Comparative Expression Analysis of Gametogenesis‐Associated Genes in Foetal and Adult Bubaline (Bubalus bubalis) Ovaries and Testes

This study was conducted to identify and analyse the expression of gametogenesis‐associated genes and proteins in foetal and adult buffalo gonads of both the sexes. Relative quantification of the genes was determined by qPCR and Western blotting. Immunohistochemistry was also performed for various gametogenesis‐associated proteins in foetal and adult gonads of both the sexes. We observed significantly (p < 0.05) increased expression of primordial germ cell‐specific, meiotic as well as genes associated with oocyte maturation and development in foetal ovaries as compared to the adult ones. However, significantly (p < 0.05) increased expression of proteins associated with oocyte maturation like GDF9 and ZP4 was found in adult ovaries, indicating temporal regulation of mRNA translation during oogenesis. Meiotic genes showed significantly (p < 0.05) increased expression in adult testes as compared to foetal testes and ovaries, indicating onset of meiosis at a later stage in spermatogenesis. In general, the expression of primordial germ cell‐associated as well as meiotic genes was higher in adult testes, indicating the increased biological activity in the organ. Immunohistochemistry revealed localized expression of gametogenesis‐associated proteins in ovarian follicles and seminiferous tubules of testes, while the surrounding somatic tissues were devoid of these proteins. The study gives an understanding of the sequential and temporal events of gene expression as well as mRNA translation during male and female gametogenesis. It could also be concluded that follicles and seminiferous tubules are the functional units of the female and male gonads, respectively, and their function could be enhanced by appropriate chemical and genetic intervention of the somatic tissue immediately surrounding them. This assumes importance in the context that buffalo attains sexual maturity at an older age of 2–3 years and have smaller ovaries with lesser number of primordial follicles in comparison with cattle, which is suggested to be the main reason of their poor breeding performance.     

Production of Wild Buffalo (Bubalus arnee) Embryos by Interspecies Somatic Cell Nuclear Transfer Using Domestic Buffalo (Bubalus bubalis) Oocytes

The objective of this study was to explore the possibility of producing wild buffalo embryos by interspecies somatic cell nuclear transfer (iSCNT) through handmade cloning using wild buffalo somatic cells and domestic buffalo (Bubalus bubalis) oocytes. Somatic cells derived from the ear skin of wild buffalo were found to express vimentin but not keratin and cytokeratin‐18, indicating that they were of fibroblast origin. The population doubling time of skin fibroblasts from wild buffalo was significantly (p < 0.05) higher, and the cell proliferation rate was significantly (p < 0.05) lower compared with that of skin fibroblasts from domestic buffalo. Neither the cleavage (92.6 ± 2.0% vs 92.8 ± 2.0%) nor the blastocyst rate (42.4 ± 2.4% vs 38.7 ± 2.8%) was significantly different between the intraspecies cloned embryos produced using skin fibroblasts from domestic buffalo and interspecies cloned embryos produced using skin fibroblasts from wild buffalo. However, the total cell number (TCN) was significantly (p < 0.05) lower (192.0 ± 25.6 vs 345.7 ± 42.2), and the apoptotic index was significantly (p < 0.05) higher (15.1 ± 3.1 vs 8.0 ± 1.4) for interspecies than that for intraspecies cloned embryos. Following vitrification in open‐pulled straws (OPS) and warming, although the cryosurvival rate of both types of cloned embryos, as indicated by their re‐expansion rate, was not significantly different (34.8 ± 1.5% vs 47.8 ± 7.8), the apoptotic index was significantly (p < 0.05) higher for vitrified–warmed interspecies than that for corresponding intraspecies cloned embryos (48.9 ± 7.2 vs 23.9 ± 2.8). The global level of H3K18ac was significantly (p < 0.05) lower in interspecies cloned embryos than that in intraspecies cloned embryos. The expression level of HDAC1, DNMT3a and CASPASE3 was significantly (p < 0.05) higher, that of P53 was significantly (p < 0.05) lower in interspecies than in intraspecies embryos, whereas that of DNMT1 was similar between the two types of embryos. In conclusion, these results demonstrate that wild buffalo embryos can be produced by iSCNT.     

Improving Potato Tuber Quality and Production by Targeted Calcium Nutrition: the Discovery of Tuber Roots Leading to a New Concept in Potato Nutrition

Calcium plays a major role in plant growth and development and in the maintenance and modulation of various cell functions, especially related to membrane structure and function and to cell wall structure. Calcium stabilizes cell membranes by bridging polar head groups of phospholipids at the membrane surface. Calcium is also an integral part of the cell wall where it provides stable intra-molecular linkages between pectin molecules, resulting in cell wall rigidity. A change in the cytosolic calcium concentration is also known to provide a cellular signal that regulates metabolism and mediates plant responses to stresses.     

A study of 1,2-dibromoethane residues in wheat and milled products

A method is described for determining residues of 1,2-dibromoethane (EDB) in milled wheat products. The residues are first extracted with benzene, then recovered azeotropically for reaction with alkali and subsequent determination of liberated bromide ion by a Volhard titration. Freshly milled flours from wheat fumigated with 65.0 and 325 mg EDB/litre at 37°C were found to contain 10.6 to 39.4 mg EDB/kg and the bran contained from 51.4 to 153 mg EDB/kg.     

Oxygen Concentration and Cysteamine Supplementation During In vitro Production of Buffalo (Bubalus bubalis) Embryos Affect mRNA Expression of BCL‐2, BCL‐XL,MCL‐1, BAX and BID

This study examined the effects of O₂ concentration (5% vs 20%) during in vitro maturation (IVM), fertilization (IVF) and culture (IVC) or supplementation of IVM and IVC media with cysteamine (50 and 100 μm , respectively; IVM, IVF and IVC carried out in 20% O₂), on blastocyst rate and relative mRNA abundance of some apoptosis‐related genes measured by real‐time qPCR in immature and in vitro‐matured buffalo oocytes and in embryos at 2‐, 4‐, 8‐ to 16‐cell, morula and blastocyst stages. The blastocyst rate was significantly higher (p < 0.05) while the percentage of TUNEL‐positive cells was significantly lower (p < 0.05) under 5% O₂ than that under 20% O₂. The mRNA expression of anti‐apoptotic genes BCL‐2 and MCL‐1 was significantly higher (p < 0.05) and that of pro‐apoptotic genes BAX and BID was lower (p < 0.05) under 5% O₂ than that under 20% O₂ concentration at many embryonic stages. Following cysteamine supplementation, the blastocyst rate and the relative mRNA abundance of BCL‐XL and MCL‐1 was significantly higher (p < 0.05) and that of BAX but not BID was lower (p < 0.05) at many stages of embryonic development, although it did not affect the percentage of TUNEL positive cells in the blastocysts significantly. The mRNA expression pattern of these genes during embryonic development was different in 5% vs 20% O₂ groups and in cysteamine supplemented vs controls. At the 8‐ to 16‐cell stage, where developmental block occurs in buffalo, the relative mRNA abundance of BCL‐2 and MCL‐1 was highest under 5% O₂ concentration and that of BAX and BID was highest (p < 0.05) under 20% O₂ concentration. These results suggest that one of the mechanisms through which beneficial effects of low O₂ concentration and cysteamine supplementation are mediated during in vitro embryo production is through an increase in the expression of anti‐apoptotic and a decrease in the expression of pro‐apoptotic genes.     

Soil and tuber calcium affecting tuber quality of processing potato (Solanum tuberosum L.) cultivars grown in Hokkaido,Japan

Calcium (Ca) nutrition for potato (Solanum tuberosum L.) is important to increase tuber Ca concentration and improve potato tuber yield and quality. High tuber Ca content among other benefits mitigates incidence of blackspot bruise through maintenance of membrane health and regulation of biochemical reactions that leads to potato tuber discoloration. However, growers avoid application of Ca fertilizer in potato production in the belief that it causes potato common scab in Hokkaido, Japan. This study was conducted in Hokkaido to determine the current status of soil Ca and tuber Ca content levels, and its effect in mitigating incidence of potato bruise. Soil and tuber samples were collected from 90 and 80 fields in Tokachi and Kamikawa districts, respectively, in 2013 and 2014. Soil samples were analyzed for base saturation, Ca saturation, and exchangeable Al. Tuber Ca content and susceptibility of tubers to bruising were also evaluated. This study found that (1) 81% and 76% of soils collected from Tokachi and Kamikawa district, respectively, were deficient in Ca level, (2) tuber Ca content was lower than the reported value (250 mg kg^?1) considered to mitigate incidence of bruise, and (3) incidence of bruise were influenced by both tuber specific gravity and Ca content. There is urgent need to apply Ca fertilizer to attain increased soil Ca levels and improve quality of tubers.     

Evaluation of the effects of climate change on forest watershed hydroecology using the RHESSys model: Seolmacheon catchment

Paddy and Water Environment - This study examines the effects of future climate changes on watershed hydroecology, including runoff, evapotranspiration, soil moisture content, gross primary...     

中成药六味地黄丸粉辐照杀菌工艺研究

采用静态堆码方式对中药六味地黄丸粉进行辐照杀菌研究。结果表明,在垂直于辐照源轴线方向不同距离处的药品堆积厚度与辐照剂量率之间呈显著负相关（ ｐ＜ ００１） ,相关系数 ｒ ＝ － ０９９４７ 。辐照工艺技术的改变会不同程度地影响杀菌效果,提高产品总体吸收剂量均匀度,是保证杀菌效果的主要因素。六味地黄丸粉中杂菌的 Ｄ１０ 值为２１７ｋ Ｇｙ 。经１００ ｋ Ｇｙ 剂量辐照后的六味地黄丸粉,没有发生质量改变,其主要药效成分含量没有变化。     

Yield and nitrogen use efficiency of wheat increased with root length and biomass due to nitrogen,phosphorus, and potassium interactions

Balanced applications of nitrogen (N), phosphorus (P), and potassium (K) are known to increase grain yield of wheat but the impact of the interactions among N, P, and K on root growth and nitrogen use efficiency (NUE) have not been proven. The aim of this study was to investigate the effect of balanced applications of N, P, and K on the rooting patterns and NUE of wheat. Two glasshouse experiments were conducted. A rhizobox study was used to assess the impact of interactions among N, P, and K fertilisers on total root length, biomass, specific root length, root length density, N use efficiency (NUE), and N uptake efficiency of the shoots (NUpE_shoot) and N nutrition index. In a separate pot study, plants were grown to maturity to confirm the effect of the observed changes in root growth on NUE, NUpE_grain, and grain/biomass yield. In the rhizobox experiment when plants were supplied with N+P+K, total root biomass increased approximately six‐fold relative to plants grown with N alone or with no fertiliser. Plants exposed to N+P+K had NUpE_shoot and NUE values that were five and ten times higher, respectively, than plants that received just fertiliser N. Plants supplied with N+P or N+P+K had N nutrition indices close to one (N‐adequate), while plants that only received N had an index of 0.62 (N‐deficient). The pot study confirmed that the changes in root length and biomass in plants exposed to N+P+K resulted in significant increases in NUE, NUpE_grain, shoot biomass, and grain yield at maturity. Interactions among fertiliser N, P, and K played a critical role in influencing root biomass and length, which was associated with increases in NUE, NUpE_shoot and NUpE_grain.