Antimicrobial activity of triterpenoids from Heliotropium ellipticum.

Seven sterols and triterpenoids have been isolated from H. ellipticum and tested for their antimicrobial activity.     

Permeability determination and pharmacokinetic study of nobiletin in rat plasma and brain by validated high-performance liquid chromatography method

In the present study, we are reporting permeability and pharmacokinetics of nobiletin in rat plasma and brain, using a validated reverse phase high performance liquid chromatographic method. Protein precipitation method was used for the extraction of nobiletin and coumarin (IS) from rat plasma and brain tissue. The system was run in isocratic mode with mobile phase consisting of potassium dihydrogen ortho-phosphate (pH 4.5; 0.04 mM) and acetonitrile in ratio of 50:50, v/v. The total chromatographic run time was 9.0 min. The method was proved to be accurate and precise at linearity range of 0.05–10 μg/mL with a correlation coefficient (r) of ≥ 0.994 in rat plasma and ≥ 0.995 in rat brain. The intra- and inter-day precision and accuracy values are found to be within the assay variability limits as per the FDA guidelines. Nobiletin was found stable in the battery of stability studies viz., bench-top, auto-sampler, freeze/thaw cycles and long term storage in a freezer at − 70 ± 10 °C. Maximum concentrations of nobiletin in both plasma and brain were observed at 1 h after single oral dosing (50 mg/kg). The maximum concentration in plasma and brain were 1.78 and 4.20 μg/mL, respectively. The AUC_0–t in plasma and brain were 7.49 and 20.66 μg·h/mL, respectively. The mean elimination half life (t_1/2) in plasma and brain were 1.80 and 11.42 h, respectively. The Parallel Artificial Membrane Permeability Assay (PAMPA) permeability of nobiletin was found to be high at both pH 4.0 and 7.0.     

Do boating and basking mix? The effect of basking disturbances by motorboats on the body temperature and energy budget of the northern map turtle

1. Basking is the primary mechanism used by many freshwater turtles to maintain their body temperature (T_b) in a range that maximizes physiological performance. Basking turtles are easily disturbed by motorboats, but the consequences of the increasingly popular use of motorboats on turtles is largely unknown.
2. In this work, predictive models built from field and laboratory data were used to assess the effects of the frequency of basking disturbance by motorboats on T_b and metabolic rate (MR) of female northern map turtles (Graptemys geographica), a species of conservation concern.
3. Simulations revealed that the effects of boat disturbance vary seasonally. In early May, a conservative estimate of the disturbance rate (0.15 per hour) resulted in a 0.34°C decrease in mean daily T_b, which translated to a 7.8% reduction in mean MR. In June, July and August, owing to warmer lake temperatures, the effect of disturbance was less marked and the observed disturbance rates (0.32, 0.96 and 1.23 per hour, respectively) reduced the mean MR of an adult female by 2.1%, 0.5%, and 0.4 %, respectively.
4. Reduction in MR decreases the rate of energy assimilation, which could translate into sublethal effects on turtles, such as reduced growth and reproductive output.
5. Motorboat usage is increasing in many areas and is probably affecting other species of freshwater turtles that use aerial basking. This study offers important insights on the implications of disturbances for species that bask.

Copyright © 2016 John Wiley & Sons, Ltd.     

Identification of a diverse mini‐core panel of Indian rice germplasm based on genotyping using microsatellite markers

Identification of a small core germplasm set representing the available genetic diversity is essential for its proper evaluation and subsequent utilization in rice improvement programmes. For constituting a small diverse mini‐core panel of Indian rice germplasm, a representative set of 6912 accessions drawn based on their geographic origin from the whole rice germplasm collection available in the National Gene Bank was genotyped using 36 microsatellite markers. Automated fragment analysis of amplicons yielded a total of 435 alleles, with an average 12.4 and range of 3–29 alleles per locus. Polymorphism information content (PIC) ranged from 0.08 (RGNMS190) to 0.86 (RM552) with an average of 0.528. Based on genotyping data, a mini‐core consisting of 98 genotypes was identified. Ninety‐four per cent of the alleles present in the core set were present in the mini‐core. The identified small but diverse panel will be useful for further intensive trait‐specific evaluation and utilization in allele mining.     

Assessment of genetic diversity within and among Basmati and non-Basmati rice varieties using AFLP,ISSR and SSR markers

Molecular markers provide novel tools to differentiate between the various grades of Basmati rice, maintain fair-trade practices and to determine its relationship with other rice groups in Oryza sativa. We have evaluated the genetic diversity and patterns of relationships among the 18 rice genotypes representative of the traditional Basmati, cross-bred Basmati and non-Basmati (indica and japonica) rice varieties using AFLP, ISSR and SSR markers. All the three marker systems generated higher levels of polymorphism and could distinguish between all the 18 rice cultivars. The minimum number of assay-units per system needed to distinguish between all the cultivars was one for AFLP, two for ISSR and five for SSR. A total of 171 (110 polymorphic), 240 (188 polymorphic) and 160 (159 polymorphic) bands were detected using five primer combinations of AFLP, 25 UBC ISSR primers and 30 well distributed, mapped SSR markers, respectively. The salient features of AFLP, ISSR and SSR marker data analyzed using clustering algorithms, principal component analysis, Mantel test and AMOVA analysis are as given below: (i) the two traditional Basmati rice varieties were genetically distinct from indica and japonica rice varieties and invariably formed a separate cluster, (ii) the six Basmati varieties developed from various indica × Basmati rice crosses and backcrosses were grouped variably depending upon the marker system employed; CSR30 and Super being more closer to traditional Basmati followed by HKR228, Kasturi, Pusa Basmati 1 and Sabarmati, (iii) AFLP, ISSR and SSR marker data-sets showed moderate levels of positive correlation (Mantel test, r = 0.42–0.50), and (iv) the partitioning of the variance among and within rice groups (traditional Basmati, cross-bred Basmati, indica and japonica) using AMOVA showed greater variation among than within groups using SSR data-set, while reverse was true for both ISSR and AFLP data-sets. The study emphasizes the need for using a combination of different marker systems for a comprehensive genetic analysis of Basmati rice germplasm. The high-level polymorphism generated by SSR, ISSR and AFLP assays described in this study shall provide novel markers to differentiate between traditional Basmati rice supplies from cheaper cross-bred Basmati and long-grain non-Basmati varieties at commercial level.The first two authors have equal contribution     

Tissue culture-derived variation in crop improvement

Tissue culture generates a wide range of genetic variation in plant species which can be incorporated in plant breeding programmes. By in vitro selection, mutants with useful agronomic traits, e.g. salt or drought tolerance or disease resistance, can be isolated in a short duration. The successful use of somaclonal variation is very much dependent on its genetic stability in the subsequent generations for which molecular markers such as RAPDs, AFLPs, SSRs and others can be helpful. The potential of somaclonal variation has yet to be fully exploited by breeders, even though a few cultivars have been developed in crops such as Brassica juncea, rice and others. This revised version was published online in July 2006 with corrections to the Cover Date.     

Protein electrophoresis as an aid to oat variety identification

Summary Ten cultivars of oats (Avena sativa L.) were investigated for their isozymes of esterase (E), leucine aminopeptidase (LAP), and anodal and cathodal peroxidase (APX and CPX).From repeated observations, a typical zymogram was prepared for each enzyme system, and the frequencies of the isozyme bands were scored for each variety separately. The results showed that: (1) most of the isozyme bands are very stable in their appearance on the gel: (2) each variety has a characteristic isozyme pattern; (3) 7 out of 10 oat cultivars showed no intravarietal isozyme variation. Some variation was found for the varieties Mesa, Rapida and Curt. This overall high level of homozygosity and homogeneity of these varieties for the isozyme bands makes this technique useful for varietal identification.     

Genetic analysis of a CSR10 (indica) × Taraori Basmati F3 population segregating for salt tolerance using ISSR markers

Segregation for salinity tolerance and ISSR markers based molecular polymorphism were investigated in a F₃ plant population raised via single-seed descent method from a cross between salt-tolerant indica rice variety CSR10 and salt-susceptible premium traditional Basmati rice variety Taraori Basmati HBC19. A total of 130 F₃plants were evaluated individually for salinity tolerance on 1–9 scale on the basis of seedling growth parameters; the average score ranged between 1.7 to 8.3. Frequency distribution curve obtained using the salinity tolerance data of F₃ population and a chi-square analysis, showed a good fit to a normal distribution. Eleven plants each in the category of salt-tolerant and salt-susceptible were selected from the segregating F₃ population for ISSR marker analysis. A total of 149 bands (4–11 bands per primer) ranging from 200 to 3530 bp were scored for the two rice varieties and the selected CSR10 × HBC19 segregating F₃ plants using 26 ISSR primers. Of these, 89 were monomorphic and 60 were polymorphic. Of the 60 polymorphic bands,36 and 20 bands were specific to CSR10 andHBC19 respectively. The remaining four bands were amplified using UBC primers 810,848, 853 and 886 and present in only some of the CSR10 × HBC19 F₃ plants. Notably, ISSR primers with dinucleotide repeat motif and 5'-anchored end amplified more number of bands (7.0 bands/primer) compared to3'-anchored dinucleotide primers (5.4bands/primer), but 3'-anchored dinucleotide primers revealed higher level of polymorphism (2.6 polymorphic bands/primer) compared to 5'-anchoreddinucleotide primers (1.43 polymorphic bands/ primer). While distribution of majority of the polymorphic bands were more or less in the expected ratios in salt-tolerant and/or salt-sensitive F₃segregating plants, but some of the bands amplified using UBC ISSR primers 823, 825,826, 849, 853, 864, 866 and 884 showed highly skewed distribution. Such polymorphic bands stand greater chances of having a linkage with the genes/ QTLs for salinity tolerance and shall be the target for further studies. This revised version was published online in July 2006 with corrections to the Cover Date.     

Effect of Water Stress on Physiological Attributes and their Relationship with Growth and Yield of Wheat Cultivars at Different Stages

The effects of water stress on physiological attributes of drought‐sensitive (Kalyansona) and drought‐tolerant (C‐306) wheat cultivars were studied in a pot experiment. Water stress was imposed by withholding irrigation at boot and anthesis stages. Leaf water potential, leaf osmotic potential and leaf turgor potential (measured with pressure chamber and osmometer), as well as leaf diffusive resistance, leaf transpiration rate and leaf‐to‐air transpiration gradient (measured with a steady‐state porometer) were measured diurnally. Growth and yield parameters were recorded after harvesting of the crop. Triplicate data were analysed using a completely randomized design and correlations amongst these parameters were computed. Water stress was found to reduce diurnal leaf water potential and leaf osmotic potential in both the genotypes but leaf osmotic potential was significantly higher in the drought‐tolerant cultivar C‐306 than in the drought‐sensitive cultivar Kalyansona. Positive turgor was recorded in both the genotypes under water stress and non‐stress conditions. Water‐stressed plants showed significantly lower turgor potential than control plants. In diurnal observations, water‐stressed plants exhibited significantly higher leaf diffusive resistance in both genotypes at both stages. The diffusive resistance of C‐306 was predominantly higher than that of Kalyansona. Water stress decreased leaf transpiration rate at both stages but the reduction was higher at the anthesis stage. The leaf‐to‐air temperature gradient was much higher in C‐306 than in Kalyansona at the boot stage but at the anthesis stage genotypic variation was non‐significant. The capacity to maintain cooler foliage was lower at the anthesis stage than at the boot stage in both the cultivars. Shoot dry weight, number of grains, test weight, grain yield, biological yield and harvest index decreased to a greater extent when water stress was imposed at the anthesis stage, while imposition of water stress at the boot stage caused a greater reduction in plant height and number of tillers. Similarly, water stress caused a smaller reduction in growth, yield and yield attributes in C‐306 than in Kalyansona. In general, the correlation coefficient of grain and biological yield with water potential and its components was positive and highly significant. Similarly, turgor potential was also correlated positively and significantly with grain yield at both the stages, but with biological yield it was significant only at the anthesis stage. A negative and significant correlation was obtained for diffusive resistance and leaf‐to‐air temperature gradient with grain yield at the boot and anthesis stages. The rate of transpiration was also positively and significantly correlated to grain and biological yields at both the stages. Amongst the yield attributes, number of leaves and number of tillers were positively correlated at the anthesis stage, whereas leaf area and shoot dry weight were significantly correlated with grain and biological yields at both the stages.     

Isozyme Banding Pattern and Estimation of Genetic Diversity among Guinea Grass Germplasm

Isozyme banding pattern was studied in Guinea grass (Panicum maximum Jacq.), a widely cultivated grass having good fodder value. Similarity among 63 accessions collected from diverse sources was worked out using five enzyme systems (SOD, GOT, ACP, Esterase and Peroxidase) following horizontal starch gel electrophoresis. Biochemical markers such as isozymes are useful supplements in identifying the genetic variation present in any crop. A total of 35 clear and unambiguous bands were used for analysis of which 8 bands were monomorphic. Polymorphism exhibited by 27 bands from all five enzyme systems indicate presence of considerable diversity in this species. The dendrogram generated after UPGMA and SAHN cluster analysis using Jaccard genetic distance showed that 63 accessions from diverse geographical locations could be grouped in three main clusters, of which two could further be divided into sub-clusters. Although the clusters comprised members from different locations, most of the accessions from similar geographical locations tended to cluster in same group.