黑胫病菌（Leptosphaeria biglobosa）在油菜叶片和茎中的侵染过程观察

为明确黑胫病菌（Leptosphaeria biglobosa）在甘蓝型油菜叶片和茎中的侵染及扩展过程，利用绿色荧光蛋 白（GFP）标记的黑胫病菌株接菌油菜叶片，利用激光共聚焦显微镜观察菌株在油菜叶片和茎中的侵染过程。结果 表明，接种油菜叶片7 h后，分生孢子萌发并长出芽管；17 h后，芽管侵入气孔；24 h后，分生孢子全部萌发；36 h后萌 发的芽管形成菌丝；120 h后，菌丝在叶片表皮细胞间隙蔓延，并侵入叶肉细胞。13 d后，菌丝侵入茎部皮层组织； 15 d后，菌丝在皮层细胞间隙蔓延，并侵染至茎表皮；21 d后，菌丝侵染至维管组织；23 d后，菌丝侵染至茎韧皮部； 25 d后，茎导管被侵染，并向木质部扩展。本研究发现的L. biglobosa 在油菜叶片和茎中的侵染过程，可为油菜与黑 胫病菌互作的研究、黑胫病致病机理及防治提供参考。     

小球藻遗传转化技术研究进展

小球藻( Chlorella )是一种单细胞真核藻类,属绿藻门、绿藻纲、绿球藻目、小球藻科、小球藻属 [1] 。作为最早开发的真核微藻之一,具有高营养价值、生长快速、结构简单、易工业化集成等显著优点。其细胞形态为球形或椭圆形,直径3~12 μm,呈单生或聚集成群状生长 [2] ,分布广泛,多见于淡水、咸水和土壤中。作为地球上最早的生命之一,小球藻基因比较稳定,至今未见有关其基因自发突变的报道。因其富含蛋白质、脂质、维生素、活性代谢产物等多种营养物质而被公认为具有高附加值和医疗保健作用,已经被广泛应用于保健食品 [3] 、水产养殖 [4] 、生物能源 [5] 等方面,关于小球藻生物技术的研究主要集中在基因组学 [6] 、分子遗传学 [7] 、代谢机理 [8] 、大规模培养 [9] 等方向。     

稻曲病菌交配型基因MAT1-2-1和MAT1-2-8的特性分析

 有性生殖在真菌的生活史和进化过程中具有重要作用,而交配型基因是控制有性生殖的关键因子。前期研究发现稻曲病菌(Villosiclava virens)MAT1-2型菌株中包含MAT1-2-1和MAT1-2-8两个交配型基因,但是它们如何调控稻曲病菌有性生殖依然不清楚。本文研究了它们在不同侵染和生长发育时期的表达模式和编码的蛋白结构特性。研究表明MAT1-2-1在侵染不同阶段一直下调表达;而MAT1-2-8在侵染早期(5 dpi)上调表达,在侵染后期下调表达。与营养菌丝阶段比较,MAT1-2-1和MAT1-2-8在有性发育过程菌核形成、菌核萌发、子座原基形成和子座成熟4个阶段的表达量都是下降的,在菌核形成阶段表达量最低。生物信息学分析显示MAT1-2-1和MAT1-2-8具有磷酸化位点,为非分泌蛋白,无明显的跨膜结构域。蛋白同源比对分析表明MAT1-2-1与香柱菌(Epichloë typhina)的MAT1-2-1同源性最高,而MAT1-2-8与绿僵菌(Metarhizium)的MBR_08192蛋白同源性最高。进一步研究发现MAT1-2-1和MAT1-2-8能够互作,并分别主要定位在细胞核和细胞基质中。通过质谱技术鉴定到MAT1-2-1的一些候选互作蛋白,如假定Ran交换因子Prp20/Pim1(KDB12229.1)、假定rRNA处理蛋白Ebp2(KDB12923.1)及组蛋白H1(KDB12711.1)等。因此,以上结果为研究稻曲病菌交配型基因MAT1-2-1和MAT1-2-8调控有性生殖的生物学功能奠定了基础。     

Characterization of bovine ileal epithelial cell line for lectin binding,susceptibility to enteric pathogens,and TLR mediated immune responses

In this study, primary and immortalized bovine intestinal epithelial cells (BIECs) were characterized for the expression of surface carbohydrate moieties. Primary BIEC-c4 cells showed staining greater than 90 % for 16 lectins but less than 50 % staining for four lectins. Immortalized BIECs showed significantly different lectin binding profile for few lectins compared to BIEC-c4 cells. BIEC-c4 cells were studied for infectivity to E. coli, Salmonella enterica, bovine rotavirus, bovine coronavirus, and bovine viral diarrhea virus. Bovine strain E. coli B41 adhered to BIEC-c4 cells and Salmonella strains S. Dublin and S. Mbandaka showed strong cell invasion. BIEC-c4 cells were susceptible to bovine rotavirus. LPS stimulation upregulated IL-10, IL-8, and IL-6 expression and Poly I:C upregulated TLR 8 and TLR 9 expression. This study provides important knowledge on the glycoconjugate expression profile of primary and immortalized BIECs and infectivity and immune responses of primary BIECs to bacterial and viral pathogens or ligands.     

Calibration and validation of optical chlorophyll‐measuring devices for use in predicting crude protein concentration in tropical grass herbage

The objective of this experiment was to evaluate the Fieldscout CM 1000 NDVI and Yara N‐Tester as easy‐to‐use and cost‐effective tools for predicting foliar chlorophylls (a, b and total) and crude protein (CP) concentrations in herbage from three tropical grass species. Optical chlorophyll measurements were taken at three stages (4, 8 and 12 weeks) of regrowth maturity in Guinea grass (Panicum maximum) and Mulato II (Brachiaria hybrid) and at 6 and 12 weeks maturity in Paspalum spp (Paspalum atratum). Grass samples were harvested subsequent to optical measurements for laboratory analysis to determine CP and solvent‐extractable chlorophylls (a, b and total) concentrations. Optical chlorophyll measurements and CP concentrations were highly correlated (Yara N‐Tester: r² = 0·77–0·89; Fieldscout CM 1000 NDVI: r² = 0·52–0·84). Crude protein prediction models from the Yara N‐Tester and Fieldscout CM 1000 NDVI accounted for 70–89% and 44–73% CP variability, respectively, in Mulato II and Guinea grass. The Yara N‐tester produced more accurate and reliable CP estimates based on very high concordance correlation coefficient [CCC (0·73–0·91)] and low rMSPE, mean and regression bias. It is concluded that the Yara N‐Tester produces more accurate and reliable CP estimates of tropical pastures.     

Utilizing a DUALEM‐421 and inversion modelling to map baseline soil salinity along toposequences in the Hunter Valley Wine district

In the oldest commercial wine district of Australia, the Hunter Valley, there is the threat of soil salinization because marine sediments underlie the area. To understand the risk requires information about the spatial distribution of soil properties. Electromagnetic (EM) induction instruments have been used to identify and map the spatial variation of average soil salinity to a certain depth. However, soils vary with depth dependent on soil forming factors. We collected data from a single‐frequency and multiple‐coil DUALEM‐421 along a toposequence. We inverted this data using EM4Soil software and evaluated the resultant 2‐dimensional model of true electrical conductivity (σ – mS/m) with depth against electrical conductivity of saturated soil pastes (EC_p – dS/m). Using a fitted linear regression (LR) model calibration approach and by varying the forward model (cumulative function‐CF and full solution‐FS), inversion algorithm (S1 and S2), damping factor (λ) and number of arrays, we determined a suitable electromagnetic conductivity image (EMCI), which was optimal (R² = 0.82) when using the full solution, S2, λ = 3.6 and all six coil arrays. We conducted an uncertainty analysis of the LR model used to estimate the electrical conductivity of the saturated soil‐paste extract (EC_e – dS/m). Our interpretation based on estimates of EC_e suggests the approach can identify differences in salinity, how these vary with parent material and how topography influences salt distribution. The results provide information leading to insights into how soil forming factors and agricultural practices influence salinity down a toposequence and how this can guide soil management practices.     

Comparative effect of advanced soy products or corn protein concentrate with porcine meal on growth,body composition,and distal intestine histology of Florida pompano,Trachinotus carolinus

The present study was designed to investigate the effects of diets containing advanced soy products (enzyme‐treated soy and fermented soy) or corn protein concentrate (CPC) in combination with porcine meal (PM) to completely replace poultry byproduct meal (PBM) on growth performance, body composition, and distal intestine histology of Florida pompano, Trachinotus carolinus. Four experimental diets were formulated to be isonitrogenous and isolipidic, to contain 400 g/kg crude protein and 80 g/kg lipid. A reference diet (PBM diet [PBMD]) contained 150 g/kg PBM and 495 g/kg soybean meal (SBM), and three test diets were formulated replacing PBM with 15 g/kg of CPC (CPC diet [CPCD]) or replacing all SBM and PBM with 535 g/kg fermented soy (fermented soybean meal diet [FSBMD]) or 451.3 g/kg enzyme‐treated soy (enzyme‐treated soybean meal diet [ESBMD]). All three test diets were supplemented with 38 g/kg of PM. Diets were fed based on a percentage of bodyweight adjusted after sampling the fish every 2 weeks to triplicate groups of Florida pompano juveniles (mean weight 8.06 ± 0.22 g). After 8 weeks of feeding, fish fed CPCD and ESBMD performed equally well in terms of final body weight, thermal growth coefficient, and percentage weight gain in comparison to fish fed PBMD. In all cases, feeding FSBMD resulted in poor feed conversion and lower feed intake compared to other treatments. Protein retention efficiency, whole‐body proximate composition, phosphorus, sulfur, potassium, magnesium, calcium, sodium, and zinc contents were not significantly influenced by the dietary treatments. The results obtained in the present histological study showed no significant differences in the thickness of serous layer, muscular layer, and submucosal layer of the intestine among treatments. Fish fed CPCD showed a significant widening of the lamina propria with an increase of cellular infiltration and higher presence of goblet cells compared to other dietary treatment. Based on these results, 451 g/kg ESBM or combination of 150 g/kg of CPC and 495 g/kg SBM supplemented with 38 g/kg PM can be utilized to develop a practical diet for juvenile Florida pompano without impacting growth, nutritive parameters, and several distal intestine health parameters.     

Carbohydrate and crude protein fractions in perennial ryegrass as affected by defoliation frequency and nitrogen application rate

The objective of this study was to evaluate the effects of defoliation frequency (either at two‐ or three‐leaf stage) and nitrogen (N) application rate (0, 75, 150, 300, 450 kg N ha^?1 year^?1) on herbage carbohydrate and crude protein (CP) fractions, and the water‐soluble carbohydrate‐to‐protein ratio (WSC:CP) in perennial ryegrass swards. Crude protein fractions were analysed according to the Cornell carbohydrate and protein system. Carbohydrate fractions were analysed by ultra‐high‐performance liquid chromatography. Sward defoliation at two‐leaf stage increased the total CP, reduced the buffer‐soluble CP fractions and decreased carbohydrate fractions of herbage (P < 0·001). The effect of defoliation frequency was less marked during early spring and autumn (P < 0·001) than for the rest of the seasons. An increase in N application rate was negatively associated with WSC, fructans and neutral detergent fibre (P < 0·001), and positively associated with CP and nitrate (N‐NO₃) contents of herbage. Nitrogen application rate did not affect CP fractions of herbage (P > 0·05). The fluctuations in CP and WSC contents of herbage resulted in lower WSC:CP ratios during early spring and autumn (0·45:1 and 0·75:1 respectively) than in late spring (1·11:1). The herbage WSC:CP ratio was greater (P < 0·001) at the three‐leaf than the two‐leaf defoliation stage and declined as the N application increased in all seasons (P < 0·001). The results of this study indicate that CP and carbohydrate fractions of herbage can be manipulated by sward defoliation frequency and N application rate. The magnitude of these effects, however, may vary with the season.