Cytological effects of different mutagens in soybean [Glycine max (L.) Merrill]

The effect of gamma rays, ethyl methane sulphonate, and their three combinations on various cytological parameters in M1 generation was studied in two soybean cultivars, Pusa-16 and PK-1042. Combined treatments exhibited high meiotic abnormalities in Pusa-16 as compared with the individual treatments of ethyl methane sulphonate and gamma rays. In PK-1042 the highest frequency of meiotic abnormalities was exhibited by gamma rays, while in Pusa-16 the frequencies of univalents were high as compared with other meiotic abnormalities. Fragments/bridges, laggards, ring and rod bivalents were other types of meiotic abnormalities that were noticed in both the cultivars. The frequencies of irregular cells in the two cultivars were high in combined treatments followed by ethyl methane sulphonate and gamma rays. The highest percentage of pollen sterility was exhibited by the combined treatments and the lowest was exhibited by gamma rays. In Pusa-16, the increase in doses of gamma rays and ethyl methane sulphonate increased the pollen sterility percentage, while in the combined treatments, the increase was noticed up to the intermediate dose and then gradually decreased at higher doses. On the other hand, in PK-1042 the increase in ethyl methane sulphonate dose increased sterility percentage, while no definite trend was noticed in gamma rays and combined treatments.     

Effect of different egg yolk-based extenders on the quality of ovine cauda epididymal spermatozoa during storage at 4°C

Cauda epididymal spermatozoa were obtained from testicles collected from abattoir(s). The pooled sperm samples were divided into four aliquots. Each aliquot was washed separately with the buffer of respective extender and finally extended with the four extenders viz. egg yolk–citrate (EYC), egg yolk–citrate–fructose (EYCF), Tris–citric acid–egg yolk–fructose (TCEYF) and egg yolk–Mcillvaine glucose (EYMG) and preserved at 4°C. The per cent sperm motility for EYC, EYCF, TCEYF and EYMG at 0 h was 50.83%, 56.67%, 75.00% and 31.67%, respectively, and at 72 h was 24.17% (EYC), 30.83% (EYCF), 51.67% (TCEYF) and 7.50% (EYMG). The corresponding figures for live sperm count at 0 h was 83.17%, 86.33%, 90.42% and 81.75% and at 72 h was 64.75%, 73.92%, 76.00% and 57.67%. The corresponding figures for mean per cent intact acrosome at 0 h was 95.33%, 95.50%, 90.92% and 97.25% and at 72 h was 86.17%, 83.92%, 77.58% and 86.33%. The sperm motility was significantly (p < 0.05) higher for TCEYF at different h of preservation from 0 h through 72 h. The sperm motility, live sperm count and per cent intact acrosome declined significantly (p < 0.05) with the advancement of storage time in all the four extenders. Our study concluded that TCEYF was best out of the extenders studied for preservation of cauda epididymal spermatozoa after double centrifugation and extension at 4°C up to 72 h of preservation. However, EYCF also has better potential for the preservation of cauda epididymal spermatozoa as viability was in close proximity and acrosomal integrity was higher compared with TCEYF extender.     

Post mortem studies on infertile buffalo bulls: anatomical and microbiological findings

Twenty-two buffalo bulls suffering from three different types of infertility were slaughtered and used for this study. Except for the reproductive system, no signs of localised or generalised disease were observed. Microbiological investigations were negative for brucellosis, vibriosis, mycoplasma and other non-specific microorganisms. Nine bulls with type 1 infertility had low bodyweights and underdevelopment of testes, accessory sex glands and endocrine glands. This picture suggests a total dysfunction of the pituitary-growth-gonadal axis. One bull of this type also showed bilateral epididymitis. Four out of 11 bulls with type 2 infertility had low bodyweights and most suffered from underdevelopment of testes, accessory sex glands and endocrine glands. Six bulls of this type had lesions of either epididymitis or orchitis or both. Two of these animals showed adhesions of periorchitis. One also showed seminal vesiculitis. In two bulls with type 3 infertility, bodyweights, reproductive organs and endocrine glands were normal. In later life, they yielded poor quality semen. Semen samples collected a few months before slaughter from nine bulls with type 2 and type 3 infertility were of poor quality and had higher percentages of abnormal spermatozoa in most cases.     

The method of cohesion determination in soil aggregates

Theoretical fundamentals and experimental methods to study the strength of water-stable bonds in soil aggregates are discussed. The strength of water-stable aggregates characterized by cohesion is proved to be one of the important structural and mechanical properties of soil. The regular variation of this parameter inthe soils under study is related to the content of humus and physical clay.     

Brassinosteroid-mediated enrichment in yield attributes, active constituents and essential oil production in Mentha arvensis L.

Recently, brassinosteroids (BRs) have emerged as a new group of growth promoting phytohormones. 28-homobrassinolide (HBR) is one of the several brassinosteroids, the role of which in enhancing growth, productivity and quality of plants, via improving various physiological processes, has been established both under stress and normal conditions. Out of a large number of essential oil bearing plants, mint (Mentha arvensis L.) constitutes the most important source of therapeutic agents used in the alternative systems of medicine. The mint plant has marvelous medicinal and aromatic values. In view of enhancing yield and quality of this medicinally important plant, a pot experiment was conducted under natural conditions of net house. The study was aimed at exploring the effect of four concentrations of HBR (10^?0, 10^?8, 10^?7 and 10^?6 M) on the performance of mint with regard to physiological attributes, herbage yield, content and yield of essential oil, and active constituents at 100 and 120 days after planting. The foliar application of HBR enhanced physiological attributes, herbage yield and the yield and content of most of the active constituents (menthol, L-methone, isomenthone and menthyl acetate) of mint at both the stages, with 10^?7 M concentration proving the best. However, the next higher concentration of HBR (10^?6 M) exhibited no further increase in the values of the attributes studied. Rather, 10^?6 M was slightly inferior to 10^?7 M concentration; but it always proved significantly better than the control.     

Effect of Hydrophilic Gums on Frozen Dough. I. Dough Quality

Disadvantages of frozen doughs are their variable performance and loss of stability over long‐term frozen storage. Changes in rheological properties of frozen doughs have been reported to be due to the physical damage of the gluten network caused by ice crystallization and recrystallization. The objective of this study was to determine the effect of hydrophilic gums on ice crystallization and recrystallization for improvement of the shelf‐life stability of frozen dough. The present research involved use of the Hard Red Spring wheat cultivar Grandin and hydrophilic gums such as carboxymethyl cellulose (CMC), gum arabic, kappa carrageenan (κ‐carrageenan), and locust bean gum at three different levels each on doughs stored frozen for up to 16 weeks. The dough characteristics were analyzed after day 0, day 1, and after 4, 8, 12, and 16 weeks of frozen storage using data from differential scanning calorimetry (DSC), water activity, extensigraph, and proof time. The ΔH value of freezable water endothermic transitions obtained using DSC increased with storage time for all treatments. However, addition of different levels of the four gums lowered the ΔH value, indicating a decrease in freezable water. Doughs with locust bean gum gave a higher peak force, measured using the Kieffer dough extensibility rig of the texture analyzer, and lower proof time, indicating better retention of baking quality. Maximum resistance to extension increased upon addition of 1 and 3%; CMC; 1 and 3%; κ‐carrageenan; and 1, 2, and 3% locust bean gum as compared with the control. The various periods of storage or gum treatments did not affect the water activity of the thawed frozen doughs. Doughs with locust bean gum gave significantly lower proof time compared with the other treatments and the control. CMC gave the second lowest values, followed by gum arabic treatment. Addition of κ‐carrageenan increased the proof time compared with the control. In summary, locust bean gum, gum arabic, and CMC improved the dough characteristics to varying degrees. κ‐Carrageenan was the only gum that showed a detrimental effect on frozen dough.     

Characterization of the Carbohydrates of Nonreduced Glutenin Fractionated by Multistacking SDS-PAGE from Two Hard Red Spring Wheat Flours

Native (nonreduced) glutenin aggregates of two hard red spring wheat flours (Len of good quality and line 205 of poor quality) containing the same high molecular weight glutenin subunits (2*, 7+9, and 5+10) were investigated for the possibility of glycosylation. Glutenins isolated by pH-precipitation were separated and purified under nonreducing conditions into five different molecular weight species by multistacking SDS-PAGE and by transfer to polyvinylidene difluoride (PVDF) membranes by electroblotting. Carbohydrate compositions of the total glutenin fraction and of the different molecular weight glutenin species separated on the stacking gels (4–12% acrylamide) were determined. More total carbohydrates were found in the total glutenin of the line 205 flour (2.33%) than in that of the Len flour (1.87%), with glucose and xylose contributing to the greater total amount in line 205. However, the total glutenin of Len had approximately twice as much of the total of arabinose, mannose, and galactose as line 205 had. After purification by electrophoresis, smaller amounts of monosaccharides (glucose, xylose, arabinose, galactose, and mannose) in the different molecular species were detected. After electroblotting to PVDF membrane to increase purification, no arabinoxylans were found. Following extraction of glutenin from membranes, the β-elimination procedure in mild base under reducing conditions was used. In that procedure, the mono- or oligosaccharide side chains are released from the protein core, and the sugars originally involved in the protein-sugar linkage are reduced to the sugar alcohol. After derivatization (with trimethylsilyl), samples were analyzed by gas chromatography and mass spectrometry. Glucose, galactose, and a small amount of mannitol were found in most of the different glutenin aggregates, except that mannitol was not found in the 8% stacking gel glutenin fraction. The content of mannitol was greater in the higher molecular weight glutenin species at the 4 and 6% origins. These results support the hypothesis that the carbohydrates and protein in the glutenin macropolymer may be covalently linked.     

Enhanced molecular identification of Botrytis spp. from New Zealand onions

Botrytis spp. associated with neck rot disease were isolated from New Zealand onions. The fungi were identified using molecular sequences of the ribosomal internal transcribed spacer (ITS) and intergenic spacer (IGS) regions, and the glyceraldehyde-3-phosphate dehydrogenase (G3PDH) gene. Analyses of the sequences showed that the majority of the isolates gathered in 2005–07 were B. aclada. A new high resolution melting analysis (HRMA) assay was developed that allowed fast and simple discrimination between B. aclada and other Botrytis spp. causing onion neck rot in New Zealand. To further verify these results, Botrytis isolates from New Zealand onions, stored in the International Collection of Microorganisms from Plants (ICMP), were also examined. Only a single isolate from the ICMP collection was B. aclada while two isolates were B. byssoidea, one B. squamosa and another closely related to Botryotinia porri. Identification of the remaining Botrytis isolates was more difficult; while IGS and ITS sequences indicated a close relationship to B. allii or B. byssoidea, a previously unreported intron insertion was observed at the 3′ end of the ribosomal small subunit gene in these isolates. No evidence of heterogeneity was observed in the G3PDH gene sequences, as might have been expected of the allodiploid B. allii, but the G3PDH sequence ruled out B. byssoidea as the identity of these isolates.     

Towards developing resistance to chickpea chlorotic dwarf virus through CRISPR/Cas9-mediated gene editing using multiplexed gRNAs

Journal of Plant Diseases and Protection - Mastreviruses are an emerging group of viruses transmitted by leafhoppers and infect both monocot and dicot plants. Chickpea chlorotic dwarf virus...