Feeding potential of <Emphasis Type="Italic">Cryptolaemus montrouzieri</Emphasis> against the mealybug <Emphasis Type="Italic">Phenacoccus solenopsis</Emphasis>

Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae) is an exotic species native to the USA, damaging cotton and other plant families. The feeding potential of different development stages of Cryptolaemus montrouzieri Mulsant, a biological control agent against mealybugs, was investigated on different development stages of P. solenopsis. Fourth instar grubs and adults of C. montrouzieri were the most voracious feeders on different instars of mealybug. The number of 1^st instar nymphs of mealybug consumed by 1^st, 2^nd, 3^rd and 4^th instar larvae and adult beetles of C. montrouzieri was 15.56, 41.01, 125.38, 162.69 and 1613.81, respectively. The respective numbers of 2^nd and 3^rd instar nymphs of mealybug consumed were 11.15 and 1.80, 26.35 and 6.36, 73.66 and 13.32, 76.04 and 21.16, 787.95 and 114.66. The corresponding figures for adult female mealybugs were 0.94, 3.23, 8.47, 12.71 and 73.40, respectively. The results indicate that C. montrouzieri has the potential to be exploited as a biocontrol agent in North India; inoculative releases of 4^th instar larvae and adults may provide instant control of P. solenopsis. Field experiments should be conducted to determine the efficiency of the ladybird on this mealybug.     

Micropropagation of Vegetable Rennet (Withania coagulans [Stocks] Dunal)—A Critically Endangered Medicinal Plant

Withania coagulans (Stocks) Dunal (Solanaceae), popularly called vegetable rennet, is a critically endangered and highly valued medicinal plant. Overexploitation and reproductive failure forced the plant species toward the verge of complete extinction. We describe here the development of a simple, rapid, and cost effective in vitro micropropagation system for W. coagulans for mass-scale production of true-to-type plantlets using nodal shoot segments. Exactly 95.5 ± 0.34% explants responded within 8–10 days (d) and produced multiple shoot buds (4.1 ± 0.10 shoots of 2.95 ± 0.15 cm length) on 0.8% agar-gelled Murashige and Skoog's (MS) basal medium supplemented with 8.88 μM 6-benzylaminopurine (BAP), 0.57 μM indole-3-acetic acid (IAA), and additives (100 mg L^?1 L-ascorbic acid, 25 mg L^?1 each citric acid, adenine sulphate, and L-arginine). The shoots in cultures were multiplied by repeated transfer on MS medium with 4.44 μM BAP, 0.57 μM IAA, and additives. Further cultures were multiplied on a large-scale through the subculturing of shoot clumps differentiated in vitro, on MS medium supplemented with 1.11 μM BAP, 0.57 μM IAA, and additives. Maximum number (19.1 ± 0.28) of healthy (6.15 ± 0.25 cm) and viable shoots differentiated on this medium. The microshoots were rooted both in vitro and ex vitro. Exactly 67.3 ± 1.01% microshoots rooted in vitro within 25–30 d on agar-gelled half-strength MS salts supplemented with 29.52 μM indole-3-butyric acid (IBA) and 200 mg L^?1 of activated charcoal (AC). Alternatively, 73.8 ± 0.65% cloned shoots rooted on sterile soilrite (soilless compost and soil conditioner) under ex vitro conditions after pulse treatment with 2.46 mM IBA for 300 s. The clones of W. coagulans were hardened in a greenhouse within 40–45 d by slow and gradual exposure of plantlets from high relative humidity (RH; 70–80%) and low (26 ± 2°C) temperature to low RH (40–50%) and high (34 ± 2°C) temperature. The hardened plantlets were transferred to soil and stored in agro-net house with more than 90% survival rate. Replacement of pure and laboratory grade sucrose with commercial grade sugar, use of less expensive commercial grade agar-agar in culture medium, higher rate of shoot proliferation, single step ex vitro rooting, and hardening of plantlets in the greenhouse are advantageous features of the protocol. The micropropagation protocol defined here is reproducible, easy to follow, and would be helpful in large-scale restoration programs through true-to-type mass-multiplication of W. coagulans.     

Pharmacokinetics following intravenous administration and pharmacodynamics of cefquinome in buffalo calves

Disposition following single intravenous injection (2 mg/kg) and pharmacodynamics of cefquinome were investigated in buffalo calves 6–8 months of age. Drug levels in plasma were estimated by high-performance liquid chromatography. The plasma concentration–time profile following intravenous administration was best described by a two-compartment open model. Rapid distribution of cefquinome was evident from the short distribution half-life (t _½α ?=?0.36?±?0.01 h), and small apparent volume of distribution (Vd_area?=?0.31?±?0.008 L/kg) indicated limited drug distribution in buffalo calves. The values of area under plasma concentration–time curve, elimination half-life (t _½β ), total body clearance (Cl_B), and mean residence time were 32.9?±?0.56 μg·h/mL, 3.56?±?0.05 h, 60.9?±?1.09 mL/h/kg, and 4.24?±?0.09 h, respectively. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration of cefquinome were 0.035–0.07 and 0.05–0.09 μg/mL, respectively. A single intravenous injection of 2 mg/kg may be effective to maintain the MIC up to 12 h in buffalo calves against the pathogens for which cefquinome is indicated.     

Waterlogging tolerance of pea at germination

Peas (Pisum sativum L.) are exposed to waterlogging at germination when grown as relay in rice‐based cropping. Ninety‐one germplasm accessions were evaluated in relay (sown in waterlogged soil), and subsequently 10 diverse genotypes compared under relay and sole cropping (conventional tillage sowing) over two seasons in Bangladesh. Contrasting genotypes, BM‐3, NL‐2 and Kaspa, were further evaluated in three waterlogging treatments (drained control, 4 and 8 days waterlogging) in the glasshouse. Conspicuous variation in waterlogging tolerance at germination was observed in the field and confirmed under controlled conditions. In relay sowing in 2011, emergence of a few genotypes was affected by waterlogging. In 2012, emergence in relay was severely affected (12 plants/m²) compared to sole sowing (37 plants/m²). Among genotypes BM‐3 had 6 plants/m² emerge, which all subsequently died, in contrast to NL‐2 in which emergence was 13 plants/m² with all plants surviving. In the glasshouse, there was 14% emergence in BM‐3, 40% in NL‐2 and 55% in Kaspa after 8 days of waterlogging. Such marked differences in waterlogging tolerance at germination in the model pea are the first reported and illustrate prospects for selection to improve adaptation to relay sowing in South Asia.     

Structural heterogeneity of yolk spheres in hierarchical follicles from hen ovary: a histochemical study

1. Four types of yolk spheres with variable structure, chemical composition and frequency of occurrence in yolk plasma of hierarchical follicles (F(4), F(3), F(2) and F(1) with diameters of 10.0, 15.5, 20.0 and 35.0 mm, respectively) of the hen ovary were identified using histochemical methods for localising lipids, carbohydrates and proteins. 2. Yolk spheres of the first type (YS(1)) had a phospholipoprotein membrane surrounding fluid matrix which stained lightly for phospholipids, proteins and acidic mucopolysaccharides. Two types of droplets were observed in the matrix of YS(1). Spheres of the second type (YS(2)) had a lipoprotein- and acidic mucopolysaccharide-rich peripheral region and a single large droplet in its fluid matrix. Droplets of YS(2), unlike YS(1), showed three regions and metachromatic staining with ninhydrin-Schiff reagent. The third type of sphere (YS(3)) had a homogeneous matrix staining for proteins, neutral lipids and florescent yellow with alcian blue and differentially with ninhydrin-Schiff reagent; it was bounded by a phospholipids- and acidic mucopolysaccharide-containing thick peripheral region. Its fluid matrix also showed toluidine-blue-positive, densely packed granules and small droplets. The fourth type (YS(4)) was seen only in bromophenol blue and Nile blue preparations, revealing the presence of proteins and neural lipids in their matrix and peripheral regions. 3. Quantitative data on the relative abundance of yolk spheres in F(4) to F(1) follicles revealed more YS(3) (51.1 to 64.7%) than YS(1) (16.2 to 28.3%) and YS(2) (19.1 to 23.2%). The percentage of YS(1) increased and that of YS(3) decreased as follicle size increased.     

Apium graveolens PI 181714 is a source of resistance to Fusarium oxysporum f. sp. apii race 4 in celery (A. graveolens var. dulce)

Celery has little genetic diversity and is highly susceptible to the new fungal pathogen Fusarium oxysporum f. sp. apii (Foa) race 4. After screening an Apium graveolens germplasm collection for resistance to Foa race 4, we crossed celery cv. 'Challenger', which is Foa race 2-resistant but Foa race 4-susceptible and A. graveolens PI 181714, which is Foa races 2- and 4-resistant but non-celery type. After selfing F₁s, we screened the F₁S₁ for race 4-resistance and celery-type and then selfed selected F₁S₁. Greenhouse and field trials indicate that three selected F₁S₂ families (76–8-4, 76–8-27 and 76–8-36) are suitable as germplasm for celery breeders for resistance to Foa race 4. A F1S3 76–8–36-124 is either fixed or nearly so for resistance to Foa races 4 and 2. Furthermore, quantitative PCR indicates that PI 181714 is resistant, rather than tolerant, to Foa races 4 and 2, and that this resistance has been introgressed into F1S3 76–8–36-124.     

Breeding for boron tolerance in lentil (Lens culinaris Medik.) using a high‐throughput phenotypic assay and molecular markers

This study describes the identification of a quantitative trait locus (QTL) in the recombinant inbred line population of ILL2024 × ILL6788 and subsequent validation of associated molecular markers. A high‐quality genetic linkage map was constructed with 758 markers that cover 1,057 cM, with an average intermarker distance of 2 cM. QTL analysis revealed a single genomic region on Lc2 to be associated with B tolerance and accounted for up to 76% of phenotypic variation (Vp). The best markers for B tolerance were assessed for their utility in routine breeding applications using validation panels of diverse lentil germplasm and breeding material derived from ILL2024. A marker generated from the dense genetic map of this study was found to be the most accurate of all markers available for B tolerance in lentil, with a success rate of 93% within a large breeding pool derived from ILL2024. However, given the number of the unrelated lines for which the marker–trait association was not conserved, B tolerance screening is still required at later stages to confirm predicted phenotypes.