Genetic tagging of Azotobacter chroococcum for colonization on wheat (Triticum aestivum) and cotton (Gossypium sp.) roots

Abstract

Azotobacter chroococcum strains E12, HT57 were genetically tagged with lac Z, gfp to study the colonization behaviour on wheat (Triticum aestivum) and cotton (Gossypium sp.) in soil under controlled conditions. 10³ – 10⁴ cfu g^?1 soil of HT57 lac Z were found to colonize roots of both cotton and wheat crops whereas 1.7 × 10⁴ – 7.2 × 10⁴ cfu g^?1 soil of E12 gfp was colonizing wheat roots and 1.6 × 10⁴ – 9.3 × 10⁴ cfu g^?1 soil of E12 gfp colonized cotton roots respectively. Tagged strains colonized mostly on root tips compared to basal roots in both the crops.     

Molecular mapping of quantitative trait loci for ascochyta blight and botrytis grey mould resistance in an inter-specific cross in chickpea (Cicer arietinum L.) using genotyping by sequencing

Ascochyta blight (AB) and botrytis grey mould (BGM) are the most devastating fungal diseases of chickpea worldwide. The wild relative of chickpea, C. reticulatum acc. ILWC 292 was found resistant to BGM whereas, GPF2 (Cicer arietinum L.) is resistant to AB. A total of 187 F₈ Recombinant Inbred Lines (RILs) developed from an inter-specific cross of GPF2 × C. reticulatum acc. ILWC 292 were used to identify quantitative trait loci (QTLs) responsible for resistance to AB and BGM. RILs along with parents were evaluated under artificial epiphytotic field/laboratory conditions for two years. Highly significant differences (P < 0.001) were observed for reaction to both pathogens in both years. Parents and RILs were genotyped-by-sequencing to identify genome wide single nucleotide polymorphism (SNPs). A total of 1365 filtered and parental polymorphic SNPs were used for linkage map construction, of which, 673 SNPs were arranged on eight linkage groups. Composite interval mapping revealed three QTLs for AB and four QTLs for BGM resistance. Out of which, two QTLs for AB and three QTLs for BGM were consistent in both years. These QTLs can be targeted for further fine mapping for deployment of resistance to AB and BGM in elite chickpea cultivars using marker-assisted-selection.     

Anticandidal activity of curcumin and methyl cinnamaldehyde

Cinnamaldehyde, its derivatives and curcumin are reported to have strong antifungal activity. In this work we report and compare anticandidal activity of curcumin (CUR) and α-methyl cinnamaldehyde (MCD) against 38 strains of Candida (3; standard, fluconazole sensitive, 24; clinical, fluconazole sensitive, 11; clinical, fluconazole resistant). The minimum inhibitory concentrations (MIC₉₀) of CUR ranged from 250 to 650 μg/ml for sensitive strains and from 250 to 500 μg/ml for resistant strains. MIC₉₀ of MCD varied between 100 and 250 μg/ml and 100–200 μg/ml for sensitive and resistant strains, respectively. Higher activity of MCD as compared to CUR was further reinforced by spot assays and growth curve studies. At their respective MIC₉₀ values, in the presence of glucose, average inhibition of H⁺-efflux caused by CUR and MCD against standard, clinical and resistant isolates was 24%, 31%, 32% and 54%, 52%, 54%, respectively. Inhibition of H⁺-extrusion leads to intracellular acidification and cell death, average pHi for control, CUR and MCD exposed cells was 6.68, 6.39 and 6.20, respectively. Scanning electron micrographs of treated cells show more extensive damage in case of MCD. Haemolytic activity of CUR and MCD at their highest MIC was 11.45% and 13.00%, respectively as against 20% shown by fluconazole at typical MIC of 30 μg/ml. In conclusion, this study shows significant anticandidal activity of CUR and MCD against both azole-resistant and sensitive clinical isolates, MCD is found to be more effective.     

Testing for Tolerance to Aphids in Indian Mustard, Brassica juncea (L.) Czern and Coss.

Brassica juncea plants were screened for aphid tolerance using three methods. Seven and 14 days old seedlings grown in wooden trays in a glasshouse were kept between infector trays with heavy aphid infestation. There were no surviving plants in the susceptible genotypes 50 days after sowing but survival rates ranging between 80 and 90 % were observed for selection 44. The progenies of the surviving plants were tested under natural infestation in the field. Plants were scored for leaves without aphids 40 days after sowing, healthy plants at harvest and seed yield in g/m² For most of these lines, the aphids’ survival, life span and fecundity (nymphs produced per adult) were recorded after introducing five newly-born nymphs on excised fifth leaves cultured in petri dishes. Selections showing greater tolerance to aphids than the check cultivars were identified in the progenies of a cross RLM-514 × T-6342. It was concluded that the excised leaf method can provide a reliable measure of the lines’ reaction to aphids.     

Antioxidant flavonoid glycosides from Evolvulus alsinoides

Oxidative damage is an established outcome of chronic stress. Thus, the present study was designed to investigate the modulatory role of ethanolic extract of Evolvulus alsinoides (EA) in terms of oxidative alterations at peripheral and central level in rats subjected to chronic unpredictable stress (CUS). CUS exposure for 7 days reduced Cu, Zn superoxide dismutase and catalase activity with increase in glutathione peroxidase activity and lipid peroxidation, while decrease in reduced glutathione level in blood plasma, frontal cortex and hippocampus regions of brain. Oral administration of EA extract at 200 mg/kg p.o. normalized these stress induced oxidative alterations with an efficacy similar to that of melatonin. Further, EA extract was taken up for detailed chemical investigation. Two new flavonol-4′-glycoside, kaempferol 4′-O-β-d-glucopyranosyl-(1 → 2)-β-d-glucopyranoside (3) and kaempferol 4′-O-α-l-rhamnopyranosyl-(1 → 6)-β-d-glucopyranoside (5) were isolated, along with eight known compounds (1, 2, 4 and 6–10). The structures of new compounds were established by detailed spectroscopic studies, while known compounds were characterized by direct comparison of their reported NMR data. All these compounds were evaluated for their in vitro antioxidant activity. Compounds 3, 5, 9 and 10 at 100 and 200 μg/ml showed significant in vitro antioxidant activity. Therefore, EA may hold great potential in preventing clinical deterioration in stress induced oxidative load and related disorders.     

Effect of Water Stress on the Photosynthesis, Productivity and Water Status of Mung Bean (Vigna radiata L. Wilczek)

The effect of water stress and its subsequent recovery on the photosynthesis, productivity and water status, was examined in three mungbean varieties ( Vigna radiata L. Wilczek) i.e. PS 16, P 105 and Pusa Baisakhi. The variety P 105 was found relatively less susceptible to water stress compared to high susceptibility in Pusa Baisakhi and PS 16. The adverse effect of stress was comparatively higher at the post-flowering and pod development stages. It was observed that variety P 105 had inherently maintained higher water status by retaining higher water potential and water content under water stress condition, thereby effectively reducing the water loss. The higher transpiration rate in this variety regulates the flow of water within the plant. This type of regulation of water flow and maintenance of high water status in P 105 helped to resist the adverse effect of water stress on photosynthesis and productivity. Such adaptation mechanism was not observed in varieties PS 16 and Pusa Baisakhi. The recover) mechanism was, however, operated efficiently in susceptible varieties PS 16 and Pusa Baisakhi particularly for recouping the vegetative growth, but its effect could not be retained for reproductive characters, wherein also variety P 105 due to its efficient partitioning system yielded better than other cultivars.     

Effect of temperature on the infectivity of Sarcocystis miescheriana cysts in pork

Sarcocysts of Sarcocystis miescheriana in the thigh muscles of pigs became non-infective to pups after heating infected pork in minute pieces at 60 degrees C for 20 min, 70 degrees C for 15 min and 100 degrees C for 5 min. Similar pieces of infected muscle tissues, when exposed to -4 degrees C for 2 days or -20 degrees C for 1 day, became non-infective to pups. The experiment suggests that pork containing sarcocysts of S. miescheriana, and possibly of S. suihominis, requires cooking at a minimum of 70 degrees C for 15 min or freezing at -4 degrees C for 2 days or -20 degrees C for 1 day for making it safe for consumption.