瓜类褪绿黄化病毒及其系统进化分析

【目的】检测新疆南疆7个县/市瓜类褪绿黄化病毒(Cucurbit chlorotic yellows virus, CCYV),进行系统发育分析,为新疆南疆CCYV预警和防控提供参考。【方法】于2019年,从中国新疆南疆7个县/市采集带有黄化特征的51份甜瓜叶片样品,RT-PCR进行CCYV的检测,进行特异性片段的克隆、测序和系统发育分析。【结果】中国新疆巴楚县、阿克苏市、莎车县、伽师县、疏勒县、疏附县均未检测到CCYV,而洛浦县的13份样本中,8份检出为CCYV阳性,检出率为61.53%,洛浦县CCYV特异性片段克隆测序获得了685 bp的核酸序列,与GenBank中的CCYV分离物外壳蛋白(Coat protein, CP)的一致性达到100%。所得序列与中国新疆(吐鲁番)、中国其他省、苏丹、日本、塞浦路斯、黎巴嫩的CP基因聚在I组(Group),沙特阿拉伯的分离物聚在Ⅱ组,伊朗的分离物聚在Ⅲ组。【结论】CCYV在中国新疆南疆洛浦县甜瓜产区发生,与中国新疆吐鲁番、中国其他省及周围国家的CCYV分离物亲缘关系很近,且群体遗传变异很小。     

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。     

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.     

油菜素内酯及配施外源钙对日光温室越冬茬番茄生长、坐果及产量的影响

为检验油菜素内酯(BR)的应用效果,采用大田试验的方法,研究不同浓度BR及其配施外源钙对日光温室越冬茬番茄生长、生理特性变化、坐果及产量的影响。结果表明,在试验浓度范围内,高浓度BR处理对番茄前期株高生长起到一定的抑制作用;适宜浓度的BR处理使株高增加。高浓度BR处理使番茄叶片MDA含量显著增高,且降低可溶性糖含量;适宜浓度的BR处理可降低番茄叶片MDA含量和相对电导率,增加番茄叶片的脯氨酸含量和可溶性糖含量,提高番茄的叶绿素含量。高浓度的BR处理会抑制番茄坐果,降低番茄第1花序的产量;适宜浓度的BR处理可提高番茄各层花序的坐果率,提高番茄产量。BR配施外源钙处理对番茄前期生长、生理指标优化、提高番茄坐果率和产量的加成效应不明显。综上,BR配施外源钙处理的效果不显著,适宜浓度的BR可以单独在日光温室越冬茬番茄生产上应用,以提高番茄的抗逆性和产量。     

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。     

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.     

寨卡病毒SYBR GreenⅠreal-time PCR方法的建立及应用

为建立一种针对寨卡病毒的快速诊断方法,本研究根据寨卡病毒的3’端保守基因序列,设计合成1对引物,建立了检测寨卡病毒的荧光定量PCR方法。结果显示:所建立的检测方法的Ct值与标准品在1.41×10^1~1.41×10^10^ copies/μL具有良好的线性关系,相关性为1,斜率为-3.502;灵敏性结果显示,该方法的检测限度为1.41×10^1 copies/μL,是普通PCR的10000倍;特异性结果显示,对CHIKV、DENV和JEV无特异性扩增,特异性强;重复性试验结果显示,组内和组间变异系数均小于1%,重复性好。本研究建立的SYBR Green I real-time PCR检测方法,可用于寨卡病毒感染的快速诊断。     

1株H1N1亚型猪流感病毒的全基因组特征及其对小鼠的致病性

旨在了解河南省猪流感病毒的流行情况及其遗传进化和基因组特征。2018年4月，从河南省某一出现疑似流感症状猪群中采集鼻拭子样品150份用于分离病毒，对分离病毒的全基因组进行序列测定和分析。同时感染6周龄BALB/c小鼠，研究其对小鼠的致病性。结果显示，获得1株H1N1亚型病毒[命名为A/swine/Henan/NY20/2018（H1N1）]。遗传进化表明，其HA和NA基因属于欧亚类禽H1N1分支，PB2、PB1、PA、NP和M基因属于2009甲型H1N1分支，NS基因属于经典H1N1分支。HA蛋白的裂解位点序列为PSIQSR↓GL，具有低致病性流感病毒的分子特征，在小鼠肺和鼻甲有效复制并能引起肺组织病理学变化。本研究分离到1株3源重排H1N1亚型病毒，对小鼠呈现一定致病力，提示应进一步加强对SIV的监测。