Development and validation of a reverse phase HPLC-DAD method for separation, detection & quantification of rutin and quercetin in buckwheat (Fagopyrum spp.).

Buckwheat has tremendous nutraceutical potential owing to its rutin and quercetin content. The aim of this study was to optimise and validate an analytical method for separating and quantifying these two flavonoids from it. Factors, such as range, linearity, precision, accuracy, limit of detection and limit of quantification, were evaluated for the two compounds using high performance liquid chromatography. On the basis of resolution and symmetry, mobile phase consisting of methanol and methanol:water:acetic acid in the ratio of (100:150:5), flow rate 1.3 ml/min and column temperature 30 °C were found to be optimal analytical conditions. Calibration curves exhibited good linearity with correlation coefficient of 0.995 & 0.9907 over the range 60-180 µg/ml & 2-10 µg/ml for rutin and quercetin respectively. LOD and LOQ values for rutin and quercetin were 6.36, 0.58 and 19.28, 1.77 µg/ml respectively. Recovery values of 96-100.8% confirmed that the method was accurate for rutin and quercetin analysis. This validated method was successfully used to analyse rutin and quercetin in leaves and seeds of buckwheat plant.

Ecology, distribution mapping and conservation implications of four critically endangered endemic plants of Kashmir Himalaya.

During the last few decades, human-driven activities have led to indiscriminate habitat destruction and exploitation of many plant species in Kashmir Himalaya. As a result, many species have become threatened and are struggling for survival. Of particular concern are the endemic and critically endangered species which have the highest risk of the extinction, hence warranting immediate conservation actions. Therefore the current study was carried out to understand the distribution, ecology and conservation implications of the four critically endangered endemic plants of Kashmir Himalaya. Habitat distribution modelling showed that the suitable potential areas for the species occurred from subalpine to alpine meadowlands with an elevational range of 1500-4600 m asl. The output of the MaxEnt model and field surveys have revealed that their highest potential distribution is in Panchari, Khrew, Ramnagar, Pahalgam, Gurez, Sonamarg, Gulmarg and Kishtwar forest ranges. Based on the field explorations and herbarium records, Saussurea costus (Falc.) Lipsch have 27 distribution areas, Gentiana kurroo Royle 18, Lilium polyphyllum D. Don 12 and Aconitum chasmanthum Stapf have 15. Precipitation of the driest month and annual mean temperature played an important role in the distribution of the studied species. The species started their lifecycle with the onset of the spring season, flowered in summer, fruit in autumn and senesce in the winter season. Under natural conditions, the number of days required for germination ranged from 180 to 210 where cold stratification played a pivotal role. Since last few decades, the populations of these species have been shrinking in their natural habitats due to over-exploitation for medicinal purposes and habitat destruction through amplified humanoid interferences like the expansion of agricultural land, road building, grazing and urbanization. Thus there is an urgent need to come up with positive strategies to save whatever is left and plan long term rescue measures not only to protect these species from extinction but also to reintroduce them along with framing the plans to supply sustained raw materials for medicine.

Pathogenesis-related proteins and peptides as promising tools for engineering plants with multiple stress tolerance.

Pathogenesis-related (PR) proteins and antimicrobial peptides (AMPs) are a group of diverse molecules that are induced by phytopathogens as well as defense related signaling molecules. They are the key components of plant innate immune system especially systemic acquired resistance (SAR), and are widely used as diagnostic molecular markers of defense signaling pathways. Although, PR proteins and peptides have been isolated much before but their biological function remains largely enigmatic despite the availability of new scientific tools. The earlier studies have demonstrated that PR genes provide enhanced resistance against both biotic and abiotic stresses, which make them one of the most promising candidates for developing multiple stress tolerant crop varieties. In this regard, plant genetic engineering technology is widely accepted as one of the most fascinating approach to develop the disease resistant transgenic crops using different antimicrobial genes like PR genes. Overexpression of PR genes (chitinase, glucanase, thaumatin, defensin and thionin) individually or in combination have greatly uplifted the level of defense response in plants against a wide range of pathogens. However, the detailed knowledge of signaling pathways that regulates the expression of these versatile proteins is critical for improving crop plants to multiple stresses, which is the future theme of plant stress biology. Hence, this review provides an overall overview on the PR proteins like their classification, role in multiple stresses (biotic and abiotic) as well as in various plant defense signaling cascades. We also highlight the success and snags of transgenic plants expressing PR proteins and peptides.

Antioxidant Potential and DNA Damage Protection by the Slate Grey Saddle Mushroom, Helvella lacunosa (Ascomycetes), from Kashmir Himalaya (India).

This study pertains to the radical scavenging potential of and DNA protection by Helvella lacunosa, an edible mushroom from Kashmir Himalaya (India). Different solvents, on the basis of their polarities, were used to extract all solvent-soluble bioactive compounds. Seven different antioxidant methods were also used to determine extensive radical scavenging activity. The mushroom ethanol extract and butanol extract showed effective scavenging activity of radicals at 95% and 89%, respectively. At 800 µg/mg, the ethanol extract was potent enough to protect DNA from degradation by hydroxyl radicals. It is evident from these findings that the presence of antioxidant substances signifies the use of H. lacunosa as food in the mountainous valleys of the Himalayan region.

Microclimatic variation in UV perception and related disparity in tropane and quinolizidine alkaloid composition of Atropa acuminata, Lupinus polyphyllus and Hyoscyamus niger.

The aim of current research was to evaluate the physiological adjustment in three medicinal herbs viz., Atropa acuminata, Lupinus polyphyllus and Hyoscyamus niger to the winter period characterised by intense UV flux in Kashmir valley across the North Western Himalaya. Quinolizidine (QA) and tropane alkaloid (TA) concentrations were analysed in these herbs thriving at two different altitudes via GC-MS and correlated by PCA analysis. This study investigated the hypothesis that UV reflectance and absorbance at low temperatures are directly related to disparity in alkaloid accumulation. Among QAs in L. polyphyllus, ammodendrine and lupanine accumulated at higher concentration and exhibited significant variation of 186.36% and 95.91% in ammodendrine and lupanine respectively in both sites. Tetrahydrohombifoline displayed non-significant variation of about 9.60% irrespective of sites. Among tropane alkaloid (TA), hyoscyamine was recorded as the most abundant constituent irrespective of the plant and site while apotropine accumulated in lesser quantity in A. acuminata than H. niger. However, apotropine demonstrated significant variation of 175% among both sites. The final concentration of quinolizidine (QA) and tropane alkaloid (TA) reflects the interplay between reflectance and absorbance of UV radiation response field. These findings suggest that spectral response of UV light contributes directly to alkaloid biosynthesis.

Cytogenetic effects of three commercially formulated pesticides on somatic and germ cells of Allium cepa.

Cytological effects of Endosri-ES (endosulfan), Nuvan-NU (dichlorvos), and Kvistin-KS (carbendazim) were evaluated on mitotic and meiotic cells of Allium cepa. Test concentrations were chosen by calculating EC50 values of formulated ES, NU, and KS, which turned to be 60, 200, and 500 ppm (parts per million), respectively. Cytological studies were undertaken on root meristem cells of A. cepa using EC50, 1/2 × EC50, and 2 × EC50 of these pesticides for 24 and 48 h. Similarly, a meiotic study was conducted by applying the pesticides at the aforesaid concentrations from seedling to bud stage. A set of onion bulbs exposed to tap water was run parallel for negative control and maleic hydrazide (112.09 ppm) as positive control. During the study period, mitotic index (MI) decreased at all the pesticide concentrations compared to the negative control. Among various chromosomal aberrations, chromatin bridges, breaks, stickiness, laggard, vagrant chromosomes, fragments, C-mitosis, multipolarity, ring chromosome as well as micronuclei were observed in mitotic preparations. In contrast, meiotic aberrations revealed comparatively less frequency of chromosomal aberrations and the most frequent were lagging chromosome, stray bivalents, secondary association, chromatin bridge, disturbed anaphase, and stickiness. Comparative analysis of the pesticides showed that NU was highly toxic to plant cells than KS, while as ES showed intermediate effects between the two. Further, our study revealed that all the three pesticides produce genotoxic effects which can cause health risks to the human populations. Graphical Abstract ᅟ.