链霉菌30702复合壳聚糖对番木瓜PRSV的防控效果

探究链霉菌30702复合壳聚糖对番木瓜环斑病毒病（papaya ringspot virus，PRSV）的防控效果，为番木瓜环斑花叶病防治提供基础。以台农二号番木瓜为材料，壳聚糖设3个浓度1.0g/L、0.5g/L、0.25g/L；30702菌液2个浓度0 cfu/L和4.2×10 6cfu/L，互相组合为6个处理，以清水处理为空白组，宁南霉素0.05 g/L为对照组，共8个处理；运用分光光度计法测量相关酶活。1.0 g/L壳聚糖复合4.2×106 cfu/L链霉菌30702的防治效果为88.6（p＜0.05），显著高于其他组；壳聚糖复合链霉菌30702组的防治效果、超氧化物歧化酶(SOD)活性、过氧化物酶（POD）活性、多酚氧化酶(PPO)活性相较于单独使用壳聚糖组有显著提升。使用1.0 g/L壳聚糖复合4.2×106 cfu/L的30702对番木瓜防控PRSV的效果最好，防治指数达到88.6%。     

脱乙酰壳聚糖保鲜水萝卜的研究

采用脱乙酰壳聚糖对水萝卜进行涂膜保鲜研究。以水萝卜失水率为指标,考察了稀醋酸浓度、脱乙酰壳聚糖浓度、浸涂时间对保鲜效果的影响,并测定保鲜前后水萝卜的呼吸强度、颜色、硬度及表面褶皱程度的变化。结果表明,最佳保鲜工艺为脱乙酰壳聚糖浓度为1.5%、醋酸浓度为1.0%、浸涂时间为30s。此工艺处理对水萝卜具有可靠的保鲜效果。     

气调包装复合保鲜技术在水产品保鲜中的应用现状

水产品营养价值高,含有多种氨基酸、不饱和脂肪酸、矿物质等营养素而脂肪含量相对较低,深受人们喜爱,也是中国居民膳食指南中的重要组成部分。水产品中由于水分、蛋白质等成分含量丰富,极易在微生物和内源酶的作用下发生蛋白质、脂肪等的降解或氧化而腐败变质,从而缩短了货架期,降低了商业价值。气调包装保鲜技术是指通过使用高阻隔性能的复合包装材料,并调整食品贮藏环境中的气体组成来改善食品贮藏品质并延长货架期的技术,因其安全性高、保鲜效果好而被广泛应用于水产品保鲜领域中,本文总结了近几年气调包装复合其他保鲜技术(低温、紫外杀菌、臭氧处理、超高压处理、化学保鲜剂、生物保鲜剂)在水产品保鲜领域中的应用,旨在为气调包装在水产品保鲜中的进一步应用提供理论依据。已有的研究表明该复合技术能有效地降低水产品贮藏末期微生物总数、挥发性盐基氮、三甲胺、生物胺等指标,并维持较高的感官评分、水分含量、鲜度值,可有效延长水产品的货架期,未来可以从气调包装复合保鲜技术的抑菌机理、理化指标变化机理、包装材料及模式等方面展开深入研究。     

转基因玉米TC1507质粒DNA标准物质的研制

标准物质具有特定量值、均匀性和稳定性三大典型特征,也是其作为测量标尺的依据。转基因生物标准物质是我国转基因产品标识制度顺利实施的关键技术支撑之一。文章以转基因玉米TC1507为对象,制备了转化体特异性的新型质粒DNA标准物质pTC1507,并对其均匀性、稳定性、量值进行了评价和测定。测试结果表明,制备的质粒DNA标准物质具有良好的瓶间和瓶内均匀性,pTC1507稳定性可靠,可以在-20 ℃稳定放置6个月以上。经过测序和实时荧光定量PCR定值以及不确定度评估,pTC1507标准物质的量值是1.01±0.053。均匀性、稳定性和量值结果表明,研制的转基因玉米TC1507质粒分子标准物质符合标准物质的典型要求,可以代替传统的基体标准物质应用于转基因玉米检测,解决传统标准物质获取困难、制备复杂、成本高等不足。     

利用QuantStudioTM 3D数字PCR分析转基因玉米MON863含量

QuantStudioTM 3D数字PCR (QuantStudioTM 3D digital PCR,3D-dPCR)是一种基于超高密度亲疏水微孔芯片实现数字PCR分液原理的新型核酸绝对定量平台,在转基因生物定量领域具有极大的应用前景.本研究基于3D-dPCR平台,以转基因玉米(Zea mays)MON863混合样品为例,建立基于单重和双重数字PCR体系的转基因生物(genetically modified organisms,GMOs)含量分析方法.与传统qRT-PCR比较发现,在缺乏样品纯度、纯合度信息的情况下,数字PCR能够较好地排除这些因素的影响,测定准确的量值.研究结果表明,QuantStudioTM 3D数字PCR是一种适用于转基因生物含量分析的精确定量方法,还可反映转基因玉米种子的基因型.本研究基于3D-dPCR建立的转基因玉米MON863单重和双重定量方法为转基因检测提供了新的方法和参考.     

链霉菌和壳聚糖对淮山土壤微生物活性与群落的影响

为探究链霉菌30702和壳聚糖对淮山土壤微生物活性与群落的影响,采用比色法和滴定法观测土壤酶活性的变化,采用高通量测序技术和Trimmomatic等软件,分析土壤微生物群落中细菌和真菌的发展变化。结果表明,链霉菌、壳聚糖、土壤原始微生物和培养时间等4个因素对淮山土壤的脲酶和过氧化氢酶活性均有显著影响,壳聚糖浓度在0~10.0 g/kg内,土壤酶活性随壳聚糖浓度的增大而提高;随时间的延长,先升再降;壳聚糖与链霉菌之间存在交互效应。链霉菌和壳聚糖能增加淮山土壤细菌的物种丰度,减少细菌的多样性,增加艾德昂菌属（Ideonella）和纤维弧菌（Cellvibrio）等有益菌属的相对丰度,而对真菌的物种丰度和多样性的影响较小,与对照之间的差异不显著,土壤优势菌属为腐质霉属（Humicola）、枝孢属（Cladosporium）、鞘氨醇单胞菌属（Sphingomonas）和uncultured_bacterium_ c_Subgroup_6。其中,链霉菌与2.5 g/kg壳聚糖复合处理的细菌物种丰度最大,细菌多样性最少。     

Minimally modified low-density lipoprotein up-regulates endothelin receptors in mouse mesenteric artery through p38 MAPK pathway

AIMTo investigate whether minimally modified low-density lipoprotein (mmLDL) affects the quantity and activity of endothelin （ET） type A (ET_A) and type B (ET_B) receptors in mouse mesenteric artery by activating p38 mitogen-activated protein kinase (MAPK) inflammatory pathway. METHODSThe KM mice were divided into normal saline (NS) group (injection of NS via caudal vein), mmLDL group (injection of mmLDL via caudal vein), LDL group (injection of LDL via caudal vein), mmLDL+SB 203580 group (injection of mmLDL via caudal vein and intraperitoneal injection of p38 MAPK pathway specific inhibitor SB 203580) and mmLDL+DMSO group (injection of mmLDL via caudal vein and intraperitoneal injection of DMSO). Mesenteric artery ring segment vasoconstriction dose-response curves affected by sarafotoxin 6c (S6c) and ET-1 were recorded by the myography system. The mRNA levels of ET_B receptor, ET_A receptor and interleukin-6 (IL-6) were detected by RT-qPCR. The protein levels of ET_B receptor, ET_A receptor, IL-6, p38 MAPK, p-p38 MAPK, NF-κB and p-NF-κB were determined by Western blot. The serum concentration of IL-6 was measured by ELISA. RESULTSThe contractile responses of the blood vessel segments to S6c and ET-1 were significantly increased by mmLDL (P<0.01). The mRNA and protein expression levels of ET_A receptor, ET_B receptor, and IL-6 significantly increased (P<0.01). The protein levels of p-p38 MAPK and p-NF-κB were significantly increased (P<0.01). The serum level of IL-6 was significantly increased (P<0.01). These effects of mmLDL were inhibited by p38 MAPK inhibitor SB 203580. CONCLUSION mmLDL increses the serum concentration of IL-6, up-regulates the expression of IL-6, ET_A receptor and ET_B receptor in mouse mesenteric artery, and enhances the vasoconstriction function medi?ated by ET_A and ET_B receptors, which is related to the activation of p38 MAPK inflammatory pathway and downstream NF-κB pathway.     

改性尿素追施对冬小麦和夏玉米季氮素挥发和淋溶的影响

以NBPT(N-丁基硫代磷酰三胺)和DMPP(3,4-二甲基吡唑磷酸盐)为复合抑制剂,利用转鼓喷涂工艺,开发出新型复合型抑制剂涂覆尿素肥料,采用扫描电镜和能谱仪分析其涂覆效果。通过田间试验系统对比评价了追施不同氮肥对调控氮素的特征效果。试验设置5个处理:(1)不施氮肥(CK);(2)农民习惯追施尿素(CU);(3)优化追施尿素(CUU);(4)优化追施抑制剂涂覆尿素(CUY1);(5)优化追施抑制剂涂覆尿素(CUY2)。在夏玉米喇叭口期、冬小麦拔节期追施氮肥后的15天内进行田间原位连续动态观测。电镜和能谱结果表明,复合抑制剂均匀涂覆于尿素表面,形成薄而致密、光滑的涂覆层,该涂覆层均匀分布有磷和硫2种元素,表明复合抑制剂与尿素已有效结合。田间试验结果表明,在同等优化施氮量下与普通尿素相比,夏玉米和冬小麦季追肥后CUY1和CUY2处理氨挥发分别降低55.19%,32.15%和52.46%,39.43%。夏玉米季追肥后,0-20 cm土层CU、CUU处理土壤硝态氮含量于第5天达到峰值,到后期已显著低于CUY1、CUY2处理,CUY2处理稳定硝态氮的效果更好。冬小麦季追肥后,0-20 cm土壤硝态氮含量CU、CUU处理分别在第5,3天达到峰值,CUY1、CUY2处理于第11天达到峰值后,硝态氮含量已显著高于相同施氮量的CUU处理。在保证产量和净收益的同时,抑制剂涂覆尿素显著降低了追施氮素的氨挥发和淋溶损失浓度,其中冬小麦季CUY1处济效益较好,夏玉米季CUY2调控氮素的效果最佳,减少向下淋溶的效果明显。     

Rapid screening of genetically modified ingredients in soybean and cotton processing by-product and waste using direct qPCR

To develop a simple and fast method for screening genetically modified ingredients from processing by-product and waste, direct quantitative PCR (qPCR) kit-Taqman which omitting multi genomic DNA preparing steps was developed in this study. A total of 18 oil crop processing by-products and wastes including 10 soybean and 8 cotton materials were collected from food processing factories. Compared with 2 commercial direct qPCR kits, conditions of DNA releasing procedure and PCR amplification were optimized. Element screening was performed at the initial step of genetically modified (GM) ingredient testing procedure via direct qPCR. GM event identification was carried out in positive samples by initial screening. Totally 5 screening elements (P–35S, T-NOS, Cp4-epsps, bar and pat) for soybean materials and 6 screening elements (P–35S, T-NOS, NPTII, Cry1Ac, bar and pat) for cotton samples were detected. In GM event identification, MON531 and MON1445 were found in cotton materials. Results were further confirmed by real-time PCR with DNA extraction and purification. The direct qPCR system proposed by this research was convenient for rapid screening and identification of GM ingredients in oil crop primary by-product and waste.