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Hyptis brevipes Poit. is a native weed species of South and Central America and has been introduced and naturalized in many parts of SE Asia. It was reported from India for the first time in 1940 from Andaman and Nicobar Islands and then in 2012 from Darjeeling Himalaya. In the present study the authors have noticed the occurrence of the species in the wild in the Dima Hasao district of Assam and the Dampa Tiger Reserve of Mizoram. The study reveals that the species has naturalized in India in some parts of the Northeast region in cultivated lands along roadsides. The detailed taxonomic account of the species along with its distribution, phenology, uses and photographs is provided to confirm its identity and to take stock on its control from its further spread to some other regions, because due to the invasive nature it may affect the growth of crop plants.

The RNA-dependent RNA polymerase (RdRp) is one of the crucial enzymes in severe acute respiratory syndrome Coronavirus-2 (SARS-CoV-2) catalysing the replication of RNA, therefore acts as a potential target for antiviral drug design. The fixation of a ligand in the active site of RdRp may alter the SARS-CoV-2 life cycle. Present work aimed at identifying novel inhibitors of the SARS-CoV-2 RdRp enzyme by performing pharmacophore-based virtual screening, molecular docking and molecular dynamics simulation (MDS). Initially, the pharmacophore model of SARS-CoV-2 RdRp was constructed and the resulting model was used to screen compounds from ChEMBL, ZINC and PubChem databases. During the investigation, 180 compounds were screened using the above model and subjected to molecular docking with RdRp. Two compounds viz. ChEMBL1276156 and PubChem135548348 showed a strong binding affinity with RdRp than its standard inhibitor, Remdesivir. Toxicity prediction of these two compounds reveals their non-toxic nature. These compounds were further subjected to MDS for 100 ns to check their stability after binding with RdRp. The MDS of RdRp-ChEMBL1276156 and RdRp-PubChem135548348 complexes show enhanced stability in comparison to the RdRp-Remdesivir complex. The average interaction energy calculated after 100 ns of MDS was -146.56 and -172.68 KJ mol-1 for RdRp-CHEMBL1276156 complex and RdRp-PubChem135548348 complex, respectively, while -59.90 KJ mol-1 for RdRp-Remdesivir complex. The current investigation reveals that these two compounds are potent inhibitors of SARS-CoV-2 RdRp and they could be tested in the experimental condition to evaluate their efficacy against SARS-CoV-2.Communicated by Ramaswamy H. Sarma.

The present study for the first time investigated the effects of Salicylic acid (SA) (50, 100 and 200 mg/L) on

The present study carried out to investigate the bioactive chemical compounds, total polyphenol content and antioxidant potential of different extracts of the

Cotton (Gossypium spp.) is a major cash crop of India and is the second-largest cotton producer in the world after China. Gossypium hirsutum and Gossypium barbadense are two tetraploid species that are majorly cultivated besides two diploid species (desi cotton). In cotton, fiber quality, drought tolerance, boll weight, boll number, and yield are essential quantitative traits with many components that are controlled by several genes present at different loci. Identifying such genes from different genomic resources of cotton using various molecular markers is necessary to accelerate the Quantitative Trait Loci (QTL) analysis. In the public domain of cotton, there is a vast number of molecular markers. However, not all are very useful for trait mapping, as most markers are away from the QTL region. Thereby, cotton improvement programs pay more attention to tightly linked markers with high predictive trait values. The present review provides an overview and updates on the comparative studies and the application of various molecular markers, i.e., RFLP, AFLP, RAPD, SSR, EST-SSR, and SNP in the cotton-breeding program. Insights gained from the study may help in successful cotton breeding and improvement.

The leaves and fruits of Aegle marmelos (L.) have antidiabetic activity. However, the mode of action and molecules having antidiabetic activity are not known. Hence, we conducted molecular docking of phytochemicals with various molecular antidiabetic targets to find the same. Docking prioritized Dipeptidyl peptidase-4 (DPP-4) as the main target for phytochemicals of Aegle marmelos. DPP-4 inactivates intestinal peptides, glucagon-like peptide-1 (GLP-1), and Gastric inhibitory polypeptide (GIP). GLP-1 and GIP stimulate a decline in blood glucose levels, but DPP-4 inhibits their functions resulting high level of glucose. Hence inhibiting the activity of DPP-4 is a well-known strategy to treat Type 2 diabetes. Therefore, to find a mechanism that may be involved to act as a natural inhibitor of DPP-4, we screened five phytochemicals out of seventy-three based on Virtual Screening, ADMET Drug-likeness analysis, and PAINS filtering. Further, all five phytochemicals, i.e. Aegeline, Citral, Marmesinin, Auraptene, β-Bisabolene, and reference compound subjected MDS for analyzing the stability of docked complexes to assess the fluctuation and conformational changes during protein-ligand interaction. The values of RMSD, RG, RMSF, SASA, and Gibbs energy revealed the good stability of these phytochemicals in the active site pocket of DPP-4 in comparison to reference. Additionally, we have done the pharmacophore analysis, which revealed many common pharmacophore features between screened phytochemicals of A. marmelos and reference molecule. Our results show that these phytochemicals are potential antidiabetic candidates and can be further modified and evaluated to develop more effective antidiabetic drugs against DPP-4 to treat Type 2 Diabetes. Communicated by Ramaswamy H. Sarma.

Introduction: Dental caries is a chronic infectious disease caused by bacteria forming a biofilm formation on the tooth surface. Drug-resistant bacteria Streptococcus mutans (S. mutans) pose a vital public health issue. To overcome this, the development of effective drugs with a novel mechanism of action is very important. Drug repurposing is considered a viable alternative approach to overcome the above issue. In the present study, we have attempted to select unique and traditional source used as in traditional medicine. To continue this effort, in this study we used phytochemical of selected to find an effective cure for dental caries. Materials and methods: Inhibition of S. mutans deoxycytidylate deaminases (SmdCDs) is a promising drug development strategy against S. mutans, responsible for biofilm formation. Since no effective drugs are available against S. mutans, we conducted a virtual screening of phytochemicals to find novel compounds against this bacterium. Hence, we created a library of 871 phytochemicals from 13 plants with reported antimicrobial activity. The phytochemical library was subjected to virtual screening against the SmdCD target. Results and discussion: Traditionally, many cultures use chewing sticks for oral hygiene maintenance. In the present work, out of 871 phytochemicals, 211 phytochemicals showed the most druggable substance with zero violation from any of the drug-likeness rules. Further, the binding energy indicates the affinity of adhesion of protein structure docked with 2 hit potential herbal compounds of which Cyclocurcumin and Androsta-1,4,6-triene-3,17-dione showed the best docking with SmdCD. Conclusions: Based on the binding energy score, we suggest that hit compounds can be used for dental caries and further could be developed as an organic mouthwash. HIGHLIGHTS Streptococcus mutans is common in the human oral cavity and contributes significantly to tooth decay mutans deoxycytidylate deaminases (SmdCDs) are responsible for biofilm formation causing drug resistance SmdCDs can be used to develop effective drug against mutans, responsible for biofilm formation Screening phytochemicals may result potential compounds which can be used to develop new drugs against mutans GRAPHICAL ABSTRACT

Recently emerged SARS-CoV-2 is the cause of the ongoing outbreak of COVID-19. It is responsible for the deaths of millions of people and has caused global economic and social disruption. The numbers of COVID-19 cases are increasing exponentially across the world. Control of this pandemic disease is challenging because there is no effective drug or vaccine available against this virus and this situation demands an urgent need for the development of anti-SARS-CoV-2 potential medicines. In this regard, the main protease (Mpro) has emerged as an essential drug target as it plays a vital role in virus replication and transcription. In this research, we have identified two novel potent inhibitors of the Mpro (PubChem3408741 and PubChem4167619) from PubChem database by pharmacophore-based high-throughput virtual screening. The molecular docking, toxicity, and pharmacophore analysis indicate that these compounds may act as potential anti-viral candidates. The molecular dynamic simulation along with the binding free energy calculation by MMPBSA showed that these compounds bind to Mpro enzyme with high stability over 50 ns. Our results showed that two compounds: PubChem3408741 and PubChem4167619 had the binding free energy of − 94.02 kJ mol−1 and − 122.75 kJ mol−1, respectively, as compared to reference X77 (− 76.48 kJ mol−1). Based on our work’s findings, we propose that these compounds can be considered as lead molecules for targeting Mpro enzyme and they can be potential SARS-CoV-2 inhibitors. These inhibitors could be tested in vitro and explored for effective drug development against COVID-19. Graphic abstract Electronic supplementary material The online version of this article (10.1007/s11030-020-10148-5) contains supplementary material, which is available to authorized users.

Vegetation cover is very effective measure to control environmental conditions. In this study, the potential of Napier grass to reduce soil erosion was evaluated. Parameters include number of stems, shoot length, number of leaves, leaf area index and experiments were performed using a hydraulic tilting flume and controlled water circulation system. Extensive experiments were carried out using (i) control (ii) whole plant plot and (iii) roots plot at 2% slope with 10.7 l/s/m discharge rate. Results summarized that Napier grass produced significantly lesser soil erosion and runoff as compared to the control. Further, it was observed that the sediment outflow and runoff generation significantly decreased with the growth stage of Napier grass. Roots were found more effective in controlling sediment rates, while shoots were more effective in reducing runoff rate. The correlation analysis indicated that runoff and sediment outflow was highly negatively correlated with the leaf area index and the number of stems, respectively.

Diabetes mellitus (DM) is a complicated metabolic disorder with several enzymes, including α-amylase and α-glycosidase. The α-amylase is responsible for postprandial glucose levels; therefore, inhi...

Phyllanthus debilis, basically a species of coastal regions in India, is reported here for the first time from Assam State of North-East India. Earlier it was known only from Upper Siang District of Arunachal Pradesh and therefore it also forms an extended distribution in North-East region. The species is described and illustrated here for easy identification.

Genetic diversity among 55 different sugarcane (Saccharum) cultivars varying in their response for red rot disease (Colletotrichum falcatum) was studied using Resistance Gene Analog Polymorphism (RGAP) markers. A total of 28 primers were used in the present study to screen 30 moderately resistant, five resistant, seven moderately susceptible, nine susceptible and four highly susceptible cultivars resulted in the generation of 87 distinct fragments with 91.9% polymorphism. Polymorphism Information Content (PIC) and Resolving Power (RP) were also calculated and the values were found to be 16.28 and 52.36 respectively. On analyzing the dendrogram, a clear distinctive grouping was observed between red rot resistant and red rot susceptible cultivars. This study, therefore clearly revealed that Resistant Gene Analog (RGA) markers showed greater genetic diversity and hence, constitute a rich source for analyzing the genetic diversity and phylogenetic relationships. The genotypes belonging to different clusters can be crossbreed to gain high variability of good combinations of characters.

A recombinant inbred line mapping population of intra-species upland cotton was generated from a cross between the drought-tolerant female parent (AS2) and the susceptible male parent (MCU13). A linkage map was constructed deploying 1,116 GBS-based SNPs and public domain-based 782 SSRs spanning a total genetic distance of 28,083.03 cM with an average chromosomal span length of 1,080.12 cM with inter-marker distance of 10.19 cM.A total of 19 quantitative trait loci (QTLs) were identified in nine chromosomes for field drought tolerance traits. Chromosomes 3 and 8 harbored important drought tolerant QTLs for chlorophyll stability index trait while for relative water content trait, three QTLs on chromosome 8 and one QTL each on chromosome 4, 12 were identified. One QTL on each chromosome 8, 5, and 7, and two QTLs on chromosome 15 linking to proline content were identified. For the nitrate reductase activity trait, two QTLs were identified on chromosome 3 and one on each chromosome 8, 13, and 26. To complement our QTL study, a meta-analysis was conducted along with the public domain database and resulted in a consensus map for chromosome 8. Under field drought stress, chromosome 8 harbored a drought tolerance QTL hotspot with two in-house QTLs for chlorophyll stability index (qCSI01, qCSI02) and three public domain QTLs (qLP.FDT_1, qLP.FDT_2, qCC.ST_3). Identified QTL hotspot on chromosome 8 could play a crucial role in exploring abiotic stress-associated genes/alleles for drought trait improvement. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12298-021-01041-y.

The present study was aimed to develop a rapid in vitro propagation procedure for Quercus serrata. Seeds, (1) without seed coat (scarified); (2) scarified seeds cut at different lengths from distal ends; and (3) with seed coat (un-scarified), were subjected to various treatments. The scarified and half cut seeds, inoculated on woody plant medium supplemented with N6benzylaminopurine (BAP: 8.88 µM) plus indole-3-acetic acid (IAA: 2.86 µM), produced significantly (p

Efficient micropropagation procedure was developed for Origanum vulgare, a high-value culinary herb, and the phytochemicals, phenolic content, antioxidant and antimutagenic activity of leaf and stem, derived from different growing stages were analyzed. The agar solidified Murashige and Skoog (MS) medium supplemented with a combination of 6-benzylaminopurine and α-naphthaleneacetic acid was optimized as best shoot-multiplication-medium. Shoots were rooted best on 1/2 strength MS medium supplemented with 50 µM indole-3-butyric acid (IBA). The plantlets were successfully acclimatized ex vitro in a soil, sand and farmyard manure mixture (2:1:1 v/v/v) with 100% survival rate in greenhouse. The total anthocyanin and total phenolic content were observed significantly higher in leaves of in vitro-raised plants. However, total tannin, flavonoid and antioxidant activity remained higher in leaves of mother plant maintained under ployhouse condition. All the plant extracts have shown significant antimutagenic activity except in vitro-growing plants. A total of 13 polyphenolic compounds were detected in different extracts using high performance liquid chromatography. Among these, catechin was detected maximum in in vitro-growing cultures and chlorogenic acid in leaves of mother plant. These findings will help the farmers, medicinal plant growers, and industries for mass multiplication and effective extraction of phytochemicals from O. vulgare.

A computational investigation was carried out to find out potential phytochemicals that could inhibit the binding of human angiotensin-converting enzyme-2 (ACE2) receptors to spike protein of SARS-CoV-2 which is an essential step to gain entry inside human cells and onset of viral infection known as Coronavirus disease (COVID-19). A library of phytochemicals was screened by virtual screening against ACE2 receptors resulting in twenty phytochemicals out of 686 which had binding energy (−11.8 to −6.9 kcal/mol). Drug-likeness gave five hits, but ADMET analysis yielded 4 nontoxic hit phytochemicals. Molecular dynamics simulation of four-hit compounds resulted in acceptable stability and good dynamics behavior. These phytochemicals are Hinokinin, Gmelanone, Isocolumbin, and Tinocordioside, from Vitis vinifera, Gmelina arborea, and Tinospora cordifolia. The above-mentioned phytochemicals may be promising ACE2 inhibitors and can prevent infection of SARS-CoV-2 by inhibiting the entry of the virus into host cells. Communicated by Ramaswamy H. Sarma

The recent outbreak of SARS-CoV-2 disease, also known as COVID-19, has emerged as a pandemic. The unavailability of specific therapeutic drugs and vaccines urgently demands sincere efforts for drug discovery against COVID-19. The main protease (Mpro) of SARS-CoV-2 is a critical drug target as it plays an essential role in virus replication. Therefore for the identification of potential inhibitors of SARS-CoV-2 Mpro, we applied a structure-based virtual screening approach followed by molecular dynamics (MD) study. A library of 686 phytochemicals was subjected to virtual screening which resulted in 28 phytochemicals based on binding energy. These phytochemicals were further subjected to drug-likeness and toxicity analysis, which resulted in seven drug-like hits. Out of seven, five phytochemicals viz., Mpro-Dehydrtectol (-10.3 kcal/mol), Epsilon-viniferin (-8.6 kcal/mol), Peimisine (-8.6 kcal/mol), Gmelanone (-8.4 kcal/mol), and Isocolumbin (-8.4 kcal/mol) were non-toxic. Consequently, these phytochemicals are subjected to MD, post MD analysis, and MM/PBSA calculations. The results of 100 ns MD simulation, RMSF, SASA, Rg, and MM/PBSA show that Epsilon-viniferin (-29.240 kJ/mol), Mpro-Peimisine (-43.031 kJ/mol) and Gmelanone (-13.093 kJ/mol) form a stable complex with Mpro and could be used as potential inhibitors of SARS-CoV-2 Mpro. However, further investigation of these inhibitors against Mpro receptor of COVID-19 is needed to validate their candidacy for clinical trials. Communicated by Ramaswamy H. Sarma

With globalisation and growing health risks of synthetic colourants, search for pigments from natural sources has increased owing to their non-toxic nature. The present study highlights the bioprospection of natural pigment from a cold adapted fungal strain of Penicillium sp. (GBPI_P155), isolated from soil of Indian Himalayan region. The fungus produced insoluble and orange-coloured pigment in liquid medium with maximum production recorded in potato dextrose (PD) broth at 15°C and 3 pH, while maximum biomass was produced at 25°C and pH 3. While examining the effect of different mineral salts, and carbon and nitrogen sources on pigment production, maximum accumulation of pigment was recorded in PD broth supplemented with 2% maltose. Following production, extraction of pigment was performed using chloroform and characterised partially by UV/vis (λmax at 495 nm and a shoulder peak at 530 nm) and Fourier Transform Infrared (FT-IR) spectroscopy. Thin layer chromatography of chloroform extract resulted in separation of pigment in three fractions with Rf values 0.911, 0.852 and 0.808, which were further analysed using Liquid Chromatography Mass Spectrometry (LC/MS). The overall approach resulted in identification of pigment as a mixture of different derivatives of carotenoids. The extracted pigment also possessed antimicrobial activity against Gram-positive and Gram-negative bacteria and actinobacteria.

Estimation of variability and genetic relationships among breeding materials is one of the important strategies in crop improvement programs. Morphological (plant height, spike length, a number of florets/spike), physiological (chlorophyll content, chlorophyll fluorescence, and rapid light curve parameters) and Directed amplification of minisatellite DNA (DAMD) markers were used to investigate the relationships among 50 Gladiolus cultivars. Cluster analysis based on morphological data, physiological characteristics, molecular markers, and cumulative data discriminated all cultivars into seven, five, seven, and six clusters in the unweighted pair-group method using arithmetic mean (UPGMA) dendrogram, respectively. The results of the principal coordinate analysis (PCoA) also supported UPGMA clustering. Variations among the Gladiolus cultivars at phenotypic level could be due to the changes in physiology, environmental conditions, and genetic variability. DAMD analysis using 10 primers produced 120 polymorphic bands with 80% polymorphism showing polymorphic information content (PIC = 0.28), Marker index (MI = 3.37), Nei's gene diversity (h = 0.267), and Shannon's information index (I = 0.407). Plant height showed a positive significant correlation with Spike length and Number of florets/spike (r = 0.729, p

The major aim of the present study was to: (i) develop an efficient clonal in vitro propagation procedure, and (ii) investigate and compare phytochemicals, antioxidant, and DNA damage protection activity in methanolic extracts of in vitro raised and mother plant leaves of Quercus serrata. The study succeeded in establishing an efficient clonal micropropagation procedure using nodal segments taken from in vivo growing mother plant. The WP medium supplemented with six-benzyleaminopurine (BAP: 8.88 μM) plus indole-3-acetic acid (IAA: 1.43μM) produced the highest number of shoots/explant having maximum average shoot length. In vitro derived shoots exhibited best rooting upon application of 100μM indole-3-butyric acid for 48 h. Among studied phytochemicals, the total phenol content was recorded higher from in vitro raised plants and total flavonoid in the mother plant. Antioxidant activities were recorded higher in case of in vitro raised plants. Also, the methanolic leaf extract of in vitro raised plants exhibited higher antimutagenic activity that of mother plant. These findings indicate Q. serrata as a promising source of natural antioxidants that could be utilized commercially for the prevention of photoaging and oxidative stress-mediated skin diseases.

We report the development of 12 novel microsatellite (SSR) markers isolated from repeat sequences of chloroplast region of Gladiolus . A total of 1188 nucleotide sequences of chloroplast regions of Gladiolus were used to search for simple sequence repeats and 108 sequences were found to contain 124 SSRs. Dinucleotide repeat SSRs were most frequent (94.4%) with the AT/AT (59.7%) repeat motif followed by TA/TA (31.5%). Assessment of genetic diversity among 62 commercially important Gladiolus cultivars was carried out using 12 SSR markers. Out of 12 SSR markers, only six showed polymorphism with allelic variation ranging from 2 to 8. These six SSR markers revealed high genetic diversity (84.6%) across different Gladiolus cultivars. The average value of Polymorphic information content (PIC = 0.32) diversity index (2.03), expected heterozygosity (0.354) and Shannon's information index (0.522) were also significantly high. These markers could be utilized for future breeding programmes, genetic diversity studies, and germplasm management of this important ornamental crop.

Gladiolus is an economically important ornamental crop, cultivated for its beautiful flowers throughout the world. The correct genotype identification of plant material is very significant for the floriculture industry. The aim of this study was to develop sequence-characterised amplified region (SCAR) markers from RAPD and ISSR fragments for identification and authentication of Gladiolus germplasm. The SCAR markers developed could be easily employed as valuable tools to identify newly developed Gladiolus cultivars. The SCAR markers, viz. ScG12, ScG34, and ScG36, are specific to the DNA from all 62 Gladiolus cultivars, as they did not amplify the DNA of other taxa of the family Iridaceae, including Iris, Amaryllis, and Narcissus. All three SCAR markers distinguished Gladiolus from other taxa of the family Iridaceae, whereas marker ScAm was specific to the ‘Amethyst’ cultivar. Our results confirmed that this particular SCAR marker distinguished the ‘Amethyst’ cultivar from the other 62 Gladiolus cu...

The plastid genome regions of two intergenic spacers, psbA-trnH and trnL-trnF, were sequenced to study the nucleotide diversity and phylogenetic relationships among Gladiolus cultivars. Nucleotide diversity of psbA-trnH region was higher than trnL-trnF region of chloroplast. We employed Bayesian, maximum parsimony (MP) and neighbour-joining (NJ) approaches for phylogenetic analysis of Gladiolus and related taxa using combined datasets from chloroplast genome. The psbA-trnH and trnL-trnF intergenic spacers of Gladiolus and related taxa-like Babiana, Chasmanthe, Crocus, Iris, Moraea, Sisyrinchium, Sparaxis and two out group species (Hymenocallis littoralis and Asphodeline lutea) were used in the present investigation. Results showed that subfamily Iridoideae have sister lineage with subfamily Ixioideae and Crocoideae. H. littoralis and A. lutea were separately attached at the base of tree as the diverging Iridaceae relative's lineage. Present study revealed that psbA-trnH region are useful in addressing questions of phylogenetic relationships among the Gladiolus cultivars, as these intergenic spacers are more variable and have more phylogenetically informative sites than the trnL-trnF spacer, and therefore, are suitable for phylogenetic comparison on a lower taxonomic level. Gladiolus cultivars are extensively used as an ornamental crop and showed high potential in floriculture trade. Gladiolus cultivation still needs to generate new cultivars with stable phenotypes. Moreover, one of the most popular methods for generating new cultivars is hybridization. Hence, information on phylogenetic relationships among cultivars could be useful for hybridization programmes for further improvement of the crop.

Genetic diversity and population structure in Bergenia stracheyi, a threatened medicinal herb in the Western Himalaya of India was analysed using directed amplification of minisatellite DNA (DAMD) and inter simple sequence repeats (ISSR) markers. A total of 41 accessions of B. stracheyi representing three populations (Khillenmarg –KLM, Jalori Pass-JLP and Rohtang-RTG) were considered in the present study. The cumulative data analysis for 26 (10 DAMD + 16 ISSR) markers revealed 87.1% polymorphism. The maximum inter-population genetic distance was found between KLM and JLP, whereas the minimum genetic distance was found between RTG and JLP populations. The analysis of molecular variance (AMOVA) revealed maximum percentage of variation among individuals within populations (75%) than among the populations (25%). Clustering pattern of the three sample populations in STRUCTURE and PCoA analyses showed high genetic variation at population level. The present study revealed that distribution patterns, high altitudinal ranges, high habitat specificity, relatively high gene flow, small and isolated population size have shaped the current population structure of B. stracheyi in the Western Himalayan region. DAMD and ISSR markers have provided significant insights into characterization of B. stracheyi populations, and facilitate selection of appropriate accessions for further utilization in conservation and bioprospecting programmes.

In vitro regeneration of sweet orange (Citrus sinensis (L.) Osbeck Family: Rutaceae) has been performed via direct and indirect organogenesis. For indirect organogenesis, callus was induced and proliferated from leaf explants derived from in vitro grown seedlings on Murashige and Skoog (MS) media containing 2,4-dichlorophenoxyacetic acid (2,4-D) alone or in combination with benzyl adenine (BA) and α-naphthalene acetic acid (NAA). For direct organogenesis, explants were placed on MS media containing BA alone or in combination of NAA and gibberellic acid (GA3). Well-developed microshoots were treated with different concentrations of NAA for rhizogenesis (a two-step procedure). Different responses to these treatments were recorded depending upon the procedure used. It appears that 4.53 μM 2,4-D in combination with 5.37 μM NAA induced 93.33% callus and proliferate 86.67% of callus into 6.93 shoots per explant. Exogenous addition of 4.44 μM BA in combination with 1.54 μM GA3 enhanced shoot multiplication rate significantly (17.73±1.69 shoots/explant) in comparison to control (0.00±0.00 shoots/explant). Microshoots were rooted best (75.00±14.43%) under the treatment 100μM NAA for 48 hrs. and rooted plantlets were transferred to soil, following acclimatization were taken to maturity in the polyhouse. Key words: Malta, Himalaya, benzyl adenine (BA), callus.

Genetic variability and relationships in 54 genotypes belonging to nine Gladiolus cultivars were determined using single primer amplification reaction methods (Random Amplified Polymorphic DNA—RAPD; Directed Amplification of Minisatellite DNA—DAMD; and Inter Simple Sequence Repeats—ISSR). A total of 30 primers (10-RAPD, 10-DAMD and 10-ISSR) resulted into 230 DNA fragments, of which 177 (75.8 %) were polymorphic in nature. The polymorphic information content (PIC = 0.18), resolving power (RP = 11.2) and marker index (MI = 11.4) was found to be highest in ISSR markers. To understand the extent of genetic variability among the Gladiolus cultivars, the cumulative dataset were used to generate UPGMA dendrogram. All the 54 genotypes clustered together in nine broad clusters, revealing that the clustering pattern of the genotypes were as per their cultivars. PCoA analyses also revealed similar clustering patterns along the axis and therefore corroborating the grouping of genotypes in UPGMA dendrogram. The STRUCTURE analysis showed the admixture of alleles and resulted into eight genetic populations. The present study demonstrates the efficiency and discriminatory potential of an individual as well as cumulative analysis of RAPD, DAMD and ISSR markers to estimate the genetic variability and inter-relationships among indigenously developed Gladiolus cultivars. Furthermore, documentation and preservation of genetically diverse Gladiolus germplasm could be an important source for improvement of this floricultural crop.

Ethnopharmacological relevance Phyllanthus emblica Linn. (Syn. Emblica officinalis Gaertn.), has been used to cure many ailments of human beings. Literature survey demonstrates that it has many pharmacological activities i.e. antidiabetic, antioxidant, anti-microbial, antifungal, antiallergic, antiviral, and anticancer properties. Aim of the study The present study aimed to identify the novel plant-derived antidiabetic compounds from P. emblica to understand the molecular basis of antidiabetic activities. Material and methods Text mining analysis of P. emblica and its disease association was carried out using server DLAD4U. Due to the highest score of P. emblica with diabetes, the virtual screening of a phytochemical library of P. emblica against three targets of diabetes was carried out. After that FAF-Drug4, admetSAR and DruLiTo servers were used for drug-likeness prediction. Additionally, pharmacophore modeling was also carried out to understand the antidiabetic activity of screened compounds. Results The docking scores, drug-likeness and pharmacophore studies found that Ellagic acid, Estradiol, Sesamine, Kaempferol, Zeatin, Quercetin, and Leucodelphinidin are potential antidiabetic compounds. Conclusions Our study shows that phytochemicals of P. emblica are very potential antidiabetic candidates. Using the modern techniques these molecules could be used to develop an effective antidiabetic drugs from a natural resource.

Berberis aristata, an endangered plant, is an important component in different medicinal formulations. The low seed germination rate coupled with fructification at high altitudes during the rainy season makes the task of seed collection difficult. In order to increase the yield of relevant plant material for medicinal and conservation purposes, an efficient in vitro regeneration protocol is important. Regeneration of endangered B. aristata from leaf-derived callus may offer an efficient alternative to natural propagation through seeds. When compared with nodal segments taken from in vitro-germinated seedlings and inoculated onto the same medium, leaf-derived callus was found to be better for shoot induction and multiplication. In optimization experiments, the rate of shoot multiplication from leaf-derived callus was increased from 17.6 ± 0.6 to 26.5 ± 0.2 shoots per callus by supplementing Woody Plant (WP) medium with 0.5 μM thidiazuron (TDZ) in place of 8.88 μM benzyladenine (BA) + 2.68 μM naphthalene acetic acid (NAA). Although shoot length was decreased when using TDZ, this could be alleviated by gibberellic acid (GA3) treatment. Rooting was achieved on half-strength WP medium supplemented with indole butyric acid (IBA) in either a one- or two-step rooting method. The two-step rooting procedure was found to be superior; under the best overall conditions, 55.6 ± 2.1% of plantlets formed roots within 25.7 ± 0.3 d. Well-rooted plantlets were successfully established in earthen pots under greenhouse conditions, with a 61.1% survival rate. This procedure is suitable for large-scale production of plants via in vitro propagation for this endangered plant species.

Genetic variability and population structure of Bergenia ciliata (Haw.) Sternb., commonly known as “ Pashanbheda ” (Stone-breaker), collected from the Western Himalayan region of India were estimated using two DNA fingerprinting methods viz ., directed amplification of minisatellite DNA (DAMD) and inter simple sequence repeats (ISSR). The cumulative data analysis of DAMD and ISSR markers for 74 accessions from eight populations showed 86.1% polymorphism. Analysis of molecular variance (AMOVA) showed highest percentage of variation within individuals of populations (73.6%) and 21.7% among populations. STRUCTURE and PCoA analyses on the hierarchical partitioning of genetic diversity showed strong admixture of individuals among the eight assumed geographical populations of B. ciliata . The data suggests that high genetic flow is one of the major factors responsible for low genetic differentiation. Preservation of genetic diversity of B. ciliata is important, both to promote adaptability of the populations to changing environment as well as to preserve a large gene pool for future prospection. The present study using DAMD and ISSR markers, therefore, provide the means of rapid characterization of accessions within the populations, and thus enable the selection of appropriate accessions for further utilization in conservation and prospection programmes.

With globalisation and growing health risks of synthetic colourants, search for pigments from natural sources has increased owing to their non-toxic nature. The present study highlights the bioprospection of natural pigment from a cold adapted fungal strain of Penicillium sp. (GBPI_P155), isolated from soil of Indian Himalayan region. The fungus produced insoluble and orange-coloured pigment in liquid medium with maximum production recorded in potato dextrose (PD) broth at 15°C and 3 pH, while maximum biomass was produced at 25°C and pH 3. While examining the effect of different mineral salts, and carbon and nitrogen sources on pigment production, maximum accumulation of pigment was recorded in PD broth supplemented with 2% maltose. Following production, extraction of pigment was performed using chloroform and characterised partially by UV/vis (λ max at 495 nm and a shoulder peak at 530 nm) and Fourier Transform Infrared (FT-IR) spectroscopy. Thin layer chromatography of chloroform extract resulted in separation of pigment in three fractions with Rf values 0.911, 0.852 and 0.808, which were further analysed using Liquid Chromatography Mass Spectrometry (LC/MS). The overall approach resulted in identification of pigment as a mixture of different derivatives of carotenoids. The extracted pigment also possessed antimicrobial activity against Gram-positive and Gram-negative bacteria and actinobacteria.

Sugarcane is the major source of sugar in Asia and Europe, grown primarily in the tropical and sub-tropical zones of the world. The main disease responsible for its low yield is red rot. Therefore, in the present study, characterization of red rot disease was performed among 55 different sugarcane cultivars varying in red rot resistance level. 18 fragments were found to be associated with red rot resistance and were identified as resistant specific markers. The resistant specific fragments were amplified by RGA169, RGA396, RGA129, RGA231, RGA251, RGA057, RGA118, RGA152, RGA327, RGA542, RGA012, RGA173, RGA184, RGA275, RGA019, RGA267, RGA281 and RGA533. 7 fragments were found to be associated with red rot susceptibility and were considered as susceptible specific markers amplified by RGA088, RGA162, RGA396, RGA231, RGA251, RGA087 and RGA275. Sequencing of five resistant fragments, viz., RGA169, RGA231, RGA251, RGA267 and RGA533 was performed and the data thus obtained showed 80-99% similarity when compared with other resistant gene sequences previously submitted in NCBI database.

An efficient micropropagation protocol has been developed for tasar oak. Nodal segments from in vitro grown seedling were used for shoot multiplication. Best shoot multiplication response, in terms of number of shoots per explant as well as shoot length, was obtained in woody plant (WP) medium supplemented with 6-benzyladenine and indole-3-acetic acid (8.88 μM BA + 1.43 μM IAA); but the establishment of cultures was difficult due to basal callus formation and necrosis in due course of time. Out of the two used growth adjuvants, casein hydrolysate (CH, 500 mg L−1) promoted shoot multiplication rate significantly in comparison to silver nitrate and also eliminated the basal callus formation problem and necrosis faced during the later stage of shoot proliferation. In vitro rooting on WP medium supplemented with 100 μM indole-3-butyric acid (IBA) when applied for 48 hr gave the best results in comparison to prolonged exposure. Well-acclimatized plantlets were transferred to the field with 80% survival rate. T...

This protocol has been developed for somatic embryogenesis in Hedychium spicatum. Simultaneously, a method has also been developed for the production of synthetic seeds by using somatic embryos. Direct somatic embryos were developed on cotyledon explants of zygotic embryos on MS supplemented with high concentration of NAA (20.0 µM). Induction of secondary embryogenesis was best in 2,4-D supplemented medium fortified with activated charcoal. Germination of somatic embryos was enhanced by using GA3. Besides this, round and semi-hard beads of somatic embryos (synthetic seeds) could be produced by using 2% Na-alginate and 100 mM calcium chloride and more than 30% germination of synthetic seeds was achieved in MS. Well acclimated plants produced via somatic embryogenesis and/or synthetic seeds were transferred to field where more than 60% survived. This simple study enabled us to obtain a number of plantlets throughout the year each cycle requiring a short period of time. Besides propagation, this study provided an ex situ method for conservation of this vulnerable Himalayan species.D. O. I.http://dx.doi.org/10.3329/ptcb.v23i2.17506Plant Tissue Cult. & Biotech. 23(2): 147-155, 2013 (December)

Microalgae have gained enormous consideration from scientific community worldwide emerging as a viable feedstock for a renewable energy source virtually being carbon neutral, high lipid content, and comparatively more advantageous to other sources of biofuels. Although microalgae are seen as a valuable source in majority part of the world for production of biofuels and bioproducts, still they are unable to accomplish sustainable large-scale algal biofuel production. Wastewater has organic and inorganic supplements required for algal growth. The coupling of microalgae with wastewater is an effective way of waste remediation and a cost-effective microalgal biofuel production. In this review article, we will primarily discuss the possibilities and current scenario regarding coupling of microalgal cultivation with biofuel production emphasizing recent progress in this area.

Dactylorhiza hatagirea (D. Don) Soo; a source of high quality ‘salep’ used as nervine tonic, has been categorized as critically endangered due to over-exploitation from the wild. This report deals with the propagation of this species using both in vitro and conventional methods. For green pod culture, four nutrient media, Knudson C (KC), Murashige and Skoog (MS), Vacin and Went (VW) and Vejsadova (VJ) were tested by adding different growth additives. MS medium supplemented with peptone (P) (1.0 g/L), morphoinoethane sulphonic acid (MES) (1.0 g/L) and activated charcoal (AC) (0.1%) was found to be the most effective medium for the development of protocorm like bodies (PLBs), development of chlorophyll and for the plantlet formation. To improve vegetative multiplication, tubers were treated with α- naphthalene acetic acid (NAA), indole-3- butyric acid (IBA) and indole acetic acid (IAA) before planting. Rooting was observed in only apical segments. Maximum rooting (38.88%) was induced with 50.0 μM IBA. This study is helpful in the propagation of this endangered orchid at lower elevation which could be a successful effort of conservation of this endangered orchid at lower elevation and this could also reduce pressure on its population in the wild. Keywords: In vitro propagation, protocorm like bodies (PLBs), vegetative propagation, conservation

A study concerning the influence of cytokinins on shoot regeneration by using different stem segments derived from in vitro raised seedlings and their subsequent rooting was conducted in banj oak (Quercus leucotrichophora L.). Cytokinins play an important role in shoot regeneration and their multiplication. In the present study, cytokinins particularly kinetin (Kn) influenced in vitro rooting and subsequent survival of these in vitro raised plants in addition to shoot multiplication. It was found that the microshoots raised via kinetin treatment rooted very well (94.44%) without any basal callus formation in comparison to microshoots raised via other cytokinin treatments (that is, BA and 2-iP). In addition to this, these in vitro raised plants showed maximum survival rate (90%) during hardening process. On the basis of available literature this is a unique and significant study regarding the comparative effect of different cytokinins on in vitro propagation study of Q. leucotrichophora by using different stem segments, particularly the influence of kinetin in vitro rooting and survival of in vitro raised plants in addition to shoot multiplication. This significant study could be useful for large scale production of successfully hardened plants so that it would be helpful in conservation of this important Himalayan forest tree species.Keywords: Quercus leucotrichophora, acclimatization, cytokinins, in vitro rooting

Mustard aphid, Lipaphis erysimi (Kalt.) is a major insect pest of rapeseed-mustard group of crops. The economic importance of this has prompted fundamental research on it including gene expression and gene silencing studies. To identify key genes regulating parthenogenetic reproduction, gene-expression pattern of five developmental genes have been studied across three developmental stages namely, nymph, third instar and adult. All the five genes showed a constitutive basal level as well as significant up-or down-regulation over and above the basal level, across the developmental stages. While LeENDO, LeSTRN and LeFAS genes showed decrease in gene expression as the insect matures to parthenogenetic mother; and LeRNH and LeSNIV showed continuous increase in transcript level peaking in alate mother. Interestingly, all the genes followed a sinusoidal pattern of gene-expression either directly or inversely related to progression of developmental stages from newborn nymph to adult parthenogenetic mother in L. erysimi. In addition, a phylogenetic analysis of these genes revealed extent of resemblance and divergence of the gene sequences with their homologs from other insects including aphid species and pollinators.