Expression of Anti-Lipopolysaccharide Factor Isoform 3 in Chlamydomonas reinhardtii Showing High Antimicrobial Activity

Antimicrobial peptides are a class of proteins with antibacterial functions. In this study, the anti-lipopolysaccharide factor isoform 3 gene (ALFPm3), encoding an antimicrobial peptide from Penaeus monodon with a super activity was expressed in Chlamydomonas reinhardtii, which would develop a microalga strain that can be used for the antimicrobial peptide production. To construct the expression cluster, namely pH2A-Pm3, the codon optimized ALFPm3 gene was fused with the ble reporter by 2A peptide and inserted into pH124 vector. The glass-bead method was performed to transform pH2A-Pm3 into C. reinhardtii CC-849. In addition to 8 μg/mL zeocin resistance selection, the C. reinhardtii transformants were further confirmed by genomic PCR and RT-PCR. Western blot analysis showed that the C. reinhardtii-derived ALFPm3 (cALFPm3) was successfully expressed in C. reinhardtii transformants and accounted for 0.35% of the total soluble protein (TSP). Furthermore, the results of antibacterial assay revealed that the cALFPm3 could significantly inhibit the growth of a variety of bacteria, including both Gram-negative bacteria and Gram-positive bacteria at a concentration of 0.77 μM. Especially, the inhibition could last longer than 24 h, which performed better than ampicillin. Hence, this study successfully developed a transgenic C. reinhardtii strain, which can produce the active ALFPm3 driven from P. monodon, providing a potential strategy to use C. reinhardtii as the cell factory to produce antimicrobial peptides.     

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。     

杏鲍菇秋葵咀嚼片配方优化与质量标准研究

为了优选杏鲍菇秋葵咀嚼片配方,以杏鲍菇粗多糖、杏鲍菇超细粉为主要原料,采用全粉末直接压片技术,选取片重差异、崩解时限、硬度及脆碎度的综合评定值为评价指标,通过响应面法确定最佳配方,并制定产品的质量指标。结果表明,杏鲍菇秋葵咀嚼片的最佳配方为杏鲍菇粗多糖38%、杏鲍菇超细粉49%和黄秋葵超细粉13%,制成的咀嚼片表面光滑、色泽均匀。综上,采用多指标综合评分法优化杏鲍菇秋葵咀嚼片配方是可行的,所得工艺配方稳定可靠,这为我国咀嚼片剂工艺创新及其产业化应用研究提供了新思路。     

菠萝蜜种子淀粉制备香草兰微胶囊的工艺研究

以菠萝蜜种子淀粉为壁材,采用饱和水溶液法制备香草兰精油微胶囊,以包埋产率为指标,采用响应面分析法对微胶囊的包埋条件进行优化探讨。结果表明:5个单因素中,影响最显著的因素为壁芯材比例、包埋温度和包埋时间。响应面优化得到香草兰精油微胶囊的最佳工艺条件为壁芯材比例为7.5∶1;包埋时间72 min;包埋温度54℃,此条件下的包埋产率为(95.46±0.2)%,包埋率为(76.35±0.6)%,载油量为(27.73±0.3)%。试验证明,此条件结果与模型预测值相吻合,此工艺条件可为菠萝蜜种子淀粉包埋香草兰精油微胶囊工艺提供理论依据。     

不同处理方式对蒸谷米复水性的影响

以稻谷为原料,对蒸谷米的复水性进行了研究。通过单因素试验,探讨超高压浸泡工序、蒸煮工序、干燥工序中相关参数的影响规律,并以蒸煮时柠檬酸添加量、蒸煮次数、微波干燥时间为变量,成品复水率为指标,通过响应面分析建立回归方程,得出蒸煮次数对复水性影响最大,柠檬酸添加量次之,干燥时间影响最小。利用模型确定了较佳参数为柠檬酸添加量1.1%,微波干燥时间6.4 min,蒸煮次数3次。     

响应面法优化超声波辅助提取薏苡仁低聚糖工艺的研究

采用响应面法优化超声波辅助提取薏苡仁低聚糖的工艺条件。在单因素试验基础上,选取液料比、超声波时间以及超声波功率3个因素结合Box-Behnken试验建立数学模型,分析考察3个因素对薏苡仁低聚糖响应值的影响程度,优化工艺参数。各因素对薏苡仁低聚糖提取率影响程度从大到小顺序依次为:超声波功率超声波时间液料比。响应面设计法优化出其最佳超声波提取条件为:超声波温度70℃,液料比33∶1(m L/g),超声波时间27 min,超声波功率450 W。在该条件下,薏苡仁低聚糖提取率为0.94%,与模型预测值0.98%接近。说明使用响应面法优化超声波辅助提取薏苡仁低聚糖的工艺条件是可行的。     

An investigation on the effect of heat-moisture treatment on baking quality of wheat by using response surface methodology

Effect of heat-moisture treatment on quality properties of two bread wheats (cvs. Tosunbey and Bayraktar) were investigated by using response surface methodology (RSM). Temperature and moisture conditions in the experimental design were in the range of 55–95 °C and 13–19%. Heat-moisture treated grains were milled into flour and quality properties were determined. The optimum moisture-temperature combination for the highest dry gluten, Zeleny sedimentation, Alveograph W and bread volume values were estimated as 14%-63 °C for Tosunbey and 19%-55 °C for Bayraktar samples. Alveograph W seems to be a good indicator of baking quality for wheats treated at higher temperatures. In order to describe the relationship between the dependent and independent variables (moisture, temperature), the response values were fitted by second order polynomial models. Significance analysis showed that the effect of both moisture and temperature on dry gluten content, sedimentation and falling number values for Tosunbey; falling number and damaged starch values for Bayraktar were significant (p < 0.05). The effect of temperature on Farinograph water absorption, W and P/G, bread volume and firmness values were significant for both cultivars (p < 0.05). It can be concluded that improvement in baking quality can be achieved and flours with different properties can be produced by heat-moisture treatments on wheat.     

Construction of recombinant capripoxviruses as vaccine vectors for delivering foreign antigens: Methodology and application

Goatpox (GTP), sheeppox (SPP) and lumpy skin disease (LSD) are three severe diseases of goat, sheep and cattle. Their typical clinical symptoms are characterized by vesicles, papules, nodules, pustules and scabs on animal skins. The GTP, SPP and LSD are caused by goatpox virus (GTPV), sheeppox virus (SPPV) and lumpy skin disease virus (LSDV), respectively, all of which belong to the genus Capripoxvirus in the family Poxviridae. Several capripoxvirus (CaPV) isolates have been virulently attenuated through serial passaging in vitro for production of live vaccines. CaPV-based vector systems have been broadly used to construct recombinant vaccines for delivering foreign antigens, many of which have been demonstrated to induce effective immune protections. Homologous recombination is the most commonly used method for constructing recombinant CaPVs. Here, we described a methodology for generation of recombinant CaPVs by the homologous recombination, and further reviewed CaPV-vectored vaccines for delivering foreign antigens.     

The role of ear environment in postharvest susceptibility of maize to toxigenic Aspergillus flavus

A kernel screening assay (KSA) was used to assess the genetic and environmental effects on the vulnerability of maize to aflatoxin accumulation. Kernels of 26 inbred lines that had been grown in seven environments, and 190 lines of the Intermated B73xMo17 (IBM) population grown in one location in the United States, were inoculated with a toxigenic strain of A. flavus and incubated in the dark at 30°C for 6 days. Percent kernel colonization (PKC), sporulation and aflatoxin were influenced by the maize genotypes (G), the location (“ear environment” or E) and the GxE interactions. Overall, low broad‐sense heritabilities were observed for PKC, sporulation and aflatoxin. PKC was significantly correlated with sporulation in all environments. Aflatoxin was positively correlated with colonization for two and with sporulation for all ear environments. Higher grain sulphur or magnesium in IBM was associated with less colonization or aflatoxin. Postharvest susceptibility of maize to aflatoxin is thus influenced by factors that are modulated by the ear environment. In a KSA, sporulation could be a proxy test for aflatoxin accumulation.     

橡胶园酸性、中性土壤交换性钙、镁测定方法研究

为了获得简单、易操作、准确度高的橡胶园酸性、中性土壤交换性钙、镁测定方法,本试验通过响应面优化试验,建立称样量、提取液体积、提取方式与交换性钙测定结果之间的数学模型。结果表明：在25℃条件下,以1 mol/L的乙酸铵为土壤浸提液,称样量为2 g、提取液体积175 mL、振荡36 min,为橡胶园酸性、中性土壤的最佳提取条件。将上述条件下得到的结果与标准方法条件下得到的结果相对比,两结果的相对误差在1.25~3.04之间,均属合理范围。利用标准样品对优化方法进行试验验证,结果与推荐值相符。改进后的方法分析成本低、操作简便、测定结果准确、稳定性好,可用于橡胶园酸性、中性土壤交换性钙、镁的测定。