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Differentially expressed transcripts from leaf and root tissue of Chlorophytum borivilianum: A plant with high medicinal value
No full textChlorophytum borivilianum is one of the important medicinal plants used for treating different health problems
such as diabetes, arthritis, physicalweakness, etc. Saponins present in C. borivilianumare the primary source of its
significant medicinal properties and are synthesized by mevalonate and non-mevalonate pathways in plants.
However, the biosynthesis of these compounds at molecular level is not studied in C. borivilianum. Cloning and
sequencing of genes involved inmetabolic processes are prerequisite to study the gene expression, their regulation
and genetic engineering experiments. Expressed sequence tags (ESTs) provide a quick insight into various
genes and their tissue specific expression. Suppression subtractive hybridization (SSH) libraries were
constructed using mRNA from leaf and root tissues of C. borivilianum. High quality non‐redundant 506 and 303
ESTswere generated fromleaf and root specific libraries respectively. These sequences were analyzed using bioinformatics
tools and grouped into different categories based on their similarity and cellular functions such as
photosynthesis, metabolism, transcription factors, cell signaling, defense, stress response etc. ESTs also showed
similarity with genes involved in saponins biosynthesis such as squalene synthase, squalene epoxidase, cytochrome
p450, glycosyltransferase, etc. Semi-quantitative analysis of some of the ESTs involved in saponins biosynthesis
confirmed their differential regulation in leaves and roots. These ESTs will provide an efficient resource to accelerate
gene discovery in C. borivilianum and will help in determining promising targets for genetic engineering of
saponins pathway
Ecological Characterization of High Altitude Himalayan Landscapes in the Upper Satluj River Watershed, Kinnaur, Himachal Pradesh, India
No full textThe Indian Remote Sensing (IRS) LISS III
satellite imagery was used to classify three landscapes
in the upper Satluj river watershed in Kinnaur region
of western Himalaya viz. Nichar, Kalpa and Pooh.
These landscapes representing different eco-climatic
zones were subjected to landscape ecological analysis
and various landscape composition, patch and edge
characteristic metrics were computed to find the
extent of fragmentation in the natural vegetation.
The natural vegetation comprised of forests (10.24%),
grasslands (31.04%) and scrublands (2.95%) as the
landscape elements (LSE). These LSEs were further
segregated into different LSE types based on the
dominant species of plant communities in the landscapes.
These LSE types were surveyed for species
diversity in the three landscapes. The total landscape
area (TLA) covered by natural vegetation in Nichar,
Kalpa and Pooh landscapes was calculated to be
651.42, 529.02 and 1,673.72 km2 making the total
area covered with natural vegetation in the district as
2,854.16 km2. The Pooh block, lying in the east of the
district, though covered the largest area of the
landscape, was poor in terms of landscape composition
as compared to the Nichar block which was rich
in landscape elements as suggested by the diversity
measures. The extent of fragmentation was found to
be highest in Nichar. The Nichar landscape was
observed to be having higher mean values of species
diversity than other blocks which is consistent with
the trends observed with patches diversity. This was
in contrast to the Pooh landscape where the patch
diversity as well as the species diversity was the
lowest. Thus, an ecological analysis utilizing landscape
metrics and biodiversity field sampling of
vegetation, was attempted for such heterogenous
landscapes in high altitudes of Himalaya, which
provided a baseline data for the management and
conservation of these virgin mountainous area
Amino Acid Composition of Five Wild Pleurotus Species Chosen from North West India
No full textAbstract: Five wild edible fungal species of Genus Pleurotus were collected from the different localities of North
West India and evaluated for the amino acid composition by using Ultra Performance Liquid Chromatography
(UPLC). Five amino acids namely aspartic acid, arginine, alanine, proline and tyrosine were detected from
the samples. Among theese five wild samples, aspartic acid was found maximum in Pleurotus pulmonarius
(0.48 %) followed by P. sajor-caju (0.44 %), P. cystidiosus (0.42 %), P. floridanus and least was found in
P. sapidus (0.37 %). Arginine was found maximum in P. sapidus (0.23 %) followed by P. floridanus (0.22 %),
P. sajor-caju (0.20 %) while P. cystidiosus and P. pulmonarius contained equal percentage of this amino acid
(0.19 %). Alanine was documented maximum in P. pulmonarius (0.19 %) followed by P. cystidiosus (0.18 %),
P. floridanus (0.17 %), P. sajor-caju (0.12 %) and minimum was documented in P. sapidus (0.11 %). Proline
was recorded maximum in P. sapidus (0.09 %) followed by P. floridanus (0.08 %), P. pulmonarius (0.06 %),
P. sajor-caju (0.05 %), while minimum was recorded in P. cystidiosus (0.02 %). Tyrosine was documented
maximum in P. sapidus (0.20 %) P. cystidiosus (0.18 %) followed by P. pulmonarius (0.17 %), P. sajor-caju
(0.15 %) and minimum was recorded in P. floridanus (0.14 %)
Morpho-Cultural, Molecular and Nutraceutical Studies on Coremiopleurotus from India
No full textCoremiopleurotus (Pleurotus cystidiosus O.K. Miller) is studied for its taxonomic, cultural, molecular
features and nutraceutical attributes and illustrated based on collection from Patiala in Punjab (India). It is a
wood decaying edible basidiomycetous mushroom, characterized by the formation of specialized anamorphic
structures called coremia. Internal transcribed spacer (ITS) region sequencing studies carried out showed
highest identity with Pleurotus cystidiosus strain ACCC 51280. The aerial mycelium of the mushroom
(anamorph) form toxocyst, which is a specialized feature of P. cystidiosus subsp. abalonus Zervakis, a hitherto
unreported variant of dhingri mushroom from India
Effect of agro-techniques on the performance of natural sweetener plant–stevia (Stevia rebaudiana) under western Himalayan conditions
No full textTwo separate field experiments were conducted during 2007, 2008 and 2009 to evaluate the effect of agrotechniques
on the performance of natural sweetener plant–stevia (Stevia rebaudiana Bertoni) under western Himalayan
conditions. In the first experiment during 2007 and 2008 two doses of farm yard manure (FYM) and
seven inorganic fertilizers were compared with untreated plot (control). Application of 50:60:50 kg NPK/ha recorded
significantly higher plant height, stem, flower and total weight/plant (dry) and consequently recorded higher
dry leaf yield as compared to other treatments. However, stevioside and rebaudioside contents were found to reduce
due to the application of manures/fertilizers. FYM application recorded higher marker compounds than inorganic
fertilizers. Owing to higher dry leaf yield, 50:60:50 kg NPK/ha recorded highest steviol glycoside (stevioside
and rebaudioside A) yield/ha than other treatments. In second experiment during 2008 and 2009, effect of land
configuration and harvesting time on growth, yield and steviol glycosides of stevia was evaluated. Planting stevia
in broad bed and furrow (BBF) recorded 31 and 23% higher leaf biomass over flat and camber bed methods, respectively.
Steviol glycosides (stevioside and rebaudioside-A) concentration was higher when stevia was planted
in camber bed as compared to flat and BBF. Significantly higher leaf biomass was obtained from stevia plants
which were harvested once at 50% flower bud stage compared with those harvested once at the 60 and 90 days
after transplanting (DAT). Rebaudioside -A content was higher when leaves were harvested at 50% flower bud
stage. Owing to higher dry leaf yield, stevia planted in BBF and harvested at 50% flower bud stage recorded highest
net return and B:C ratio than other treatments
Increase in flavan-3-ols by silencing flavonol synthase mRNA affects the transcript expression and activity levels of antioxidant enzymes in tobacco
No full textFlavonoids are plant secondary metabolites widespread throughout the plant kingdom
involved in many physiological and biochemical functions. Amongst the flavonoids,
flavan-3-ols (catechin and epicatechin) are known for their direct free radical
scavenging activity in vitro, but studies on their antioxidant potential and interaction
with antioxidant enzymes in vivo are lacking. Here, the flavonoid pathway was
engineered by silencing a gene encoding flavonol synthase (FLS) in tobacco to direct
the flow of metabolites towards production of flavan-3-ols. FLS silencing reduced
flavonol content 17–53%, while it increased catechin and epicatechin content 51–
93% and 18–27%, respectively. The silenced lines showed a significant increase in
expression of genes for dihydroflavonol reductase and anthocyanidin synthase, a
downstream gene towards epicatechin production, with no significant change in
expression of other genes of the flavonoid pathway. Effects of accumulation of flavan-
3-ols in FLS silenced lines on transcript level and activities of antioxidant
enzymes were studied. Transcripts of the antioxidant enzymes glutathione reductase
(GR), ascorbate peroxidase (APx), and catalase (CAT) increased, while glutathione-
S-transferase (GST), decreased in FLS silenced lines. Enhanced activity of all the
antioxidant enzymes was observed in silenced tobacco lines. To validate the affect
of flavan-3-ols on the antioxidant system, in vitro experiments were conducted with
tobacco seedlings exposed to two concentrations of catechin (10 and 50 lm) for
2 days. In vitro exposed seedlings produced similar levels of transcripts and activity
of antioxidant enzymes as FLS silenced seedlings. Results suggest that flavan-3-ols
(catechin) might be increasing activity of GR, Apx and CAT by elevating their
mRNAs levels. Since these enzymes are involved in scavenging of reactive oxygen
species, this strategy would help in tailoring crops for enhanced catechin production
as well as making them tolerant to oxidative stresses
Densitometric HPTLC Analysis of Juglone, Quercetin, Myricetin, Rutin, Caffeic Acid, and Gallic Acid in Juglans regia L.
No full textAsensitive, selective, and precise high-performance thin-layer chromatography
(HPTLC) method was developed for simultaneous
analysis of six bioactive phenolic compounds, i.e., juglone,
quercetin, myricetin, rutin, caffeic acid, and gallic acid in methanol
extract and its fractions (hexane, chloroform, ethyl acetate, and
butanol fractions) from bark of Juglans regia. Good separation was
achieved on RP-18 F254S TLC plate using methanol–water–formic
acid–acetic acid (48.8:46.4:2.4:2.4, v/v). The densitometric determination
of the compounds was carried out at 254 nm in reflectance/
absorbance mode. The method was validated in terms of linearity,
sensitivity, accuracy, precision, robustness, and specificity. The linear
regression data for the calibration plots of the reference compounds
showed a good linear relationship with higher correlation
coefficient (r2 ≥ 0.997). Accuracy of the method was evaluated in
terms of average percent recovery, which ranged from 98.63 to
101.06%. HPTLC results revealed qualitative and quantitative differences
in the phenolic compounds in the extract and fractions. The
ethyl acetate fraction contained gallic acid followed by myricetin,
rutin, quercetin, and caffeic acid in higher amount in comparison to
the extract and fractions. The present method can be used for routine
quality control of J. regia extracts
In Vitro Expression and Production of Antibody Against Cymbidium mosaic virus Coat Protein
No full textPolyclonal rabbit antisera were produced using
coat protein of Cymbidium mosaic virus (CymMV) Indian
isolate expressed in E. coli as GST fusion. The expressed
protein was purified by GST-fusion protein purification kit
for use as an immunogen in rabbits. Antisera prepared in
this manner reacted in double antibody sandwich enzymelinked
immunosorbent assay (DAS ELISA) with extract
from CymMV-infected tissue. The results indicate that
polyclonal antisera prepared from expressed CymMV coat
proteins were useful for the detection of CymMV in an
array of assays. The detection system developed is highly
effective for detection of Indian strain of the virus in
comparison to kits available in the international market
Optimising parameters for biolistic gun-mediated genetic transformation of tea [Camellia sinensis (L.) O. Kuntze]
No full textGenetic improvement of tea through breeding is difficult.Therefore, transgenic tea plants expressing the osmotin gene
from Nicotiana tabacum were produced using parameters optimised for biolistic-gun mediated transformation. During
optimisation, a total of 4,500 somatic embryos were bombarded using nine combinations of variable target distances
and burst pressures, while keeping the gap distance (0.6 cm) and macrocarrier flight distance (16 mm) constant. A total
of 90 independent, PCR-positive lines were generated. Southern hybridisation confirmed integration of the osmotin
gene in 26 out of 27 PCR-positive lines (three independent lines from each of the nine parameter combinations were
selected at random). Statistical analysis revealed that the efficiency of transgene integration was significantly affected
by target distance. Only those lines derived from somatic embryos bombarded with 1.0 μg plasmid DNA using a 7.58
MPa burst pressure and 9-cm target distance showed osmotin expression. This was evident from strong northern
hybridisation and RT-PCR signals. Leaves of 4-year-old transgenic plants growing in a contained polythene tunnel
showed improved osmotic adjustment in response to osmotic stress imposed by NaCl. The osmotic potentials of
transgenic leaves immersed in 100 mM or 200 mM NaCl solutions were more negative than those of non-transformed
control leaves
