鸡Ii结合Rab5a和Rab7b分子的分析

本研究旨在探明鸡恒定链（invariant chain，Ii）与内吞体转运蛋白Rab5a和Rab7b结合的结构域和在细胞内共定位的特征。首先，用PCR和基因突变技术将Ii胞浆区与跨膜区[Ii_{（Cyt-Tra）}]、Ii CLIP （class Ⅱ-associated invariant chain peptide）-三聚体区[Ii_{（CLIP-TRIM）}]和Ii突变体[Ii_{（M81-87aa）}、Ii_{（M91-99aa）}和Ii_{（M81-99aa）}]分别插入pET-32a和pEGFP-C1构建相应的原核和真核重组质粒。其次，将构建的含有绿色荧光蛋白的重组质粒与实验室保存的含有红色荧光Rab5a和Rab7b的重组质粒共转染至人胚胎肾细胞系293 T，观察它们的共定位。将构建的原核重组质粒进行表达和纯化，最后用拉下法和免疫印迹检测Ii与Rab5a和Rab7b的结合域。结果表明，成功构建Ii结构域及Ii突变体的重组质粒。Ii_{（Cyt-Tra）}及Ii突变体均能与Rab5a和Rab7b在细胞内共定位，而Ii_{（CLIP-TRIM）}与空载体却不能。Ii的胞浆区和跨膜区是与Rab5a和Rab7b结合的功能结构域，而不是CLIP与三聚体区。综上所述，鸡Ii与Rab5a和Rab7b共定位和结合的区域是其胞浆区和跨膜区，而不是内质网腔区。这些结果提示Rab分子参与了Ii在胞内细胞器的转运机制，为进一步研究Ii及其载体在细胞内的转运机制和功能提供了新的途径。     

马尾松叶表微生物多样性

为发掘对林木生长发育有利的优良微生物资源,并筛选适合叶表微生物的收集方法,以马尾松针叶为试验材料,分别用悬摇法和超声波法收集马尾松叶表微生物,用扩增子高通量测序技术、MUSCLE和Qiime软件研究马尾松叶表微生物的多样性。结果表明:扫描电镜观测结果显示,马尾松针叶表面定殖有大量微生物,包括真菌(菌丝及孢子)和细菌。扩增子高通量测序结果表明,马尾松叶表微生物物种丰富,包含细菌运算分类单位(OTUs)490个,真菌OTUs 1273个。马尾松叶表细菌以未分类的蓝细菌属(unidentified_Cyanobacteria)(36.53%)、未分类的拜叶林克氏菌属(unidentified_Beijerinckia)(28.60%)、鞘氨醇单胞菌属(Sphingomonas)(2.35%)为优势属;叶表真菌以枝孢属(Cladosporium)(2.45%)、拟盘多毛孢属(Pestalotiopsis)(0.92%)、无头孢菌属(Capnobotryella)(0.91%)为优势属。针对叶表细菌多样性的研究表明,悬摇法和超声波法均有较高的物种检出度;在叶表真菌多样性的研究中超声波法优于悬摇法,但超声波法样品间数据变异性较大,测定结果不稳定。     

黑果枸杞茎叶生长及其生态化学计量特征对灌水施肥的响应

在石羊河中游田间条件下,通过灌水和施肥调节黑果枸杞生长。测定不同时期黑果枸杞茎、叶生长量及其化学计量学特征变化,分析器官水平生长速率与化学计量学特征的关系,验证生态化学计量学理论"生长速率假说"。灌水施肥显著促进了茎长、基径和叶片长、宽及叶干重的生长(P<0.05),而茎长、基径、叶面积和叶干重的相对生长速率与对照之间无显著差异。各处理下黑果枸杞新梢C含量及C∶N、C∶P随生育期进程呈增加趋势,而N、P及N∶P呈降低趋势;灌水和施肥处理后茎C含量及C∶N、C∶P、N∶P低于对照,茎N、P含量高于对照。各处理叶片C、N、P含量在生育期内呈降低趋势,而C∶N、C∶P及N∶P呈增加趋势;灌水和施肥后叶片C含量及C∶N、C∶P、N∶P低于对照,叶片N、P含量高于对照。茎C含量及C∶N、C∶P显著高于叶片(P<0.05),而N、P含量及N∶P显著低于叶片(P<0.05)。生长速率假说认为,生物个体的生长速率与体内的N∶P、C∶P具有负相关关系,与N、P含量呈显著的正相关关系。各处理黑果枸杞茎、叶的生长速率与其N、P含量及C∶P、N∶P总体相关性不显著。表明施肥灌水调节下黑果枸杞茎叶生长及化学计量学特征不支持生长速率假说。     

Replication of neurovirulent equine herpesvirus type 1 (EHV-1) in CD172a+ monocytic cells

Equine herpesvirus type 1 (EHV-1) is responsible for respiratory disorders, abortion and myeloencephalopathy (EHM) in horses. Two pathotypes of EHV-1 strains are circulating in the field: neurovirulent (N) and non-neurovirulent (NN). For both strains, CD172a⁺ monocytic cells are one of the main carrier cells of EHV-1 during primary infection, allowing the virus to invade the horse’s body. Recently, we showed that EHV-1 NN strains showed a restricted and delayed replication in CD172a⁺ cells. Here we characterize the in vitro replication kinetics of two EHV-1 N strains in CD172a⁺ cells and investigate if the replication of these strains is similarly silenced as shown for EHV-1 NN strains. We found that EHV-1 N replication was restricted to 7–8% in CD172a⁺ cells compared to 100% in control RK-13 cells. EHV-1 N replication was not delayed in CD172a⁺ cells but virus production was significant lower (10^3.0 TCID₅₀/10⁵ inoculated cells) than in RK-13 cells (10^8.5 TCID₅₀/10⁵ inoculated cells). Approximately 0.04% of CD172a⁺ cells produced and transmitted infectious EHV-1 to neighbour cells compared to 65% of RK-13 cells. Unlike what we observed for the NN strain, pretreatment of CD172a⁺ cells with histone deacetylases inhibitors (HDACi) did not influence the replication of EHV-1 N strains in these cells. Overall, these results show that the EHV-1 replication of N strains in CD172a⁺ cells differs from that observed for NN strains, which may contribute to their different pathogeneses in vivo.     

Determination of critical nitrogen dilution curve based on leaf area index for winter wheat in the Guanzhong Plain,Northwest China

Excessive use of nitrogen(N) fertilizers in agricultural systems increases the cost of production and risk of environmental pollution. Therefore, determination of optimum N requirements for plant growth is necessary. Previous studies mostly established critical N dilution curves based on aboveground dry matter(DM) or leaf dry matter(LDM) and stem dry matter(SDM), to diagnose the N nutrition status of the whole plant. As these methods are time consuming, we investigated the more rapidly determined leaf area index(LAI) method to establish the critical nitrogen(N_c) dilution curve, and the curve was used to diagnose plant N status for winter wheat in Guanzhong Plain in Northwest China. Field experiments were conducted using four N fertilization levels(0, 105, 210 and 315 kg ha-1) applied to six wheat cultivars in the 2013–2014 and 2014–2015 growing seasons. LAI, DM, plant N concentration(PNC) and grain yield were determined. Data points from four cultivars were used for establishing the N_c curve and data points from the remaining two cultivars were used for validating the curve. The N_c dilution curve was validated for N-limiting and non-N-limiting growth conditions and there was good agreement between estimated and observed values. The N nutrition index(NNI) ranged from 0.41 to 1.25 and the accumulated plant N deficit(N_(and)) ranged from 60.38 to –17.92 kg ha~(-1) during the growing season. The relative grain yield was significantly affected by NNI and was adequately described with a parabolic function. The N_c curve based on LAI can be adopted as an alternative and more rapid approach to diagnose plant N status to support N fertilization decisions during the vegetative growth of winter wheat in Guanzhong Plain in Northwest China.     

Identification and quantification of anthocyanins in different coloured cultivars of ornamental kale (Brassica oleracea L. var. acephala DC)

Anthocyanins are responsible for the colour of many fruits, vegetables, flowers, and coloured-leaved trees. Ornamental kale (Brassica oleracea L. var. acephala DC) is widely cultivated for its colourful inner leaves. To investigate the relationship between the degree of colouration and anthocyanin distribution, content, and composition in ornamental kale, the authors studied the pigment characteristics of five cultivars with different coloured leaves (white, pink, red, purple, and purple-black). Microscopy observation, spectrophotometer, and high-performance liquid chromatography-mass spectrometry (HPLC-MS) analysis of fresh inner leaves revealed that pink, red, and purple colourations were associated with high levels of anthocyanin, while purple-black was the result of the combination of anthocyanins and chlorophyll. In the coloured cultivars, anthocyanins were abundant mainly in the first and second cell layers below the epidermis in both the hypocotyls and inner leaves. No anthocyanin was found in the white-leaved phenotype cultivar. Anthocyanin content increased as leaf colour deepened from pink, red, to purple cultivars, which had little chlorophyll and carotenoid. The authors identified eight anthocyanins in the four coloured cultivars, including one non-acylated, four monoacylated, and three diacylated cyanidin glycosides. Cyanidin-3-(sinapoyl)(feruloyl)-diglucoside-5-glucoside was the most abundant anthocyanin in the four coloured cultivars followed by cyanidin-3-(sinapoyl)-diglucoside-5-glucoside. The analysis of anthocyanin accumulation characterisation provides important information on evaluating colouration patterns in coloured plants, and will be helpful for breeding desired leaf colours in ornamental kale.     

11个葡萄品种在乐山地区的花芽分化规律

研究了11个葡萄品种3年生树在四川乐山地区避雨栽培条件下的花芽分化表现。结果表明,结果母枝的花芽分化率,魏可、维多利亚、黑芭拉多、金手指、金优2号和巨玫瑰6个品种的都高于50%,较易形成花芽;户太8号、巨峰、夏黑的在43%~48%;美人指和红指较难形成花芽。结果母枝不同节位的花芽分化率,多数品种在第3节位≥50%;结果枝上花穗着生节位在第3.8~4.9节,其中双花率以巨玫瑰、巨峰和户太8号较高,分别为83.45%、73.61%和73.06%。以结果母枝的花芽分化率≥50%节位作为冬剪的最低节位,则金优2号、户太8号、巨玫瑰、金手指、维多利亚5个品种在第3~4节,黑芭拉多和巨峰2个品种在第2~3节,魏可在1~2节。夏黑表现花芽分化率较低,为43.33%,可能与夏黑不耐弱光有关,设施栽培时应加强透光性。     

宁南霉素等药剂对番茄黄化曲叶病毒病的田间防效

[目的]评价宁南霉素、艾德拉和碧护3种药剂混合使用防治番茄黄花曲叶病毒病的效果。[方法]通过田间试验研究8%宁南霉素、艾德拉绿(花、果)与碧护组成的药剂对番茄黄化曲叶病毒病的防治效果。[结果]8%宁南霉素、艾德拉绿(花、果)与碧护组成的试验药剂对番茄黄化曲叶病毒病具有很好的田间防治效果,第3次施药后10、20、30 d防效均在82%以上,与对照药剂番茄黄化曲叶病毒灵差异达极显著水平,且其产量比清水对照增加25 747.5 kg/hm~2,保产率为103.90%。[结论]宁南霉素、艾德拉和碧护混合使用对番茄黄花曲叶病毒病具有较好的防治效果,为番茄黄花曲叶病毒病的有效防治提供了理论依据。     

浙江省油菜花期降水量风险评估

对浙江省66个县市(区)1971—2014年油菜花期降水量及油菜产量资料进行统计分析,并基于信息扩散的风险评估模型,对浙江省油菜花期降水量在各级降水量下的超越概率进行计算分析,评估各地油菜花期面临的旱涝风险;同时分析了浙江省油菜花期各级降水量风险值与油菜产量变化之间的关系。结果表明:油菜花期降水量严重过剩的风险高值区主要位于浙西南地区,降水量短缺风险高值区主要位于浙江省沿海岛屿及宁波的部分县市;浙江省油菜花期降水量短缺基本可以通过灌溉得到改善,但油菜减产与花期降水量过多显著相关,油菜花期降水量300 mm是油菜是否减产的阈值。该结果可为油菜种植合理布局及农业保险政策提供科学依据。