Genetic variation and gene flow estimation of <Emphasis Type="Italic">Nepenthes khasiana</Emphasis> Hook. F- A threatened insectivorous plant of India as revealed by RAPD markers

Random Amplified Polymorphic DNA (RAPD) markers were utilized for determination of diversity within and among the three populations of Nepenthes khasiana Hook f., a threatened insectivorous plant of Meghalaya (India). A total of 90 bands were generated from 10 random amplification polymorphic DNA (RAPD) primers of which 71 were found to be polymorphic (78.89%). Nei’s gene diversity (h) ranged between 0.124–0.201 with overall diversity of 0.228 while Shannon’s information index I) values recorded between 0.187–0.308 with an average value of 0.352. The values of gene flow (Nm = 1.284) and the diversity among populations (0.280) recorded demonstrates higher genetic variation within the population. AMOVA analysis revealed a low level of genetic variation (21.96%) among the populations. This study indicates that some variation still exists within and between the existing populations of N. khasiana, thus, these populations could provide materials for re-establishing of this important rare and threatened species.     

Genomic characterization of drought tolerance-related traits in spring wheat

Drought tolerance was investigated in ‘C306’, one of the most drought tolerant wheat cultivars bred in India in the 1960’s. An intervarietal mapping population of recombinant inbred lines of the cross ‘C306’ × ‘HUW206’ was evaluated for drought tolerance components, namely potential quantum efficiency of photosystem (PS) II (F_v/F_m), chlorophyll content (Chl), flag leaf temperature (Lt), and grain yield per plant (Gyp) under stress. Three independent experiments were conducted under well-watered and water-stressed conditions in greenhouses and growth chambers at Kansas State University (USA). Five hundred and sixty microsatellite markers covering the entire genome were screened for polymorphism between the parents. A QTL (QLt.ksu-1D) for Lt (low flag leaf temperature under stress) on the short arm of chromosome 1D between markers Xbarc271 and Xgwm337 at LOD 3.5 explained 37% of the phenotypic variation. A QTL for F_v/F_m (QF _v /F _m .ksu-3B) and Chl (QChl.ksu-3B) controlling quantum efficiency of PS II and chlorophyll content under stress were co-localized on chromosome 3B in the marker interval Xbarc68–Xbarc101 and explained 35–40% of the phenotypic variation for each trait. A QTL (QGyp.ksu-4A) for Gyp on chromosome 4A at a LOD value of 3.2 explained 16.3% of the phenotypic variation. Inconsistent QTLs were observed for F_v/F_m on chromosomes 3A, 6A, 2B, 4B, and 4D; for Chl on 3A, 6A, 2B and 4B; and for Lt on 1A, 3A 6A, 3B and 5B. The identified QTLs give a first glimpse of the genetics of drought tolerance in C306 and need to be validated in field experiments using the marker-phenotype linkages reported here.     

Natural compounds enhancing growth and survival of rhizobial inoculants in vermicompost-based formulations

The present study demonstrates the usefulness of natural microbial growth-promoting compounds for improving the stability and life of vermicompost-based (both granular and its aqueous extract) bioformulations. Granular vermicompost maintained the number of cells of Rhizobium meliloti Rmd 201 up to 5.9 × 10⁸ after 180 days at 28°C compared with 2.1 × 10⁸ in charcoal (powdered), while aqueous extract of the vermicompost supported the 5.6 × 10⁷ rhizobia numbers even after 270 days. The addition of 25 μL/mL cow urine and 0.01 mM calliterpinone, a natural plant growth promoter, increased the rhizobia number significantly in granular vermicompost and its aqueous extract, respectively.     

Molecular characterization and phylogeny of phytoplasma associated with bunchy top disease in its new host Okra (<Emphasis Type="Italic">Abelmoschus esculentus</Emphasis>) in India reveal a novel lineage within the 16SrI group

Okra plants with bunchy top disease were found to be prevalent during the period of August–October 2009 in New Delhi, India. The common symptoms observed were shortening of internodes, aggregation of leaves at the apical region, reduced leaf lamina, stem reddening, fruit bending, phyllody and stunting of plants. The disease incidence ranged from 2–60% accompanied by significant reductions in production of both flowers and seeds. Nested polymerase chain reaction targeting phytoplasma specific 16S rDNA and rp genes revealed all symptomatic plants to be positive for phytoplasma. Homology searches depicted its closest identity to phytoplasmas of 16SrI ‘Candidatus Phytoplasma asteris’, like the Sugarcane yellows and Periwinkle phyllody phytoplasmas. Profiles for 16S rDNA obtained with 10 restriction endonucleases, differed in TaqI sites for two phytoplasma isolates (BHND5 & 10) from the standard pattern of 16SrI-B subgroup, the latter was seen in the case of isolate BHND1. Restriction fragment analysis of rp genes with AluI, Tsp509I matched with patterns of the rpI-B phytoplasmas. Phylogenetic reconstruction of rp genes revealed okra bunchy top phytoplasma (BHND1) as a divergent isolate, the subsequent sequence analysis of which showed the presence of a novel BslI site. These significant differences suggest that multiple phytoplasma strains are affecting okra, one of which is a diverging lineage within the 16SrI-B group while others represent a new 16SrI subgroup not reported so far. Additionally, this is the first report of a phytoplasma associated disease in okra plants worldwide.     

Thermal stability of alpha-amylase from malted jowar (Sorghum bicolor)

Malted cereals are rich sources of alpha-amylase, which catalyzes the random hydrolysis of internal alpha-(1-4)-glycosidic bonds of starch, leading to liquefaction. Amylases play a role in the predigestion of starch, leading to a reduction in the water absorption capacity of the cereal. Among the three cereal amylases (barley, ragi, and jowar), jowar amylase is found to be the most thermostable. The major amylase from malted jowar, a 47 kDa alpha-amylase, purified to homogeneity, is rich in beta structure ( approximately 60%) like other cereal amylases. T(m), the midpoint of thermal inactivation, is found to be 69.6 +/- 0.3 degrees C. Thermal inactivation is found to follow first-order kinetics at pH 4.8, the pH optimum of the enzyme. Activation energy, E(a), is found to be 45.3 +/- 0.2 kcal mol(-)(1). The activation enthalpy (DeltaH), entropy (DeltaS*), and free energy change (DeltaG) are calculated to be 44.6 +/- 0.2 kcal mol(-)(1), 57.1 +/- 0.3 cal mol(-)(1) K(-)(1), and 25.2 +/- 0.2 kcal mol(-)(1), respectively. The thermal stability of the enzyme in the presence of the commonly used food additives NaCl and sucrose has been studied. T(m) is found to decrease to 66.3 +/- 0.3, 58.1 +/- 0.2, and 48.1 +/- 0.5 degrees C, corresponding to the presence of 0.1, 0.5, and 1 M NaCl, respectively. Sucrose acts as a stabilizer; the T(m) value is found to be 77.3 +/- 0.3 degrees C compared to 69.6 +/- 0.3 degrees C in the control.     

Sublethal Toxicity of the Herbicide Butachlor on the Earthworm Perionyx sansibaricus and its Histological Changes (5 pp)

Goal, Scope and Background   With the advent of the Green Revolution, there has been a quantum leap in the use of synthetic herbicides and pesticides throughout the world to sustain high-yielding crop varieties. Continuous use of these synthetic chemicals leads to loss of soil fertility and soil organisms. Histopathological studies may signal a damaging effect of organisms resulting from prior or ongoing exposure to toxic agents. A large number of studies have reported general histological changes in earthworms. A fewer studies have reported more specific types of histopathological studies in Eisenia foetida, Dendrodrilus rubidus, Lumbricus terrestris, Lumbricus rubellus and Octolasium transpandanum. However, no reports are available on Perionyx sansibaricus. The aim of the present study is (i) to determine the sublethal effects of the herbicide Butachlor on Perionyx sansibaricus on growth and cocoon production, (ii) to evaluate the histological changes in the exposed worms.Methods   The test substrate, namely powdered, dried cow dung, was contaminated with 0.0657, 0.1315 and 0.1973 mg kg–1 of Butachlor, and a control was maintained. Four groups of each 10-test species of Perionyx sansibaricus were used per concentration level (control, 0.0657, 0.1315 and 0.1973 mg kg–1). Various life history parameters such as biomass, cocoon production and histological changes were measured.Results   Worm growth was observed at various exposures over 60 days. While exposing the earthworm to the herbicide, no mortality was observed, not even at a higher dose. At the end of the experiment, the control group had a mean biomass of 0.0831±0.00 mg and, in the exposed group, at herbicide concentrations of 0.1973, 0.1315 and 0.0657 mg kg–1, the mean biomass was found to be 0.0497±0.00 mg, 0.0628±0.00 mg and 0.0781±0.00 mg, respectively. The mean earthworm biomass was found to be decreased with increasing herbicide concentration. Similarly, cocoon production was also reduced by the increasing herbicide concentration. All earthworms in the exposed group were found to have glandular cell enlargement and to be vacuolated. Conclusion   The results clearly indicate that Butachlor can retard the growth and cocoon production in Perionyx sansibaricus. The effect of the herbicide Butachlor can be assessed by the histological observation of the intestinal region, as evidenced by glandular cell enlargement at all the exposed concentrations, which may massively affect food intake and which in turn may indirectly inhibit the earthworm reproductive capacity.Recommendation and Outlook   Risk assessment is normally aimed at the protection of human health and the ecosystem, and the interrelationship of these two areas of protection is easy to perceive. The use of earthworms in risk assessment is to obtain more information on environmental quality and ensure environmental safety. There is an urgent need to test the chemicals causing toxicity to earthworms, because earthworms play a major role in soil fertility as well as acting as a transferring route from the soil to the terrestrial ecosystem.     

Serological evidence of Trichinellosis in local pigs of Nepal

In Nepal, animal husbandry is a major source of income. Pig husbandry is practiced in rural, peri-urban, and urban communities. Free ranging "back yard" pigs and the practice of feeding offal is a very common management practice which potentially allows for the transmission of trichinellosis; however, this zoonosis has never been reported from this region. A total of 425 serum samples were collected from local pigs. These were initially screened by ELISA after which positive samples were examined by Western blot. This procedure identified two samples which had clear specific bands for Trichinella; however, muscle samples tested by HCL-pepsin digestion were found to be negative. If these highly specific serological analyses are confirmed, this would be the first report of trichinellosis in Nepal and a prevention program should be initiated to limit the access of pigs to open garbage dumps which exist both in towns and on farms.     

Role of bioinoculants in development of salt-resistant saplings of Morus alba (var. sujanpuri) in vivo

Pot experiments were conducted for the development of salt-resistant saplings of Morus alba (var. sujanpuri) involving bioinoculants, namely arbuscular mycorrhizal (AM) fungi, Azotobacter and indole butyric acid (IBA). The IBA and sodium chloride (NaCl) concentrations were optimized prior to the experiments. By using both low and high concentrations, 15 ppm IBA and 0.05% NaCl (w/v) were found to be optimum in acting synergistically with AM fungi and Azotobacter and also for increasing all the growth parameters and microbial count in the rhizosphere. For in vivo development of salt-resistant saplings, the optimal concentration of IBA, along with AM fungi and Azotobacter in different combinations, was applied. The saplings were irrigated regularly with 0.05% NaCl water. Although growth parameters such as AM infection percentage, AM spores per 100 g soil and Azotobacter cells/g soil were affected by NaCl watering, the inoculation of both bioinoculants significantly enhanced survival percentage of saplings from 25 to 50% under salt stress. Maximum survival (55%) of saplings was found with IBA (15 ppm)+AM fungi+Azotobacter.     

Effect of progesterone on the magnum proteins during primary stimulation of chick oviduct

1. The effect of progesterone on the secretion of protein by the magnum of 5‐d‐old, female chicks was determined.

2. The supernatant prepared by centrifuging an homogenate of the magnum at 105 000g was found, by immunodiffusion, to contain an antigenic component which precipitated the antisera for conalbumin 1, conalbumin 2 and ovalbumin after 5 d treatment with progesterone: there was no reaction to ovomucoid, lysozyme and avidin antisera.

3. Disc‐electrophoresis of the homogenate revealed two bands at the site of ovalbumin.

4. Incorporation of ³H‐lysine into the magnum proteins of progesterone‐treated chicks did not differ from that of controls.

5. The secretion available in the magnum may be only a transudate from the serum and not a true secretory product. Progesterone behaved qualitatively as oestrogen in this study although the action is much less pronounced and was delayed.     

Therapeutic management of copper deficiency in buffalo heifers: Impact on immune function

To evaluate the magnitude of copper deficiency in Northern India and to examine the various haematobiochemicals, enzymes, vitamins and immune functions affected by copper deficiency, and to identify the parameters which can be of diagnostic importance in copper deficiency, a survey was conducted in 12 districts of Northern India. Significant deficiency of copper was observed in soil, fodder and serum samples of buffalo heifers. Fifty hypocuperaemic buffalo heifers were selected from these areas and were randomly divided into two groups, A and B. The heifers in group A were provided with mineral mixture containing copper sulphate and in group B without copper sulphate. Significant (p < 0.01) improvement in serum ceruloplasmin level was observed within 30 days of treatment, while significant (p < 0.01) improvement in monoamine oxidase and liver cytochrome oxidase was observed at the 60th day of treatment in group A animals. Significant improvement was observed in T₃ and T₄, in the animals of group A within 60 days of treatment. The values of vitamin A and E showed significant (p < 0.01) improvement within 30 days of treatment. The phagocytic activity of neutrophils against Candida albicans significantly (p < 0.01) improved in group A within 60 days of treatment. Similarly, significant improvement in superoxide dismutase activity in red blood cells was observed at the 30th day, and in total leukocytes and whole blood at the 60th day in group A animals. Significant improvement in liver copper level was observed at the 30th day of treatment, while in group B the liver copper was significantly (p < 0.01) depleted at the 60th day of experimentation. Additional copper supplementation improved growth performance significantly in group A.