Heterologous expression of the ThIPK2 gene enhances drought resistance of common wheat

Th IPK2 is an inositol polyphosphate kinase gene cloned from Thellungiella halophila that participates in diverse cellular processes. Drought is a major limiting factor in wheat(Triticum aestivum L.) production. The present study investigated whether the application of the Th IPK2 gene could increase the drought resistance of transgenic wheat. The codon-optimized Th IPK2 gene was transferred into common wheat through Agrobacterium-mediated transformation driven by either a constitutive maize ubiquitin promoter or a stress-inducible rd29 A promoter from Arabidopsis. Molecular characterization confirmed the presence of the foreign gene in the transformed plants. The transgenic expression of Th IPK2 in wheat led to significantly improve drought tolerance compared to that observed in control plants. Compared to the wild type(WT) plants, the transgenic plants showed higher seed germination rates, better developed root systems, a higher relative water content(RWC) and total soluble sugar content, and less cell membrane damage under drought stress conditions. The expression profiles showed different expression patterns with the use of different promoters. The codon-optimized Th IPK2 gene is a candidate gene to enhance wheat drought stress tolerance by genetic engineering.     

Selective inhibition of endothelin receptor A improves white matter lesions in hypertensive rats

AIM:To observe whether selective inhibition of endothelin receptor A (ETRA) improves white matter lesions (WMLs), and explore the mechanism. METHODS:Sprague-Dawley rats (n=33) were randomly divided into sham operation group (n=9), treatment group[stroke-prone renovascular hypertensive rats-modified 2 vessel occlusion (RHRSP-modified 2VO) + ambrisentan (n=12)] and placebo group[RHRSP-modified 2VO + vehicle (n=12)]. Drug and vehicle administration was performed from 17th to 20th week and monitoring of systolic arterial pressure was performed weekly. Morris water maze test was conducted to evaluate the function of cognition. The protein levels of endothelin-1 (ET-1) in the cortex, corpus callosum and caudate putamen were quantitatively analyzed respectively. The severity of WMLs and the relationship between ET-1 and vessels were observed by the method of histopathology. RESULTS:The difference of systolic arterial pressure between treatment group and placebo group was not significant. The animals in treatment group exhibited shorter escape latency (P<0.05), more times of crossing platform (P<0.05), lower level of ET-1 in corpus callosum and caudate putamen (P<0.05), respectively, improved WMLs severity (P<0.05) and lower binding level of ET-1 to vessels compared with the placebo group. CONCLUSION:Selective inhibition of endothelin receptor A improves the severity of WMLs and ameliorates the cognitive function.     

黏虫CYP9A113基因的克隆及外源物质对基因表达的诱导效应

黏虫是我国作物上最重要的害虫之一。细胞色素P450能够参与昆虫外源物质代谢。本研究采用RACE技术克隆了一条编码黏虫P450基因的cDNA序列,并通过Real-time PCR技术,检测了4种外源物质对该基因表达的诱导效应。该基因被国际P450命名委员会命名为CYP9A113,GenBank登录号为KY436739。利用2.5%高效氯氟氰菊酯乳油的LD_(50)处理黏虫3 h,LD_(10)、LD_(30)和LD_(50)处理12 h和24 h,可诱导表达CYP9A113基因;20%氯虫苯甲酰胺悬浮剂的LD_(10)处理黏虫12、24和48 h,LD_(30)和LD_(50)处理24 h,CYP9A113基因表达呈诱导效应;0.1和0.5 mg/mL香豆素处理6、12、24和48 h,CYP9A113基因表达均呈诱导效应;0.1和0.5 mg/mL吲哚-3-甲醇处理3、6、12、24和48 h,CYP9A113基因表达均呈诱导效应。     

一个潜在催化莱鲍迪D苷合成的新型糖基转移酶

尿苷二磷酸糖基转移酶(uridine diphosphate glycosyltransferases,UGTs)催化糖基转移反应,与植物次生代谢密切相关。本研究根据甜叶菊(Stevia rebaudiana)转录组数据库,克隆到一个催化莱鲍迪D苷(rebaudioside D,RD)合成的新型糖基转移酶候选基因,对其开展生物信息学分析。结果表明,该基因开放阅读框长1380 bp,编码459个氨基酸,等电点(pI)预测为5.54,理论分子量约49.66 kD,系统发育分析表明该基因与向日葵中的UGT89A2同源,故将其命名为SrUGT89A2。构建pET28a-SrUGT89A2原核表达载体,并在大肠杆菌(BL21(DE3))中诱导表达得到重组蛋白,HPLC检测表明粗酶液能催化甜叶菊提取液形成一个新的色谱峰,该峰保留时间与莱鲍迪D苷一致。经进一步纯化UGT89A2蛋白,添加不同甜菊糖苷标准品为催化底物,但未鉴定出该蛋白催化的具体糖苷。该潜在催化甜菊糖RD苷合成的新型糖基转移酶基因SrUGT89A2的发现,为RD苷的生物合成和甜菊糖苷的生物途径研究提供新的理论依据。     

基于变量优选与机器学习的干旱区湿地土壤盐渍化数字制图

土壤盐渍化是导致土壤退化和生态系统恶化的主要原因之一，对干旱区的可持续发展构成主要威胁。为了尽可能精确地监测土壤盐渍化的空间变异性，该研究收集新疆艾比湖湿地78个典型样点，其中选取54个样本作为训练集，24个样本作为独立验证集。基于Sientinel-2 多光谱传感器（Multi-Spectral Instrument，MSI）、数字高程模型（Digital Elevation Model，DEM）数据提取3类指数（红边光谱指数、植被指数和地形指数），经过极端梯度提升（Extreme Gradient Boosting，XGBoost）算法筛选有效特征变量，构建了关于土壤电导率（Electrical Conductivity，EC）的随机森林（Random Forest，RF）、极限学习机（Extra Learning Machine，ELM）和偏最小二乘回归（Partial Least Squares Regression，PLSR）预测模型，并选择最优模型绘制了艾比湖湿地盐渍化分布图。结果表明：优选的红边光谱指数基本能够预测EC的空间变化；红边光谱指数与植被指数组合建模效果总体上优于其与地形指数的组合，3类指数组合的建模取得了较为理想的预测精度，其中RF模型表现最优（验证集R2=0.83，RMSE=4.81 dS/m，RPD=3.11）；在整个研究区内，中部和东部地区土壤盐渍化程度尤为严重。因此，XGBoost所筛选出的环境因子结合机器学习算法可以实现干旱区土壤盐渍化的监测。     

典型喀斯特洼地植被恢复过程中土壤碳氮储量动态及其对极端内涝灾害的响应

西南喀斯特地区是我国主要的生态脆弱区之一,石漠化严重,旱涝灾害频发。植被恢复是提升脆弱生态系统土壤碳氮固持的有效方式,但该区不同植被恢复方式土壤碳氮动态监测的研究还很缺乏。本研究以典型喀斯特峰丛洼地为对象,选取人工林、牧草地、人工林+牧草地、撂荒地自然恢复4种最主要的植被恢复方式为研究对象,以耕地作为对照,对比分析退耕前(2004年)、退耕10年(2014年)和13年后(2017年)土壤碳氮储量动态变化特征。其中2004—2014年研究区未发生极端内涝灾害, 2014—2017年连续发生2次极端内涝灾害事件。研究结果表明,退耕10年后, 4种恢复方式下土壤有机碳(SOC)储量均显著增加,但退耕13年后,除撂荒地SOC持续增加外,其他3种恢复方式下SOC表现出下降趋势。植被恢复后土壤全氮(TN)储量提升相对缓慢,退耕10年仅牧草地显著增加,退耕13年后人工林+牧草和撂荒地TN增加,且撂荒地在退耕后呈持续增加趋势。相关性分析结果表明,土壤交换性Ca~(2+)与SOC、TN均呈显著正相关关系,且与2014年相比, 2017年不同植物恢复方式下土壤交换性Ca~(2+)均显著下降,这可能与研究区2015年和2016年连续内涝灾害有关。以上结果说明,不同恢复方式均能显著提升喀斯特地区土壤碳氮固持,并以自然恢复最佳,其生态系统能有效抵御极端气候灾害带来的负面影响。     

PBD和RSM联用优化聚酯型儿茶素A化学合成技术参数

为探明化学合成条件对聚酯型儿茶素A（Theasinensin A,TSA）得率的影响,通过单因素试验和Plackett-Burman Design（PBD）确定TSA化学合成关键因子,然后采用响应面法（Response surface methodology,RSM）,进一步优化TSA化学合成技术参数。结果表明,氯化铜用量、甲醇体积分数、温度对TSA得率的影响差异极显著,主因素效应为甲醇体积分数>氯化铜用量>温度,最优条件为氯化铜用量43%,甲醇体积分数26%,温度15℃,聚酯型儿茶素得率为59.12%,与模型预测值59.34%接近。PBD和RSM联用优化TSA化学合成工艺可行,预测性较好,可为其他种类儿茶素氧化聚合物的高效化学合成提供借鉴和理论依据。     

A pilot study to investigate the measurement of immunoglobulin A in Welsh Cob and Welsh Pony foals’ faeces and their dam’s milk

ABSTRACT

Aims: To determine if an ELISA for measurement of IgA in equine serum could be used to measure concentrations of IgA in foal faeces and to determine correlations with concentrations in the milk of the dam.

Methods: Faeces from 20 Welsh Cob and Welsh Pony foals and milk from their dams were collected within 12?hours (Day 0) and at 6 days after parturition (Day 6). On Day 6, faeces could not be collected from 2/20 foals, and milk samples could not be collected from 3/20 mares. An equine IgA ELISA validated for serum and plasma was used to measure concentrations of IgA in all samples in triplicate. The precision of the assay for each sample type was determined using modified CV.

Results: IgA was not detectable in 7/20 Day 0 faecal samples and in 2/18 Day 6 faecal samples. For samples with detectable IgA, the mean modified CV was 10.5 (95% CI?=?6.0–15.0)% for Day 0 faecal samples, and was 6.8 (95% CI?=?4.3–9.4)% for Day 6 faecal samples. Median concentrations of IgA in faeces on Day 0 were lower than concentrations on Day 6 (0.7?mg/g vs. 37?mg/g dry matter; p?=?0.003). Concentrations of IgA in milk and faeces on Day 6 were statistically correlated (r?=?0.59; p?=?0.006).

Conclusions and clinical relevance: The IgA ELISA showed acceptable precision when used to estimate concentrations of IgA in foal faeces during the first week of life, but IgA could not be detected in 37% of meconium samples collected on Day 0. This assay may be useful for investigation of the role of maternal milk IgA in the gastrointestinal tract of neonatal foals, but further assessment of both accuracy and precision of the ELISA is required.     

Serum amyloid A (SAA) mRNA expression in chicken and quails in response to bacterial stress

Monitoring of acute phase proteins such as serum amyloid A at gene expression level may provide quick information about immune status of the host and its susceptibility towards common infections. Present study was carried out to evaluate and compare the mRNA expression of SAA gene in Rhode Island Red chicken (RIR) and Japanese quails using real time PCR analysis in response to inactivated Salmonella gallinarum culture. The results showed that expression of SAA gene was approximately 17–33 folds higher in case of birds administered with bacterial culture when compared to un-inoculated controls and expression was higher and quicker in case of quails than RIR chicken. The SAA genes from chicken and quail were cloned and upon sequence analysis it was observed that deduced amino acid sequence of SAA from chicken and quails were having approximately seven percent variation which might have significance in function of this protein in these species.