异硫氰酸丙烯酯毒杀松材线虫作用机制初步研究

对异硫氰酸丙烯酯毒杀松材线虫作用机制进行了初步研究。测定了其对线虫虫体形态、运动行为、体内总糖和蛋白质含量的影响,以及处理前后线虫体内过氧化氢酶(CAT)、乙酰胆碱酯酶(AChE)和谷胱甘肽-S-转移酶(GST)活性变化。结果表明:经异硫氰酸丙烯酯处理后的松材线虫虫体形态严重扭曲,运动形态出现由正常的S形变化为多节弯曲和螺旋状卷曲等异常状况,线虫行动随时间的推移变得迟缓,最终呈C形、新月形或直接死亡。随着异硫氰酸丙烯酯处理质量浓度的增大,线虫体内总糖和蛋白质含量显著上升,糖和蛋白质的代谢发生紊乱;线虫体内CAT、AChE和GST活性显著降低,且均低于对照组,表明线虫体内氧化和抗氧化作用失衡;细胞受到极大的影响,神经系统遭到破坏,解毒能力明显降低。研究结果可为研究利用异硫氰酸丙烯酯作为植物源杀线虫剂提供依据。     

异硫氰酸荧光素标记鳗弧菌条件优化及应用条件对荧光信号的影响

为了实现对目标细菌的有效示踪,实验以异硫氰酸荧光素(FITC)为标记物,分别设置FITC标记浓度0.2、2、20、50、100、200μg/mL,标记时间1、15、30、60、120、240 min,研究了FITC标记浓度与标记时间对鳗弧菌标记效果的影响,同时对在不同温度(25、4、-20、-70℃)及不同时间(24、48、96、192 h)存储条件下鳗弧菌标记荧光的稳定性进行了比较,对标记鳗弧菌经甲醛灭活后的荧光变化进行了观察,研究了不同光照条件(0、150、1 500、25 000 lx)对标记鳗弧菌的荧光衰减影响,以及在-20和-70℃条件下反复冻融对标记鳗弧菌荧光强度的影响。结果表明,FITC的最适标记浓度范围为20~100μg/mL,最适标记时间范围60~120 min,储藏在-20和-70℃条件下,标记后的鳗弧菌荧光信号比储藏在4和25℃条件下稳定,标记后的鳗弧菌经甲醛灭活后的荧光信号稳定性优于对照组,不同强度自然光照对标记鳗弧菌的荧光衰减效果不明显,标记细菌经3次冻融,发现在-70℃条件下冻存的细菌荧光信号稳定性保持良好。本研究结果为FITC用于标记海洋细菌提供了实用的技术参数。     

扩展青霉基因组DNA五种提取方法比较

为寻找一种适合于扩展青霉基因组DNA的提取方法,比较了固体培养、液体静置培养和液体振荡培养3种培养方式的提取效果,分别采用氯化苄法、蜗牛酶法、CTAB法、SDS法及异硫氰酸胍法提取扩展青霉菌基因组DNA,经DNA质量浓度测定及电泳分析,比较5种方法的差异,并利用扩展青霉的多聚半乳糖醛酸酶(Polygalacturonase)基因内一段保守序列,设计PCR引物,扩增大小为288 bp的特异目的基因,检测其灵敏性.结果表明:固体培养的方式较适合扩展青霉基因组DNA的提取,另外5种提取方法均能提取出扩展青霉菌基因组DNA,扩增出目的条带,其中氯化苄法和蜗牛酶法所提DNA的纯度均较好,但蜗牛酶法所提DNA量较少.因此,氯化苄法是5种提取方法中最可靠的DNA提取方法,所提DNA浓度高、纯度好、结果稳定,可作为PCR反应的模板进行特异性扩增,且该方法的最低检测限大约为1.01×10-1μg/mL.     

魔芋叶片中4种总RNA提取方法的比较

本研究分别利用TRIzol法、异硫氰酸胍法、改良的CTAB法及试剂盒这4种方法提取魔芋叶片的总RNA,并用紫外光谱和琼脂糖凝胶电泳对各个提取效果进行鉴定。反复试验的结果表明:TRIzol法难以提取魔芋叶片总RNA,异硫氰酸胍法提取的总RNA浓度低、杂质较多且有部分降解,这两种方法不适宜用于魔芋叶片总RNA提取;试剂盒方法提取的总RNA产率和纯度较高,但稳定性差、成功率低且成本较高;改良的CTAB法利用PVP和β-巯基乙醇去除酚类物质,水饱和酚-氯仿去除蛋白质,醋酸钾去除多糖,该方法提取得到的总RNA的纯度较高,OD260/OD280值为1.893,而且效果稳定,有良好的可重复性。因此,我们认为改良的CTAB法是一种简便、快速且经济的提取魔芋叶片总RNA的方法。     

辣根硫代葡萄糖苷水解工艺比较及其水解产物的结构鉴定

以辣根为原料,筛选不同的水解工艺,研究硫代葡萄糖苷水解为异硫氰酸酯(ITCs)的转化率.结果表明,在温度40℃、pH值7、料液比1∶20(g∶mL)的条件下,水中加入Vc0.2%(以辣根粉计),水解2h,水蒸气蒸馏法得到的异硫氰酸酯质量分数最高(88.71%),经GC-MS、IR、1HNMR、13CNMR和MS鉴定为苯乙基异硫氰酸酯,经检索该化合物在辣根中首次分离鉴定.而采用二氯甲烷和水为1∶2的体积比的水解液水解辣根粉得到的异硫氰酸酯为苯乙基异硫氰酸酯(54.89%)和烯丙基异硫氰酸酯(5.18%).采用黑芥子酶酶解硫代葡萄糖苷法,水解产物中苯乙基异硫氰酸酯含量最低(10.67%).     

Characterization of the host range and sensitivity to fungicides of Trichothecium spp. associated with fruit rot in the field and in storage

During 2017–2019, we observed Trichothecium spp. causing fruit rot in the field and in storage. This study was conducted to examine morphological differences of the species from different hosts, reveal the species’ potential host range, and evaluate the efficacy of five fungicides. Six strains of Trichothecium spp. isolated from nectarine, peach and walnut were selected. Although the colony morphology, mycelial growth rate and spore size differed among hosts, phylogenetic analysis based on the internal transcribed spacer and part of the 5ʹ end of the β-tubulin gene showed that all tested strains belonged to Trichothecium roseum. For its host range, 23 kinds of fruit were examined using T. roseum strain YT-1 as an inoculum; 10 kinds of fruit, including pear, apple, mango, Chinese chestnut, pepino melon, fig and durian, were susceptible to T. roseum, with minimum inoculation concentrations ranging from 10⁴ to 10⁵ spores/ml. The fungicides that most effectively inhibited the six isolates were fluazinam and fludioxonil, with EC₅₀ values of 0.07–0.1 and 0.01–0.04 μg/ml, respectively, followed by difenoconazole (0.81–2.96 μg/ml), boscalid (5.43–13.51 μg/ml) and azoxystrobin (9.18–27.25 μg/ml). Improvement of the shelf life of nectarines held in plastic trays was explored using allyl isothiocyanate (AITC) against T. roseum YT-1. The application of 10 μl/L AITC significantly suppressed disease symptoms. The findings provide useful information for future disease emergency management in the field and for food preservation.     

萝卜肉质根辣味定量分析

用４甲硫基３丁烯异硫氰酸盐（ＭＴＢ－ＩＴＣ）作为辣味的定量指标，用气相色谱法测定其含量。在生食较辣的萝卜榨汁液中ＭＴＢ－ＩＴＣ含量为１００～２００μｍｏｌ／１００ｍＬ。肉质根生长初期其含量较低，定桩期增长最快，近露肩期达最大值，然后逐步下降；在整个生长过程中肉质根不同部位的辣味物质变化趋势一致；辣味物质在萝卜皮（韧皮部）的含量大于萝卜肉（木质部），萝卜肉中，真根部的顶端含量最高，向根头部的方向逐步减少。辣味强弱与播种期早晚有关。     

In vitro effect of recombinant human tumor necrosis factor-alpha on canine neutrophil apoptosis

Neutrophils (polymorphonuclear leukocytes: PMNs) are essential for the host defense against various infections and are often injurious to the host, causing inflammatory diseases where tumor necrosis factor-alpha (TNF-alpha) is suggested to play an important role. Since an effect of TNF-alpha on canine PMN apoptosis has not been studied, canine PMNs were stimulated with recombinant human (rh)TNF-alpha in the present study to investigate the effect of TNF-alpha on canine PMN apoptosis. PMN apoptosis and function to produce ROS were assessed by flow cytometry. Delayed apoptosis was observed in the PMNs treated with rhTNF-alpha at 100 ng/ml, accompanied by retention of capability to produce ROS. However, PMN apoptosis was accelerated by rhTNF-alpha combined with cycloheximide. Therefore, it is indicated that TNF-alpha is able to activate anti- and pro-apoptotic pathways in PMNs and that the inhibition of PMN apoptosis by TNF-alpha requires protein synthesis in the PMNs.     

用改进的异硫氰酸胍一步法提取完熟水果RNA

以完熟水果为材料提取RNA,除了要最大限度地抑制内源RNA酶活性,严格杜绝外源RNA酶污染外,选择合适的试验材料也是试验成功的关键。选取健壮植株上发育正常的果实,剖取果皮或近果皮果肉组织,在Chomczynski硫氰酸肌一步法的基础上,进行了2次变性,反复抽提,无菌操作等技术改进,获得了满意的纯化效果。