Coexpression of 16.8 kDa antigen of Mycobacterium avium paratuberculosis and murine gamma interferon in a bicistronic vector and studies on its potential as DNA vaccine

Paratuberculosis or Johne’s disease is a chronic gastric disease of ruminants. For this disease there is no effective treatment or preventive measure available. 16.8 kDa protein is an immunogenic protein of Mycobacterium avium paratuberculosis and can be an ideal candidate for developing a DNA vaccine construct. In present study a bicistronic DNA vaccine construct pIR16.8/IFN was developed using eukaryotic vector pIRES 6.1. Two genes MPT (expressing 16.8 kDa protein) and murine IFNγ were cloned, expressed and immunoreactivity was studied in murine model. Immunoreactivity was also compared with monocistronic construct pIR16.8 expressing 16.8 kDa protein. Both pIR16.8 and pIR16.8/IFN showed eukaryotic expression of respective proteins in BHK21 cells. The expressed proteins also showed immunoreactivity when reacted with hyperimmune sera raised against recombinant 16.8 kDa protein in western blot assay and immunofluorence assay. Both constructs were used as DNA vaccine in murine model and immunogenecity was studied by DTH, lymphocyte proliferation assay and NO determination. DTH reaction was significantly high in pIR16.8/IFN than pIR16.8 group, similarly lymphocyte proliferation and NO release was higher in pIR16.8/IFN group than pIR16.8 group. This indicated T cell epitopic nature of 16.8 kDa protein. The study also showed that co-expression of IFNγ with mycobacterial gene can enhance immunogenecity of DNA vaccine and can be used as immunoadjuvant.     

Effects of germination and cooking on polyphenols and in vitro protein digestibility of horse gram and moth bean

Changes in polyphenols and in vitro protein digestibility due to germination and cooking were studied in horse gram and moth bean. Horse gram had higher amounts of polyphenols than moth bean. Polyphenol content in horse gram decreased from 1.6% to 1.1% at 48h germination, but in moth bean there was a reduction in polyphenol content from 1.3% to 1.0% at 24 h germination with a significant increase from 1.0% to 1.7% at 36 h germination. Seeds germinated for 48 h also exhibited higher amounts of polyphenols than in raw moth bean. The germination had no effect on total protein content, but a progressive increase in free amino acid content was observed in both legumes. In the ungerminated form, horse gram protein was more digestible than moth bean protein. The in vitro protein digestibility of both the legumes increased during germination. Cooking decreased the protein digestibility of horse gram either in ungerminated or germinated form, but such a decrease was noticed in moth bean at only 36 h germination. Moth bean either in raw form or germinated for 24 h did not exhibit a decrease in in vitro protein digestibility after cooking. Cooking decreased polyphenol content by 20%–50%. The apparent decrease in polyphenols in moth bean and horse gram due to cooking has been attributed to a decrease in assayable polyphenols.     

Soil moisture levels affect mycorrhization during early stages of development of agroforestry plants

Plants respond differentially to different arbuscular mycorrhizal fungi (AMF) as well as to the different soil moisture levels. Based on this background, the present study was carried out to investigate the effects of different levels of soil moisture and AMF inoculations on mycorrhization and growth of important agroforestry plants, viz., Phaseolus mungo, Triticum aestivum, Eucalyptus tereticornis, and Albizia procera. The experiments consisted of main treatment, i.e., three levels of soil moisture [field capacity (FC?=?16 %), half-field capacity (FC/2?=?8 %) and double-field capacity (2×FC?=?32 %)] and four subtreatments (mycorrhizal inoculations), viz., Acaulospora scrobiculata, Glomus cerebriforme, Glomus intraradices, and un-inoculated (control). AMF inoculations significantly (P?<?0.05) increased growth and P uptake, in all tested plant species. In P. mungo, maximum AMF efficiency was observed at FC while in other plants, AMF were equally effective at FC/2 and 2×FC. Different inoculants were effective at different moisture levels. Furthermore, mycorrhization was the highest at FC. AMF inoculations were more important than soil moisture (explaining 33–97 % variation in growth) in P. mungo, T. aestivum, and A. procera (forward selection method), whereas soil moisture was more important for growth of E. tereticornis. Thus, it may be stated that depending upon soil moisture, inoculation of plants with suitable AMF consortium can be beneficial.     

Quick-cooking beans (Phaseolus vulgaris L.): I. Investigations on quality

Soaking three beans cultivars (Phaseolus vulgaris L.: Great Northern, kidney, and pinto) in mixed salt solution (sodium chloride 2.5% + sodium bicarbonate 1.5% + sodium tripolyphosphate 1.0% + sodium carbonate 0.5%) resulted in 80%–85% reduction in cooking time over corresponding controls. Irradiation (-rays) at 500 krads of soaked and dehydrated beans caused a reduction of nearly 50% in cooking time. Water uptake and leaching losses for each treatment during soaking at 22°, 37°, and 45°C were investigated. High temperature (37° and 45°C) and pH (9.0) caused greater water imbibition and total solid loss than at room temperature (22°C). Organoleptic evaluation revealed that quick-cooking Great Northern beans appear to be more acceptable than kidney and pinto beans. Quick-cooking cooked beans had better in vitro protein digestibility than conventionally cooked beans. Phenolic content was found to be inversely related to in vitro digestibility.Utah Agricultural Experiment Station Journal article no. 2481.     

Interrelationships between certain physical and chemical properties of dry bean (Phaseolus vulgaris L.)

The interrelationships among certain physical and chemical properties of ten cultivars of dry beans (Phaseolus vulgaris L.) were investigated. The length/breadth ratio was similar (1.51–1.65) except for kidney beans (>2.0) and sanilac (1.37) cultivars. The breadth/thickness ratio ranged from 1.17–1.65. The 100-bean weight indicated a wide variation of 15.03–50.33 g. The density, bulk density, and porosity characteristics were within a narrow range of 1.18–1.36 g/cc, 68–75 g/100 cc, and 40.7–48.5%, respectively. Water uptake rates during the first 6 h of soaking at room temperature (21 °C) were characteristic of the cultivar. At the end of 24-h soaking, however, all cultivars had absorbed similar amounts of water (approximately 1 g/g bean). Leaching losses (g solids leached/100 g beans) had characteristic trends and ranged from 0.54 for cranberry to 3.46 for sanilac cultivars after 24-h soaking. Most correlations between selected chemical constituents and physical characteristics of the dry bean were relatively low.Journal Paper No. 2751 of Utah Agricultural Experiment Station and a contribution of Western Regional Project W-150.     

Changes in phytate phosphorus and minerals during germination and cooking of horse gram and moth bean

The phytate phosphorus in horse gram and moth bean seeds accounted for 57% and 55%, respectively, of the total phosphorus. During germination, there was a continuous decrease in the proportion of phytate phosphorus. In 48 h germinated seeds, the phytate phosphorus accounted for only 20% and 26%, respectively of the total phosphorus. A significant amount of calcium was leached out during germination. Other minerals were not affected significantly. Cooking decreased phytate phosphorus in ungerminated and germinated seeds of both the legumes. Calcium content in ungerminated and 12 h germinated moth bean seeds decreased significantly during cooking. Such a decrease was noticed in ungerminated seeds of horse gram. No significant changes were noticed in magnesium, iron, manganese, and zinc contents of both the legumes due to cooking.     

Molecular genetic diversity analysis in emmer wheat (Triticum dicoccon Schrank) from India

Emmer wheat (Triticum dicoccon Schrank) is still largely cultivated in India, and highly appreciated for the preparation of traditional dishes. Moreover, its nutritional characteristics could justify a development of its cultivation. The perspective of genetic improvement however requires a good knowledge of the genetic diversity existing within the eco-geographic group of Indian emmer wheats. A set of 48 emmer wheat accessions from India including 28 from a local collection and 20 Indian accessions obtained from CIMMYT, Mexico, was assessed for genetic variability using 47 microsatellite (SSR) markers, distributed over all the 14 chromosomes. The number of alleles per locus ranged from 2 to 9, with an average of 3.87 alleles per locus. A total of 201 alleles were detected at 52 loci with average polymorphic information content of 0.35 per locus and a mean resolving power of 1. The pair-wise similarity coefficients calculated from binary data matrix based on presence or absence of alleles varied from 0.15 to 0.98, but was greater than 0.5 for most accessions, indicating a high level of similarity. A cluster analysis based on the similarity matrix identified nine distinct accessions and three clusters. All the recently developed commercial varieties were distinctly different from the clusters. Based on the analysis, it appears that Indian emmer wheats are not very diverse. Consequently, there is a need to increase the diversity within the Indian emmer wheat eco-geographic group, by introducing diversity from other eco-geographic groups, or even from other wheat species.     

Potential of extracts of the tropical plant Balanites aegyptiaca (L) Del. (Balanitaceae) to control the mealy bug, Maconellicoccus hirsutus (Homoptera: Pseudococcidae)

The effects of extracts of different parts of the perennial tropical plant Balanites aegyptiaca (L) Del., including various solvent extracts of roots, methanol extracts from leaves, fruits, flowers and roots, partially purified saponins obtained from its roots and a standard saponin were studied on the life cycle (adult longevity, number of eggs, crawlers, adults, weight of adults and % wax content) of a laboratory-reared parthenogenic line of the mealy bug, Maconellicoccus hirsutus (Homoptera: Pseudococcidae). Extracts derived from various parts of B. aegyptiaca (leaves, fruits, flowers, and roots in methanol) affected the life cycle of M. hirsutus with a methanol root extract being the most effective at a concentration of 500 μg ml⁻¹. Partially purified saponin of B. aegyptiaca and the commercial bark saponin extract (Sigma) from Quillaja saponaria at a concentration of 500 μg ml⁻¹ were effective in reducing the longevity of M. hirsutus. Significant reductions in oviposition by M. hirsutus were found for all the extracts at a concentration of 500 μg ml⁻¹. Extracts also affected the number of emerging crawlers, number of adults as well as the weight and wax content of emerging adults. These studies suggest that B. aegyptiaca plant extracts and saponins can be useful botanical insecticides for the protection of crops from mealy bugs.