A rapid,simple, and sensitive immunoagglutination assay with silica nanoparticles for serotype identification of Pseudomonas aeruginosa

An agglutination test based on colored silica nanoparticles (colored SiNps) was established to detect serotypes of Pseudomonas aeruginosa. Monodisperse colored SiNps were used as agglutination test carriers. The colored SiNps were prepared through reverse microemulsion with reactive dyes, sensitized with 11 kinds of mono-specific antibodies against P. aeruginosa, and denoted as IgG-colored SiNps. Eleven kinds of IgG-colored SiNps were individually mixed with P. aeruginosa on a glass slide. Different serotypes of P. aeruginosa could be identified by agglutination test with evident agglutination. The P. aeruginosa could be detected in a range from 3.6 × 10⁵ to 3.6 × 10¹² cfu mL^?1. This new agglutination test was confirmed to be a specific, sensitive, fast, easy-to-perform, and cost-efficient tool for the routine diagnosis of P. aeruginosa.     

串番茄新品种红星302 的选育

红星302 是以抗病、耐低温弱光材料01-61 为母本，以01-23 为父本育成的串番茄一代杂种，无限生长类型，果实串收，果色亮红，果实椭圆形，酸甜适口，品质佳，VC 含量为364 mg · kg^-1，可滴定酸0.44%，可溶性糖4.25%，平均单果质量30.79 g，硬度为11.40×10⁵ Pa。单株结果数约为53个，产量4 900 kg ·（ 667 m²）^-1 左右，高抗蕨叶病毒病、青枯病，适宜河北地区冬季日光温室和早春保护地栽培。     

不同水稻种质中渗透胁迫抗性基因DREB2A的遗传多样性分析

本研究旨在分析转录因子DREB2A基因在不同水稻种质中的遗传多样性,以期为水稻耐渗透胁迫遗传改良提供分子工具。利用单倍型分析、系统进化树、遗传距离和密码子偏好性分析,对85份不同类型水稻种质中DREB2A基因的功能性核苷酸序列变异及遗传多样性进行了研究。共鉴定出55个核苷酸变异位点,其中12个位于编码区,43个位于非编码区;鉴定出12个DREB2A等位基因型,其中来自非洲栽培稻(Oryza glaberrima)的3个等位基因型表现大片段变异;根据DREB2A基因的序列变异鉴定出39个单倍型,其中33个为新单倍型;将所有单倍型分为三组（Group I、II和III）,其中非洲栽培稻(Oryza glaberrima)中的10个单倍型单独分为一组(Group III);系统进化树、遗传距离和密码子偏好性分析均表明Group III与其他两组具有较大差异。对85份不同类型水稻种质中DREB2A基因的序列分析表明,非洲栽培稻(Oryza glaberrima)中的DREB2A等位基因在序列变异、系统进化关系和密码子偏好性方面均明显不同于其他种质材料中的等位基因。     

龙井茶适制茶树品种多性状综合评价——基于多元统计分析

确定适宜加工龙井茶的茶树品种的多性状评价指标,建立龙井茶适制品种农艺性状的综合评价体系,为鉴定筛选优良龙井茶适制品种或资源提供技术支撑。对浙江省15份主要栽培茶树品种（系）的芽叶长宽比、百芽重、展叶角度、亮度值(L)、红绿属性(a)、黄蓝属性(b)、色彩饱和度(C)、氨基酸、茶多酚、水浸出物、咖啡碱、酚氨比、外形、汤色、香气、滋味和叶底17项农艺性状进行综合比较,并联合使用主成分分析、聚类分析和层次分析对茶树品种（系）的龙井茶适制性进行综合评价。17项农艺性状的平均变异系数为12.05%,百芽重的变异系数最大(28.15%),叶底的变异系数最小(0.86%),适制龙井茶农艺性状变量间既相对独立又密切相关,应用主成分分析法,将17个评价指标压缩成6个综合指标,经过分析6个主成分函数式中的17项农艺性状系数,可以将17项农艺性状分为外形因子、内质因子、感官因子3个主要指标,这3个主要指标可以较准确地评价龙井茶适制品种,其中芽叶长宽比、a值、茶多酚、酚氨比、外形、香气和汤色是主要性状。采用聚类分析,可以将15个品种分为2个大的类群,采用层次分析可以将茶树品种采制龙井茶的适宜性进行综合排序。采用多元统计分析中的主成分分析和聚类分析对15份茶叶品种的17项农艺性状进行龙井茶适制性综合评价是可行的,结合层次分析可以得到更加准确的排序,可以从不同角度进行较全面客观评价,为龙井茶适制品种的选育提供参考。     

甘肃、青海地区小麦条锈菌监测及群体遗传多样性分析

【目的】了解甘肃和青海小麦条锈菌（Puccinia striiformis f. sp. tritici）春季流行传播路线、群体遗传多样性和生殖模式,明确春季流行期两省小麦条锈菌的传播关系及菌源交流规律,进而为两省小麦条锈病的预测预报、确定越夏初始菌源来源和有效治理提供理论依据。【方法】选择条锈病常发生的地区作为调查和研究区域。甘肃省4个试验点：陇南市文县、陇东平凉市崆峒区、中部麦区定西市临洮县、临夏州临夏县;青海省2个试验点：西宁市城北区、海东市互助县。2017年秋季,在甘肃和青海省6个试验点内根据当地小麦播种适期依次种植82份变异观察圃材料。2018年4—8月,对试验点82份变异观察圃材料进行田间病害调查,并采集到551份小麦条锈菌标样,使用15对引物进行SSR分子标记分析。利用GenAlEx和POPPR v2.5.0软件对数据进行相关分析, 不显著的rbarD值表示连锁平衡,用于推断群体是否发生有性重组。【结果】82份变异观察圃材料在甘肃地区发病比青海地区严重。15对引物组合共扩增出81个位点,每对引物组合产生的多态性位点为2—12个。551份样本克隆矫正后,共鉴定出505个多位点基因型（MLG）,其中仅有32个MLG被克隆并进行了2—6次重新采样。甘肃和青海群体总的基因型多样性（G=0.917）较高,其中,甘肃平凉群体的最高,青海互助群体次之,甘肃临洮群体最低。小麦条锈菌的遗传变异主要在各群体内部个体之间。春季流行期,菌源在各群体之间交流频繁,青海东部（互助和西宁）群体与甘肃（平凉和临夏）群体之间的基因流高于青海（互助和西宁）群体与甘肃文县群体之间的基因流。最小时空网络图（MSN）和非参数主成分分析（DAPC）表明青海互助和西宁的群体与来自于甘肃平凉和临夏的群体之间菌源关系最密切,差异最小;与临洮群体遗传距离相对较远且临洮群体相对独立;文县群体则是一个完全独立的群体,与其他5个群体之间的差异最大。连锁不平衡分析表明,甘肃文县、临夏和青海西宁群体存在不显著的rbarD值表示连锁平衡,是有性生殖群体,其中文县群体（rbarD=0.0139,P=0.186）显示出明显的有性重组特征。【结论】小麦条锈病春季流行期,甘肃地区与青海东部地区的传播路线以甘肃平凉、临夏到青海的传播为主,甘肃文县到青海的传播为辅。甘肃文县、临夏和青海西宁3个群体存在有性生殖现象,对甘肃、青海地区条锈菌丰富的遗传多样性的形成具有一定作用。     

Effect of pidotimod on renal function and inflammatory response in rat IgA nephropathy model

AIM:To explore the effect of pidotimod on the renal function in IgA nephropathy (IgAN) rat model, and to further study whether this effect is related to the inhibition of inflammatory response. METHODS:The SD rats (n=36) were randomly divided into control group, IgAN model group, IgAN with prednisone treatment group and IgAN with pidotimod treatment group, with 9 rats in each group. The IgAN model was induced by consecutive oral administration of bovine gamma globulin (BGG) for 8 weeks followed by injection of BGG through tail vein for 3 d. After the IgAN model was established, the drug was continuously used for 4 weeks. At the end of the treatment, the urine protein, serum creatinine and blood urea nitrogen were examined by an automated analyzer. IgA deposition in the renal tissues was observed by immunofluorescence staining. The mRNA expression levels of renal fibrosis markers transforming growth factor-β1 (TGF-β1) and fibronectin 1 in the renal tissues were detected by RT-qPCR. The mRNA and protein levels of pro-inflammatory cytokines interleukin-1β (IL-1β) and IL-6 in the renal tissues were determined by RT-qPCR and Western blot, respectively. RESULTS:No significant difference of the body weight was observed in different groups. Compared with control group, the content of urine protein, serum creatinine and blood urea nitrogen were significantly increased (P<0.01), whereas those were reversed by pidotimod treatment. The results of immunofluorescence staining showed that pidotimod inhibited IgA deposition in the IgAN rats. Pitomod treatment inhibited the mRNA expression levels of renal fibrosis markers TGF-β1 and fibronectin 1, and the mRNA and protein levels of pro-inflammatory cytokines IL-1β and IL-6 in the renal tissues of IgAN rats. CONCLUSION:Pidotimod alleviates IgAN progression in rats by inhibition of inflammatory response.     

Regulatory effect of astragaloside IV on SDF-1α/CXCR4 in EPCs

AIM: To investigate the effects of astragaloside IV (AS-IV) on chemokine receptor 4 (CXCR4) and stromal cell-derived factor 1α (SDF-1α) in endothelial progenitor cells (EPCs) and its mechanism. METHODS: Rat bone marrow-derived EPCs were cultured in vitro. The proliferation, adhesion, migration, apoptosis and tube formation capacity of EPCs treated with AS-IV and AMD3100, a specific blocker of CXCR4, were observed. The effects of AS-IV on the expression of SDF-1α/CXCR4 at mRNA and protein levels and the protein level of p-CXCR4 in the EPCs were determined. RESULTS: AS-IV significantly enhanced the proliferation, adhesion, migration and tube formation abilities of EPCs, reduced the apoptosis of EPCs, and up-regulated the mRNA and protein expression of SDF-1α and CXCR4 and the p-CXCR4 protein level in the EPCs. On the other hand, AMD3100 blocked the up-regulating effect of AS-IV on the mRNA and protein expression of CXCR4 and the p-CXCR4 protein level in the EPCs, but did not affect the effect of AS-IV on the expression of SDF-1α. CONCLUSION: AS-IV might enhance the biological function of EPCs by regulating the expression of SDF-1α/CXCR in EPCs.     

Effect of DNA methylation of miR-30a-5p promoter region on hepatic injury induced by homocysteine

AIM: To explore the role of DNA methylation of microRNA-30a-5p(miR-30a-5p) promoter region in hepatic injury. METHODS: Four-week-old normal mice and cystathionine β-synthase (CBS) single gene knockout mice were used and divided into normal (CBS^+/+, n=12) group and single gene knockout (CBS^+/-, n=12) group, and the mice were fed with high methionine diet for 8 weeks. HL-7702 hepatic cells were routinely cultured in vitro and divided into control group, homocysteine (Hcy) group and Hcy+5-azacytidne (AZC) group. Serum Hcy, alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured by automatic biochemical analyzer. The levels of ALT and AST in the cells culture medium were determined by the microplate method. Hepatic injury in the mice were observed with HE staining. Cell viability staining was used to measure the viability of hepatocytes. RT-qPCR was used to detect the expression of miR-30a-5p in the liver tissues and hepatocytes. The correlation between the expression of miR-30a-5p and serum ALT and AST levels was analyzed by Pearson correlation analysis. DNA methylation level of miR-30a-5p promoter region in the liver tissues and hepatocytes was detected by nested landing methylation-specific PCR (nMS-PCR). RESULTS: Compared with the CBS^+/+ mice, the serum levels of Hcy, ALT and AST in the CBS^+/- mice were significantly increased (P < 0.05). HE staining showed the hepatocyte swelling and nuclear fragmentation and dissolution. The expression level of miR-30a-5p in the liver tissues was decreased (P < 0.01). Besides, the expression level of miR-30a-5p in the mice was negatively correlated with serum ALT and AST levels (r²=0.4557, P=0.0003, r²=0.4626, P=0.0003), and the DNA methylation of miR-30a-5p promoter region was increased (P < 0.01). In the HL-7702 cells, compared with control group,the ALT and AST levels were increased in Hcy group (P < 0.05, P < 0.01), and the cell viability was remarkablely decreased. DNA methylation of miR-30a-5p promoter region was increased (P < 0.01), which decreased after treated the cells with AZC (P < 0.05), while the expression level of miR-30a-5p in the cells was increased (P < 0.05). CONCLUSION: Hypermethylation of miR-30a-5p promoter region may play an important role in hepatic injury.     

基于叶绿体DNA分析的楸子种质遗传多样性研究

利用4对叶绿体DNA引物扩增49份楸子[Malus prunifolia（Willd.）Borkh.]种质资源的4个叶绿体DNA基因间区trnH-psbA、trnS-trnG spacer + intron、trnT-5′trnL和5′trnL-trnF序列，基于4个叶绿体DNA基因间区的序列变异，从母系遗传的角度评价楸子的遗传多样性水平。结果显示：4个叶绿体DNA基因间区序列经测序、拼接、比对和合并之后的片段长度为3 790 bp，共有173个多态性变异位点，其中包含2个单一突变位点、20个简约信息位点和151个插入/缺失位点。在49份楸子种质中，trnH-psbA、trnS-trnG spacer + intron、trnT-5′trnL和5′trnL-trnF区域的变异位点的数量分别为26个、25个、120个和2个，单倍型数量分别为9个、7个、8个和3个，合并之后的叶绿体DNA片段的单倍型有14个。核苷酸多样性和单倍型多样性最高的区域均为trnH-psbA（Hd = 0.775，Pi = 0.02143），最低的为5′trnL-trnF（Hd = 0.481，Pi = 0.00072）。49份楸子种质4个叶绿体DNA区域合并后的遗传多样性较高（Hd = 0.854，Pi = 0.00949）。Tajima’s D检验中，4个叶绿体DNA区域在P > 0.10水平上均不显著，楸子的4个叶绿体DNA区域在进化上遵循中性进化模型。楸子的遗传变异主要存在于群体内部，不同居群间基因交流频繁，多数居群间遗传分化较少，与地理距离不完全相关。     

羊角椒新品种寿研梦扬的选育

寿研梦扬是由母本CSVHPM1407001和父本CSVHPF1407002配制而成的羊角椒一代杂种。植株长势旺盛,连续坐果能力强;果实长羊角形,果长25～30 cm,果肩宽3～4 cm;青熟果黄绿色,老熟果红色;外表光亮,商品性好,耐贮运;单果质量100 g左右,辣味浓,宜鲜食;田间对病毒病、炭疽病和疫病的抗性强于对照喜洋洋。保护地栽培鲜椒产量可达12 840 kg·(667 m~2)~(-1)左右。适宜山东、河北等地区早春、秋延保护地种植。