Forty-two years impact of chemical fertilization on soil phosphorus partition and distribution under rice-based cropping systems.

Understanding of soil phosphorus (P) transformation is crucial to minimize its edge-of-field loss associated with ecosystem disservices. A sequential chemical extraction procedure was used to assess the impact (42 years) of organic and chemical fertilizations on soil P partition and distribution under subtropical rice based cropping systems. Experimental treatments were control, N, NP, NK, NS, NZn, NPK, NSZn, NPKSZn, and N+FYM (farmyard manure). Composite soils were collected from 0-5, 20-25 and 40-45 cm depths, extracted, and analyzed for soluble P, NaHCO3-P (inorganic and organic), NaOH-P (inorganic and organic), acid soluble (H2SO4), and residual P fractions. The NPKSZn significantly increased the concentration of soil inorganic P compared to other treatments. When FYM was applied together with N fertilizer, the organic P concentration increased, which was statistically identical to NPKSZn and NPK treatments. While the labile (NaHCO3-Pi, NaOH-Po), residual, and total P concentrations were stratified at 0-5 cm depth, the concentration of NaHCO3-Po, NaOH-Pi, and acidic P fractions increased with soil depth. The soluble, NaHCO3 (Pi and Po), NaOH-Pi and NaOH-Po, acidic, and residual P fractions constituted about 0.4, 6.6, 1.7, 21.3, 37.7, and 8.3%, respectively, of the total P. A higher concentration of the labile P at the surface soil indicated that the impact of chemical fertilization stratified the available P for plant uptake or susceptible to edge-of-field loss. The NPKSZn and N+FYM both had higher NaHCO3-Po and NaOH-Po concentrations within 40-45 cm and 0-25 cm depths, suggesting that N+FYM could promote the transformation of non-labile P into labile P pool, by reducing P fixation by soil and transport them at 20-45 cm depth. It is concluded that long-term fertilization increased the concentration of P pools especially labile P by saturating the soil adsorption sites especially in surface soil.

First Report of 'Candidatus Phytoplasma solani' (16SrXII-A) Associated with Fox Nut (Euryale ferox Salisb.) Stolbur Disease in India.

Fox nut also known as Gorgon nut, Makhana (Euryale ferox Salisb.) is a high value aquatic crop belonging to the family Nymphaeaceae. In India, it is generally grown in flood prone areas of North Bihar, lower Assam, parts of West Bengal, Odisha, Manipur and Tripura (Jana et al., 2018). India contributes nearly 70-80% of the global fox nut production. During September 2021, a phytoplasma like symptom was noticed on fox nut leaves at Basudeopur Farm of Research Centre for Makhana, Darbhanga, Bihar, India (23° 9' N and 65° 53' E). The characteristic symptom was that some portion of leaf lamina deformed along the veins with wrinkled and raised overgrowth or hypertrophy. The veins were thickened and reddened in the infected leaf area. The infection occurs in petiole as well as in flower stalk. The disease incidence was found as high as 30% which caused severe yield loss which was calculated to be 40% in that particular field. Total of 20 sampled fox nut plants, 10 symptomatic and 10 asymptomatic ones, were collected and tested for the presence of phytoplasma. A nested PCR assay using the phytoplasma universal 16S rRNA primer pairs: P1/P7 followed by R16F2n/R16R2 (J. Jovic et al. 2011) amplified the expected ~1.2-kb 16S rDNA fragment in all 10 symptomatic samples. No amplification was detected from asymptomatic samples. One of the ten 1.2-kb nested 16S rDNA PCR products was gel purified, cloned into the pGEM-T-easy plasmid vector (Promega, Madison, WI), and sequenced and was deposited in NCBI under the Accession no.OL873590. BLAST analysis showed that the sequence of the PCR 16S rDNA product was 100% identical to several GenBank sequences of Ca. P. solani (16SrXII Stolbur group) viz. KF907506. Furthermore, analysis by iPhyClassifier software showed that the virtual restriction fragment length polymorphism (RFLP) pattern of the sequenced PCR 16S rDNA product is identical (similarity coefficient 1.00) to the reference pattern of the 16SrXII-A subgroup. Identification of 'Ca. P. solani' was conducted following the STOL11 stolbur-specific protocol (Radonjic et al. 2009). Sequencing of tuf gene (Elongation factor Tu) was performed by using tuf marker genes (Cvrkovic et al. 2014) from 10 symptomatic and 10 asymptomatic samples. Sequence of the amplified gene (896 bp) was deposited in GenBank under Accession number OM174272. The presence of 'Ca. P. solani' was detected in all symptomatic samples, while all control plants tested negative. The RFLP analysis of tuf gene nested PCR products using HpaII endonuclease (Fermentas) revealed uniform tuf-b type in all positive samples. Nucleotide blast analyses showed that the tuf gene was 100% identical to STOL11 strain of C. P. solani subgroup 16SrXII-A (Accession No JQ797670). For developing a suitable management strategy, identification of the vector is essential. Leaf hoppers visiting the infected plants as well as nearby crop fields will be tested for presence of the phytoplasma. To the best of our knowledge, this is the first report of Candidatus Phytoplasma solani' (16SrXII-A) infecting Fox nut (Euryale ferox Salisb.) in India. References Cvrkovic et al. 2014. Plant Pathol. 63:42. https://doi.org/10.1111/ppa.12080 Jana, B. R., et al. 2018. Int. J. Curr. Microbiol. App. Sci. 7(12): 578-587. https://doi.org/10.20546/ijcmas.2018.712.072 Jovic, J. et al. 2011. B. Insectol. 64:S83. ISI Radonjic, S. et al. 2009. J. Phytopathol. 157:682. https://doi.org/10.1111/j.1439- 0434.2009.01560.

Work Ability among Nurses in a Public Medical College Hospital.

This cross sectional study was carried out 197 nurses of Shaheed Suhrawardy Medical College Hospital (ShSMCH), Dhaka, Bangladesh from 1st July 2015 to 30th June 2016. Purpose of the study was to assess work ability and its association with sociodemographic characteristics & work related variables which affect work ability among nurses in a Public Medical College Hospital. Sampling method was simple random sampling. By face to face interview data was collected with the help of semi-structured questionnaire and all data were analyzed with SPSS software version 21. For descriptive statistics means, SD and range were calculated as required. Data were presented in frequency table, pie diagram. Statistical test chi-square was used and p<0.05 was consider to be statistically significant. It was found that 136(69%) respondents were between the age of 21 to 39 years and mean age was 35.95 Years, 187(94.9%) were female, 156(79.2%) were Muslim, 166(84.3%) had Diploma, 180(91.4%) family income were equal to and more than 40,000 Tk. 189(95.5%) had done clinical work; 49(24.9%) worked at medicine. Majority 183(92.9%) respondents had good to excellent work ability and 14(7.1%) had less good work ability. In this study significant association (p<0.05) was found between work ability and gender, educational status and type of job. This research provides an initial step in understanding the work ability of nurses in a Public Medical College Hospital. The present study showed that nurses' work ability is at the good to excellent level. This study also reveals significant association between work ability and gender, educational status and type of job.

Identification of genetic markers of resistance to echinocandins, azoles and 5-fluorocytosine in Candida glabrata by next-generation sequencing: a feasibility study.

OBJECTIVES: Multi-antifungal drug resistance in Candida glabrata is increasing. We examined the feasibility of next-generation sequencing (NGS) to investigate the presence of antifungal drug resistance markers in C. glabrata. METHODS: The antifungal susceptibility of 12 clinical isolates and one ATCC strain of C. glabrata was determined using the Sensititre YeastOne® YO10 assay. These included three isolate pairs where the second isolate of each pair had developed a rise in drug MICs. Single nucleotide polymorphisms (SNPs) in genes known to be linked to echinocandin, azole and 5-fluorocytosine resistance were analysed in all isolates through NGS. RESULTS: High-quality non-synonymous SNPs in antifungal resistance genes such as FKS1, FKS2, CgCDR1, CgPDR1 and FCY2 were identified. For two of three isolate pairs, there was a >60-fold rise in MICs to all echinocandins in the second isolate from each pair; one echinocandin-resistant isolate harboured a mutation in FKS1 (S629P) and the other in FKS2 (S663P). Of the third pair, both the 5-fluorocytosine-susceptible, and resistant isolates had a mutation in FCY2 (A237T). SNPs in CgPDR1 were found in pan-azole-resistant isolates. SNPs in other genes linked to azole resistance (CgCDR1, ERG9 and CgFLR1) were present in both azole-susceptible and azole-resistant isolates. SNPs were also identified in Candida adhesin genes EPA1, EPA6, PWP2 and PWP5 but their presence was not associated with higher drug MICs. CONCLUSIONS: Genome-wide analysis of antifungal resistance markers was feasible and simultaneously revealed mutation patterns of genes implicated in resistance to different antifungal drug classes.

A method of direct PCR without DNA extraction for rapid detection of begomoviruses infecting jute and mesta.

UNLABELLED: We have developed a simple method of direct PCR (dPCR) without time-consuming procedures of DNA extraction by directly using the leaf bits for rapid detection of begomoviruses in jute and mesta. The leaf bits were treated with a lysis buffer for 35 min, and the lysate was used as PCR template. Different components and their concentration in lysis buffer systems were optimized and the optimal buffer system composed of 20 mmol l(-1) tris (hydroxymethyl aminomethane (Tris)-Cl (pH 8·0), 1·5 mmol l(-1) ethylenediaminetetraacetic acid (EDTA) (pH 8·0), 1·4 mol l(-1) NaCl and 200 µg/mL Proteinase K. Further, 3% PVP (w/v) and ß-marcaptoethanol (1% v/v) were additionally added into the buffer in case of jute. Under optimized PCR conditions, both viral DNA as well as plant (jute and mesta) genomic DNA were amplified from the lysate. dPCR required fewer reagents and less incubation time reducing both time and cost of detection. SIGNIFICANCE AND IMPACT OF THE STUDY: Identification of begomoviruses by serology is not suitable due to difficulty in preparing high titre and specific antisera. Begomoviruses are routinely detected by PCR-based techniques using universal or specific primers. However, it is a prerequisite to isolate pure DNA from the samples before PCR. DNA extraction from some plants such as jute, mesta is very difficult due to the presence of mucilage and other impurities. Therefore, we have developed a method of direct PCR without DNA extraction for detection of begomoviruses from these crops. It is the first report of a direct PCR method in jute and mesta.

First Report of a 16Sr I-B Phytoplasma Associated with Phyllody and Stem Fasciation of Flax (Linum usitatissimum) in India.

Flax or linseed is grown as a fiber or oilseed crop in tropical and temperate regions. It is commercially cultivated in many countries of the world including Canada, China, India, the United States, Ethiopia, Pakistan, Russia, Poland, and Argentina (1). In December 2013, symptoms suggestive of phytoplasma infection were noticed on flax in different experimental fields of Central Research Institute for Jute and Allied Fibres (CRIJAF) research farm, Barrackpore, India, and the incidence was less than 2%. Because incidence of phytoplasma diseases are increasing worldwide, occurrence of a phytoplasma in a new geographical area poses an imminent threat. The infected plants showed floral virescence, phyllody, and stem fasciation (flattened stem). Floral malformation was very conspicuous with abnormal structures replacing normal flowers. All the floral parts, including petals, turned into green leaves. Total DNA was extracted from leaf mid veins of three symptomatic and three asymptomatic plants using a DNeasy Plant Mini Kit (Qiagen). PCR was carried out with the phytoplasma-specific universal P1/P7 primer set followed by nested primer pair R16F2n/R16R2 (2), resulting in DNA amplicons that were 1.8 kb and 1.2 kb, respectively, in all symptomatic samples tested. No amplification was observed with DNA from symptomless samples. This suggested association of a phytoplasma with the disease. The five purified nested PCR products were cloned in a pGEM-T Easy vector (Promega) and sequenced. One of the sequences that proved to be identical to the others was deposited in GenBank (Accession No. KJ417660). The consensus sequence was analyzed by NCBI BLAST and found to share 99% similarity with the 16Sr DNA sequence of the 'Candidatus Phytoplasma asteris' reference strain (GenBank HQ828108), which belongs to 16SrI group. The phylogenetic tree based on 16SrDNA sequence of phytoplasmas belonging to group 16SrI and other distinct phytoplasma groups also showed that the phytoplasma clustered with members of group 16SrI (3). The nested PCR product of R16F2n/R16R2 was digested using restriction enzymes AluI, BfaI, BstU, HhaI, HpaI, KpnI, MseI, and RsaI. The RFLP patterns were compared with those of known phytoplasma strains (2) and they matched the patterns for aster yellows subgroup B (16Sr I-B). Subsequently, the iPhyClassifier 16Sr group/subgroup classification based on similarity (4) analyses showed that the studied strain had 16SrDNA sequences in the 16SrI-B group with a similarity coefficient of 1.00. To the best of our knowledge, this is the first report of 16SrI-B phytoplasma associated with flax in India. References: (1) K. P. Akhtar et al. Phytoparasitica 41:383, 2013. (2) I. M. Lee et al. Int. J. Syst. Evol. Microbiol. 54:337, 2004. (3) N. Saitou and M. Nei. Mol. Biol. Evol. 4:406, 1987. (4) Y. Zhao et al. Int. J. Syst. Evol. Microbiol. 59:2582, 2009.

First Report of Potato leaf roll virus (PLRV) Naturally Occurring on Jute (Corchorus olitorius) in India.

Jute (Corchorus olitorius L.) is an important bast fiber crop that is mainly grown in the Southeast Asian countries like India, Bangladesh, Nepal, China, Indonesia, Thailand, Myanmar, and a few South American countries. In June 2013, symptoms suggestive of a viral disease were noticed on jute (cv. JRO524) in an experimental field of the CRIJAF research farm, Barrackpore, India, and the incidence of the disease was less than 2%. The infected plants showed stunted growth and short height. Mostly the upper leaves elongated with curling and coiling of lamina. Puckering and shoe string effect were also noticed. Petioles and stipules of the affected leaves were exceptionally longer. Although initially the incidence was low, it may spread to larger areas in subsequent years. Because the jute fiber is extracted from the stem, stunted growth and short height would badly affect the fiber yield and quality. Ten symptomatic and ten asymptomatic healthy looking samples were collected from the field. Corchorus golden mosaic begomovirus is common in jute; therefore, all the samples were tested by PCR using JMFL-AF/JMFL-AR, DNA-A component specific primer pair and JMFL-BF/JMFL-BR, DNA-B component specific primer pairs (1). However, there was no amplification. Because the aphid Aphis gossypii was often noticed in the jute field, all the samples were tested by double-antibody sandwich (DAS)-ELISA for common aphid transmitted viruses, e.g., Cucumber mosaic virus, Bean common mosaic virus, Cowpea mosaic virus, Papaya ring spot virus, Potato leaf roll virus (PLRV), Potato virus Y, and Watermelon mosaic virus using commercial diagnostic kits (Agdia). The symptomatic samples showed positive reaction only for PLRV. Five ELISA-positive samples and five asymptomatic healthy samples were used for RNA extraction. Total RNA was extracted by using QIAGEN RNeasy mini kit. RT-PCR was carried out with PLRV CP gene specific primer pair (3) which generated a cDNA amplicon of 627 bp in all ELISA-positive symptomatic samples. PLRV was not detected in symptomless samples. The five purified cDNA products were cloned in a pGEM-T Easy vector (Promega) and were sequenced. One of the five identical sequences was deposited in GenBank (Accession No. KF233880). The consensus sequence was analyzed by NCBI BLAST and found to share 99% similarity with the coat protein sequence of PLRV reference strain (S77421). Nucleotide span and ORF finder (NCBI) analysis indicated the 627-bp PCR amplicon coded part of a coat protein gene that had 100% identity with translated gene product (Protein ID AAB33483). PLRV is a small isometric RNA virus with worldwide distribution belonging to the family Luteoviridae whose natural host range is mainly restricted to solanaceous plants and few plants of other families (2,4). To the best of our knowledge, this is the first report of PLRV naturally occurring on jute (C. olitorius). References: (1) R. Ghosh et al. J. Virol. Methods 159:34, 2009. (2) S. Guyader and D. G. Ducray. J. Gen. Virol. 83:1799, 2002. (3) M. A. Mayo et al. J. Gen. Virol. 70:1037, 1989. (4) K. Mukherjee et al. Virus Genes 26:247, 2003.

First Report of a 16SrV-C Phytoplasma Causing Little Leaf and Bunchy Top of Tossa Jute (Corchorus olitorius) in India.

Jute is the most important phloem fiber crop of the world, and is mainly grown in the South East Asian countries of India, Bangladesh, Nepal, China, Indonesia, Thailand, and Myanmar, and few South American countries. The fiber is used in making sacks, ropes, bags, carpets, shoes, geo-textiles, and home decorations. There are two kinds of jute: tossa jute (Corchorus olitorius L.) and white jute (C. capsularis). In June 2012, symptoms suggestive of phytoplasma infection (little leaf and bunchy top) were noticed on tossa jute in different experimental fields of the CRIJAF research farm, Barrackpore, India, and the incidence of the disease varied from 5 to 20%. The infected plants showed profuse lateral branching with a bushy appearance. In many plants, branching at the apical portion developed a bunchy top symptom with tufts of smaller leaves. Leafy stem was also common in many plants with main stems covered with numerous little leaves. Total DNA was extracted from leaf midveins of 15 symptomatic and 5 asymptomatic plants by using an improved salt concentration and simple sodium acetate CTAB method (1). PCR was carried out with universal P1/P7 primer set followed by nested primer pair R16F2n/R16R2 (3), resulting in DNA amplicons that were 1.8 kb and 1.2 kb, respectively, in all symptomatic samples tested. Phytoplasma was not detected in symptomless samples. The five purified nested products were cloned in a pGEM-T Easy vector (Promega) and sequenced. One of the sequences that proved to be identical was deposited in GenBank (Accession No. KF501045). The consensus sequence was analyzed by NCBI BLAST and found to share 99% similarity with the 16Sr DNA sequence of the alder yellows phytoplasma reference strain (GenBank Accession No. AY028789), which belongs to the 16SrV group. The phylogenetic tree based on the 16SrDNA sequence of phytoplasmas belonging to group 16SrV and other distinct phytoplasma groups also showed that the phytoplasma clustered with members of subgroup 16SrV (4). Subsequently, in silico RFLP analysis of the nested PCR product with the pDRAW32 program using AluI and TruI restriction site used for 16SrV subgroups A, B, C, D, and E indicated that the 16SrV Corchorus strain belonged to subgroup C. RFLP patterns from all symptomatic C. olitorius samples were identical to the 16SrV-C pattern (2). The vector species transmitting the concerned phytoplasma in C. olitorius still needs to be identified. The leaf hopper, Amrasca biguttula biguttula, may be a potential vector as it is often noticed in jute fields. To the best of our knowledge, this is the first report of 16SrV-C phytoplasma associated with tossa jute (C. olitorius) in India. Initiative has to be taken to manage this disease; otherwise, branching of the main stems would badly affect the fiber quality as well as yield. References: (1) C. Biswas et al. Lett. Appl. Microbiol. 56:105, 2012. (2) B. Duduk et al. J. Phytopathology 152:575, 2004. (3) I. M. Lee et al. Int. J. Syst. Evol. Microbiol. 54:337, 2004. (4) N. Saitou and M. Nei. Mol. Biol. Evol. 4:406, 1987.

Interleukin-15 administration increases graft-versus-tumor activity in recipients of haploidentical hematopoietic SCT.

Utilizing a clinically relevant haploidentical (HI) murine transplant model, lethally irradiated B6D2F1 (H2K(b/d)) mice were transplanted with T cell-depleted (TCD) BM from B6CBAF1 (H2K(b/k)) mice. We found that administration of IL-15 significantly increases the numbers of CD8+ T and natural killer (NK) cells in spleen and BM after transplantion without GVHD. Graft-versus-tumor (GVT) potency of the graft was evaluated upon tumor challenge using P815 tumor cells (H2(d)). IL-15 administration without T-cell infusion did not result in any survival improvement. However, IL-15 in combination with very low-dose T-cell infusion (1 × 10(4)) significantly increased GVT activity and improved survival in recipients of HI hematopoietic SCT (HSCT). This effect was observed when IL-15 was given at a later time point, rather than immediately following transplantation. IL-15 administration also specifically increased slow-proliferative CD8+ T-cell proliferation and IFN-γ secretion in CD8+ T cells in recipients of CFSE (carboxyfluorescein succinimidyl ester)-labeled HI T-cell infusion, whereas there was no effect on CD4+ T-cell proliferation, suggesting the critical effect of IL-15 on CD8+ T-cell homeostasis in HI host. We conclude that IL-15 can be used for enhancing antileukemia effect of HI-HSCT, which requires presence of donor-derived T cells.

Haploidentical hematopoietic SCT increases graft-versus-tumor effect against renal cell carcinoma.

Allogeneic hematopoietic SCT (HSCT) has been shown to be an effective treatment option for advanced renal cell cancer (RCC). However, tumor resistance/relapse remains as the main post transplant issue. Therefore, enhancing graft-versus-tumor (GVT) activity without increasing GVHD is critical for improving the outcome of HSCT. We explored the GVT effect of haploidentical-SCT (haplo-SCT) against RCC in murine models. Lethally irradiated CB6F1 (H2K(b/d)) recipients were transplanted with T-cell-depleted BM cells from B6CBAF1 (H2K(b/k)) mice. Haplo-SCT combined with a low-dose haploidentical (HI) T-cell infusion (1 × 10(5)) successfully provided GVT activity without incurring GVHD. This effect elicited murine RCC growth control and consequently displayed a comparative survival advantage of haplo-SCT recipients when compared with MHC-matched (B6D2F1CB6F1) and parent-F1 (B6CB6F1) transplant recipients. Recipients of haplo-SCT had an increase in donor-derived splenic T-cell numbers, T-cell proliferation and IFN-γ-secreting donor-derived T-cells, a critical aspect for anti-tumor activity. The splenocytes from B6CBAF1 mice had a higher cytotoxicity against RENCA cells than the splenocytes from B6 and B6D2F1 donors after tumor challenge. These findings suggest that haplo-SCT might be an innovative immunotherapeutic platform for solid tumors, particularly for renal cell carcinoma.

A multiplex nested PCR assay for simultaneous detection of Corchorus golden mosaic virus and a phytoplasma in white jute (Corchorus capsularis L.).

A multiplex nested PCR assay was developed by optimizing reaction components and reaction cycling parameters for simultaneous detection of Corchorus golden mosaic virus (CoGMV) and a phytoplasma (Group 16Sr V-C) causing little leaf and bunchy top in white jute (Corchorus capsularis). Three sets of specific primers viz. a CoGMV specific (DNA-A region) primer, a 16S rDNA universal primer pair P1/P7 and nested primer pair R16F2n/R2 for phytoplasmas were used. The concentrations of the PCR components such as primers, MgCl2 , Taq DNA polymerase, dNTPs and PCR conditions including annealing temperature and amplification cycles were examined and optimized. Expected fragments of 1 kb (CoGMV), 674 bp (phytoplasma) and 370 bp (nested R16F2n/R2) were successfully amplified by this multiplex nested PCR system ensuring simultaneous, sensitive and specific detection of the phytoplasma and the virus. The multiplex nested PCR provides a sensitive, rapid and low-cost method for simultaneous detection of jute little leaf phytoplasma and CoGMV. Based on BLASTn analyses, the phytoplasma was found to belong to the Group 16Sr V-C.

A simple method of DNA isolation from jute (Corchorus olitorius) seed suitable for PCR-based detection of the pathogen Macrophomina phaseolina (Tassi) Goid.

A simple method was developed for isolating DNA from jute seed, which contains high amounts of mucilage and secondary metabolites, and a PCR protocol was standardized for detecting the seedborne pathogen Macrophomina phaseolina. The cetyl trimethyl ammonium bromide method was modified with increased salt concentration and a simple sodium acetate treatment to extract genomic as well as fungal DNA directly from infected jute seed. The Miniprep was evaluated along with five other methods of DNA isolation in terms of yield and quality of DNA and number of PCR positive samples. The Miniprep consistently recovered high amounts of DNA with good spectral qualities at A260/A280. The DNA isolated from jute seed was found suitable for PCR amplification. Macrophomina phaseolina could be detected by PCR from artificially inoculated as well as naturally infected jute seeds. The limit of PCR-based detection of M. phaseolina in jute seed was determined to be 0·62 × 10(-7) CFU g(-1) seed.

First Report of Bacterial Leaf Spot Caused by Xanthomonas campestris pv. olitorii on Jute Grown for Seed in India.

Jute (Corchorus olitorius L.) is the second most important fiber crop after cotton in terms of global production (3). In November 2011, symptoms suggestive of bacterial infection were observed on a seed crop of jute at the CRIJAF research farm, Barrackpore, West Bengal, India. The disease appeared as small, brown, circular spots, usually less than 5 mm in diameter on the leaves and some of the spots were surrounded by a yellow halo. The lesions on the stems were elongated and in some cases were found to girdle the stem. In the later stages of disease, brown sunken spots were found on the green capsules. Disease incidence varied from about 20% to 90% of the total plants in different affected fields at the CRIJAF research farm. Bacterial leaf spot of jute with similar symptoms was reported in 1957 from Sudan (4). Five symptomatic and three asymptomatic leaf samples were collected from different jute fields. Bacterial colonies isolated on nutrient agar medium from infected young leaves were Xanthomonas-like and pale yellow cream in color. Total DNA was extracted from symptomatic as well as asymptomatic leaf samples by using an improved salt concentration and simple sodium acetate CTAB method (2). Single bacterial colonies were transferred to nutrient agar (NA) medium plates and incubated at 28°C for 48 h. Pure colonies from plates were used directly for DNA extraction using the QIAGEN DNeasy Blood and Tissue kit. PCR was carried out with Xanthomonas campestris specific primers NZ8F3/NZ85R3 (1), which generated an amplicon of 530 bp from all the symptomatic leaf samples as well as pure cultures of the isolated bacteria. No amplification was obtained from asymptomatic leaves. The amplicons from the five symptomatic samples collected from the field were sequenced and showed 100% identity with one another, and one sequence (strain JB-CO-13) was deposited in GenBank (Accession No. KC342185). The BLASTn analysis revealed that bacterial strain JB-CO-13 had 100% identity with X. campestris pv. olitorii (EU285213). Nucleotide span and ORF finder (NCBI) analysis indicated the 530-bp PCR amplicon coded part of a gyrase B gene that had 100% identity with a translated gene product (Protein ID: ABX84334). Three leaves of five 1-month-old jute plants (cv. JRO 204) in pot culture were infiltrated each with a separate bacterial strain using suspensions (1 × 105 CFU/ml) in distilled water. The negative control consisted of leaves infiltrated with sterile distilled water. The plants were kept in a greenhouse with mean maximum and minimum temperatures of 28.96 and 21.8°C, respectively. The plants were covered with plastic bags to maintain high relative humidity (>80%). Typical bacterial lesions were recorded on all the inoculated plants after 1 week. No lesions were seen on the negative control. To the best of our knowledge, this is the first report of bacterial leaf spot on C. olitorius caused by X. campestris pv. olitorii from India. References: (1) J. Adriko et al. Plant Pathol. 61:489, 2012. (2) C. Biswas, et al. Lett. Appl. Microbiol. 56:105, 2013. (3) Food and Agriculture Organization of the United Nations. Agricultural Commodities: Profiles and Relevant WTO Negotiating Issues. Online: http://www.fao.org/docrep/006/Y4343E/y4343e03.htm , 2003. (4) K. A. Sabet. Ann. Appl. Biol. 45:516, 1957.

Electronic and magnetic properties of quasifreestanding graphene on Ni.

For the purpose of recovering the intriguing electronic properties of freestanding graphene at a solid surface, graphene self-organized on a Au monolayer on Ni(111) is prepared and characterized by scanning tunneling microscopy. Angle-resolved photoemission reveals a gapless linear pi-band dispersion near K[over] as a fingerprint of strictly monolayer graphene and a Dirac crossing energy equal to the Fermi energy (EF) within 25 meV meaning charge neutrality. Spin resolution shows a Rashba effect on the pi states with a large (approximately 13 meV) spin-orbit splitting up to EF which is independent of k.

Argon nanobubbles in Al(111): a photoemission study.

Two fundamental manifestations of Al conduction electron response to Ar atom core hole in the final state of photoemission have been studied in implanted Ar bubbles in Al(111). Ar 2p binding energy and the Doniach-Sunjic asymmetry of the core-level line shape vary systematically as functions of Ar+ implantation energy and number of ions bombarded (fluence). The observations are explained by relating the strength of Al conduction electron screening to the size of the Ar nanobubbles.

Threshold retinopathy of prematurity: ocular changes and sequelae following cryotherapy.

PURPOSE: To determine ocular changes and sequelae following cryotherapy for threshold retinopathy of prematurity (ROP). METHODS: This is a retrospective study of 49 eyes of 26 premature babies with threshold ROP treated with cryotherapy between 1995 and 1998. All eyes included in the study had favourable structural outcome after cryotherapy. Follow-up examination of all babies was done 12 - 62 months (average 28 months) after cryotherapy. Visual axis, fixation pattern, anterior segment examination, cycloplegic refraction and dilated fundus examination with indirect ophthalmoscopy were undertaken in all eyes during follow-up. RESULTS: Posterior pole retinal residuae observed following cryotherapy were tortousity of blood vessels in 32 (65.3%), narrow temporal arcade in 22 (44.89%), temporal crescent in 17 (34.69%), disc drag in 13 (26.53%) and macular heterotopia in 7 (14.28%) eyes. Myopia was observed in 20 (40.82%) eyes and strabismus in 5 (19.23%) babies. The significant risk factor for ocular changes was ROP with more clock hours of involvement (p < 0.05). Higher period of gestation was associated with posterior pole changes (p< 0.05). CONCLUSIONS: All premature babies with threshold ROP treated with cryotherapy require frequent and long-term follow up to look for retinal residuae, refractive status, and ocular motility disorders.