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In India, nearly 29% of adults (over 42% of men and 14% of women) aged 15 or above use tobacco daily or occasionally.1 Out of 100 million smokers, nearly 40 million smoke cigarettes. ITC (British American Tobacco’s India affiliate) dominates the Indian-manufactured cigarette market with 77% market share,2 while Godfrey Phillips India (Phillip Morris International’s India affiliate) controls 13% of the market.3 In terms of volume, however, bidi (also beedi, a forest tree leaf hand-rolled indigenous cigarillo using sun-cured flaked tobacco) dominates the smoking landscape. In 2014, it is estimated that nearly 550 billion4 bidis were made by women and children in the poorest parts of India.5 In addition, nearly 200 million adults use smokeless tobacco (SLT).1 SLT is the most widely consumed tobacco product in the country that is homemade, largely unregulated, untaxed and sold locally. About 30% of the SLT market consists of commercially manufactured products, where Dharampal Satyapal Group (DS Group) enjoys a large market share.6 Historically, the Indian tobacco industry has spent large sums in aggressively marketing and advertising its products.7 With the coming into force of the Cigarettes and Other Tobacco Products Act, 2003 (COTPA), however, all forms of direct and indirect tobacco advertising were prohibited in India.8 In particular, section 5, clause 3, subclause (b) of COTPA prescribed that: > No person, shall, under a contract or otherwise promote or agree to promote the use or consumption of…(b) any trade mark or brand name of cigarettes or any other tobacco product in exchange for a sponsorship, gift, prize or scholarship given or agreed to be given by another person.8 Article 13 of the WHO Framework Convention on Tobacco Control (WHO FCTC)9 considers corporate social responsibility (CSR) activities to be a form of tobacco advertising …

Background Indian Premier League (IPL) cricket tournament - a shorter version of the sport which is played between assorted company-owned or individual owner teams - has grown immensely popular over the last decade. There is long history of tobacco advertisement, promotion and sponsorship (TAPS) in cricket. While direct and indirect TAPS is completely prohibited under the Cigarettes and Other Tobacco Products Act (COTPA), 2003 the nexus between tobacco industry and cricket in India continues. We monitored any tobacco product and allied brand or surrogate advertisement, promotion, sponsorship (TAPS) or placement during every match played in the IPL-13 season in the year 2020. Methods Using an investigative approach, 60 match highlights during IPL-13 (during September 19 to November 10, 2020) were monitored for TAPS (both on-site i.e. sportswear and/or in stadia) shown on the official website ( www.iplt20.com ). All sponsors and advertisers were validated using brand names or logos from their respective official website. Results Two IPL teams (Rajasthan Royals and Royal Challengers Bangalore) were sponsored by major tobacco industry owned brands. Violations by a major smokeless tobacco company promoting their brand extension product i.e. Vimal Elaichi, on the official telecast websites ( www.espncricinfo.com and www.cricbuzz.com ) and television channels ( www.Zeenews.india.com/Gujarati ) was observed. Tobacco companies i.e. Kamla Pasand and ITC Limited were also listed as associate sponsors for the 2020 championship. Conclusion Despite comprehensive ban under India's tobacco control law (COTPA, 2003), sponsorship by tobacco industry was found widespread in IPL-13. Advertisement and logos of tobacco companies are persistent during the telecast and promotion of the IPL events, in clear violation of India's tobacco control legislation and Article 13 of WHO's Framework Convention on Tobacco Control (WHO-FCTC) and its guidelines. Given that India and South Asia comprise the largest audience for televised cricket, strong exclusion criteria is needed by the international cricket authority and national cricket associations to ensure compliance with the national law and global good practice, making the cricketing world truly tobacco free.

A novel e-waste-degrading strain, PE08T, was isolated from contaminated soil collected from a paper mill yard in Lalkuan, Uttarakhand, India. Strain PE08T was Gram-stain-negative, rod-shaped, aerobic, oxidase-positive and catalase-positive. Optimum growth was observed at 30 °C (range, 5–40 °C), with 1–2 % NaCl (range, 0–3 %) and at pH 7 (range 6–11). The phylogeny based on 16S rRNA gene sequences delineated strain PE08T to the genus Pseudomonas and showed highest sequence similarity to Pseudomonas furukawaii KF707T (98.70 %), followed by Pseudomonas aeruginosa DSM 50071T (98.62 %) and Pseudomonas resinovorans DSM 21078T (97.93 %). The genome of strain PE08T was sequenced and had one scaffold of 6056953 bp, 99.84 % completeness and 182× coverage were obtained. The G+C content in the genome was 64.24 mol%. The DNA–DNA hybridization and average nucleotide identity values between strain PE08T and its closely related type strain, P. resinovorans DSM 21078T were below 34.8 % and 87.96 %, respectively. The phylogenetic analysis based on whole-genome sequence and concatenated GyrB and RpoB proteins revealed that strain PE08T forms a district clade in the family Pseudomonadaceae . The predominant fatty acids were summed feature 8 (C18 : 1ω7c and/or C18 :1 ω6c), summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c), C16 : 0 and C12 : 0. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. The phenotypic, chemotaxonomic and genetic analysis, including overall genome relatedness index values, indicated that strain PE08T represents a novel species of the genus Pseudomonas , for which the name Pseudomonas lalkuanensis sp. nov. is proposed. The type strain is PE08T (=MCC 3792=KCTC 72454=CCUG 73691).

We performed whole-genome sequencing of two phytoplasmas associated with sugarcane grassy shoot (SCGS) and Bermuda grass white leaf diseases. These are the first draft genomes of SCGS phytoplasma (strain SCGS) and ‘Candidatus Phytoplasma cynodontis’ (strain LW01) and may help to delineate these phytoplasmas at a finer taxonomic level.

Red kidney bean (RKB) is an important cash crop of the Western Indian Himlayan (WIH) regions. Despite of having a long cultivation history, very little has known about its rhizospheric microbial diversity, which is becoming important for employing sustainable agricultural plans. In this perspective, the bacterial diversity and community structure of Himalayan RKB rhizosphere were studied and compared across the altitudinal zones of WIH. Metagenomic analysis revealed that Proteobacteria (31%) was the major phylum with the dominance of the genera Sphingomonas in the Garhwal while Pseudomonas in the Kumaun regions of Western Indian Himalaya (WIH), followed by Bacteroidetes (20%), Fermicutes (9%), Acidobacteria (7%), Actinobacteria (6%), Chloroflexi (5%), Gemmatimonadetes (3%) and Planctomyces (2%). Furthermore, the Garhwal soil was found to harbor more bacterial diversity than the Kumaun with unique genera Conexibacter, Ornithobacterium, Lacibacter, Salinibacter, Fervidicola, Haliangium, Pirellula, Caloramator, Desulfitobacterium, Edaphobacter, Holophaga and Massilia. The principal component analysis revealed that the bacterial diversity was associated with the content of “total solids” as well as “volatile acids” present in the respective soils. Additionally, large number of unclassified clone sequences suggesting the need of culture dependent studies to trap “yet not cultivated” bacterial species. Conclusively, the present study provides an overview of RKB associated bacterial diversity which must be explored for sustainable hill agricultural plans.

Leucas aspera (Wild.) Linn. (Family: Lamiaceae) is a commonly found weed throughout India, known for its pharmacological properties. Its white flowers and leaves are used in many Ayurvedic formulations for the treatment of chronic rheumatism, psoriasis, snake bites and skin eruptions (Prajapathi et al., 2010). During a survey of commercial flower crop fields in May 2018, a few L. aspera plants, growing as unwanted weeds in the fields and surrounding agricultural wastelands with the symptoms of phyllody, virescence and little leaves were observed in Emmekoppalu (12.2106, 76.2511; n= 1/26 plants) and Beerihundi (12.1630, 76.3225; n= 2/59 plants) localities of Mysuru district, and Srirangapatna in Mandya district (12.2541, 76.411; 1/67 plants), Karnataka- India(Figure 1). 'n' denotes the symptomatic/ asymptomatic samples observed. The disease incidence in the surveyed localities ranged less than four per cent. The total genomic DNA was extracted from the leaf midrib tissues of three representative symptomatic and two asymptomatic samples using the CTAB method. The phytoplasma 16S rRNA gene was amplified in nested PCR assay by P1/P7 followed by R16F2n/R16R2 primers using Long Amplification (LA) Taq polymerase (Takara, Japan). Additionally, the PCR assays were performed for the amplification of phytoplasma secA gene using the primers SecAfor1/SecArev3 and SecAfor2/SecArev3 (Hodgetts et al., 2008). The DNA templates from all the symptomatic samples generated amplicons of approximately 1.25kb (16S rRNA gene) and 480 bp (secA gene) revealing the association of phytoplasma strains. No amplifications were observed for the asymptomatic L. aspera samples. The obtained 16S rRNA gene sequences (MN223676, MT807111 and MZ093053) showed 97.96, 98.37 and 98.18 % sequence identity, respectively; with the 'Candidatus Phytoplasma aurantifolia', strain 'WBDL (U15442) using EzBiocloud database. The NCBI-BLAST analysis revealed maximum identity to various Peanut witches' Broom (PWB) phytoplasma strains. The virtual RFLP tool, iPhyClassifier delineated the Leucas phyllody phytoplasma strains (MN223676, MT807111 and MZ093053) to group 16SrII (PWB, Peanut Witches' broom group) subgroup D with the similarity coefficient 1.0 (Zhao et al. 2009). The obtained secA gene sequences (MZ151944, MZ151945 and MZ151946) were 98.15 to 100 % similar to the strain sequences of PWB phytoplasma strains. Further, the clustering pattern in the phylogenetic trees (16S rRNA and secA genes) constructed using MEGA 7 confirmed that the Leucas phyllody phytoplasma sequences were closely related to PWB strains. To the best of our knowledge, this is the first report on the association of 16SrII-D subgroup phytoplasma with the phyllody disease of L. aspera. In India, many weeds and wild plants serve as alternative hosts of PWB phytoplasmas and aid in the emergence of related diseases in economically important crops (Thorat et al., 2016; Thorat et al., 2017). The close genetic association of phytoplasma strains found in L. aspera and many other crops indicates the presence of common insect vector(s) transmitting these phytoplasmas (Yadav et al. 2015). This report is an addition to the catalogue of the weed species harboring phytoplasma strains associated with economically important crop plants (Rao et al., 2017). The screening of phytoplasma strains in weeds, alternate hosts and known/ unknown insect vectors is therefore essential to develop management strategies and effective management of phytophagous insect vectors feeding on both weeds and crop plants.

Two aerobic, Gram-stain variable, catalase and oxidase-positive, spore-forming, motile rods of strain UniB2T and UniB3T, were isolated from digestive syrup containing fungal diastase (10 mg/ml), pepsin (2 mg/ml) and sugar base containing polyethene glycol. Based on 16S rRNA gene sequence analysis, strain UniB2T has the highest sequence similarity with Paenibacillus humicus NBRC 102415T (98.30 %) and strain UniB3T showed the highest sequence similarity with Niallia circulans DSM 11T (98.95 %). The DNA G+C content of UniB2T was 63.74 mol %. The dDDH and ANI values between the strain UniB2T and its phylogenetically close relative were T were C16:00 (13.9 %), C15:0 anteiso (39.7 %), C17:0 anteiso (15.5 %). The DNA G+C content of UniB3T was 35.6 mol%. The dDDH and ANI values between the strain UniB3T and its close relatives were T were C16:0 (13.54 %), C15:0 anteiso (40.09 %) and C17:0 anteiso (16.03 %). Major polar lipids for both strains were Diphosphotidylglycerol and Phosphatidylethanolamine. Additionally, both the strains showed unique carbon utilization and assimilation pattern that differentiated them from their phylogenetically related neighbours. These phenotypic, genotypic and chemotaxonomic characters indicated the strains UniB2T and UniB3T represent two novel species for which the names Paenibacillus albicerus sp. nov. (Type strain UniB2T =MCC 3997T =KCTC 43095T =JCM34513T) and Niallia alba sp. nov. (Type strain UniB3T = MCC 3998T =KCTC 43235T =JCM 34492T) are proposed.

ObjectiveAreca nut is one of the most widely consumed substances globally, after nicotine, ethanol and caffeine and classified as carcinogenic to humans. This study examines the disparity and determinants of areca nut consumption with and without tobacco in India.DesignNationally representative cross-sectional study.ParticipantsWe used the nationally representative Global Adult Tobacco Survey 2016–2017. The analytical sample size was 74 037 individual’s aged 15 years and above with a response rate of 92.9%.MeasuresCurrent consumption of areca nut without tobacco and with tobacco.MethodWe examined determinants of areca nut consumption (without tobacco and with tobacco) using multinomial logistic regression, accounting for the survey design.ResultsAbout 23.9% (95% CI 23.1 to 24.8) of the adult population consume areca nut, that is, approximately 223.79 million people in India; majority of users (14.2%–95% CI 13.5 to 14.9) consumed areca nut with tobacco. When compared with women, men were more likely to consume areca nut (with tobacco relative risk (RR)=2.02; 95% CI 1.85 to 2.21 and without tobacco RR=1.13; 95% CI 1.07 to 1.20). Age, marital status, education, occupation, caste, religion and region were significantly associated with areca nut consumption. However, the direction and magnitude of association differ with respect to the areca nut consumption with and without tobacco.ConclusionThe ongoing tobacco control efforts would not address the majority of areca nut users until greater attention to areca nut consumption with and without tobacco is reflected in health policies in India.

Background: Pigeonpea [Cajanus cajan (L.) Millsp.] is an important legume crop from the Family Fabaceae. Pigeonpea commonly known as ‘Arhar’ or ‘Tim’ is mainly consumed in the form of split pulse as ‘dal’. Pigeonpea is cultivated as an annual or semi-perennial crop, usually in mixed cropping systems. Pigeonpea is attacked by insect-pests right from sowing to harvesting and also during the storage. Gram pod borer, H. armigera has been a major pest in most parts of the country. Therefore, keeping these views in mind, the present study was conducted.Methods: Present investigations were carried out during Kharif season of 2013 and 2014 to evaluate some insecticides and biopesticides against Helicoverpa armigera in various modules on short duration pigeonpea.Result: The mean larval population of H. armigera after spray was minimum in M3 (1.15 larvae plant-1) followed by M2 (1.17 larvae plant-1) and maximum in module M5 (1.61 larvae plant-1) during Khaif 2013. During Khaif 2014 mean larval population of H. armigera after spray was minimum in M2 (1.11 larvae plant-1) followed by M3 (1.12 larvae plant-1) and maximum in module M5 (1.58 larvae plant-1). Hence these modules (M2 and M3) can be adopted by the farmers for effective and economic management of H. armigera.

Polyhouses are utilized in an incredible degree to cover present-day society’s fresh food needs, and the assortment, quality, and quantity of items are critical variable factors responsible for the popularization of polyhouses. A modern set of polyhouse provides ideal conditions for plant development concerning the external climate conditions. For this reason, heating measurement gadgets like a thermometer, humidity, and CO2 controllers are utilized. A polyhouse of 460 m3 volume installed for experimental purposes in DEI, Agra. The heating load during December 2019 is determined to develop the ideal condition within the polyhouse for the optimum growth of the plant. The heating load is of fluctuating nature during the observed period that develops the need to load fluctuating the control devices for smother operation the system. Warming load is analyzed on an hourly base for one month, corresponding to sun based radiation, encompassing air temperature and humidity to have a, however, much as could be possible closer to reality information on load fluctuation. In that manner, amassed results are keeping away from, which on account of the day by day, count base would conceal the concurrence of warming and cooling needs around the same time.

Sugarcane Grassy Shoot (SCGS) disease is known to be related to Rice Yellow Dwarf (RYD) phytoplasmas (16SrXI-B group) which are found predominantly in sugarcane growing areas of the Indian subcontinent and South-East Asia. The 16S rRNA gene sequences of SCGS phytoplasma strains belonging to the 16SrXI-B group share 98.07 % similarity with ‘Ca. Phytoplasma cynodontis’ strain BGWL-C1 followed by 97.65 % similarity with ‘Ca. P. oryzae’ strain RYD-J. Being placed distinctly away from both the phylogenetically related species, the taxonomic identity of SCGS phytoplasma is unclear and confusing. We attempted to resolve the phylogenetic positions of SCGS phytoplasma based on the phylogenetic analysis of 16S rRNA gene (>1500 bp), nine housekeeping genes (>3500 aa), core genome phylogeny (>10 000 aa) and OGRI values. The draft genome sequences of SCGS phytoplasma (strain SCGS) and Bermuda Grass White leaf (BGWL) phytoplasma (strain LW01), closely related to ‘Ca. P. cynodontis’, were obtained. The SCGS genome was comprised of 29 scaffolds corresponding to 505 173 bp while LW01 assembly contained 21 scaffolds corresponding to 483 935 bp with the fold coverages over 330× and completeness over 90 % for both the genomes. The G+C content of SCGS was 19.86 % while that of LW01 was 20.46 %. The orthoANI values for the strain SCGS against strains LW01 was 79.42 %, and dDDH values were 22. Overall analysis reveals that SCGS phytoplasma forms a distant clade in RYD group of phytoplasmas. Based on phylogenetic analyses and OGRI values obtained from the genome sequences, a novel taxon ‘Candidatus Phytoplasma sacchari’ is proposed.

The management of electronic waste (e-waste) becomes a global issue in this digital era. Existing conventional practices are harmful for dealing with the e-waste, therefore, indigenous soil bacteria were explored for e-waste treatment through enrichment culture approach followed by screening, identification and their bioformulation used for in situ investigation. Soil bacteria were enriched in the presence of e-waste after 30 days of incubation under standard laboratory conditions. This finding was established through λmax values that were higher in the case of soil enrichment than the control. The biodegradation of e-waste by the selected strains during enrichment and in situ experiment was confirmed by FTIR, TG-DTG-DTA and SEM analysis. The FTIR spectra evidently support that microbial communities present in the enriched soil has affected the C-chain and used as carbon source for their growth. This chemical structural degradation of e-waste was further substantiated by thermal and SEM analysis. Thermograms experimentally show that the decomposition of the treated samples achieved comparatively at very low temperature than the control sample, while SEM micrographs revealed the surface morphology with distinct disintegrations. These result authenticated the biodegradation process carried out by the soil bacteria. Furthermore, bacterial community analysis confirmed that the used strains were persisting in the experimental pits throughout the trial period. Thus, this study besides providing direct and standardized protocol for screening and selection of efficient e-waste utilizing bacteria is also demonstrating potential consortia which are ready to be used.

Present investigations were carried out during Kharif season of 2013-14 and 2014-15 at the Agricultural Research Farm, Banaras Hindu University, Varanasi to monitor the population of Helicoverpa armigera (Hubner) by using pheromone traps and to start appropriate control measures in time. The maximum numbers of moth catches were recorded during 44th (16.8 pheromone trap-1) and 45th (19.5 pheromone trap-1) standard week in all the modules during Kharif 2013 and 2014. Relationship between pheromone trap catches and egg population was found significant and positively correlated in all modules except M3 during 2013. In the succeeding year 2014, the correlation between pheromone trap catches and egg population was positive and significant only in case of M5. The correlation between pheromone trap catches and larval population was positive in all modules but significant in case of M1 during 2013. In the succeeding year 2014, the correlation between pheromone trap catches and larval population was positive in all modules but significant only in M5.

Symptoms of excessive shoot proliferation were observed in the Njallani cultivar of small cardamom accompanied by stunting of stalks with fewer degenerated capsules at Nedumkandam Panchayat of Idukki district of Kerala in 2017. Five symptomatic Elettaria cardamomum shoot proliferation (ECSP) plant samples were collected and processed for DNA extraction and PCR assays utilizing universal phytoplasma 16S ribosomal-specific primers pair, P1/P7 followed by R16F2n/R16R2. Sequence comparison analysis of the R16F2n/R16R2 region of 16SrRNA gene showed 100% sequence identity with the ‘Candidatus Phytoplasma australasia’- related strain. Phylogeny and virtual RFLP analyses of 16S rRNA gene sequences confirmed the association of ‘Ca. P. australasia’ strain subgroup D with ECSP disease. The association of 16SrII group was further established and validated by amplifying phytoplasma-specific multilocus candidate genes by utilizing specific primers of secA, secY, SAP11, and tuf genes. The multilocus gene sequence comparison analysis again confirmed the association of ‘Ca. P. australasia’ with the ECSP phytoplasma isolate. This is the first report of phytoplasma association with small cardamom. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s13205-019-1944-9) contains supplementary material, which is available to authorized users.

Bacterial strains with poly-3-hydroxybutyrate (PHB) degradation potential were isolated from waste yard soil samples of selected industrial sites in Uttarakhand, India, and two microbial consortia were developed, i.e. Consortium I comprises Pseudomonas sp. strain Rb10, Pseudomonas sp. strain Rb11 and Bacillus sp. strain Rb18, and Consortium II is composed of Lysinibacillus sp. strain Rb1, Pseudomonas sp. strain Rb13 and Pseudomonas sp. strain Rb19. The current study involved enrichment selection via liquid and semi-solid media, followed by isolation and screening of bacterial strains using PHB pellets and films. Furthermore, the identification and characterisation was done by triphasic approach. The utilisation of PHB by the characterised strains was confirmed by Fourier transform infrared spectroscopy and scanning electron microscopy. Moreover, the minimum inhibitory concentration of solubilised PHB was found to be 2.5 mg/mL, which was detected through ‘clear zone assay’. Further, the selection a...

Weeds and wild plants as hosts of phytoplasmas play an important role in the epidemiology and emergence of phytoplasma diseases of economically important crops. In this chapter phytoplasmas detected in weeds and wild plants, their geographic origins, symptoms, identification, and their role in natural dissemination of phytoplasmas are described.

Phytoplasmas are associated with diseases in several hundred plant species, including many economically important industrial crops like sugarcane, sugar beet, cassava, and cotton. A number of phytoplasma diseases are associated with sugarcane. Originally restricted to Asian countries, they are spreading rapidly to newer locations with the help of infected seed material and leafhopper vectors. In cassava, the two phytoplasma diseases causing serious yield losses are cassava frog skin in Latin America and cassava witches’ broom in Asia. Because of unreliable and nonspecific symptoms, the identification and characterization of the phytoplasmas associated with these and other industrial crops at an early stage of plant growth are challenging. Here the progress made in understanding biology, economic importance, symptomatology, diagnosis, epidemiology, and control of phytoplasmas infecting sugarcane, sugar beet, cassava, and cotton crops are summarized.

Agriculture is an important livelihood activity in the Himalayan regions. Our previous studies revealed the presence of diverse diazotrophic assemblage in indigenous red kidney bean (RKB) rhizospheric soil from two different locations of Western Indian Himalaya, namely S1 (Chhiplakot, 30.70° N/80.30° E) and S2 (Munsyari, 30.60° N/80.20° E), selected on the basis of real-time PCR analysis. In this study, two 16S rRNA gene clone libraries (SB1 and SB2, respectively) were constructed using the same rhizospheric soil samples for assessing the total bacterial diversity and their community structure. A total of 760 clones were obtained, with ≈54–59% of these sequences belonging to the phylum Proteobacteria. While sequences belonging to Bacteroidetes, Chloroflexi, Acidobactria, Planctomycetes, Firmicutes, Nitrospira, Gemmatimonadetes, Cyanobacteria, Verrucomicrobia, OD1, OP11 and Actinobacteria were encountered in both the soils, sequences belonging to bacteria from the classes Chlorobi and BRC1 were only detected in the S1 soil. Both the libraries showed similar bacterial community compositions, with Pseudomonas (≈33–34%) as predominant genus. Phylogenetic analysis revealed that all the clone sequences were clustered in different bacterial groups as per their resemblance with their respective phylogenetic neighbours. Major clusters were formed by Gammapreoteobacteria followed by Bacteroidetes and Alphapro-teobacteria. A good fraction of the clone sequences has no resemblance with existing groups, thereby suggesting the need of culture-dependent studies from Himalayan regions. To the best of our knowledge, this study is the first major metagenomic effort on Himalayan RKBs rhizobacteria revealing fundamental information that needs to be explored for functional studies.

Symptoms of stunting (shortening of internodes), twisting and flat stem (the fasciation of a stem), discoloration of petals, deformed flowers, and witches' broom were recorded on an ornamental plant, plumed cockscomb (Celosia argentea L., fam: Amaranthaceae). The survey conducted at Indian Agricultural Research Institute (IARI) campus, New Delhi and Karnal region, Haryana, India, during September 2014 to March 2015 revealed disease incidence of 40 and 10%, respectively. The 16S rRNA gene sequence comparison and phylogenetic relationships of Celosia phytoplasma strains under study confirmed that they were associated with two different phytoplasma groups ('Candidatus Phytoplasma australasia' and 'Ca. P. asteris'). Virtual RFLP analysis of 16S rRNA gene sequences allowed further classification of the Celosia phytoplasma strains into the 16SrI-B and 16SrII-D subgroups. Notably, the detection of 'Ca. P. asteris' phytoplasma was reported in seeds of C. argentea by nested PCR assays; however, no evidence of phytoplasma presence was detected in seedlings raised from these seeds. This observation is the first record of the association of 16SrI-B and 16SrII-D subgroups of phytoplasmas with flat stem and witches' broom disease of C. argentea anywhere from the world.

Leaf yellows and declining symptoms were observed on garlic and onion plants at Pune and Nasik region of Maharashtra, and Gorakhpur region of Uttar Pradesh in March-April 2016. The disease incidence varied from 12% to 30% in different surveyed regions. After nucleic acid extraction and nested PCR analysis using phytoplasma specific primer pairs (P1/P7 followed by 3F/3R) of a 16S ribosomal gene an expected DNA fragments of 1.3 kb were amplified in two symptomatic onion samples (from Gorakhpur and Narayangaon) and one symptomatic garlic sample (from Pune). Pair wise sequence comparison of 16S rRNA gene of the onion leaf yellows phytoplasma (GKP strain) showed 100% sequence identity with a strain of phytoplasma associated with sugarcane grassy shoot disease (KP746932) and 99% sequence identity with phytoplasma strain associated with Jasminum sambac little leaf disease (KF728950) and Ca. P. oryzae (AB052873) belonging to rice yellow dwarf (16SrXI) group. However, another strain of onion leaf yellows phytoplasma (Narayangaon strain) and garlic leaf yellows and declining phytoplasma (Pune strain) showed 99% sequence identity with phytoplasma strains of grapevine yellows (KX011516), eggplant big bud (JX464672) and sesame phyllody (JX464672) belonging to Ca. P. phoenicium (16SrIX) group. Phylogenetic analysis of 16Sr RNA gene sequences of phytoplasma strains from onion and garlic in the present study confirmed their close relationships with phytoplasma strains of 16SrXI and 16SrIX groups. This is the first report of association of 16SrXI and -IX groups of phytoplasma causing leaf yellows and declining disease of onion and garlic in India.

Cashew nut (Anacardium occidentale L.), an evergreen perennial tree, is an economically important cash crop extensively used in many Indian desserts and cuisines. Cashew nuts are used for their nutritional value and are a major source of protein, fats, vitamins, oleic acid, and palmitic acid. Cashew nut cultivation is undertaken in 17 states of India covering an area of 1.04 Mha. India is the second-largest cashew nut producer in the world, producing about 817,000 MT of raw cashew nuts per annum (https://cashew.icar.gov.in). During a survey in June 2018 of cashew nut orchards in Nagicherra (latitude N 23° 48′ 37.6″; longitude E 91° 19′ 29.7″), West Tripura, India, cashew nut plants were found exhibiting deformed nuts and inflorescence decline (CaID) with prevalence of 5.6% (in a 1-ha area), which were suggestive of phytoplasma infection. Three symptomatic and asymptomatic cashew nut samples were collected, and DNA was extracted from tender shoots using CTAB. To detect the association of phytoplasma with symptomatic cashew nut samples, the isolated genomic DNA samples were assayed for amplification of 16S rRNA region using phytoplasma-specific primer pairs, namely, P1/P7 followed by nested PCR primers R16F2n/R16R2. The 16S rRNA gene of phytoplasma was detected in all collected symptomatic CaID samples, but no amplification was observed in asymptomatic samples. The obtained nested PCR fragments of ∼1.25 kb were sequenced directly (MK076296, MK729150) which showed 99.8% sequence identity with ‘Candidatus Phytoplasma australasia’, strain ‘Carica papaya’ (Y10097). The virtual restriction fragment length polymorphism analysis of 16S rRNA gene sequences delineated the CaID strain to 16SrII-D subgroup with a similarity coefficient of 1.0 using iPhyClassifier. The association of CaID phytoplasma strain with symptomatic cashew nut samples was further established by amplifying phytoplasma-specific partial secA gene (Hodgetts et al. 2008) and tuf gene (Al-Subhi et al. 2018). The phylogenetic analysis of the obtained sequences of 16S rRNA, secA (MK726365, MK726366), and tuf (MK726367, MK726368) genes confirmed the close association of CaID phytoplasma isolates with ‘Ca. P. australasia’ 16SrII-D subgroup member strains. To the best of our knowledge, this is the first report of the association of ‘Ca. P. australasia’ (16SrII-D) with inflorescence decline symptoms of cashew nut in the world. This is significant because phytoplasma strains of 16SrII-C and D have widespread host range including several fruit trees all across India (Rao et al. 2017). These strains are also known to be associated with diseases in weeds, acting as alternate hosts, spreading phytoplasmas through polyphagous insect vectors (Thorat et al. 2016). Likewise, the cashew plants are intercropped in coconut plantations and with plants like maize, groundnut, cowpea, millets, pineapple, and cassava, which are known to be susceptible to phytoplasma-related diseases, posing a threat to cashew nut plantations (Rao et al. 2017; Visalakshi et al. 2015).

Present study done during kharif 2013–14 and 2014–15 at the Agricultural Research Farm, Banaras Hindu University, Varanasi evaluates the egg and larval population of Helicoverpa armigera in pigeonpea, as influenced by selected IPM modules. The least egg population was observed with the module M2 (pheromone traps @ 20 + bird perches @ 20 + Bacillus thuringiensis subsp. kurstaki @ 1.0 kg + spinosad 45SC @ 73 g/ha. As regards larval population, the module M4 (pheromone traps @ 20 + bird perches @ 20 + indoxacarb 14.5 SC @ 60 g a.i. + imidacloprid 17.8 SL @ 20 g a.i. + indoxacarb 14.5 SC @ 60 g a.i./ha) was the one with least numbers. The relationship between egg and larval counts was positive and highly significant in all modules during both the years.

The forest of Southern India, especially in the states of Karnataka, Tamil Nadu and Kerala are recognised for natural populations of sandalwood contributing 90% of its distribution in India. The sandal spike disease (SSD) of sandalwood is known to be associated with the presence of aster yellows (‘Candidatus Phytoplasma asteris’, 16SrI-B). The recent survey and collection of the symptomatic sandalwood samples from the Marayoor sandalwood reserve of Kerala state confirmed the presence of this phytoplasma; however, also 16SrXI-B phytoplasmas were identified. These phytoplasmas were present in single or in mixed infection in the collected sandalwood, sugarcane and Indian gooseberry samples from this area. This study is the first report of 16SrXI-B phytoplasmas in sugarcane in Kerala of witches’ broom disease in Indian gooseberry associated with aster yellows phytoplasmas and the first report of 16SrXI-B phytoplasma presence in SSD plants. These findings unveil serious threat to existing sandalwood and sugarcane cultivation in the Marayoor sandalwood reserve area, and demand a thorough investigation on phytoplasma insect vectors to efficiently manage these diseases associated with phytoplasmas.

Marigold plants with suspected symptoms of phytoplasma infection were collected during field surveys from three different districts of Karnataka where the incidence of these symptoms was ranging from 1 to 11%. The presence of phytoplasmas in the samples showing phyllody and virescence symptoms was confirmed by nested PCR assays using P1/P7 and R16F2n/R16R2 phytoplasma specific primer pairs. The nucleotide sequence identity and phylogenetic analyses of the 16S rRNA gene sequence indicated the presence of a ‘Candidatus Phytoplasma australasia’ -related strain in the diseased marigold samples. The phylogenetic analysis of secA gene sequences of this phytoplasma strain showed its clustering with strains enclosed in the 16SrII-D subgroup, confirming the phytoplasma identification. To the best of our knowledge, this constitutes the first report of 16SrII phytoplasmas associated with marigold phyllody from the Karnataka state. As the most widely cultivated commercial flower of Karnataka, marigold has a significant role in the national and international flower trade. Phyllody and associated symptoms have a direct negative impact on the quality and market value of these flowers.

Red kidney beans (RKBs) are one of the major components in the human diet of Western Indian Himalaya (WIH). Their cultivation in these habitats is strongly influenced by various biotic and abiotic stresses and therefore, there must be a selection of RKB associated microorganisms that are adapted to these harsh conditions. Seven cold adaptive diazotrophs from the same rhizosphere were isolated in our previous study to reveal the low-temperature associated proteins and mechanisms. However, the diversity and phylogenetic affiliations of these rhizosphere diazotrophs are still unknown. In this study, RKB rhizospheric soil from two different agro-ecosystems of WIH namely S1 (Chhiplakot, 30.70°N/80.30°E) and S2 (Munsyari, 30.60°N/80.20°E) were explored for the assessment of nitrogenase reductase gene (nifH) diversity by plating respective clone libraries SN1 and SN2. The RKB rhizosphere diazotroph assemblage was very diverse and apparently consists mainly of the genera Rhizobium, followed by unknown diazotrophic microorganisms. Deduced amino acid sequence analysis revealed the presence of diverse nifH sequences, affiliated with a wide range of taxa, encompassing members of the Proteobacteria, Actinobacteria and Firmicutes. Members of cyanobacteria, methanotrophs and archaea were also detected. To the best of our knowledge, this is the first major metagenomic effort that revealed the presence of diverse nitrogen-fixing microbial assemblages in indigenous RKB rhizospheric soil which can further be explored for improved crop yield/productivity. Electronic supplementary material The online version of this article (doi:10.1007/s13205-014-0238-5) contains supplementary material, which is available to authorized users.

Nitrogen (N) deficiency and low temperature conditions are the prominent facet of Western Himalayan agro-ecosystems. A slight change in the environment alters the protein expression of the microorganisms. Therefore, proteomes of the two psychrotrophs Dyadobacter psychrophilus B2 and Pseudomonas jessenii MP1 were analyzed using two dimensional electrophoresis and MALDI-TOF-MS, to determine the physiological response of altitudinally different but indigenous microorganisms in response to cold stress under N depleting conditions. Functional assessment of 150 differentially expressed proteins from both the psychrotrophs revealed several mechanisms might be involved in cold stress adaptation, protein synthesis/modifications, energy metabolism, cell growth/maintenance, etc. In both the proteomes, abundance of the proteins related to energy production and stress were significantly increased while, proteins related to biosynthesis and energy consuming processes decreased. ATP synthase subunit alpha, beta, ATP-dependent Clp protease, Enolase, groL HtpG and N(2)-fixation sustaining protein CowN proteins were found to be expressed in both B2 and MP1, similarly to previously studied diazotrophs under low temperature N2 fixing conditions and therefore, can be considered as a biomarker for monitoring the nitrogen fixation in cold niches. Nevertheless, in both the diazotrophs, a good fraction of the proteins were related to hypothetical proteins which are still uncharacterized, thereby, suggesting the need for in-depth studies on cold adapted diazotrophs and their adaptive mechanisms.

The ε-caprolactam is the monomer of the synthetic non-degradable nylon-6 and often found as nonreactive component of nylon-6 manufacturing waste effluent. Environmental consequences of its toxicity to natural habitats and humans pose a global public concern. Soil samples were collected from three designated solid waste dumpsites, namely, Abule-Egba, Olusosun and Isheri-Igando in Lagos State, Nigeria. Sixteen bacteria isolated from these samples were found to utilize the ε-caprolactam as a sole source of carbon and nitrogen at concentration of ≤20 g l(-1). The isolates were characterized using their 16S rRNA gene sequence and showed similarity with Pseudomonas sp., Proteus sp., Providencia sp., Corynebacterium sp., Lysinibacillus sp., Leucobacter sp., Alcaligenes sp. and Bordetella sp. Their optimal growth conditions were found to be at temperature range of 30 to 35 °C and pH range of 7.0-7.5. High Performance liquid chromatography analysis of the ε-caprolactam from supernatant of growth medium revealed that these isolates have potential to remove 31.6-95.7 % of ε-caprolactam. To the best of our knowledge, this study is first to report the ability of Proteus sp. and Bordetella sp. for ε-caprolactam utilization.

A novel phytoplasma subgroup strain SC-Phy385 (GenBank accession number HF586648) was found associated with Sugarcane Grassy Shoot (SCGS) disease in India. Sugarcane (Saccharum officinarum L.) leaf samples with 'grassy shoot' symptoms were collected from the western region of Maharashtra state. The analysis based on restriction fragment length polymorphism (RFLP) profiles and 16S rRNA gene sequences indicated that most of the phytoplasma strains belong to the 16SrXI-B group. However, SCGS phytoplasma strain SC-Phy385 showed unique RFLP profiles with a similarity coefficient of 0.87 using online interactive phytoplasma classification tool, iPhyClassifier. The actual and virtual RFLP profiles of SC-Phy385 using amplified products of the 16S rRNA partial gene and its multiple sequence alignment with other RYD group phytoplasma sequences revealed that this strain was different from phytoplasma groups described earlier, representing new subgroup 16SrXI-F. The 16S rRNA gene-based phylogenetic analysis of reported RYD and BGWL group phytoplasmas showed that they are a genetically diverse group of phytoplasmas.

A novel bacterial strain, MG7, with high cellulase activity was isolated and identified by morphological characteristics and molecular phylogeny analysis as Paenibacillus barcinonensis. Maximum production of cellulase by MG7 was observed at pH 7.0 and 35°C. The enzyme was purified with a specific activity of 16.88 U/mg, the cellulase activity was observed in a zymogram, and its molecular mass (58.6 kDa) was confirmed by SDS-PAGE. The purified enzyme showed maximum activity at pH 6.0 and 65°C and degraded cellulosic substrates such as carboxy methyl cellulose (CMC), Avicel, filter paper, and beta-glucan. The enzyme showed stability with 0.5% concentration of various surfactants. The K(m) and V(max) of cellulase for CMC and Avicel were found to be 0.459 mg/ml and 10.46 mg/ml/h, and 1.01 mg/ml and 10.0 mg/ml/h, respectively. The high catalytic activity and its stability to temperature, pH, surfactants, and metal ions indicated that the cellulase enzyme by MG7 is a good candidate for biotechnological applications.

Genetic improvement of sugarcane variety CoC 671 was carried out through somaclonal variation. The somaclones were obtained by developing callus on 0.8M EMS and 13.57M 2–4.D containing medium. All the 310 regenerated plants were hardened in green house and planted in field for preliminary selection. On the basis of biometric and biochemical parameters 17 somaclones were evaluated in rod row trial and then for smut resistance supplementing with smut inoculum and detection by PCR. The somaclonal variants TC 906 was resistant and TC 922 was moderately resistant to smut. Somaclone TC 922 (2.17 kg, 165 t/ha, 3.34 cm) and TC 906 (2.07 kg, 144.11 t/ha, 3.25 cm) were superior in single cane weight, cane yield and cane diameter over their donor CoC 671 (1.72 kg, 128 t/ha, 2.74 cm). The morphological variations viz. stem colour, internodal alignment, internodal waxiness and leaf width were distinct and stable. The present study reports beneficial traits of the somaclones derived from CoC 671 and their field performance, especially for smut resistance and yield traits and early maturity.

Altitude is the major factor affecting both biodiversity and soil physiochemical properties of soil ecosystems. In order to understand the effect of altitude on soil physiochemical properties and bacterial diversity across the Himalayan cold desert, high altitude Gangotri soil ecosystem was studied and compared with the moderate altitude Kandakhal soil. Soil physiochemical analysis showed that altitude was positively correlated with soil pH, organic matter and total nitrogen content. However soil mineral nutrients and soil phosphorus were negatively correlated to the altitude. RT-PCR based analysis revealed the decreased bacterial and diazotrophic abundance at high altitude. Metagenomic study showed that Proteobacteria, Acidobacteria and Actinobacteria were dominant bacteria phyla at high altitude soil while Bacteroidetes and Fermicutes were found dominant at low altitude. High ratio of Gram-negative to Gram positive bacteria at Gangotri suggests the selective proliferation of Gram negative bacteria at high altitude with decrease in Gram positive bacteria. Moreover, Alphaproteobacteria was found more abundant at high altitude while the opposite was true for Betaproteobacteria. Abundance of Cytophaga, Flavobacterium and Bacteroides (CFB) were also found comparatively high at high altitude. Presence of many taxonomically unclassified sequences in Gangotri soil indicates the presence of novel bacterial diversity at high altitude. Further, isolation of bacteria through indigenously designed diffusion chamber revealed the existence of bacteria which has been documented in unculturable study of WIH (Western Indian Himalaya) but never been cultivated from WIH. Nevertheless, diverse functional free-living psychrotrophic diazotrophs were isolated only from the high altitude Gangotri soil. Molecular characterization revealed them as Arthrobacter humicola, Brevibacillus invocatus, Pseudomonas mandelii and Pseudomonas helmanticensis. Thus, this study documented the bacterial and psychrophilic diazotrophic diversity at high altitude and is an effort for exploration of low temperature bacteria in agricultural productivity with the target for sustainable hill agriculture.

The influence of dried cell powder and culture filtrates of endophytic fungi on production of inophyllum in cell suspension cultures of leaf- and stem-derived callus of Calophyllum inophyllum was investigated. Two fungi, Nigrospora sphaerica and Phoma spp., endophytic to C. inophyllum, were isolated from leaf tissues, and were identified by both 18S rRNA gene amplification and sequencing. Elicitation of suspension cultures of both callus types of C. inophyllum with dried cell powder and culture filtrates of both fungi consistently elicited production of inophyllum A, B, C, and P. In comparison to stem-derived callus, suspension cultures of leaf-derived callus enhanced production of most inophyllum. Of the four inophyllum studied, the highest production of inophyllum A, C, and P was achieved in elicited suspension cultures of leaf-derived callus. Suspension cultures of stem-derived callus enhanced production only of inophyllum B. When suspension cultures of leaf-derived callus were elicited with 40 mg dried cell powder of Phoma spp., a level of 751-fold (6.84 mg/100 g elicited biomass) of inophyllum A was produced, compared to control. Whereas, a level of 414-fold (6.22 mg/100 g elicited biomass) of inophyllum B was produced when suspension cultures of stem-derived callus were elicited with 20 mg dried cell powder of N. sphaerica. When compared to control, a 10% culture filtrate of N. sphaerica in suspension cultures of leaf-derived callus elicited inophyllum C and P production by 928-fold (7.43 mg/100 g elicited biomass) and 750-fold (1.5 mg/100 g elicited biomass), respectively.

The sandal spike disease of sandalwood is known to be associated with aster yellows (AY) phytoplasmas and is a prime threat to sandalwood cultivation in India. The Marayoor Sandalwood Reserve (Kerala state) has an excellent collection of naturalised and matured sandalwood trees and therefore sandalwood forest were surveyed and symptomatic samples were collected from this area. The study confirmed the disease association with AY phytoplasmas and allow to verify the presence of a novel AY phytoplasma strains indicating that phytoplasma strains are differentiating with their plant host species and geographic regions.

Tamarind is an economically important tree species; its fruit pulp is widely used in kitchen in India. A symptomatic tamarind tree with yellow leaf symptoms found in the western part of India was assayed for phytoplasma presence. The 16S rDNA sequence of the detected phytoplasma (TM01) was found clustering with sequences in the elm yellows (16SrV) phytoplasma group. The virtual RFLP analysis of obtained 16S rDNA sequence TM01 revealed that it could be tentatively classified into a novel phytoplasma subgroup, 16SrV-L since its AluI, HaeIII and MseI restriction profiles of R16F2n/R16R2 sequenced amplicons generated were unique as compared to the earlier reported profiles of this phytoplasma subgroup. This is the first phytoplasma report in Tamarindus indica.

Agriculturally important microorganisms mainly comprise bacteria, fungi, cyanobacteria, phytoplasmas and other groups like viruses. Most of the bio-inoculant technology for plant growth promotion (PGP) and biological control of plant diseases is based on bacteria, fungi and cyanobacteria. Plant Pathogenic organisms like Phytoplasmas cause serious diseases in economically important plants. Rapid and authentic identification of agriculturally important microorganisms is imperative before their use in bio-inoculant formulation as well as for diagnosis of pathogens to prevent the crops from damage. Many agricultural microbiologists and plant pathologists are still using traditional approaches of identification of an organism resulting into poor resolution of its taxonomic status. Polyphasic approach of microbial classification using the phenetic, chemotaxonomic and genotypic methods in combinations is the recent approach in microbial taxonomy. In current chapter, we discussed the recent advances in the taxonomy of bacteria (including cyanobacteria and phytoplasma) and fungi. We appeal agricultural microbiologists and plant pathologist for the use of polyphasic approach for better delineation of organism in focus in addition to traditional approaches.

Nasare, K., Yadav, A., Singh, A. K., Shivasharanappa, K. B., Nerkar, Y. S., and Reddy, V. S. 2007. Molecular and symptom analysis reveal the presence of new phytoplasmas associated with sugarcane grassy shoot disease in India. Plant Dis. 91:1413-1418. A total of 240 sugarcane (Saccharum officinarum) plants showing phenotypic symptoms of sugarcane grassy shoot (SCGS) disease were collected from three states of India, Maharashtra, Karnataka, and Uttar Pradesh. Phytoplasmas were detected in all symptomatic samples by the polymerase chain reaction (PCR) amplification of phytoplasma-specific 16S rRNA gene and 16S-23S rRNA spacer region (SR) sequences. No amplification was observed when DNA from asymptomatic plant samples was used as a template. Sixteen samples were selected on the basis of phenotypic symptoms and geographic location, and cloning and sequencing of the 16S rRNA and spacer regions were performed. Multiple sequence alignments of the 16S rRNA sequences revealed that they share very high sequence similarity with phytoplasmas of rice yellow dwarf, 16SrXI. However, the 16S-23S rRNA SR sequence analysis revealed that while the majority of phytoplasmas shared very high (>99%) sequence similarity with previously reported sugarcane phytoplasmas, two of them, namely BV2 (DQ380342) and VD7 (DQ380343), shared relatively low sequence similarity (79 and 84%, respectively). Therefore, these two phytoplasmas may be previously unreported ones that cause significant yield losses in sugarcane in India.

Parthenium weed (Parthenium hysterophorus, Asteraceae) is a common, invasive weed found throughout the Indian subcontinent, Africa, America and Australia. The observed disease, Parthenium Witches’ Broom (PWB), characterised by phylloidy, stunting and prolific branching was observed during a field survey conducted between September 2014 to June 2015. PWB phytoplasma was detected in all symptomatic plant samples and in the previously reported insect vector, Orosius albicinctus belonging to family Cicadellidae (subfamily Deltocephalinae) and other Hemipteran insect samples by amplifying its 16S rRNA gene. To our knowledge, this is the first report of a ‘Ca. Phytoplasma aurantifolia’- related strains associated with witches’ broom disease of P. hysterophorus and insect vector Orosius albicinctus.

The bottle gourd (Lagenaria siceraria [Mol] Standl.), member of Cucurbitaceae family, is a common and very popular vegetable grown in most parts of tropical and subtropical India and consumed in almost every household throughout the year. In India, 1.81 million tons of bottle gourd production was recorded from a cultivation area of 103.23 thousand hectares in 2015. Bottle gourd plants with typical virescence and phyllody symptoms, less vigor and stunted growth, were observed in fields in Pune district of Maharashtra state, during surveys in 2012, 2013, and 2014 (March to May). Affected plants showed the peduncle bearing floral parts abnormally elongated with no fruit setting. The observed symptoms were reminiscent of a phytoplasma infection. The average visual disease incidence was 0.51% in the surveyed areas, covering approximately 12,000 plants. To detect the presence of phytoplasma, 25 mg of leaf tissue from 26 representatives symptomatic and four asymptomatic plants were used for total DNA extraction using CTAB method. A phytoplasma 16S rRNA gene (1.25 kb) was detected in all selected symptomatic plant samples when tested using PCR assay with primers P1/P7 followed by nested PCR using R16F2n/R16R2 primers. Additionally, 480 bp of phytoplasma partial secA gene (Hodgetts et al. 2008) and 440 bp of the partial tuf gene (Makarova et al. 2012) was amplified, purified, and sequenced to confirm the phytoplasma presence. The 16S rRNA, secA, and tuf genes did not amplify by PCR in asymptomatic bottle gourd plant. The obtained 16S rRNA gene sequences (LT594117 and LT594118) from symptomatic plants showed 99.6% identity with a reference strain OAY (‘Ca. Phytoplasma asteris,’ M30790) when compared using the EzTaxon 16S rRNA database (Kim et al. 2012). The virtual RFLP pattern derived using iPhyClassifier from the R16F2n/R16R2 fragment of 16S rRNA sequences was found most similar to the reference pattern of strain BTSpLT138_IX, Papaya Bunchy top phytoplasma (16SrI-X, JF781308), with a pattern similarity coefficient of 0.98 (Zhao et al. 2009). The virtual RFLP patterns with AluI and MseI were confirmed by actual RFLP analysis. Furthermore, the partial sequences of secA (LT617882 and LT617883) genes showed 88% sequence identity with Chrysanthemum yellows (AJS09571) and New Jersey aster yellows (AJS09572), confirming the association of ‘Ca. P. asteris’ (16SrI) with symptomatic bottle gourd plants using NCBI-BLAST. The tuf gene sequences (LT617884 and LT617885) showed a sequence identity of 100% with Eggplant dwarf phytoplasma (BAC76339) reported from Japan. Previously, the 16SrIII-Y phytoplasma strain of ‘Ca. P. pruni’ was reported to be associated with L. siceraria yellows and little leaf in Brazil (Montano et al. 2015). This study is the first record of subgroup 16SrI-X of ‘Ca. P. asteris’ phytoplasma strain associated with virescence and phyllody disease symptoms in bottle gourd in India. This disease may become a serious threat to bottle gourd cultivation as it causes 100% yield loss due to floral abnormality and sterility. Also, bottle gourd is a regular and widely cultivated crop for a large community of farmers in India, making it more vital to study epidemiology of the new disease.

Since 2011, chlorosis with defined chlorotic streak symptoms, yellowing, stunting, and drying of infected clumps of 155 durum (Triticum durum L.) and two bread wheat (T. aestivum L.) genotypes (30- to 45-day-old seedlings) were regularly observed at Regional Station of ICAR - Indian Agricultural Research Institute, Indore (Madya Pradesh), India. The disease incidence was greater in durum (1.7 to 20%) compared with bread wheat (1.3 to 10.4%). Ten randomly selected symptomatic leaf samples were analyzed for the presence of virus particles using leaf dip electron microscopy, PCR, and rolling circle amplification, but no virus particles viz. Wheat streak mosaic virus, Barley yellow dwarf virus, Wheat spindle streak mosaic virus, and Soilborne wheat mosaic virus were detected, nor positive serological relationships established with the antibodies of these viruses. To detect the association of phytoplasma with symptomatic wheat samples, genomic DNA from three representative durum wheat genotypes (Is-1: H18638; Is-2: PDW245; Is-3: MACS3063) and one bread wheat genotype (Is-271: HW2004) were assayed for amplification of 16S rRNA gene using primers P1 and P7 followed by nested PCR using primers R16F2n and R16R2 (Gundersen and Lee 1996). The 16S rRNA gene of phytoplasma was detected in all symptomatic samples, and no amplification was observed in asymptomatic wheat samples. The obtained PCR fragments of 1.25 kb were sequenced directly using primers 343R, 704F, 907R, and 1028F. The 1,245-kb sequences were deposited in GenBank (LT594113–16), which showed 100% sequence identity with each other and 98.16% identity with reference phytoplasma strain BGWL-C1 (Candidatus Phytoplasma cynodontis, AJ550984) when analyzed using EzTaxon database. The MEGA6 constructed neighbor joining phylogenetic analysis and virtual RFLP analysis using 17 restriction enzymes of 16SrRNA gene sequences allowed affiliating the wheat phytoplasma strains with 16SrXI-B subgroup strain with a similarity coefficient of 1 (Zhao et al. 2009). The association with the 16SrXI-B group was further established by amplifying

Phytoplasmas have been found associated with 129 plant species in India including vegetables, legumes, spices, medicinal plants and ornamental plants, cash and oil crops, palms, fruit trees and weeds, where ten 16Sr groups of phytoplasmas have been identified so far. Sesame phyllody, brinjal little leaf, sugarcane grassy shoot, sandal spike, coconut root wilt, areca nut yellow leaf and many diseases in ornamental plants are causing the most severe economic losses in the country. Identified phytoplasmas are related to ‘Candidatus Phytoplasma asteris’, ‘Ca. P. pruni’, ‘Ca. P. ziziphi’, ‘Ca. P. trifolii’, ‘Ca. P. solani’, ‘Ca. P. cynodontis’, ‘Ca. P. oryzae’, ‘Ca. P. phoenicium’, ‘Ca. P. australasia’ and ‘Ca. P. pini’. Among them ‘Ca. P. asteris’-related strains (aster yellows, 16SrI group) are the prevalent phytoplasmas identified associated with 62 diseases followed by phytoplasmas classified in 16SrII, 16SrVI, 16SrXIV and 16SrXI groups. At least seven insect vectors, belonging to leafhoppers and planthoppers species, have been reported as putative or natural vectors for important phytoplasma diseases. The majority of phytoplasma disease reports are from north and south part of India. Little attempt has been made to genomics, epidemiology, host phytoplasma interaction and management aspects of these diseases. Presently the suggested effective management practices in India are growing resistant varieties, control of insect vectors, weed species as alternative hosts and use of healthy planting materials.

Disease symptoms characterized by phyllody, yellowing of leaves, stunting, and virescence were observed in economically important fodder crops like alfalfa and “bajra” x napier grass hybrid grass. Similar symptoms were found in bell pepper or “shimlamirch”, a common vegetable grown all over India; “bajra” a significant widespread cereal crop in Maharashtra and Karnataka and in the nuisance weed Parthenium hysterophorus collected in the vicinity of above fodder growing fields. The successful amplification of specific 16S rRNA gene in all collected symptomatic plants confirmed the presence of phytoplasmas and their sequences analysis revealed their close relationship with strains of ‘ Candidatus Phytoplasma australasia’. Using EzTaxon, a microbial 16S rRNA sequence database, and i PhyClassifier- an online phytoplasma taxonomy tool, group 16SrII, subgroup D was identified for the classification of these sequences. This is the first report of 16SrII-D phytoplasmas associated with phyllody and yellowing diseases of “bajra” and bell pepper in India (PDF) First report of 16SrII-D phytoplasmas associated with fodder crops in India. Available from: https://www.researchgate.net/publication/322687543_First_report_of_16SrII-D_phytoplasmas_associated_with_fodder_crops_in_India [accessed Dec 24 2018].

Himalayas are considered as a reservoir of diversified and dynamic gene pool. This study describes the response of a Himalayan psychrophilic diazotroph to low temperature diazotrophy. Seven cold adaptive N2 fixing bacteria were isolated and identified as Bacillus sp., Arthrobacter sp., Rhodococcus sp., Pseudomonas sp., etc. In order to examine the physiological response to low temperature diazotrophy, differential proteomic analysis of Pseudomonas migulae S10724 strain was carried out using two dimensional electrophoresis and MALDI–TOF–MS. Functional assessment of 66 differentially expressed proteins revealed several mechanisms thought to be involved in low temperature adaptation and nitrogen fixation, including general stress adaptation, protein and nucleic acid synthesis, energy metabolism, cell growth/maintenance, etc. Major fraction of the upregulated proteins was stress proteins, while majority of the downregulated proteins were related to cell division. Furthermore, MALDI–TOF–MS-based identification of randomly selected peptides encountered two exclusively expressed proteins: NifU family SUF system FeS assembly protein and membrane protein, suppressor for copper-sensitivity B precursor which might have a crucial role at low temperature nitrogen fixation. To the best of our knowledge, this is the first report of the isolation and differential proteomic analysis of psychrophilic diazotroph from Himalayan high altitude rhizospheric soil.

Soybean (Glycine max L. Merr., Fabaceae), a native plant of South Asia, is an important source of vegetable oil and proteinaceous feed for cattle, poultry, and humans worldwide. Soybean is also considered a good source of omega-3 fatty acids, vitamin B12, amino acids, minerals, isoflavones, and probiotics in the form of fermented soy products. In India, soybean is a commercially important crop as 11.94 million metric tons of seeds and 1.69 million metric tons of edible oil were produced in 2013 according to FAOSTAT (Food and Agriculture Organization, Statistical Division), United Nations. Maharashtra is the second largest producer of soybean seeds in India and Vidarbha is a hotspot for soybean cultivation. Leaves of symptomatic soybean plants showing typical witches’-broom symptoms, which included stunting, shortened internodes, and intense proliferation of leaf and flower buds, were collected from fields of Akola district (Vidarbha region), Maharashtra, during the monsoon season (August and September) of 2015. A total of 197 symptomatic soybean samples of cultivar JS 335 were collected from 26 fields of Akola district. To estimate disease incidence, ∼70 to 100 plants from 10 to 15 randomly selected rows in each surveyed field were visually recorded for disease symptoms. Disease incidence was ranged from 2 to 10% in these fields. To confirm the association of phytoplasma, 25 mg of leaf tissue from 10 representative symptomatic and one asymptomatic plant were used for total DNA extraction using MO BIO PowerPlant Pro DNA Isolation Kit (Carlsbad, CA). The phytoplasma 16S rRNA gene was detected in all 10 symptomatic samples using PCR assay with phytoplasma universal primers P1 and P7 (Smart et al. 1996). No amplification was observed in the DNA isolated from the asymptomatic plant. All 10 PCR fragments were sequenced directly using primers 343R, 704F, 907R, 1028F, and 1492R (Baker et al. 2003). The obtained 16S rRNA gene sequences were identical to each other which showed 100% homology with a strain PpYc (Candidatus Phytoplasma australasia, Y10097, 16SrII D) when compared using the EzTaxon 16S rRNA database. The representative sequence was deposited in GenBank with accession no. LT558766. The virtual RFLP (Zhao et al. 2009) pattern derived from the R16F2n/R16R2 part of 16S rRNA sequence (LT558766) was found identical to the reference pattern of the 16Sr group II, subgroup D (Y10097), with a pattern similarity coefficient of 1. Earlier, 16SrI group phytoplasma reported from New Delhi, India, was identified to be associated with soybean witches’-broom disease (Kumar et al. 2014). However, peanut witches’-broom group phytoplasma (Ca. P. aurantifolia, 16SrII-C) was recorded on soybean only from Malawi and Mozambique (Lava Kumar et al. 2011). To our knowledge, this is the first report of 16SrII-D phytoplasma strain associated with witches’-broom disease of soybean from India. Soybean is the number one oilseed crop in terms of both area and production and plays a significant role in improving the socioeconomic conditions of small and marginal farmers of central India, making this disease study more significant.

Transcription control of gene expression depends on a variety of interactions mediated by the core promoter region, sequence specific DNA-binding proteins, and their cognate promoter elements. The prominent group ofcisacting elements in plants contains an ACGT core. Theciselement with this core has been shown to be involved in abscisic acid, salicylic acid, and light response. In this study, genome-wide comparison of the frequency of occurrence of two ACGT elements without any spacers as well as those separated by spacers of different length was carried out. In the first step, the frequency of occurrence of theciselement sequences across the whole genome was determined by using BLAST tool. In another approach the spacer sequence was randomized before making the query. As expected, the sequence ACGTACGT had maximum occurrence inArabidopsis thalianagenome. As we increased the spacer length, one nucleotide at a time, the probability of its occurrence in genome decreased. This trend continued until an unexpectedly sharp rise in frequency of (ACGT)N25(ACGT). The observation of higher probability of bigger size motif suggests its directed evolution inArabidopsis thalianagenome.

The investigations on efficacy of different newer insecticides action (Acetamiprid 20 SP @ 20 g a. i./ha, spinosad 45% SC @ 73 g a. i./ha, rynaxypyr 18.5 SC 30 g a. i/ha, dimethoate 30 EC @ 600 g a. i./ha, profenophos+DDVP 20 ml+0.5 ml/l, acephate 75 SP @ 750 g a. i./ha) against pod fly (Melanogromyza obtusa Malloch) and pod borer (Helicoverpa armigera Hubner) on long duration pigeonpea variety “Bahar” were carried out at Agricultural Research Farm of Banaras Hindu University during the kharif seasons of 2012–13 and 2013–14. Among the insecticides tested, the lowest pod and grain damage by H. armigera was recorded in the plot treated with rynaxypyr 18.5 SC @ 30 g a. i./ha (1.66 & 2.67% and 0.53 & 2.05%, respectively) which was at par with spinosad 45% SC @ 73 g a. i./ha (2.00 & 3.33% and 0.57 & 2.51%, respectively). While in case of pod fly, the lowest pod and grain damage was recorded in the plot treated with spinosad 45% SC @ 73 g a. i./ha (22.66 & 23.00% and 11.55 & 7.74%, respectively) which was at par with acetamiprid 20 SP @ 20 g a. i./ha (26.66 & 24.33 and 11.78 & 8.68%, respectively).

The disease symptoms characterised by little leaf, phyllody, stunting and branch proliferation were observed during a field survey conducted from June 2015 to January 2016 on common invasive weeds Cleome viscosa (tick weed), Trichodesma zeylanicum (cattle bush) and Tephrosia perpurea (wild indigo), which were found infected with peanut witches’ broom phytoplasmas (16SrII group) strains. Phylogenetically similar phytoplasma strains were also detected in symptomatic samples of Sesamum indicum (sesame), Vigna unguiculata (cow pea), Phaseolus vulgaris (French bean), Dendrocalamus strictus (bamboo) and Carica papaya (papaya) from the same or adjacent fields. This is the first report of 16SrII-D group phytoplasmas (‘Candidatus Phytoplasma aurantifolia’-related strains) associated with witches’ broom disease of C. viscosa, D. strictus and yellow stunting disease of C. papaya. The association of 16SrII-C group phytoplasmas with witches’ broom disease of T. zeylanicum and T. purpurea is also reported for the first time. From the obtained results, it can be inferred that the weed hosts harbors 16SrII group phytoplasma strains associated with little leaf and witches’ broom diseases of economically important grain legume crop species like S. indicum, V. unguiculata and P. vulgaris.

Wild indigo (Tephrosia purpurea (L.) Pers.) grows as a common weed throughout the Indian subcontinent. The plant has pinnate leaves, white or purplish flowers, and flat hairy pods, and is cultivated as a green manure crop. The plant extracts contain compounds such as tephrosin, an aromatic ester, prenylated flavonoid, and sesquiterpene (2) that have medicinal properties. The newly recognized disease, Tephrosia purpurea witches' broom (TPWB), was characterized by chlorosis, stunting, and proliferative branching, which were suggestive of phytoplasma infection during a field survey conducted in November 2013. To determine the presence of phytoplasma, 2 g of compound leaves from three symptomatic and asymptomatic plants were used for total DNA extraction using the CTAB method. The phytoplasma 16S rRNA gene was detected in all three symptomatic plants using nested PCR with universal phytoplasma primer pairs, P1/P7 followed by R16F2n/R16R2 (4). No amplification was observed in DNA isolated from asymptomatic plants. PCR fragments (1,246 bp in length) generated from symptomatic T. purpurea plants were sequenced directly using five different primers viz. 343R, 536F, 704F, 907R, and 1103F. TPWB phytoplasma 16S rRNA gene sequence (GenBank Accession No. HG792252) showed 99.12% homology with a ‘Candidatus Phytoplasma aurantifolia’ strain WBDL (U15442) when compared using the EzTaxon 16S rRNA database (3). Virtual restriction fragment length polymorphism (RFLP) analysis was carried out on the obtained sequence using iPhyClassifier (5). The virtual RFLP pattern derived from the HG792252 sequence was different to the reference patterns of previously established 16Sr groups and subgroups. The reference pattern of the 16Sr group II, subgroup C (AJ293216) was most similar with a similarity coefficient of 0.92, which placed it in a new subgroup, 16Sr II-M (1). Furthermore, virtual RFLP results were confirmed by digesting R16F2n/R16R2 amplicon with BstUI, DraI, HinfI, HpaI, and MseI restriction enzymes according to manufacturer's instructions. To our knowledge, this is the first report of a ‘Ca. P. aurantifolia’-related strain associated with witches'-broom disease of T. purpurea in India. References: (1) H. Cai et al. Int. J. Syst. Evol. Microbiol. 58:1448, 2008. (2) A. K. Khalafalah et al. Pharmacognosy Res. 2:72, 2010. (3) O.-S. Kim et al. Int. J. Syst. Evol. Microbiol. 62:716, 2012. (4) C. Smart et al. Appl. Environ. Microbiol. 62:2988, 1996. (5) Y. Zhao et al. Int. J. Syst. Evol. Microbiol. 59:2582, 2009.