Assessment of genetic diversity in Solanum trilobatum L., an important medicinal plant from South India using RAPD and ISSR markers

Solanum trilobatum L. is an Indian medicinal plant containing rich amount of steroidal glyco-alkoloids that can be used as precursor for commercial steroid production. Two efficient marker systems such as Random Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeats (ISSR) were used for the first time to assess the genetic diversity across 14 S. trilobatum accessions obtained from five South Indian states. Twenty out of 60 RAPD primers generated 189 distinct bands of which 160 were polymorphic with an average of 8 polymorphic bands per primer. A maximum of up to 15 fragments were amplified with an average of 9.45 bands per primer and the amplicons varied in size between 100 and 3,000 bp. The percentage of polymorphism ranged from 55.5 to 100, with an average of 84.6. ISSR profiling using 7 out of 20 primers amplified 83 bands and the number of amplified fragments varied from 2 to 16 with a size range of 200–1,800 bp. Totally 72 polymorphic bands were obtained using 7 ISSR primers at an average of 10.28 polymorphic bands per primer. Polymorphism percentage varied from 50 to 100 among the selected accessions resulting in an average percentage of polymorphism of 86.7. The PIC values ranged from 0.49 to 0.93 for RAPD and 0.16 to 0.90 for ISSR primers. The study pointed out that ISSR markers were more efficient than RAPD markers in evaluating the degree of genetic variation in S. trilobatum. The UPGMA cluster analysis grouped all Tamil Nadu accessions in one cluster and other state accessions in another cluster. The Principal component analysis also substantiates this clustering pattern. Thus the phylogenetic relationship and a high genetic variation revealed in the present study could provide a baseline data for conservation and improvement of this plant in future. Also the molecular markers identified in this study will be helpful in authentication of this species to prevent adulteration in herbal medicine.     

Sprouted Sorghum Extract Elicits Coleoptile Emergence,Enhances Shoot and Root Acclimatization,and Maintains Genetic Fidelity in indica Rice

The high growth-stimulating effect of plant extract has urged the plant biotechnologists to use natural supplements in the culture media instead of synthetic phytohormones. We advocated the effect of sprouted sorghum extract(SSE) on emergence, in vitro acclimatization, and genetic fidelity in coleoptile derived callus of indica rice variety ADT36. The use of SSE with Murashige Skoog medium efficiently acclimatized the root and shoot apical systems. A higher mat and seminal roots(3.4 g biomass) with an efficient shoot primordium elongation were observed with an increase in the concentration of SSE. Seeds treated with SSE medium showed higher germination and earlier coleoptile maturation about 48 h compared to untreated seeds, and there was a higher expression of e EF-1α with an increase in coleoptile length. B5 medium was effective on inducing embryogenic and nodular callus from 3-day-old coleoptile with 3.0 mg/L 2,4-dichlorophenoxyacetic acid and further proliferated effectively with 0.8 mg/L kinetin with a fresh weight of 180 mg. Highly significant regeneration was observed with combination of 2.5 mg/L 6-benzylamino purine and 3.0 mg/L α-naphthaleneacetic acid. The metabolic and genetic profiles of in vitro and directly cultivated plants were the same, examined through Fourier-transform infrared spectroscopy, random amplified polymorphic DNA(RAPD), inter-simple sequence repeat(ISSR) and R-ISSR(combination of RAPD and ISSR) markers, respectively, and thus confirming the significant efficacy of the SSE incorporated medium. Disarmed T-DNA was transformed to coleoptile derived callus through Agrobacterium tumefaciens LBA4404 and confirmed by GUS assay. The T-DNA integration was confirmed by DNA blot analysis using DNA from transient GUS-expressed explants. Thus, SSE can be used as a natural and organic supplement for organogenesis and efficient acclimatizations of shoot and root apical meristems in regenerated plants.     

Modeling the Fibrillation of Kevlar® KM2 Single Fibers Subjected to Transverse Compression

In this work, fibrillation is introduced as an energy absorbing mechanism in the modeling of Kevlar® KM2 single fibers subjected to quasi-static transverse compression. Fibrillation is simulated using a finite element model of the fiber cross-section containing discrete fibrils connected by interfibrillar cohesive zones. Model predictions of nominal stress-strain response for an assumed bilinear cohesive traction-separation interfibrillar behavior are compared to experimental data. Analysis shows that modeling of the microstructural fibril network, represented by a distribution of strong cohesive interactions, is necessary to capture the experimental response. The model provides valuable insight into the unique deformation mechanisms governing fiber fibrillation under transverse compression.     

Fingerprinting in cucumber and melon (<Emphasis Type="Italic">Cucumis</Emphasis> spp.) Genotypes using morphological and ISSR markers

In the present investigation, 13 Cucumis genotypes from different geographical areas of India were screened for genetic diversity using 19 morphological traits and 15 ISSR primers. The analysis of morphological traits grouped the accessions into six clusters. Cluster V contained the maximum number of genotypes namely Kanivellari, Long Green, Andaman Local, Perundurai Local, and Sempatti Local. Clusters I and VI contained the minimum number of genotypes. Among all the characters, the highest mean value was observed in fruit length (23.38) and the lowest mean value was observed in stripes on the blossom end (1.31). The 15 ISSR primers generated 109 polymorphic alleles. The average number of ISSR alleles generated was 8.3 per primer and the level of polymorphism was 87.20%. The ISSR primer UBC 825 was highly informative with a PIC value of 0.8934. The 13 genotypes were grouped into six clusters based on ISSR markers. Cluster III contained the maximum number of genotypes, namely Kanivellari, Sankagiri Local, Perundurai Local, Long Melon, and Sempatti Local, while Clusters I, II, IV, and V (Karur Local, Andaman Local, Edapaddi Local, and N 78, respectively) contained the minimum number of genotypes. The ISSR profile generated genotypes specific allele namely, UBC 812_700bp and UBC 812_1000bp for Cluster VI which contained Cucumis genotypes collected from the northern part of India. Similarly, UBC 808 produced specific allele UBC 808_650bp formed in Cluster III which contained genotypes collected from Tamil Nadu and Kerala.     

7-O-(6-O-benzoyl-beta-D-glucopyranosyl)-rutin from leaves of Canthium dicoccum

A new flavonol glycoside, characterized as 7-O-(6-O-benzoyl-beta-D-glucopyranosyl)-rutin (1), has been isolated from the leaves of Canthium dicoccum. Detailed 1H COSY, HSQC, HMBC and TOCSY NMR as well as positive and negative electrospray MS and MS/MS data have been provided for the new compound.     

Phenolic compounds and antioxidant capacity of Georgia-grown blueberries and blackberries

Blueberries and blackberries grown at various locations in Georgia were collected and analyzed for flavonoids, total anthocyanins, total polyphenols, and Trolox-equivalent antioxidant capacity (TEAC). Each sample was analyzed for phenolic acids (gallic acid, p-hydroxybenzoic acid, caffeic acid, ferulic acid, and ellagic acid) and flavonoids (catechin, epicatechin, myricetin, quercetin, and kaempferol). A high-performance liquid chromatographic (HPLC) method with photodiode array detection was used for analysis. Compounds were analyzed as aglycons after acid hydrolysis with 1.2 M HCl. Identification of each compound was based on retention time and UV spectra by comparison with pure commercial standards. Phenolic acids ranged from 0.19 to 258.90 mg/100 g fresh weight (FW), and flavonoids ranged from 2.50 to 387.48 mg/100 g FW. Total polyphenols ranged from 261.95 to 929.62 mg/100 g FW, and total anthocyanins ranged from 12.70 to 197.34 mg/100 g FW. TEAC values varied from 8.11 to a maximum of 38.29 microM/g FW. A linear relationship was observed between TEAC values and total polyphenols or total anthocyanins. The data indicate that blueberries and blackberries are rich sources of antioxidants.     

SPAR Markers-Assisted Assessment of Genetic Diversity and Population Structure in Finger Millet (<Emphasis Type="Italic">Eleusine Coracana</Emphasis> (L.) Gaertn) Mini-Core Collection

Finger millet is an important staple food crop of semi-arid tropics also known as “super cereal” and has a higher calcium content than any other crops. Thousands of germplasm are being maintained and its genetic characterization is essential for further utilization in crop improvement. This research was performed to estimate the diversity and population genetic structure in the mini-core collection of finger millet by using SPAR markers, namely RAPD, ISSR, and DAMD markers. Altogether, 32 primers were used in this study, which produced 408 bands among which 344 were polymorphic. Analysis by combining all three marker systems revealed 84.31% of polymorphism among 90 genotypes of finger millet. Average polymorphism information content (PIC) produced by the ISSR, RAPD, and DAMD markers were 0.79, 0.81, 0.62, and average Rp values were 12.84, 8.17, and 8.53, respectively. The Jaccard's similarity value ranged from 0.233-0.861. IE 6059 and IE 5870 genotypes showed the highest Jaccard's similarity value of 0.861 in UPGMA analysis. Neighbor joining-based phylogenetic analysis produced two major clusters and the genotypes were grouped based on their geographical region of origin. Principal component analysis and principal coordinates analysis also confirmed the results. In population STRUCTURE analysis, the genotypes were divided into two subpopulations (P1and P2). These results confirmed that the genotypes we have assessed were genetically diverse and were clustered based on their geographic region of origin. The information obtained from this study will be useful in population management strategies and selection of genotypes for an effective breeding program in the future.     

Aurantoside K, a new antifungal tetramic acid glycoside from a Fijian marine sponge of the genus Melophlus

A new tetramic acid glycoside, aurantoside K, was isolated from a marine sponge belonging to the genus Melophlus. The structure of the compound was elucidated on the basis of spectroscopic analysis (1H NMR, 1H-1H COSY, HSQC, and HMBC, as well as high-resolution ESILCMS). Aurantoside K did not show any significant activity in antimalarial, antibacterial, or HCT-116 cytotoxicity assays, but exhibited a wide spectrum of antifungal activity against wild type Candida albicans, amphotericin-resistant C. albicans, Cryptococcus neoformans, Aspergillus niger, Penicillium sp., Rhizopus sporangia and Sordaria sp.     

Phenolic content and antioxidant capacity of muscadine grapes

Fruits of 10 cultivars of muscadine grapes (five bronze skin and five purple skin) grown in southern Georgia were separated into skin, seed, and pulp. Each fruit part and the leaves from the corresponding varieties were extracted for HPLC analysis of major phenolics. Total phenolics were determined colorimetrically using Folin-Ciocalteu reagent. Total anthocyanins were determined according to a pH-differential method, using a UV-visible spectrophotometer. Antioxidant capacity was determined by the Trolox equivalent antioxidant capacity (TEAC) assay. Gallic acid, (+)-catechin, and epicatechin were the major phenolics in seeds, with average values of 6.9, 558.4, and 1299.4 mg/100 g of fresh weight (FW), respectively. In the skins, ellagic acid, myricetin, quercetin, kaempferol, and trans-resveratrol were the major phenolics, with respective average values of 16.5, 8.4, 1.8, 0.6, and 0.1 mg/100 g of FW. Contrary to previous results, ellagic acid and not resveratrol was the major phenolic in muscadine grapes. The HPLC solvent system used coupled with fluorescence detection allowed separation of ellagic acid from resveratrol and detection of resveratrol. Reported here for the first time are the phenolic content and antioxidant capacity of muscadine leaves. Major phenolics in muscadine leaves were myricetin, ellagic acid, kaempferol, quercetin, and gallic acid, with average concentrations of 157.6, 66.7, 8.9, 9.8, and 8.6, respectively. Average total phenolics were 2178.8, 374.6, 23.8, and 351.6 mg/g gallic acid equivalent in seed, skin, pulp, and leaves, respectively. Total anthocyanin contents were 2.1 and 132.1 mg/100 g of FW in the skins of bronze and purple grapes, respectively, and 4.3 and 4.6 mg/100 g of FW in seeds and pulps, in that order. Antioxidant capacity values were, on average, 2.4, 12.8, 281.3, and 236.1 microM TEAC/g of FW for pulps, skins, seeds, and leaves, respectively.