Highly specific assays to detect isolates of Pseudomonas syringae pv. actinidiae biovar 3 and Pseudomonas syringae pv. actinidifoliorum directly from plant material

Pseudomonas syringae pv. actinidiae (Psa) is responsible for bacterial canker of kiwifruit. Biovar 3 of Psa (Psa3) has been causing widespread damage to yellow‐ and green‐fleshed kiwifruit (Actinidia spp.) cultivars in all the major kiwifruit‐producing countries in the world. In some areas, including New Zealand, P. syringae pv. actinidifoliorum (Pfm), another bacterial pathogen of kiwifruit, was initially classified as a low virulence biovar of Psa. Ability to rapidly distinguish between these pathovars is vital to the management of bacterial canker. Whole genome sequencing (WGS) data were used to develop PCR assays to specifically detect Psa3 and Pfm from field‐collected material without the need to culture bacteria. Genomic data from 36 strains of Psa, Pfm or related isolates enabled identification of areas of genomic variation suitable for primer design. The developed assays were tested on 147 non‐target bacterial species including strains likely to be found in kiwifruit orchards. A number of assays did not proceed because although they were able to discriminate between the different Psa biovars and Pfm, they also produced amplicons from other unrelated bacteria. This could have resulted in false positives from environmental samples, and demonstrates the care that is required when applying assays devised for pure cultures to field‐collected samples. The strategy described here for developing assays for distinguishing strains of closely related pathogens could be applied to other diseases with characteristics similar to Psa.     

Discovery of a sooty mould scavenging beetle, <Emphasis Type="Italic">Leiochrinus nilgirianus</Emphasis> Kaszab (Coleoptera: Tenebrionidae) on coconut palms infested by the invasive rugose spiralling whitefly, <Emphasis Type="Italic">Aleurodicus rugioperculatus</Emphasis> Martin (Hemiptera: Aleyrodidae)

Scavenging action of Leiochrinus nilgirianus Kaszab (Coleoptera: Tenebrionidae) on sooty mould deposition on coconut palms due to heavy honeydew production by the invasive rugose spiralling whitefly, Aleurodicus rugioperculatus Martin (Hemiptera: Aleyrodidae), is reported for the first time from Kerala, India. Adult beetles, averaging 2.07?±?0.9 / leaflet, with a maximum of five beetles per leaflet, and immature stages were generally confined to the abaxial of the palm leaves during daytime, but were found feeding on sooty mould during morning hours in damp conditions. The specially aided adaptive leg features of L. nilgirianus with bristle-like hairs on the undersurface of tarsomeres probably aid in proper adhesion on sooty mould laden plant surface and well developed tarsal claws that perhaps assist swift movement on fungus-laden leaflets. On an average one adult beetle could clear 1–2 mm² sooty mould laden area in a period of one minute. In situ habitat conservation of L. nilgirianus would help to reduce the sooty mould at no cost in the most natural and eco-friendly manner avoiding chemical management options and other expensive methods.     

Establishing Soil Silicon Test Procedure and Critical Silicon Level for Rice in Louisiana Soils

Calibration of crop responses to applied silicon (Si) serves as a basis for developing Si fertilizer recommendation guidelines. A greenhouse experiment was set up in a randomized complete block design with five replications, two sources of Si (wollastonite and slag) and four Si rates (0, 170, 340 and 680 kg ha^?1) to calibrate plant-available Si for growing rice in Louisiana soils. Silicon concentrations were determined in soils using seven different extraction procedures. Based on a quadratic model (p < 0.05), the estimated soil Si critical level using 0.01 M calcium chloride (CaCl₂) for Sharkey clay soil was 110 mg kg^?1 while for Crowley silt loam and Commerce silt loam, levels were 37 and 43 mg kg^?1, respectively. These results suggest that suitability of an extractant that gives the best estimate of plant-available Si could considerably depend on soil type and it is unlikely that there is a universal extractant for all soils.     

Agronomic evaluation of mulching and iron nutrition on productivity,nutrient uptake,iron use efficiency and economics of aerobic rice-wheat cropping system

Field experiments were carried out during rainy (kharif) and winter (rabi) seasons (June–April) of 2008–2010 at Indian Agricultural Research Institute (IARI), New Delhi, to study the productivity, nutrients uptake, iron (Fe) use-efficiency and economics of aerobic rice-wheat cropping system as influenced by mulching and Fe nutrition. The highest yield attributes, grain and straw yields (5.41 tonnes ha^?1 and 6.56 tonnes ha^?1, respectively) and nutrient uptake in rice was recorded with transplanted and puddled rice (TPR) followed by aerobic rice with Sesbania aculeata mulch. However, residual effect of aerobic rice with wheat straw mulch was more pronounced on yield attributes, grain and straw yields (4.20 and 6.70 tonnes ha^?1, respectively) and nutrient uptake in succeeding wheat and remained at par with aerobic rice with Sesbania mulch. Application of iron sulfate (FeSO₄) at 50 kg ha^?1 + 2 foliar sprays of 2% FeSO₄ was found to be the best in terms of all the yield attributes, grain and straw yield (5.09 and 6.17 tonnes ha^?1, respectively) and nutrient uptake and remained at par with 3 foliar sprays of 2% FeSO₄. Although residual effect of iron application failed to increase the yield attributes, yield and nutrient uptake nitrogen, phosphorus and potassium (N, P, K) except Fe. The highest system productivity, nutrient uptake, gross returns, net returns, B: C ratio and lowest cost of cultivation were recorded with aerobic rice with wheat straw and Sesbania aculeata mulch. Application of FeSO₄ at 50 kg ha^?1 + two foliar sprays of 2% FeSO₄ was found better in respect of system productivity, nutrient uptake, gross returns, net returns, B:C ratio and cost of cultivation in aerobic rice-wheat cropping system. The Fe use efficiency values viz. partial factor productivity (kg grain kg^?1 Fe), agronomic efficiency (kg grain increased kg^?1 Fe applied), agrophysiological efficiency (kg grain kg^?1 Fe uptake), physiological efficiency (kg biomass kg^?1 Fe uptake), apparent recovery (%) utilization efficiency and harvest index (%) of applied Fe were significantly affected due to methods of rice production and various Fe nutrition treatments in aerobic rice and aerobic rice-wheat cropping system.     

A high-throughput, low cost gel-based SNP assay for positional cloning and marker assisted breeding of useful genes in cereals

Several SNP (single nucleotide polymorphism) genotyping methods have been developed in the past most of which require sophisticated instrumentation and large initial investments. We describe here a high-throughput SSCP (single strand conformation polymorphism) system on our HEGS (high efficiency genome scanning) platform, which is simple, accurate, cost effective and requires neither restriction digestion of the amplification products nor elaborate post-PCR processing detection. Several parameters critical to SSCP analysis were optimized viz., gel matrix and concentration, gel running temperature, buffer composition, running conditions and PCR primer design so as to identify SNPs in amplicons ranging from 100 to 750 bp in size. A simple post-PCR processing system was developed using fluorescent dye for quick and easy detection of SNP polymorphism. HEGS-SSCP was also found to be useful in uncovering simple sequence repeat differences between different genotypes that differ by one or few di/tri nucleotide repeats. The practical utility of this system is illustrated with two successful efforts towards construction of high resolution linkage maps of a lesion mimic locus on chromosome 7 and a major quantitative trait locus conditioning field blast resistance on chromosome 4 in rice.     

Aflatoxin B1 induced alterations in the stability of the lysosomal membrane in Oreochromis mossambicus (Peters, 1852)

This article elucidates the effect of crude dietary aflatoxin B₁ (AFB₁) on the stability of the lysosomal membrane in tilapia (Oreochromis mossambicus), and the analysis was made at the biochemical and cellular level. The liver lysosomal acid phosphatase activity was used to evaluate the functional integrity of the lysosome. Three experimental groups were fed on diets containing 0.375, 2.5 and 6 mg kg^?1 of AFB₁ for a time period of 6 weeks, whereas a fourth group fed on the semi‐purified diet formed the control. Significant changes were observed in the stability of the lysosomal membrane in aflatoxin exposed groups. This is the first report, so far, of changes induced by AFB₁ on the stability of lysosomal membrane in tilapia (O. mossambicus).     

Effect of aromatase inhibitors on the ovarian development of stunted yearlings of rohu (Labeo rohita): a preliminary study

In India, many of the fish farmers stock 1-year-old stunted fishes (stunted yearlings) of Indian major carps (IMC) for enhancing fish production through compensatory growth, but many of them observed problems of early maturation in these fishes. Application of aromatase inhibitors for deceleration of ovarian maturation is one of the probable solutions to mitigate this issue. In the present study, a synthetic aromatase inhibitor letrozole [25 (L₂₅) and 50 (L₅₀) mg kg^?1 feed] and a plant-derived aromatase inhibitor, grape seed extract [100 (G₁₀₀) and 200 (G₂₀₀) mg kg^?1 feed], were fed to stunted yearlings of rohu (Labeo rohita) for 45 days well before the onset of breeding season. Maturation indices such as gonadosomatic index (GSI) and serum oestradiol (E) levels indicated a dose-dependent inhibition of ovarian development in the aromatase-inhibitor-treated fish. Higher dose of letrozole (GSI, 15.12 ± 0.18; E, 3.19 ± 0.42) and grape seed extract (GSI, 16.90 ± 0.40; E, 3.60 ± 0.75) were found to be more effective since control fish showed further advancement in maturation (GSI, 21.20 ± 1.10; E, 7.33 ± 0.74) during the peak breeding season (15th June). Histological observations also confirmed the results revealing a delayed initiation of ovarian development in the case of higher doses of letrozole and grape seed extracts. These results indicate the possible use of aromatase inhibitors in arresting the early maturation process in IMC.     

Investigation of transferrin polymorphism in Garole sheep

The aim of this study was to determine the genetics of polymorph systems of Transferrin in Garole sheep breed. The present study was conducted on 95 adult Garole sheep comprising 52 ewes and 43 rams, maintained at Sheep and Goat Breeding Farm of West Bengal University of Animal and Fishery Sciences, West Bengal, during the period from April–September, 2009. The polymorphism of transferrin was determined through SDS-Polyacrylamide gel electrophoresis technique. It was found that the transferrin type was controlled by five codominant alleles (TfA, TfB, TfC TfD and TfE) in Garole sheep. These five alleles, because of co-dominant nature of inheritance, determined the occurrence of nine transferrin genotypes in the analyzed flock. Four (TfAA, TfBB, TfCC and TfDD) of these were homozygous and the remaining five (TfAD, TfBC, TfBD, TfCD and TfDE) heterozygous. It was found that the TfDD genotype (0.263) was predominant while TfDE genotype (0.042) was least common in the analyzed flock. Frequencies of other genotypes were as: TfCD(0.242), TfBD(0.126), TfCC(0.084), TfBB(0.074), TfAA(0.063), TfAD and TfBC (0.053 for each genotype ) in whole population. From the result it was found that in whole population combined, the heterozygotic genotypic frequency (0.516) was more than that of homozygotic genotypic frequency (0.484). Considerable variations were recognized in the frequencies of transferrin alleles. In the whole population frequencies of transferrin alleles were found to be TfA = 0.089, TfB = 0.163, TfC = 0.232, TfD = 0.495 and TfE = 0.021. Transferrin system has shown an absence of genetic equilibrium among the analyzed herd (χ2 value = 51.31). In conclusion, there were polymorphism in Transferrin types and the presence of differences among the frequencies of the five alleles by categories could be a source of genetic variation in Garole sheep.