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.     

Cyanobacteria mediated plant growth promotion and bioprotection against Fusarium wilt in tomato

Cyanobacteria - phytopathogenic fungi - tomato plant interactions were evaluated for developing suitable biological options for combating biotic stress (Fusarium wilt) and enhancing plant vigour. Preliminary evaluation was undertaken on the fungicidal and hydrolytic enzyme activity of the cyanobacterial strains (Anabaena variabilis RPAN59, A. laxa RPAN8) under optimized environmental/nutritional conditions, followed by amendment in compost-vermiculite. Such formulations were tested against Fusarium wilt challenged tomato plants, and the Anabaena spp. (RPAN59/8) amended composts significantly reduced mortality in fungi challenged treatments, besides fungal load in soil. Cyanobacteria amended composts also led to an enhancement in soil organic C, nitrogen fixation, besides significant improvement in growth, yield, fruit quality parameters, N, P and Zn content. The tripartite interactions also enhanced the activity of defence and pathogenesis related enzymes in tomato plants. A positive correlation (r?=?0.729 to 0.828) between P content and pathogenesis/defense enzyme activity revealed their role in enhancing the resistance of the plant through improved nutrient uptake. Light and scanning electron microscopy (SEM) revealed cyanobacterial colonization, which positively correlated with reduced fungal populations. The reduced disease severity coupled with improved plant growth/ yields, elicited by cyanobacterial treatments, illustrated the utility of such novel formulations in integrated pest and nutrient management strategies for Fusarium wilt challenged tomato crop.     

Resistance to blast (Magnaporthe grisea) in a mini-core collection of finger millet germplasm

Blast caused by Pyricularia grisea [teleomorph: Magnaporthe grisea] is an economically important and widespread disease of finger millet in the world. Host resistance is the most economical and effective means of combating this disease as finger millet is predominantly grown by resource-poor and marginal farmers. At the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT), we evaluated a finger millet mini-core collection of 80 germplasm accessions (about 1 % of the total germplasm collection representing major trait variability) for blast resistance both in the field and greenhouse. Field evaluation was done using a refined screening technique that included new improved rating scales for leaf, neck and finger infection. Sixty six of the 80 accessions showed combined resistance to leaf, neck and finger blast in two seasons (2009 and 2010) of field screening. A highly significant and positive correlation was found between neck and finger blast ratings (r?=?0.92), whereas small but significant correlations were found between leaf blast and neck blast (r?=?0.25) and between leaf blast and finger blast (r?=?0.30). These accessions were also screened for leaf blast resistance in the greenhouse by artificial inoculation of seedlings to confirm field observations. Fifty-eight of the 80 accessions were resistant to leaf blast in the greenhouse screen as well. These resistant accessions represented one wild (africana) and four cultivated races (vulgaris, plana, elongate and compacta) of finger millet that originated from 13 countries in Asia and Africa and exhibited considerable diversity for agronomic traits, such as maturity period, plant height and panicle type. These blast resistant accessions from the mini-core collection would be useful in finger millet disease resistance breeding programs.     

Quality of potato tubers as affected by freezing: II. lipids and minerals

The effect of freezing on the chemical composition of Katahdin and Atlantic potatoes was studied. Tubers were exposed to a temperature of ?20°C for 24 hrs., then thawed for 1 hr. at 20°C prior to analysis. Frozenthawed tubers were compared with unfrozen controls for total lipid, phospholipid, fatty acid and mineral content. Following freezing, a significant (p<.01) decrease was observed in crude lipid and phospholipid content of both cortex and pith sections. Generally, an increase in palmitic acid and a decrease in linolenic acid occurred in the phospholipid fractions of both varieties. A decrease in both macro and micro mineral content also occurred following freezing.     

Dual Inoculation of Arbuscular Mycorrhizal and Phosphate Solubilizing Fungi Contributes in Sustainable Maintenance of Plant Health in Fly Ash Ponds

Fly ash is one of the residues produced during combustion of coal, and its disposal is a major environmental concern throughout coal-based power-generated counties. Deficiencies of essential nutrients, low soil microbial activity, and high-soluble salt concentrations of trace elements are some of the concerns for reclamation of fly ash ponds. The effect of fly-ash-adapted arbuscular mycorrhizal (AM) fungi and phosphate solubilizing fungus Aspergillus tubingensis was studied on the growth, nutrient, and metal uptake of bamboo (Dendrocalamus strictus) plants grown in fly ash. Co-inoculation of these fungi significantly increased the P (150%), K (67%), Ca (106%), and Mg (180%) in shoot tissues compared control plants. The Al and Fe content were significantly reduced (50% and 60%, respectively) due to the presence of AM fungi and A. tubingensis. The physicochemical and biochemical properties of fly ash were improved compared to those of individual inoculation and control. The results showed that combination of AM fungi and A. tubingensis elicited a synergetic effect by increasing plant growth and uptake of nutrients with reducing metal translocation.     

The effect of phosphorus fertilization on the chemical quality of Katahdin potatoes

The influence of phosphorus fertilization on the content of ascorbic acid, lipids, total nitrogen, protein, and non-protein nitrogen of Katahdin potatoes was investigated. Phosphorus in the form of mono ammonium phosphate was applied at rates of 0, 56, 112, and 168 Kg/ha (0, 50, 100, and 150 lb/acre). Ascorbic acid, total nitrogen, protein and non-protein nitrogen content were significantly increased (p<.01) and the phospholipid content was significantly reduced (p<.01) by phosphorus application.