苹果赤霉素信号转导因子MdGAMYB的克隆和表达分析

以‘长富2号’苹果为试验材料,从其短枝顶芽中克隆得到1个赤霉素信号转导因子MdGAMYB,对其进行生物信息学和表达分析。结果表明,MdGAMYB的开放阅读框(ORF)长度为1 656bp,编码551个氨基酸,蛋白质分子量为59.741 kD。生物信息学分析表明MdGAMYB编码的蛋白存在多个糖基化位点和磷酸化位点;序列分析表明,Md GAYMB和其他物种的GAMYB蛋白有很高的相似性,均含有保守的R2R3 DNA结合域和GAMYB家族所特有的Box1,Box2和Box3保守区域;系统进化分析表明,Md GAYMB与梨、梅花、草莓、枣和葡萄等的GAMYB蛋白具有较高的同源性。实时荧光定量PCR分析表明,Md GAYMB具有组织表达特异性,在叶片、花和芽中的表达量较高。外源GA3处理抑制了花芽孕育和翌年成花,抑制MdGAMYB的表达。在易成花品种‘烟富6号’中的表达量高于难成花品种‘长富2号’。     

Foliage‐applied sodium nitroprusside and hydrogen peroxide improves resistance against terminal drought in bread wheat

Terminal drought is threatening the wheat productivity worldwide, which is consumed as a staple food by millions across the globe. This study was conducted to examine the influence of foliage‐applied stress signalling molecules hydrogen peroxide (H₂O₂; 50, 100, 150 μm ) and nitric oxide donor sodium nitroprusside (SNP; 50, 100, 150 μm ) on resistance against terminal drought in two bread wheat cultivars Mairaj‐2008 and BARS‐2009. These stress signalling molecules were applied at anthesis stage (BBCH 61); drought was then imposed by maintaining pots at 35% water holding capacity. Terminal drought caused significant reduction in grain yield of both tested bread wheat cultivars; however, foliage application of both stress signalling molecules at either concentration improved the performance of both bread wheat cultivars. Maximum improvement in 100‐grain weight (12.2%), grains per spike (19.7%), water‐use efficiency (WUE; 19.8%), chlorophyll content index (10.7%), total soluble phenolics (21.6%) and free leaf proline (34.3%), and highest reduction in leaf malondialdehyde contents (20.4%) was recorded when H₂O₂ was foliage‐applied at 100 μm . Foliage application of SNP enhanced the grains per spike, 100‐grain weight and grain yield by 14.9%, 11.3% and 20.1%, respectively, than control. The foliage‐applied stress signalling molecules improved the accumulation of soluble phenolics, proline and glycine betaine with simultaneous reduction in malondialdehyde contents, which enabled wheat plants to sustain the biological membranes under stress resulting in better stay green (high chlorophyll contents) under drought. This helped improving the grain number, grain weight, grain yield, WUE and transpiration efficiency. In crux, foliage‐applied H₂O₂ and SNP, at pre‐optimized rate, may be opted to lessen the drought‐induced yield losses in bread wheat in climate change conditions.     

Molecular characterisation of Theileria equi and risk factors associated with the occurrence of theileriosis in horses of Punjab (Pakistan)

Theileria equi (T. equi) is an obligate intra- and extra-erythrocytic parasite that causes equine theileriosis (ET) in equids. Equine theileriosis is considered a notifiable disease of global significance, a major constraint to the international movement of horses, and endemic in many countries. This disease may be difficult to diagnose, as it can produce variable and nonspecific clinical signs. A cross-sectional study was designed for the molecular characterisation of T. equi and to investigate the associated risk factors of ET accompanied by its consequences on haematological and sero-biochemical parameters. A convenience sampling of 500 blood samples were collected from ET suspect horses from January to December 2017. PCR was performed on all blood samples targeting the 18S rRNA gene of T. equi followed by sequencing; 9% animals tested positive with confirmed sequences. The isolates of this study showed high homology with Cuban, Russian and Brazilian isolates of T. equi (accession numbers KY111762.2 , MG551915.1 and KY952237.1 , respectively). Based on multivariate analysis, the principal risk factors consisted of absence of dogs on the premises and presence of tick infestation. The haemato-biochemical parameters showed a decrease in granulocytes and erythrocytes, and an increase in lymphocytes, monocytes, mean corpuscular volume, mean corpuscular haemoglobin, mean platelet volume, glucose, phosphorus and aspartate aminotransferase in positive horses. This is the first study which identified ET in Punjab (Pakistan) using molecular techniques and risk factors together with the haemato-biochemical variations in horses.     

Development of a monoclonal antibody-based co-agglutination test to detect enterotoxigenic Escherichia coli isolated from diarrheic neonatal calves

Escherichia coli (E. coli) strains were collected from young diarrheic calves in farms and field. Strains that expressed the K99 (F5) antigen were identified by agglutination tests using reference antibodies to K99 antigen and electron microscopy. The K99 antigen from a selected field strain (SAR-14) was heat-extracted and fractionated on a Sepharose CL-4B column. Further purification was carried out by sodium deoxycholate treatment and/or ion-exchange chromatography. Monoclonal antibodies to purified K99 antigen were produced by the hybridoma technique, and a specific clone, NEK99-5.6.12, was selected for propagation in tissue culture. The antibodies, thus obtained, were affinity-purified, characterized and coated onto Giemsa-stained Cowan-I strain of Staphylococcus aureus (S. aureus). The antibody-coated S. aureus were used in a co-agglutination test to detect K99+ E. coli isolated from feces of diarrheic calves. The specificity of the test was validated against reference monoclonal antibodies used in co-agglutination tests, as well as in ELISA. Specificity of the monoclonal antibodies was also tested against various Gram negative bacteria. The developed antibodies specifically detected purified K99 antigen in immunoblots, as well as K99+ E. coli in ELISA and co-agglutination tests. The co-agglutination test was specific and convenient for large-scale screening of K99+ E. coli isolates.     

Farmers' attitudes towards techniques for improving oestrus detection in dairy herds in South West England

Unidentified heats contribute to declining fertility rates in English dairy herds. Several techniques have been advocated to improve heat detection rates. Despite demonstrable technical efficacy and cost-effectiveness, uptake is low. A study in South West England used the Theory of Reasoned Action (TORA) to explore dairy farmers' attitudes and beliefs towards heat detection techniques. Few farmers were convinced that following prescribed observation times, milk progesterone testing and using pedometers would fit their system or improve on their current heat detection practices. Perceived difficulty of using a technique was not a constraint on adoption. Without promotion that addresses identified barriers and drivers to adoption, little change in current practice can be expected.     

Glycinebetaine Improves Chilling Tolerance in Hybrid Maize

Plant growth and development is hampered by various environmental stresses including chilling. We investigated the possibility of improving chilling tolerance in hybrid maize by glycinebetaine (GB) seed treatments. Maize hybrid (Hycorn 8288) seeds were soaked in 50, 100 and 150 mg l^?1 (p.p.m.) aerated solution of GB for 24 h and were dried back. Treated and untreated seeds were sown at 27 °C (optimal temperature) and at 15 °C (chilling stress) under controlled conditions. Germination and seedling growth was significantly hindered under chilling stress. Moreover, chilling stress significantly reduced the starch metabolism and relative water contents (RWC), and increased the membrane electrolyte leakage. However, activities of antioxidants (catalase, superoxide dismutase and ascorbate peroxidase) were increased under stress conditions. Seed treatments with GB improved the germination rate, root and shoot length, seedling fresh and dry weights, leaf and root scores, RWC, soluble sugars, α‐amylase activity and antioxidants significantly compared with untreated seeds under optimal and stress conditions. Induction of chilling tolerance was attributed to maintenance of high tissue water contents, reduced membrane electrolyte leakage, and higher antioxidant activities and carbohydrate metabolism. Seed treatment with 100 mg l^?1 GB was the best treatment for improving the performance of hybrid maize under normal and stress conditions compared with control and other levels used.     

Remobilization of Nitrogen and Carbohydrate from Stems of Bread Wheat in Response to Heat Stress during Grain Filling

When wheat (Triticum aestivum L.) is grown under heat-stress conditions during grain filling, preanthesis stored total non-structural carbohydrates (TNC) and nitrogen (N) could serve as alternative source of assimilates. This study was performed to evaluate wheat genotypes for their ability to accumulate and remobilize TNC and N stored in their stem to support grain filling under heat stress. Eighteen genotypes were used for N remobilization study while nine of them were used for TNC remobilization study. They were grown in pots and placed in a vinyl house with the maximum temperature kept below 30 °C. Five days after anthesis (5DAA), half of the pots were taken to phytotrons where temperature was gradually increased and the maximum was set at 38 °C. Grain yield and grain weight decreased by about 35 % under heat stress. Significant differences were found among genotypes in percentage reduction in grain yield, grain weight, grain filling duration and harvest index because of heat stress. The N and TNC concentrations of the stem at 5DAA were significantly different among genotypes. Heat stress significantly reduced the N remobilization efficiency of most of genotypes. However, heat stress significantly increased TNC remobilization efficiency and significant variation were observed among genotypes. N remobilization efficiency across treatments significantly correlated with grain yield, grain weight, harvest index and grain filling duration. TNC at 5DAA negatively correlated with N at 5DAA and harvest index, but the TNC remobilization efficiency under heat stress positively correlated with mainstem grain yield, grain weight and harvest index. The rate of chlorophyll loss from flag leaf positively correlated with N and TNC remobilization efficiencies under heat stress suggesting a link between leaf senescence and remobilization efficiency. The results indicate that heat stress negatively affected grain yield, its components and N remobilization while it increased TNC remobilization because of the increasing demand for resources.