were transferred to cotton via whitefly-mediated transmission. Cotton plants containing CLCuBuV-1, CLCuBuV-2 or CLCuMV-2 together with satellites showed curling and stunting, whereas the plants having CLCuMV-1 and the satellites showed only mild and indistinguishable symptoms. CLCuBuV-1 (intact TrAP) showed severe symptoms in comparison to CLCuBuV-2 (mutant TrAP). The present study reveals that two types of CLCuBuV, one with an intact TrAP and the other with a mutant TrAP, exist in natural infection of cotton in India. Additionally, CLCuMuV-1, which has a recombinant origin, induces mild symptoms in comparison to the other CLCuMV isolates.

Background: Blood is an important component of the human body that is responsible for complete human identity. The ABO and Rh groups are very useful for blood transfusion, organ transplantation, biomedical research, and anthropological study. The blood group incompatibility has been associated with some diseases. Objective: To examine the relationship between blood group and infertility in men and to know the frequency of different blood groups among infertile men who were referred from different infertility clinics and centers. Materials and Methods: A total number of 88 infertile men along with 88 fertile men as controls were evaluated using the antigen–antibody agglutination test. Result: The ABO blood group distribution among the infertile men was 40.90%, 29.54%, 19.31%, and 10.22% for blood groups O, A, B, and AB, respectively, and the prevalence of ABO blood group in fertile were 44.31%, 27.27%, 22.72%, and 05.68% for groups O, A, B, and AB, respectively. Conclusion: This preliminary study showed that the ABO blood group has no statistically significant association with male infertility.

A begomovirus and its associated alpha- and betasatellite were detected in tomato plants affected with leaf curl disease. Based on a nucleotide sequence identity of 99 %, this begomovirus was designated an isolate of cotton leaf curl Burewala virus (CLCuBuV). The alphasatellite exhibited 93 % sequence identity to cotton leaf curl Burewala alphasatellite (CLCuBuA) and is hence referred to here as a variant of CLCuBuA. The detected betasatellite was recombinant in nature and showed 70 % sequence identity to the known betasatellites. Inoculation of healthy tomato with CLCuBuV plus betasatellite, either in the presence or the absence of alphasatellite, led to typical leaf curling, while inoculation with CLCuBuV in the absence of betasatellite resulted in mild symptoms. This confirmed the role of the betasatellite in expression of disease symptoms. We propose to name the newly detected betasatellite tomato leaf curl Hajipur betasatellite (ToLCHJB).

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