Comparative proteomics analysis of maize (Zea mays) leaves infected by small brown planthopper (Laodelphax striatellus)

Maize rough dwarf disease (MRDD) is a viral disease caused by brown planthopper infestation, and leads to great yield loss, especially in China. Comparative proteomics was performed using maize inbred line Zheng 58 and LN 287. MRDD pathogen was detected as rice black-streaked dwarf virus (RBSDV) by quantitative real time PCR (qRT-PCR) in Shandong Province, China. The modified trichloroacetic acid (TCA)/acetone method was used for soluble protein extraction from leaves. Two-dimensional electrophoresis (2-DE) analysis was performed on 24-cm long, pH 4–7 linear immobilized pH gradient (IPG) strips, and gels were stained with silver and coomassie brilliant blue. We identified 944 proteins expressed in RBSDV infected maize leaves by proteomics approaches. Among these, 44 protein spots that revealed a 1.5-fold difference in intensity were identified by mass spectrometry between mock-inoculated and RBSDV infected samples. Among these, 17 and 26 spots were up-regulated, and 27 and 18 spots were down-regulated in the virus infected samples of Zheng 58 and LN 287, respectively. Differential protein spots were analyzed by mass spectrometry identification, which could be divided into six categories. Furthermore, the expression of stress-related proteins was detected and confirmed by qRT-PCR. This study lays the foundation for further investigations, enabling the enhancement of MRDD resistance in maize.     

粪肠球菌生长曲线的测定及其对小鼠脑组织的影响

为测定致羔羊脑炎粪肠球菌（Enterococcus faecalis）的生长曲线，寻求一种快速而准确的方法测定不同生长时期粪肠球菌数量，并客观评价其毒性强弱及其对小鼠脑组织的影响，试验采用平板菌落计数法和OD-Monitor振荡比浊法（D_λ值法）测定粪肠球菌的生长曲线，探究该菌在合适时间段内的吸光值（D_{600 nm}）与平板菌落计数法测定的活菌数（CFU）的关系。用粪肠球菌感染小鼠，观察记录小鼠的死亡情况，最后采用Karber法计算粪肠球菌感染小鼠的半数致死量（LD₅₀）。用LD₅₀的剂量感染小鼠，及时采集死亡小鼠脑组织，未死亡的小鼠72 h后全部剖杀取脑组织，一部分做涂片染色，制作病理切片，观察病理变化；一部分进行培养，用于PCR方法进行细菌的回收鉴定。结果显示，用两种方法测定此株粪肠球菌的生长曲线基本一致，在2~8 h生长迅速，为对数生长期，8~14 h生长缓慢，为稳定期，14 h之后死亡数增加，进入衰亡期；对12 h粪肠球菌D_{600 nm}与CFU的关系进行探讨，成功建立回归方程：y=20.769x-1.3422，R²=0.997；其感染小鼠的LD₅₀为7.77×10¹¹个活菌。以此剂量感染小鼠，脑组织涂片染色和培养染色，均能看到革兰氏阳性球菌；PCR结果显示，均出现了大小为112 bp的条带。对脑组织进行病理学观察发现该菌可导致脑组织充血、出血、形成微血栓，脑膜充血。通过生长曲线和其D_{600 nm}与CFU关系的建立，可实时监测粪肠球菌数量，为后期更深入研究粪肠球菌穿越血脑屏障的机制奠定重要的理论基础。     

干湿交替灌溉对抗旱性不同水稻品种产量的影响及其生理原因分析

【目的】旨在阐明全生育期干湿交替灌溉对抗旱性不同水稻品种产量的影响。【方法】以抗旱性差异显著的4个水稻品种(籼稻扬稻6号和两优培九,粳稻旱优8号和镇稻88)为材料,以常规水层灌溉(CI)为对照,在盆栽条件下研究了轻干湿交替灌溉(WMD)和重干湿交替灌溉(WSD)对水稻产量、根系、叶片及籽粒部分生理特性的影响。【结果】与CI相比,WMD处理下抗旱性较强品种扬稻6号和旱优8号产量分别提高6.90%和7.45%,抗旱性较弱品种两优培九和镇稻88产量分别降低7.28%和8.10%。WSD处理下,4个水稻品种的产量均显著下降,抗旱性较弱的品种产量降幅远高于抗旱性较强的水稻品种。WMD处理下,扬稻6号和旱优8号复水后根系氧化力、根系与叶片细胞分裂素(玉米素+玉米素核苷)含量、叶片光合速率和籽粒中蔗糖-淀粉代谢途径关键酶的活性均较CI有不同程度提高,而两优培九和镇稻88上述指标则与CI持平或有不同程度降低。WSD处理下,4个品种上述指标均较CI不同程度降低。【结论】轻干湿交替灌溉条件下,根系活性强、叶片细胞分裂素含量和光合速率高、籽粒中蔗糖-淀粉代谢途径关键酶活性强是抗旱性较强水稻品种的基本生理特征。     

Impact of sampling and storage technique,and duration of storage,on the composition of fresh grass when analysed using near‐infrared reflectance spectroscopy

The objective of this study was to examine the effect of sampling technique (pluck or cut), storage duration (immediate analysis, 24‐h or 48‐h), storage temperature (ambient or chilled) and storage conditions (air present, air excluded or breathable) on the composition of fresh grass sampled from a sward managed to simulate grazing. Treatments were repeated across four sampling dates, with grass samples stored in grip seal bags prior to analysis using near‐infrared reflectance spectroscopy. Grass sampled by ‘pluck’ had a higher crude protein and ME content, and a lower acid detergent fibre (ADF) content, compared to that sampled by ‘cut’. Grass stored for 48 h had a lower water soluble carbohydrate (WSC) and ME content and a higher ADF content than for immediate analysis. Samples stored for 24 h did not differ from immediate analysis. Grass stored at ambient temperature had a lower WSC and ME content compared to immediate analysis. Grass stored under ‘breathable’ conditions had a lower ME content and higher ADF content than immediate analysis or samples stored with air present or air excluded. It is recommended that grass for analysis should be sampled by cutting, stored chilled (4°C) in a sealed bag to minimize exposure to oxygen and analysed within 24 h of harvest.     

Polymerisation of gluten proteins in developing wheat grain as affected by desiccation

The breadmaking quality of wheat is affected by the composition of gluten proteins and the polymerisation of subunits that are synthesised and accumulated in developing wheat grain. The biological mechanisms and time course of these events during grain development are documented, but not widely confirmed. Therefore, the aim of this study was to monitor the accumulation of gluten protein subunits and the size distribution of protein aggregates during grain development. The effect of desiccation on the polymerisation of gluten proteins and the functional properties of gluten were also studied. The results showed that the size of glutenin polymers remained consistently low until yellow ripeness (YR), while it increased during grain desiccation after YR. Hence, this polymerisation process was presumed to be initiated by desiccation. A similar polymerisation event was also observed when premature grains were dried artificially. The composition of gluten proteins, the ratios of glutenin to gliadin and high molecular weight-glutenin subunits to low molecular weight-glutenin subunits, in premature grain after artificial desiccation showed close association with the size of glutenin polymers in artificially dried grain. Functional properties of gluten in these samples were also associated with polymer size after artificial desiccation.     

Disulphide structure of high-molecular-weight (HMW-) gliadins as affected by terminators

This study aimed at elucidating SS-bonds of HMW-gliadins (HGL) from wheat with the focus on terminators of glutenin polymerisation. HGL from wheat flour extracts non-treated or treated with the S-alkylation reagent N-ethylmaleinimide (NEMI) were compared. HGL from wheat flour Akteur were isolated, hydrolysed with thermolysin and the resulting peptides pre-separated by gel permeation chromatography and analysed by liquid chromatography/mass-spectrometry using alternating electron transfer dissociation/collision-induced dissociation. Altogether, 22 and 28 SS-peptides from samples without and with NEMI treatment, respectively, were identified. Twenty-six peptides included standard SS-bonds of α- and γ-gliadins, high-molecular-weight and low-molecular-weight glutenin subunits. Eleven SS-bonds were identified for the first time. Fifteen peptides unique to HGL contained cysteine residues from gliadins with an odd number of cysteines (ω5-, α- and γ-gliadins). Thus, gliadins with an odd number of cysteines, glutathione and cysteine had acted as terminators of glutenin polymerisation. Decisive differences between samples without and with NEMI treatment were not obvious showing that the termination of polymerisation was already completed in the flour. The two HGL samples, however, were different in the majority of ten peptides that included disulphide-linked low-molecular-weight (LMW) thiols such as glutathione and cysteine with the former being enriched in the non-treated HGL-sample.     

Effect of dietary GroBiotic®‐A supplementation as a prebiotic on the intestinal microflora,growth performance,haemato‐serological parameters,survival rate and body composition in juvenile beluga (Huso huso Linnaeus, 1754)

The objectives of the present study were to investigate of GroBiotic^®‐A (GBA) on growth, autochthonous intestinal microbiota and haemato‐serological parameters of beluga juvenile. A total of 180 fish (40.82 ± 5.81 g) were fed diets containing graded levels of GBA (0, 5, 10 and 20 g kg^?1 diet) for 8 weeks. No significantly differences in body composition, total viable aerobic bacteria, mean cell volume, mean cell haemoglobin, mean cell haemoglobin concentration, neutrophils, eosinophils, monocytes, albumin, glucose, triglyceride, cholesterol, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and lactate dehydrogenase were detected between treatment groups. The fish fed on 10 and 20 g kg^?1 GBA significantly showed higher lactic acid bacteria, final weight, body weight increases, weight gain and lower feed conversion ratio compared with the control and 5 g kg^?1 groups. The group fed 20 g kg^?1 GBA showed a highly significant difference in condition factor, survival rate, final length, total red and white blood cells, lymphocytes, haematocrit, haemoglobin, total protein, total immunoglobulin and lysozyme activity. The specific growth rate of the treatment groups was significantly elevated compared with the control groups. These results indicated that GBA at level 20 g kg^?1 improved growth, welfare and survival of beluga juvenile.     

土壤有机碳对沼渣或牛粪施用后温室气体排放潜力的影响—盆栽试验结果

A change in the European Union energy policy has markedly promoted the expansion of biogas production.Consequently,large amounts of nutrient-rich residues are being used as organic fertilizers.In this study,a pot experiment was conducted to simulate the high-risk situation of enhanced greenhouse gas (GHG) emissions following organic fertilizer application in energy maize cultivation.We hypothesized that cattle slurry application enhanced CO2 and N2O fluxes compared to biogas digestate because of the overall higher carbon (C) and nitrogen (N) input,and that higher levels of CO2 and N2O emissions could be expected by increasing soil organic C (SOC) and N contents.Biogas digestate and cattle slurry,at a rate of 150 kg NH4+-N ha-1,were incorporated into 3 soil types with low,medium,and high SOC contents (Cambisol,Mollic Gleysol,and Sapric Histosol,termed Clow,Cmedium,and Chigh,respectively).The GHG exchange (CO2,CH4,and N2O) was measured on 5 replicates over a period of 22 d using the closed chamber technique.The application of cattle slurry resulted in significantly higher CO2 and N2O fluxes compared to the application of biogas digestate.No differences were observed in CH4 exchange,which was close to zero for all treatments.Significantly higher CO2 emissions were observed in Chigh compared to the other two soil types,whereas the highest N2O emissions were observed in Cmedium.Thus,the results demonstrate the importance of soil type-adapted fertilization with respect to changing soil physical and environmental conditions.     

玉米北方炭疽病研究进展

 玉米北方炭疽病是一种世界性玉米病害,也是中国北方玉米产区的主要病害之一。本文主要就该病害名称由来、田间症状特征与鉴定、致病菌生物学特性、致病力分化、遗传转化与靶基因敲除方法、影响发病条件、品种抗病性鉴定与抗病性遗传和防治方法等方面进行了综述,以期为玉米北方炭疽病防控技术提供参考。     

干旱胁迫对不同种源云南松幼苗生物量与根系形态的影响

通过盆栽控水，采用双因素随机区组设计研究了9个不同地理种源云南松幼苗生物量分配和根系形态对干旱胁迫的响应，比较了干旱胁迫下幼苗生物量及其分配以及根系形态在种源间的差异，并通过隶属函数法对不同种源幼苗干旱适应性进行了初步评价。结果表明:1）干旱胁迫下，各种源云南松幼苗地径下降明显，苗高未发生显著变化，各器官生物量和总生物量多呈下降趋势，干旱胁迫并没有导致幼苗根生物量分配比和根冠比的增加，但根系长、比根长、根表面积和比根面积在干旱胁迫下增加显著。2）不同种源云南松幼苗地径、苗高、生物量及其分配比和根系形态在干旱处理下均存在显著差异，其中地径和苗高分别以西昌和双柏种源最大，各器官生物量和总生物量以双柏、西昌和册亨种源较高，察隅种源各器官和总生物量较低，但该种源具有最大的根生物量分配比和根冠比；会泽种源根长、根表面积、比根长和比根面积均最大。研究发现，在土壤水分限制条件下，云南松幼苗以根系形态变化代替根系生物量增加的策略来适应干旱，各种源云南松幼苗生长对干旱胁迫响应的差异可能是由种源地理气候因子长期影响所形成的不同适应性策略所导致。经平均隶属函数值法初步评价了不同种源云南松幼苗对干旱适应性的强弱，结果显示，西藏察隅、云南会泽和四川西昌种源对土壤干旱的适应性较强，云南双柏、贵州册亨、云南云龙种源居中，而云南新平、永仁和禄丰种源适应性较弱。