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Plants are the major source of natural flavour ingredients reported for their wide applications in food and pharmaceuticals, oral care and wellness products, etc. We have investigated the water-soluble fractions (WSF) of basil tetraploid (O. basilicum L.) for their toxicity and biological potential against Salmonella Typhimurium, a pathogen causing around one million cases of illnesses in the United States every year. The WSF obtained using a Clevenger-type apparatus was further divided into two equal parts, one each for in-vivo toxicity evaluation and quality assessments, respectively. The proportions of major phenylpropanoid identified as meta-eugenol in the WSF were found in the range of 42.8-57.9 %, which was substantially in higher proportion as compared to essential oil (20.9-23.0 %). Based on sub-acute oral toxicity data, WSF has not shown any adverse effect with levels as high as 500 μL/25 g body weight in Swiss albino mice. Besides, the WSF also exhibited a maximum reduction in bacterial load in mice infected with Salmonella Typhimurium in a dose-dependent manner. We have shown the biological potential of basil water-soluble fraction as an effective bacterial load-suppressing agent for the prevention of Salmonella infections in animal model., (© 2024 Wiley-VHCA AG, Zurich, Switzerland.)

Background: Telepharmacy is a method utilized in pharmacy practice that delivers pharmaceutical care services via telecommunication technology. In the Philippines, the current process for patients to avail of telepharmacy services utilizes a variety of existing applications or websites instead of a single application. Objectives: This study aimed to assess patient feedback on the feasibility of the newly developed telepharmacy mobile application, YourRx. Specifically, it evaluated the application's functionality, usability, security, and performance. Methods: The study had 3 phases: (1) the design and development phase, (2) the implementation phase, wherein the pharmacists and patients were oriented beforehand with the use of YourRx mobile application, and (3) the evaluation phase, where the mobile application was evaluated for its functionality, usability, security, and performance by the patients through the use of a survey questionnaire and an interview. Results: YourRx application was developed and evaluated successfully. It is available for Android users and has primary features, including video calls, sharing, and setting an appointment. A total of 46 patients used the YourRx. Most of the patients were very satisfied with the navigation, service acquisition, and overall design of the YourRx. They expressed convenience in acquiring telepharmacy services because the application was easy to understand, use, and navigate. Conclusion: YourRx is a pioneering telepharmacy mobile application in the Philippines. The results of this study substantiate that YourRx is a user-friendly platform that provides patients convenient access to telepharmacy services with less time and effort thus improving patient health outcomes., Competing Interests: Declaration of Conflicting InterestsThe author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

The newly released interspecific hybrid variety CIM-Shishir, resulting from a cross between Ocimum basilicum and Ocimum kilimandscharicum claims to be a multicut, lodging resistant, cold tolerant, high essential oil yielding with linalool rich variety. It has a purple-green stem and has a unique feature and advantage of better survival in the winter season than other O. basilicum varieties, illustrating its physiological mechanisms for cold tolerance. In this study, we subjected both the CIM-Shishir variety and a control plant to cold stress to investigate the impact of low temperatures on various physiological, trichome developments, secondary metabolite constitution aspects related to essential oil production, and gene expression. The analysis revealed a significantly higher density and altered morphology of trichomes on the leaf surface of the variety subjected to low temperatures, indicating its adaptation to cold conditions. Furthermore, when comparing the treated plants under low-temperature stress, it was observed that the relative electrolyte leakage and Malondialdehyde (MDA) contents substantially increased in the control in contrast to the CIM-Shishir variety. This finding suggests that CIM-Shishir exhibits superior cold tolerance. Additionally, an increase in proline content was noted in the variety exposed to low temperatures compared to the control. Moreover, the chlorophyll and anthocyanin content gradually increased with prolonged exposure to low-temperature stress in the newly developed variety, indicating its ability to maintain photosynthetic capacity and adapt to cold conditions. The activities of superoxide dismutase (SOD) also increased under low-temperature conditions in the CIM-Shishir variety, further highlighting its cold tolerance behaviour. In our research, we investigated the comprehensive molecular mechanisms of cold response in Ocimum. We analyzed the expression of key genes associated with cold tolerance in two plant groups: the newly developed hybrid variety known as CIM-Shishir Ocimum, which exhibits cold tolerance, and the control plants susceptible to cold climates that include WRKY53, ICE1, HOS1, COR47, LOS15, DREB5, CBF4, LTI6, KIN, and ERD2. These genes exhibited significantly higher expression levels in the CIM-Shishir variety compared to the control, shedding light on the genetic basis of its cold tolerance. The need for climate-smart, resilient high-yielding genotype is of high importance due to varied climatic conditions as this will hit the yield drastically and further to the economic sectors including farmers and many industries that are dependent on the bioactive constituents of Ocimum., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Plants are known to be the natural factory for the production of flavor chemicals. Essential oils comprised of aldehyde as a functional group are potent in deciphering flavor effects in beverages and fresh and prepared food products. In the majority, these are manufactured through synthetic routes, resulting in high product carbon footprints or CO 2 equivalents in total greenhouse gas emission. FDA has banned some of the synthetic flavor chemicals due to the health hazards associated with them. However, consumer's preference for natural is at stake due to the absence of quantitative traceability tools. The accelerator mass spectrometer (AMS) analysis revealed a distinction between natural and fossil-derived citral and its blends in Cymbopogon essential oils. The plant-derived citral contained a percent modern carbon (pMC) value in the range of 99-100 %. In contrast, the fossil fuel-derived citral showed zero pMC. Similarly, blends of Cymbopogon oils with 30-50 % (w/w) of fossil origin citral contained pMC equivalent to the proportions of modern carbon. These results showed the usefulness of AMS in quantifying the amount of 14 C associated with flavor ingredients. Besides, acute oral toxicity data revealed Cymbopogon oils as the safe flavoring substance at the highest 2000 mg/kg body weight dose in Swiss albino mice., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Herbal medicines are popular natural medicines that have been used for decades. The use of alternative medicines continues to expand rapidly across the world. The World Health Organization suggests that quality assessment of natural medicines is essential for any therapeutic or health care applications, as their therapeutic potential varies between different geographic origins, plant species, and varieties. Classification of herbal medicines based on a limited number of secondary metabolites is not an ideal approach. Their quality should be considered based on a complete metabolic profile, as their pharmacological activity is not due to a few specific secondary metabolites but rather a larger group of bioactive compounds. A holistic and integrative approach using rapid and nondestructive analytical strategies for the screening of herbal medicines is required for robust characterization. In this study, a rapid and effective quality assessment system for geographical traceability, species, and variety-specific authenticity of the widely used natural medicines turmeric, Ocimum , and Withania somnifera was investigated using Fourier transform near-infrared (FT-NIR) spectroscopy-based metabolic fingerprinting. Four different geographical origins of turmeric, five different Ocimum species, and three different varieties of roots and leaves of Withania somnifera were studied with the aid of machine learning approaches. Extremely good discrimination ( R 2  > 0.98, Q 2  > 0.97, and accuracy = 1.0) with sensitivity and specificity of 100% was achieved using this metabolic fingerprinting strategy. Our study demonstrated that FT-NIR-based rapid metabolic fingerprinting can be used as a robust analytical method to authenticate several important medicinal herbs., Competing Interests: The authors declare that there are no conflicts of interest., (© 2023 The Author(s).)

CIM-Saumya is an improved, methyl chavicol rich variety of Ocimum basilicum (Family-Lamiaceae), developed by Council of Scientific and Industrial Research-Central Institute of Medicinal and Aromatic Plants. This plant possesses analgesic, anti-ulcerogenic, anti-inflammatory, anti-oxidant, cardiac stimulant, Central Nervous System depressant, hepatoprotective and immunomodulator activities due to the presence of various phytoconstituents. Among them rosmarinic acid, caffeic acid and ferulic acid are the three major phenolic compounds responsible for its therapeutic utility. These compounds are produced in very low amounts in the in vivo plants. Therefore, the present study has been conducted for establishment of cell suspensions, optimization of inoculums size, growth kinetics and screening of elicitor and precursors for the accumulation of cell biomass and the production of the three important phenolic compounds in cell suspension of O. basilicum (CIM-Saumya). Leaf derived friable callus was used for establishing the cell suspension in liquid Murashige and Skoog's medium fortified with 1 g/L casein hydrolysate + 2.26 µM 2,4-dichlorophenoxyacetic acid + 0.465 µM kinetin + 2.68 µM naphthalene acetic acid. The growth kinetic analysis pattern of cell suspension revealed the maximum biomass increments (% BI = 486.7) and production of RA 8.086 mg/g dry weight was found in 30th day harvested cells. Whereas, the other two phenolic compounds i.e. ferulic acid (0.0125 mg/g dry weight) and caffeic acid (0.38 mg/g dry weight) was recorded highest on 25th day of growth cycle. In the present study, one biotic elicitor i.e. yeast extract and three precursors [peptone, tryptone and lactalbumin hydrolysate] were tested, among them, lactalbumin hydrolysate (100 mg/L; added at 16th day) treated cells recorded highest estimated phenolic compounds yield (251.5 mg/L; 6.81 fold compared to the control) and biomass increments i.e. % BI = 1207 with 1.85 fold compared to the control. The highest rosmarinic acid content i.e. 25.47 mg/g DW (4.4 fold compared to the control) and 24.42 mg/g dry weight (4.1 folds compared to the control) was noticed in 30th day harvested cells treated with yeast extract (1 g/L on 0 day) and lactalbumin hydrolysate (100 mg/L added on 16th day), respectively. While caffeic acid content (0.91 mg/g dry weight) showed 2.9 folds higher compared to the control in cells treated with peptone 200 mg/L added on 16th day of culture cycle. All the treated cells showed enhanced phenylalanine ammonia-lyase enzyme activity with highest specific activity in lactalbumin hydrolysate followed by tryptone, peptone, and yeast extract., Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-023-01316-6., Competing Interests: Conflict of interestThe authors declare that they have no conflict of interest., (© Prof. H.S. Srivastava Foundation for Science and Society 2023. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Identification of plant species is a crucial process in natural products. Ocimum , often referred to as the queen of herbs, is one of the most versatile and globally used medicinal herbs for various health benefits due to it having a wide variety of pharmacological activities. Despite there being significant global demand for this medicinal herb, rapid and comprehensive metabolomic fingerprinting approaches for species- and variety-specific classification are limited. In this study, metabolomic fingerprinting of five Ocimum species ( Ocimum basilicum L., Ocimum sanctum L., Ocimum africanum Lour. , Ocimum kilimandscharicum Gurke. , and Hybrid Tulsi) and their varieties was performed using LC-MS, GC-MS, and the rapid fingerprinting approach FT-NIR combined with chemometrics. The aim was to distinguish the species- and variety-specific variation with a view toward developing a quality assessment of Ocimum species. Discrimination of species and varieties was achieved using principal component analysis (PCA), partial least squares discriminate analysis (PLS-DA), data-driven soft independent modelling of class analogy (DD-SIMCA), random forest, and K-nearest neighbours with specificity of 98% and sensitivity of 99%. Phenolics and flavonoids were found to be major contributing markers for species-specific variation. The present study established comprehensive metabolomic fingerprinting consisting of rapid screening and confirmatory approaches as a highly efficient means to identify the species and variety of Ocimum, being able to be applied for the quality assessment of other natural medicinal herbs.

Prishanparni (Uraria picta Desv.), a critically endangered annual shrub belonging to the family 'Papillionaceae'. It is widely distributed throughout India, Sri Lanka, Bangladesh, Tropical Africa, Malay Islands, and the Philippines. The consistent performances of U. picta accessions based on ten economic traits studied were identified as P-12, P-16, P-21, P-22, P-31, P-47, and P-48. These accessions could be used for commercial cultivation in northern Indian plains. Among the total twenty-three studied accessions P-50, P-21, P-48, and P-47 were found superior for rhoifolin content in their aerial as well as root part, which may have various therapeutic potentials used in traditional and modern systems of medicines. These accessions can be exploited for commercial cultivation or in a hybridization program for further crop improvement. Wide range cultivation of the selected accessions in the Indo-Gangetic plains will fit in the existing cropping systems of this region, resulting in comparatively better supplementation of herb to the pharmaceutical and herbal drug industries and reducing the pressure on the wild populations., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

To understand the altered developmental changes and associated gene expression in inter-genomic combinations, a study was planned in two diverse yet closely related species of Ocimum, targeting their hybrid F1 and amphidiploids. The existing developmental variations between F1 and amphidiploids was analyzed through phenotypical and anatomical assessments. The absence of 8330 transcripts of F1 in amphidiploids and the exclusive presence of two transcripts related to WNK lysine-deficient protein kinase and geranylgeranyl transferase type-2 subunit beta 1-like proteins in amphidiploids provided a set of genes to compare the suppressed and activated functions between F1 and amphidiploids. The estimation of eugenol and methyleugenol, flavonoid, lignin and chlorophyll content was correlated with the average FPKM and differential gene expression values and further validated through qRT-PCR. Differentially expressed genes of stomatal patterning and development explained the higher density of stomata in F1 and the larger size of stomata in amphidiploids. Gene expression study of several transcription factors putatively involved in the growth and developmental processes of plants clearly amalgamates the transcriptome data linking the phenotypic differences in F1 and amphidiploids. This investigation describes the influence of interspecific hybridization on genes and transcription factors leading to developmental changes and alleviation of intergenomic instability in amphidiploids.

The positive effect of herbal supplements on aging and age-related disorders has led to the evolution of natural curatives for remedial neurodegenerative diseases in humans. The advancement in aging is exceedingly linked to oxidative stress. Enhanced oxidative stress interrupts health of humans in various ways, necessitating to find stress alleviating herbal resources. Currently, minimal scientifically validated health and cognitive booster resources are available. Therefore, we explored the impact of plant extracts in different combinations on oxidative stress, life span and cognition using the multicellular transgenic humanized C. elegans , and further validated the same in Mus musculus , besides testing their safety and toxicity. In our investigations, the final product-the HACBF (healthy ageing cognitive booster formulation) thus developed was found to reduce major aging biomarkers like lipofuscin, protein carbonyl, lipid levels and enhanced activity of antioxidant enzymes. Further confirmation was done using transgenic worms and RT-PCR. The cognitive boosting activities analyzed in C. elegans and M. musculus model system were found to be at par with donepezil and L-dopa, the two drugs which are commonly used to treat Parkinson's and Alzheimer's diseases. In the transgenic C. elegans model system, the HACBF exhibited reduced aggregation of misfolded disease proteins α-synuclein and increased the health of nicotinic acetylcholine receptor, levels of Acetylcholine and Dopamine contents respectively, the major neurotransmitters responsible for memory, language, learning behavior and movement. Molecular studies clearly indicate that HACBF upregulated major genes responsible for healthy aging and cognitive booster activities in C. elegans and as well as in M. musculus . As such, the present herbal product thus developed may be quite useful for healthy aging and cognitive boosting activities, and more so during this covid-19 pandemic., Supplementary Information: The online version contains supplementary material available at 10.1007/s13237-022-00407-1., Competing Interests: Conflict of interestThe authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© The Author(s) under exclusive licence to Archana Sharma Foundation of Calcutta 2022, Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

Mentha arvensisL. (corn mint) is well known for the production of menthol, a used commodity in flavouring industries, and provides natural fragrances. Glandular trichomes are responsible for producing specific secondary metabolites in vascular plants having species chemistry. Ten cultivars/varieties of M. arvensis, namely, Saksham, Kosi, Himalaya, Gomti, Sambhav, Kalka, Damroo, Kushal, and Shivalik, were used to study the developmental regulation of trichomes, essential oil yield, chemical constituents of essential oil and morphological parameters were estimated with gene expression using a randomized block design. Simultaneously, RNA sequence-based transcriptome analysis was done to reveal the transcription factors and differential gene analysis, which are responsible for the biosynthesis of essential oil as well as trichome development. Plant growth showed the maximum transition between 35 and 50 days stage, while essential oil and its metabolite bioconversion was observed in between 70 and 100 days stage. Glandular trichomes were maximally increased between 50, 70 days, and 100 days stage in var. Kosi followed by var. Saryu which has rapid growth in oil content. Menthol reductase activity was found to be a regulatory element during development, as it follows the inverse trend of menthol content and leads to menthol accumulation in subcuticular spaces. Transcriptional factors, cog, and nonredundant novel genes were identified. The composition of mintessential oils is regulated at multiple levels, including transcript abundance, catalytic properties of enzyme catalysts, and cell type-specific epigenetic processes., (Copyright © 2021 Elsevier Masson SAS. All rights reserved.)

Studies on genetic diversity could enhance taxonomic authentication and evolutionary relationship among the species of Ocimum. Therefore, diversity among 36 Ocimum accessions representing species from different regions of world were analyzed using Start Codon-Targeted Polymorphism (SCoT) and inter-simple sequences repeat (ISSR) marker. Marker systems used in this study was potentially targeted the different regions of the genome and included 18 SCoT and 15 ISSR primers, which showed successful amplification profile for Ocimum. Between these two, SCoT revealed the highest mean value of percentage of Polymorphism (84.6%), polymorphic information content (PIC, 0.65), and resolving power (Rp, 8.80), which were higher than ISSR. A total of 140 and 111 amplicons were obtained with SCoT and ISSR marker. The Mantel test indicted a significant correlation (r 2  = 0.44) between ISSR and SCoT, which suggested a common genetical background among the accessions. The principal coordinate study showed the selection of different Ocimum genotypes by the cluster analysis. This study will help and support identification, genetic mapping, and molecular ecology to enhance the breeding program's efficiency for developing elite varieties to meet industrial demand globally. The present study is the first report of the genetic diversity, and relationship determination with SCoT-based molecular marker among Ocimum accessions.

Purpose: To increase the size of the flowers for easy plucking, flower yield, pyrethrins content (%), and elite mutant selection in pyrethrum., Materials and Methods: To increase pyrethrum production and acclimatize in north Indian plain condition, a genetic improvement program was undertaken to widen the range of variations for size and yield of flowers and pyrethrins content (%) in pyrethrum crop. Pyrethrum seeds of the variety Avadh were irradiated with gamma rays at 20 to 300 Gy doses in Gamma chamber 5000 (cobalt-60 research irradiator)., Results: Observations gathered visually in M 1 based on vigor, synchronization of flowering, and flower's size. Out of 90 M 2 families, 20 mutants were raised in M 3 along with the check-in preliminary evaluation trial. The four promising mutants, 1 (20 Gy-3), 7 (40 Gy-5), 10 (40 Gy-8), 14 (60 Gy19-10) was grown for four years in a bench-scale trial (randomized block design, replicated thrice) to test the yield performance and selection of high yielding elite mutant (s). It has been found that pyrethrum is sensitive to gamma rays irradiation and produced a high range of qualitative and quantitative variations. After massive screening over four years, two promising mutants for high dry flower yield and pyrethrins content, namely 7 (40 Gy-5), and 10 (40GY-8) were isolated., Conclusions: The mutagenesis changed traits mean in positive or negative directions. Pyrethrum plant is highly sensitive to gamma irradiation and produced a high range of variability in the qualitative and quantitative traits. The mutagenesis changed the mean of traits in both positive and negative directions. Due to mutagenic efficacy, two mutants 7 (40 Gy-5), and 10 (40GY-8) were expressed high performance for pyrethrin percent i.e., 87.23 and 59.78% improvement over the check variety 'Avadh', with synchronous flowering. These two mutants are in the pipeline for release as a variety for cultivation in the North Indian plains.

Purpose: The nature and kind of some successful mutations achieved through gamma radiation at CSIR-CIMAP, Lucknow are reviewed and described in this article. Medicinal and aromatic crops (MACs) came under the mutation-breeding program very late - perhaps during the 1980s in India. Nevertheless, successes have been glaring and unique too. The plant breeders of the Institute concerted attempts were made for the genetic restructuring of the plant-frame in Hyoscyamus niger L. (black henbane) and Cymbopogon martinii (palmarosa), enhanced alkaloid biosynthesis in henbane, weak, or lack of latex biosynthesis in Papaver somniferum L. (Opium poppy), increased productivity of seeds in Plantago ovata F. (isabgol), and new/increased formation of essential oils in Chamomila recutita [L.] Rauschert (Chamomile) and Cymbopogon winterianus (Java citronella). Several qualitative macro mutations were generated for commercial exploitation, e.g. an unbranched (Ub) and physiologically most efficient variety Aekla and high tropane alkaloid bearing variety Aela of H. niger , opium less oil-seed variety Sujata of opium poppy, a dwarf mutant and a male sterile (ms) line of palmarosa, and new compound yielding variety of chamomile and geranium., Conclusions: The ample quantitative variation was also created by reshuffling the polygenic background in both seed and vegetatively propagated MACs, and subsequently, with applying mutation breeding approach superior varieties were evolved and released after a rigorous screening in the field-evaluation or under pipeline for release. These varieties are Niharika and Mayuri of psyllium; Vallary, CIMAP Sammohak, CIM Ujjwala, and YEL (yellow) of German chamomile and Manjari and CIM Jeeva of Java citronella.

microRNAs are small non-coding RNA molecule that plays an important role in metabolism. Chrysopogon zizanioides (L.) Roberty is an important aromatic plant used in perfumery industries, soil, water conservation, and agricultural practices. In this study, the transcriptomic sequence of vetiver leaf and root was subjected to miRNA identification by the computational methods. miRNA identification was carried out using a homology-based method by C-mii software with several other online tools. A total of 80 miRNA were identified from both leaf and root sequences. Target identification was done by identified miRNA sets. A total of 25 and 31 miRNA families were identified in both leaf and root, respectively, with ten common families involve in different ontological function. miR169 and miR5021 regulate most of the target in leaf and root. In vetiver, many primary and secondary metabolism elements are regulated by miRNA as photo-system, transcription factor, terpenoid metabolism, etc. Here is the first in silico study revealing the specific miRNAs and their target genes for corresponding root and leaf tissues respectively in C. zizanioides.

Purpose: To develop elite mutants in chamomile ( Chamomilla recutita [L.] Rauschert) for increasing the quantity and quality of essential oil rich in acetylenic compound (2 Z ,8 Z )-matricaria acid methyl ester by applying γ -rays irradiation. Molecular and chemical analysis was performed for ithe dentification/differentiation of mutant genotype. Materials and methods: Chamomile ( Chamomilla recutita [L.] Rauschert) variety Vallary seeds were irradiated by applying γ -rays irradiation at 100, 200, 300, 400, 500, 600, 700, 800, 900 and1000 Gy doses at a dose rate of 55 Gy/min and mutants were isolated and analyzed for the quantity and quality of essential oil. The oil was found to be rich in acetylenic compound (2 Z ,8Z)-matricaria acid methyl ester and the results obtained were validated using gas chromatography (GC) coupled with either Flame Ionization detection (GC-FID) or mass spectrometer (GC-MS), and nuclear magnetic resonance (NMR). Results: The selected mutant SELM-1 (Selection Mutant-1) showed the production potential of 7.00-7.50 q ha -1 dry flowers and 6.00-6.50 kg ha -1 essential oil yield. Essential oil of mutant SELM-1 contained in [(2 Z ,8 Z )-matricaria acid methyl ester] (76-80%) useful in cosmetic, perfumery, and pharmaceutical industries. Conclusion: γ -rays irradiation method is a very efficient mutation breeding method for chamomile crop. GC-FID or GC-MS and NMR methods are found to be the most powerful methods for screening of essential oil chemical compounds isolated from the mutants. The novel mutant (SELM-1) is very promising in terms of high flower and essential oil yield rich in acetylenic compound (2 Z ,8 Z )-matricaria acid methyl ester (76-80%), hence, it was released as variety in Council of Scientific & Industrial Research (CSIR)-Central Institute of Medicinal and Aromatic Plant (CIMAP), Lucknow U.P. (India) named as CIM-Ujjwala for commercial cultivation.

Main Conclusion: Transcriptome analysis and biochemical characterization of the putative l-3,4-dihydroxyphenylalanine (l-DOPA) pathway in Mucuna pruriens (L.) DC have been performed. Spatio-temporal quantification of the putative l-DOPA biosynthetic pathway genes and its correlation with respective metabolites was established. l-tyrosine, l-DOPA, and dopamine from all plant parts were quantified. The de novo transcriptome analysis was performed using leaves of the selected M. pruriens mutant T-IV-9 during maturity. The putative L-DOPA pathway and its regulatory genes were retrieved from transcriptome data and the L-DOPA pathway was biochemically characterized. The spatial and temporal gene expression for the L-DOPA pathway was identified with respect to the chemical constituents. L-tyrosine, L-DOPA, and dopamine contents were highest in leaves during maturity (about 170-day-old plants). The polyphenol oxidase (PPO) was highly expressed in tender stems (230-fold higher as compared to seeds) as well as a high L-DOPA content. The PPO gene was highly expressed in leaves (3367.93 in FPKM) with a 79-fold increase compared to control plants during maturity. L-DOPA was found in every part with varied levels. The highest L-DOPA content was found in mature dried seed (3.18-5.8%), whereas the lowest amount was recorded in mature and dried leaves. The reproductive parts of the plant had a higher amount of L-DOPA content (0.9-5.8%) compared to the vegetative parts (0.2-0.91%). Various amino acid transporters and permeases were expressed in M. pruriens. The transcripts of dopa decarboxylase (DDC) were found in almost all parts of the plant, but its higher content was limited to the leaf.

In traditional, herbal medicine, and aromatherapy, use of essential oils and their aroma compounds have been known since long, for the management of various human diseases. The essential oil is a mixture of highly complex, naturally occurring volatile aroma compounds synthesized by medicinal and aromatic plants as secondary metabolites. Essential oils widely used in pharmaceutical, cosmetic, sanitary, food industry and agriculture for their antibacterial, antiviral, antifungal, antiparasitic, insecticidal, anticancer, neuroprotective, psychophysiological, and anti-aging activities. Moreover, volatile aroma compounds comprise a chemically diverse class of low molecular weight organic compounds with significant vapor pressure. However, aroma compounds produced by plants, mainly attract pollinators, seed dispersers and provide defense against pests or pathogens. However, in humans, about 300 active olfactory receptor genes are involved to detect thousands of different aroma compounds and modulates expression of different metabolic genes regulating human psychophysiological activity, brain function, pharmacological signaling, and therapeutic potential. Keeping in mind this importance, present database, namely, AromaDb (http://bioinfo.cimap.res.in/aromadb/) covers information of plant varieties/chemotypes, essential oils, chemical constituents, GC-MS profile, yield variations due to agro-morphological parameters, trade data, aroma compounds, fragrance type, and bioactivity details. The database includes 1,321 aroma chemical structures, bioactivities of essential oil/aroma compounds, 357 fragrance type, 166 commercially used plants, and their high yielding 148 varieties/chemotypes. Also includes calculated cheminformatics properties related to identification, physico-chemical properties, pharmacokinetics, toxicological, and ecological information. Also comprises interacted human genes affecting various diseases related cell signaling pathways correlating the use of aromatherapy. This database could be a useful resource to the plant's growers/producers, an aroma/fragrance industrialist, health professionals, and researchers exploring the potential of essential oils and aroma compounds in the development of novel formulations against human diseases.

Background: The study was carried out to assess the genetic variability present in ashwagandha and to examine the nature of associations of various traits to the root yield of the plant., Methods: Fifty-three diverse genetic stocks of ashwagandha ( Withania somnifera ) were evaluated for 14 quantitative characteristics. Analysis of variance, correlation, and path coefficient analysis were performed using the mean data of 2 years., Results: Analysis of variance revealed that the genotypes differed significantly for all characteristics studied. High heritability in conjunction with high genetic advance was observed for fresh root weight, 12 deoxywithastramonolide in roots, and plant height, which indicated that selection could be effective for these traits. Dry root weight has a tight linkage with plant height and fresh root weight. Further, in path coefficient analysis, fresh root weight, total alkaloid (%) in leaves, and 12 deoxywithastramonolide (%) in roots had the highest positive direct effect on dry root weight., Conclusion: Therefore, these characteristics can be exploited to improve dry root weight in ashwagandha genotypes and there is also scope for the selection of promising and specific chemotypes (based on the alkaloid content) from the present germplasm.

Purpose: To develop elite genotypes in Mucuna pruriens (L.) DC with high L-DOPA (L-3, 4 dihydroxyphenylalanine) yields, with non-itching characteristics and better adaptability by applying γ-irradiation. Molecular and chemical analysis was performed for screening based on specific characteristics desired for developing suitable genotypes., Materials and Methods: Developed, mutant populations were analyzed for L-DOPA % in seeds through TLC (thin layer chromatography), and the results obtained were validated with the HPLC (High performance liquid chromatography). The DNA (Deoxyribonucleic acid) was isolated from the leaf at the initial stage and used for DNA polymorphism. RNA (Ribonucleic acid) was isolated from the leaf during maturity and used for expression analysis., Results: The selected mutant T-I-7 showed 5.7% L-DOPA content compared to 3.18% of parent CIM-Ajar. The total polymorphism obtained was 57% with the molecular marker analysis. The gene expression analysis showed higher fold change expression of the dopadecarboxylase gene (DDC) in control compared to selected mutants (T-I-7, T-II-23, T-IV-9, T-VI-1)., Conclusion: DNA polymorphism was used for the screening of mutants for efficient screening at an early stage. TLC was found suitable for the large-scale comparative chemical analysis of L-DOPA. The expression profile of DDC clearly demonstrated the higher yields of L-DOPA in selected mutants developed by γ-irradiation in the seeds of the control.

Mentha arvensis (corn mint) is well known for the production of menthol, a widely used commodity in pharma and flavoring industries and provides natural fragrances and products. Glandular trichomes are specialized hairs found on the aerial surface of vascular plants species producing specific secondary metabolite chemistry. Correlations were established among trichomes, oil yield, and major secondary metabolites. Nine improved, elite cultivars representing different M. arvensis genotypes were used for analysis. Phenotypic coefficient of variation (PCV) and genotypic coefficient of variation (GCV) were estimated; results indicated the presence of considerable amount of genetic variability, thereby emphasizing wide scope of selection. Positive and significant associations were found among glandular trichomes, oil yield, essential oil constituents, and leaf morphology itself, whereas morphological parameters of leaf show positive and negative correlations to average number of trichome and essential oil constituents. Average number of glandular, non-glandular trichomes, their ratios, menthol content, and trichome number showed a good heritability. Trichomes were studied microscopically in leaf parts in all varieties for analyzing their distribution pattern. The trichome number variations showed significant correlation throughout the genotypes with essential oil yield and monoterpenoid constituents. Differential changes were analyzed for Glutathione S-transferases, Glutathione reductase, Malondialdehyde, phenolics, and chlorophyll content. Gene expressions were analyzed for biosynthesis genes and selected transcription factors TRANSPARENT TESTA GLABRA 1 (TTG1), ENOLASE 1, GLABRA 3, GTL 1, NUCLEAR TRANSCRIPTION FACTOR Y SUBUNIT B-6, WRKY transcription factor 22, putative WRKY 33, WRKY 17, WRKY 1, and WRKY 65-like for harnessing their relation with trichome development in M. arvensis genotypes.

Cobalt nitrate, nickel sulphate, hydrogen peroxide, sodium nitroprusside, and culture filtrates of Pseudomonas monteili, Bacillus circularans, Trichoderma atroviridae, and Trichoderma harzianum were tested to elicit ginsenoside production in a cell suspension line of Panax quinquefolius. Abiotic elicitors preferentially increased panaxadiols whereas biotic elicitors upregulated the panaxatriol synthesis. Cobalt nitrate (50 μM) increased total ginsenosides content by twofold (54.3 mg/L) within 5 days. It also induced the Rc synthesis that was absent in the control cultures. Elicitation with P. monteili (2.5 % v/v, 5 days) also supported 2.4-fold enhancement in saponin yield. Elicitation by T. atroviridae or hydrogen peroxide induced the synthesis of Rg3 and Rh2 that are absent in ginseng roots. The highest ginsenosides productivity (3.2-fold of control) was noticed in cells exposed to 1.25 % v/v dose of T. atroviridae for 5 days. Treating cells with T. harzianum for 15 days afforded maximum synthesis and leaching (8.1 mg/L) of ginsenoside Rh1.

Ocimum (Lamiaceae) is an important source of essential oils and aroma chemicals especially eugenol, methyl eugenol, linalool, methyl chavicol etc. An elite evergreen hybrid has been developed from Ocimum kilimandscharicum and Ocimum basilicum, which demonstrated adaptive behavior towards cold stress. A comparative molecular analysis has been done through RAPD, AFLP, and ISSR among O. basilicum and O. kilimandscharicum and their evergreen cold-tolerant hybrid. The RAPD and AFLP analyses demonstrated similar results, i.e., the hybrid of O. basilicum and O. kilimandscharicum shares the same cluster with O. kilimandscharicum, while O. basilicum behaves as an outgroup, whereas in ISSR analysis, the hybrid genotype grouped in the same cluster with O. basilicum. Ocimum genotypes were analyzed and compared for their trichome density. There were distinct differences on morphology, distribution, and structure between the two kinds of trichomes, i.e., glandular and non-glandular. Glandular trichomes contain essential oils, polyphenols, flavonoids, and acid polysaccharides. Hair-like trichomes, i.e., non-glandular trichomes, help in keeping the frost away from the living surface cells. O. basilicum showed less number of non-glandular trichomes on leaves compared to O. kilimandscharicum and the evergreen cold-tolerant hybrid. Trichomes were analyzed in O. kilimandscharicum, O. basilicum, and their hybrid. An increased proline content at the biochemical level represents a higher potential to survive in a stress condition like cold stress. In our analysis, the proline content is quite higher in tolerant variety O. kilimandscharicum, low in susceptible variety O. basilicum, and intermediate in the hybrid. Gene expression analysis was done in O. basilicum, O. kilimandscharicum and their hybrid for TTG1, GTL1, and STICHEL gene locus which regulates trichome development and its formation and transcription factors WRKY and MPS involved in the regulation of plant responses to freezing and cold. The analysis showed that O. kilimandscharicum and the hybrid were very close to each other but O. basilicum was more distinct in all respects. The overexpression of the WRKY coding gene showed high expression in the hybrid as compared to O. kilimandscharicum and O. basilicum and the transcription factor microspore-specific (MPS) promoter has also shown overexpression in the hybrid for its response against cold stress. The developed evergreen interspecific hybrid may thus provide a base to various industries which are dependent upon the bioactive constituents of Ocimum species.

Background: Ocimum sanctum L. (O. tenuiflorum) family-Lamiaceae is an important component of Indian tradition of medicine as well as culture around the world, and hence is known as "Holy basil" in India. This plant is mentioned in the ancient texts of Ayurveda as an "elixir of life" (life saving) herb and worshipped for over 3000 years due to its healing properties. Although used in various ailments, validation of molecules for differential activities is yet to be fully analyzed, as about 80 % of the patents on this plant are on extracts or the plant parts, and mainly focussed on essential oil components. With a view to understand the full metabolic potential of this plant whole nuclear and chloroplast genomes were sequenced for the first time combining the sequence data from 4 libraries and three NGS platforms., Results: The saturated draft assembly of the genome was about 386 Mb, along with the plastid genome of 142,245 bp, turning out to be the smallest in Lamiaceae. In addition to SSR markers, 136 proteins were identified as homologous to five important plant genomes. Pathway analysis indicated an abundance of phenylpropanoids in O. sanctum. Phylogenetic analysis for chloroplast proteome placed Salvia miltiorrhiza as the nearest neighbor. Comparison of the chemical compounds and genes availability in O. sanctum and S. miltiorrhiza indicated the potential for the discovery of new active molecules., Conclusion: The genome sequence and annotation of O. sanctum provides new insights into the function of genes and the medicinal nature of the metabolites synthesized in this plant. This information is highly beneficial for mining biosynthetic pathways for important metabolites in related species.

Sweet basil (Ocimum basilicum), a member of the Lamiaceae, is used as an ornamental as well as a culinary herb. It is a rich source of the phenolic compound methyl chavicol and is used as a traditional medicinal plant in India, where the crop is grown on ~2,500 ha annually (4). The species is native to India, where it has been cultivated for >5,000 years. During the rainy season, August of 2013, a severe leaf blight was observed on 30- to 45-day-old sweet basil plants in experimental fields (approximately 5 ha) at the CSIR-CIMAP and adjoining areas in Lucknow. Initial symptoms comprised small, irregular, necrotic lesions that coalesced into a leaf blight. Infected parts of the leaves turned black during wet and humid conditions. The incidence of symptoms ranged from 20 to 30%. Infected leaf samples were cut into small pieces and surface-sterilized with 1% sodium hypochlorite for 1 min, followed by two rinses in sterilized, distilled water. The leaf pieces were then blotted dry with sterilized filter paper, placed onto potato dextrose agar (PDA), and incubated at 28°C for 3 to 5 days. Blackish-brown fungal colonies developed. Microscopic examination revealed the presence of brown conidiophores that were cylindrical, septate, unbranched, and straight or geniculate near the apex. Conidia were three-septate, mostly curved at the third cell from the base, which was usually larger and darker than the other cells; intermediate cells were brown or dark brown; terminal cells were subhyaline or pale brown and 16 to 23.5 × 8.5 to 11.5 μm (the average size of 100 conidia was 19.9 × 10.18 μm). On the basis of these characteristics, the fungus was identified as Cochliobolus lunatus (anamorph Curvularia lunata (Wakk.) Boedijin) (1,2). The identification was confirmed by sequencing the internal spacer (ITS) region of ribosomal DNA (rDNA). Genomic DNA was extracted from five fungal isolates, using the 5 Prime Archive Pure DNA Cell/Tissue kit, and subjected to a polymerase chain reaction (PCR) assay with the universal primers ITS1 and ITS4 (5). The amplified product was cloned and sequenced. An NCBI-BLASTn search showed greatest homology (98% similarity) with the ITS sequence of C. lunatus (GenBank Accession No. DQ836800). The sequence was deposited in Genbank (KM272001). A pathogenicity test was carried out using 10, 30-day-old sweet basil (cv. CIM Soumya) plants in pots, by spraying a spore suspension (10 5 spores/ml) onto the leaves of each plant. Five plants treated similarly with sterilized, distilled water served as a control treatment. The plants were kept at 27 ± 2°C and 85 ± 3% RH for 8 to 10 days. Small, irregular, necrotic lesions appeared after 4 days on all inoculated leaves, while leaves of control plants remained asymptomatic. Fungi re-isolated from inoculated leaves resembled C. lunatus on the basis of microscopic and sequence data, fulfilling Koch's postulates. The fungus was not re-isolated from the control plants. C. guatemalensis has been reported to cause a leaf spot on sweet basil in Korea (3). To our knowledge, this is the first report of a sweet basil leaf blight caused by C. lunatus in India. C. lunatus has the potential to reduce the yield of sweet basil. References: (1) L. M. Liu et al. Plant Dis. 98:686, 2014. (2) D. S. Manamgoda et al. Fungal Divers. 56:131, 2012. (3) J. H. Park et al. Plant Dis. 96:580, 2012. (4) H. A. A. Taie et al. Not. Bot. Hort. Agrobot. Cluj. Napoca 38:119, 2010. (5) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, San Diego, 1990.

Background: Ocimum L. of family Lamiaceae is a well known genus for its ethnobotanical, medicinal and aromatic properties, which are attributed to innumerable phenylpropanoid and terpenoid compounds produced by the plant. To enrich genomic resources for understanding various pathways, de novo transcriptome sequencing of two important species, O. sanctum and O. basilicum, was carried out by Illumina paired-end sequencing., Results: The sequence assembly resulted in 69117 and 130043 transcripts with an average length of 1646 ± 1210.1 bp and 1363 ± 1139.3 bp for O. sanctum and O. basilicum, respectively. Out of the total transcripts, 59648 (86.30%) and 105470 (81.10%) from O. sanctum and O. basilicum, and respectively were annotated by uniprot blastx against Arabidopsis, rice and lamiaceae. KEGG analysis identified 501 and 952 transcripts from O. sanctum and O. basilicum, respectively, related to secondary metabolism with higher percentage of transcripts for biosynthesis of terpenoids in O. sanctum and phenylpropanoids in O. basilicum. Higher digital gene expression in O. basilicum was validated through qPCR and correlated to higher essential oil content and chromosome number (O. sanctum, 2n = 16; and O. basilicum, 2n = 48). Several CYP450 (26) and TF (40) families were identified having probable roles in primary and secondary metabolism. Also SSR and SNP markers were identified in the transcriptomes of both species with many SSRs linked to phenylpropanoid and terpenoid pathway genes., Conclusion: This is the first report of a comparative transcriptome analysis of Ocimum species and can be utilized to characterize genes related to secondary metabolism, their regulation, and breeding special chemotypes with unique essential oil composition in Ocimum.

Papaver somniferum produces therapeutically useful benzylisoquinoline alkaloids (BIAs) like papaverine, thebaine, codeine, and morphine that accumulate in its capsular latex. Morphine is a potent analgesic but is also abused as a narcotic, which has increased the demand for non-narcotic thebaine that can be converted into various analgesics. To curtail the narcotic menace, many distinct genotypes of the plant have been developed that are deficient in morphine and/or latex. Sujata is one such latex-less low alkaloid-producing variety developed from the alkaloid-rich gum harvest variety Sampada. Its utility for gene prospecting and studying differential gene regulation responsible for its low alkaloid, nutritive seed oil, and latex-less phenotype has been exploited in this study. BIA profiling of Sujata and Sampada capsules at the early and late stages indicated that except for thebaine, Sujata had a depressed alkaloid phenotype as compared to Sampada. Comparative transcript-based analysis of the two genotypes was carried out in the early stage capsule (higher thebaine) using subtractive hybridization and microarray. Interrogation of a P. somniferum array yielded many differentially expressing transcripts. Their homology-based annotation classified them into categories--latex related, oil/lipid related, alkaloid related, cell wall related, and others. These leads will be useful to characterize the highly sought after Sujata phenotype.

Centella asiatica germplasm collected from north, north-eastern and southern parts of India was compared for biomass and centellosides productivity under uniform agro-climatic conditions of the Indo-Gangetic plains at Lucknow. The highest biomass accumulation (411.9 g FW/m2 area) was recorded in accession A from north India, followed by 284.0, 135.7 and 29.2 g FW/m2 in accessions M, B and E from southern, eastern and north-eastern regions, respectively. Accession M possessed the highest asiaticoside content (52.1 mg/gDW) that was 1.58, 2.34 and 21.7 folds more than accessions A, B and E, respectively. The madecassoside level in leaves of accessions B and M was comparable (28.9 and 25.7 mg/gDW) and two folds more than accession A (13.9 mg/gDW). The madecassic and asiatic acid content in leaf tissue of all four accessions remained low in Lucknow. Amplified fragment length polymorphism (AFLP) analysis with 23 primers yielded 696 fragments, 563 of which were polymorphic. Accession M out-grouped with genetic dissimilarity indices of 83, 85 and 95% from accessions A, E and B, respectively. Commercial cultivation of accessions M and A through a four months growth cycle (June to September) in agro-climatic conditions of the Indo-Gangetic plains is suggested.

Hydroxylase/acetyltransferase elicitors and cyclooxygenase inhibitor along with various precursors from primary shikimate and secoiridoid pools have been fortified to vincamine less hairy root clone of Vinca minor to determine the regulatory factors associated with vincamine biosynthesis. Growth kinetic studies revealed that acetyltransferase elicitor acetic anhydride and terpenoid precursor loganin significantly reduce the growth either supplemented alone or in combination (GI = 140.6 ± 18.5 to 246.7 ± 24.3), while shikimate and tryptophan trigger biomass accumulation (GI = 440.2 ± 31.5 to 540.5 ± 40.3). Loganin also downregulates total alkaloid biosynthesis. Maximum flux towards vincamine production (0.017 ± 0.001 % dry wt.) was obtained when 20-day-old hairy roots were fortified with secologanin (10 mg/l) along with tryptophan (100 mg/l), naproxen (8.4 mg/l), hydrogen peroxide (20 μg/l), and acetic anhydride (32.4 mg/l). This was supported by RT PCR (qPCR) analysis where 2- and 3-fold increase in tryptophan decarboxylase (TDC; RQ = 2.0 ± 0.09) and strictosidine synthase (STR; RQ = 3.3 ± 0.36) activity, respectively, was recorded. The analysis of variance (ANOVA) for growth kinetics, total alkaloid content, and gene expression studies favored highly significant data (P < 0.05-0.01). Above treated hairy roots were also up-scaled in a 5-l stirred-tank bioreactor where a 40-day cycle yielded 8-fold increase in fresh root mass.

A genotype 'Sujata' developed earlier at CSIR-CIMAP from its parent 'Sampada' is considered to be the latex-less variety of Papaver somniferum. These two genotypes are contrasting in terms of latex and stem strength. Earlier we have carried out microarray analysis to identify differentially expressing genes from the capsules of the two genotypes. In this study, the peduncles of the two genotypes were compared for the anatomy revealing less number of laticifers in the cortex and vascular bundles. One of the important cell wall-related genes (for laccase) from the microarray analysis showing significantly higher expression in 'Sampada' capsule was taken up for further characterization in the peduncle here. It was functionally characterized through transient overexpression and RNAi suppression in 'Sujata' and 'Sampada'. The increase in acid insoluble lignin and total lignin in overexpressed tissue of 'Sujata', and comparable decrease in suppressed tissue of 'Sampada', along with corresponding increase and decrease in the transcript abundance of laccase confirm the involvement of laccase in lignin biosynthesis. Negligible transcript in phloem compared to the xylem tissue localized its expression in xylem tissue. This demonstrates the involvement of P. somniferum laccase in lignin biosynthesis of xylem, providing strength to the peduncle/stem and preventing lodging., (© 2013 Scandinavian Plant Physiology Society.)

Growth and in vitro asiaticoside accumulation in multiple shoot cultures of Centella asiatica (L.) Urban was studied as a function of nutrient manipulations in the culture media. Shoot cultures raised in liquid Murashige and Skoog medium supplemented with 2.5 mg/l kinetin attained a growth index (GI) of 6.06 along with the highest asiaticoside content of 3.8 mg/g dry weight on the 35th day of the culture cycle. The shoot growth and asiaticoside accumulation were found to be influenced by the relative proportions of NH(4)(+)-N:NO(3)(-)-N or Cu(2+) concentration in the medium. Asiaticoside content in shoots increased from 5.3 to 8.9 and 8.7 mg/g dry weight when total nitrogen concentration of 60 mM in the control medium was reduced to 50 and 40 mM with a corresponding change in NH(4) (+):NO(3)(-) ratio from 20:40 to 20:30 or 20:20, respectively. Total nitrogen level higher than 60 mM drastically reduced the asiaticoside concentration in these in vitro shoot cultures. Medium devoid of Cu(2+) significantly favored higher asiaticoside accumulation in the cultured tissue (7.05 mg/g dry weight) along with an improved biomass production (GI = 7.7) when compared with shoots reared on the control medium with 0.10 μM Cu(2+) (GI = 5.8; asiaticoside content = 4.4 mg/g dry weight). Carbohydrate enrichment of the medium by increasing the sucrose concentration from 3.0 to 5.0 or 7.0% was also beneficial for biomass and asiaticoside production with GI = 17.1 and 16.9 and asiaticoside content = 7.2 and 5.2 mg/g dry weight, respectively, in comparison to control cultures maintained on medium containing 3.0% sucrose. The procedure described here provides a viable production platform for generating clean and quality material from Centella with high bioactive content.

Velvet bean (Mucuna pruriens) seeds contain the catecholic amino acid L-DoPA (L-3,4-dihydroxyphenylalanine), which is a neurotransmitter precursor and used for the treatment of Parkinson's disease and mental disorders. The great demand for L-DoPA is largely met by the pharmaceutical industry through extraction of the compound from wild populations of this plant; commercial exploitation of this compound is hampered because of its limited availability. The trichomes present on the pods can cause severe itching, blisters and dermatitis, discouraging cultivation. We screened genetic stocks of velvet bean for the trichome-less trait, along with high seed yield and L-DoPA content. The highest yielding trichome-less elite strain was selected and indentified on the basis of a PCR-based DNA fingerprinting method (RAPD), using deca-nucleotide primers. A genetic similarity index matrix was obtained through multivariant analysis using Nei and Li's coefficient. The similarity coefficients were used to generate a tree for cluster analysis using the UPGMA method. Analysis of amplification spectra of 408 bands obtained with 56 primers allowed us to distinguish a trichome-less elite strain of M. pruriens.

A thin, profusely branched, fast growing hairy root line of Panax quinquefolium (American ginseng) was established by co-culturing epicotyl explants with a wild type strain of Agrobacterium rhizogenes. The transformed roots grew by over 10-fold from the initial inoculum within 8 weeks. The crude ginsenosides content in the roots was about 0.2 g/g dry wt level up to the 10th week of culture. Ginsenosides Rb2, Rd, Re, Rf and Rg1 constituted 47-49% of the crude saponin fraction between 6 and 8 weeks of growth whereas, Rc ginsenoside was accumulated only after 9th weeks when the biomass started receding. PCR amplification analysis of the hairy roots confirmed their transgenic nature by showing the presence of Ri-TL DNA with rolA, rolB and rolC genes in their genome.

Plantago ovata, popularly known as isabgol, has great commercial and medicinal importance due to thin rosy white membranous seed husk. Isabgol seeds and husks have emollient, demulcent and laxative properties. We used both biometric and molecular techniques to assess the genetic variability and relatedness of 80 germplasm accessions of Plantago spp (P. ovata, P. lanceolata, and P. major) collected both from India and abroad. The range of D2 values (2.01-4890.73) indicated a very high degree of divergence among the accessions. Based on the degree of divergence, 80 accessions/genotypes were grouped into seven clusters. Thirty-six accessions were analyzed through RAPD profiling for similarity and genetic distances, using 20 random primers. Intraspecific differences in all three species were smaller [range for P. ovata (2-17%), P. lanceolata (3-15%), P. major (2-11%)] than interspecific diversity. These highly divergent lines could be used to produce superior hybrids.

Isabgol is a medicinal plant known for its high-quality dietary fiber. The genetics and inheritance of economic characters, such as number of panicles, panicle length, seed yield, and swelling factor, were measured by diallel analysis of F(1) progenies from seven parents. The additive component of genetic variance was significant for days to flowering, plant height, branches/plant, peduncle length, panicle length, days to maturity, and swelling factor, whereas the dominance component of genetic variance was significant for all the characters except panicles/plant. Additivity was not significant for all the characters, indicating absence of non-allelic interactions (epistasis) in controlling gene expression. Heritability in the narrow sense was very high for panicle length, days to flowering, and plant height, and moderate for branches/plant, panicles/plant, days to maturity, seed yield, husk yield, and swelling factor. However, the degree of genetic improvement was only high for panicle length, seed yield and husk yield. We conclude that hybridizations, isolation of superior genotypes by sib selection and recurrent selection, and exploitation of hybrid vigor in specific parental-cross combinations are good strategies for isabgol crop improvement.

Two macromutants [unbranched (Ub) and restricted branched (Rb)], one point mutation [yellow flowerd ('Ada')] and two flowering variants (early and late) were isolated in the M2 generation of gammairradiated progenies of black henbane (H. niger) - a medicinal plant belonging to family Solanaceae. These major gene changes were coupled with simultaneous changes in several polygenic systems conditioning continuous variation for flowering time, plant height, numbers of shoots and leaves, biomass and crude drug content. Such quantitative variation in qualitative mutants were measured for both per se means and cross-bred performance in terms of: (1) shifts in univariate means, (2) changes in pooled divergence, (3) alterations in gca effects, and (4) changes in heterotic capacity. The mutants were significantly divergent from the parental base for all the six metric traits, both individually as well as collectively. Divergence among mutants was wider (D(2) = 1491 to 11 469) than that for mutants versus source parent (D(2) = 117 to 7120). Conspicuous alterations in both the nature and the magnitude of the gca effects associated with all mutants, except L-2013 (late flowering), relative to the parental base were observed for all of the metric traits examined (gca ∼- per se means). Similarly, the mutants manifested greater heterotic potential than the parental base. This is even more strongly reflected in mutant x mutant hybrids for all of the characters. The amount of economic heterosis for diverse traits followed the pattern: crude drug > biomass > flowering time ∼- plant height > number of shoots ∼- number of leaves.

The nature of gene action for biomass yield and tropane alkaloid content was ascertained through diallel analysis in Egyptian henbane, HYOSCYAMUS MUTICUS L. Dominance variation was preponderant over additive genetic variance, dominance being in the overdominance range for both traits. On an overall basis, recessive alleles had a positive (i.e. increasing) effect on both the characters. However, the positive or negative effects of recessive or dominant alleles were genotype-specific. Hence, transgressive breeding was suggested for isolating segregants with dominant alleles for herb yield and with recessives for alkaloid content so as to achieve overall high yields of tropane alkaloid per unit area.

Based on numerical and graphic analyses of 6 x 6 diallele cross progenies (n (2) = 36) over two environments, dominance was found to be most prevalent (in overdominance range without being inflated by non-allelic interactions) in parents for latex yield, and morphine, codeine, thebaine, and narcotine contents. However, the additive component was also significant for the former three traits. Dominant alleles mostly with positive effects were, by and large, asymmetrically distributed in parents except for narcotine content. Heritability estimates were moderate and corresponded with low genetic advance estimates for morphine, codeine, and thebaine contents. Ample genetic diversity among parents was reflected for all traits except narcotine content. On an overall basis, heterosis breeding was suggested to be a rewarding proposition for genetic improvement of the opium poppy ( PAPAVER SOMNIFERUM L.).

The efficacy of enzyme-linked immunosorbent assay (ELISA) in the diagnosis of gut tuberculosis was evaluated. The study included 95 patients with gut tuberculosis, 40 with pulmonary tuberculosis, 25 with non-tubercular intestinal diseases and 40 healthy controls. Mantoux response and antibody titre against PPD using ELISA were studied. The ELISA sensitivity in gut tuberculosis was 80% and in pulmonary tuberculosis 90%. It was also positive in 12% of patients with non-tubercular intestinal diseases and 5% of healthy controls. The sensitivity of Mantoux test in gut (77%) and pulmonary (84%) tuberculosis was comparable to that of ELISA. However, Mantoux test lacked diagnostic significance due to high positivity in non-tubercular disease patients (55%) and healthy controls (56%), whereas ELISA was highly specific (93%) for the diagnosis of gut tuberculosis.