粪肠球菌生长曲线的测定及其对小鼠脑组织的影响

为测定致羔羊脑炎粪肠球菌（Enterococcus faecalis）的生长曲线，寻求一种快速而准确的方法测定不同生长时期粪肠球菌数量，并客观评价其毒性强弱及其对小鼠脑组织的影响，试验采用平板菌落计数法和OD-Monitor振荡比浊法（D_λ值法）测定粪肠球菌的生长曲线，探究该菌在合适时间段内的吸光值（D_{600 nm}）与平板菌落计数法测定的活菌数（CFU）的关系。用粪肠球菌感染小鼠，观察记录小鼠的死亡情况，最后采用Karber法计算粪肠球菌感染小鼠的半数致死量（LD₅₀）。用LD₅₀的剂量感染小鼠，及时采集死亡小鼠脑组织，未死亡的小鼠72 h后全部剖杀取脑组织，一部分做涂片染色，制作病理切片，观察病理变化；一部分进行培养，用于PCR方法进行细菌的回收鉴定。结果显示，用两种方法测定此株粪肠球菌的生长曲线基本一致，在2~8 h生长迅速，为对数生长期，8~14 h生长缓慢，为稳定期，14 h之后死亡数增加，进入衰亡期；对12 h粪肠球菌D_{600 nm}与CFU的关系进行探讨，成功建立回归方程：y=20.769x-1.3422，R²=0.997；其感染小鼠的LD₅₀为7.77×10¹¹个活菌。以此剂量感染小鼠，脑组织涂片染色和培养染色，均能看到革兰氏阳性球菌；PCR结果显示，均出现了大小为112 bp的条带。对脑组织进行病理学观察发现该菌可导致脑组织充血、出血、形成微血栓，脑膜充血。通过生长曲线和其D_{600 nm}与CFU关系的建立，可实时监测粪肠球菌数量，为后期更深入研究粪肠球菌穿越血脑屏障的机制奠定重要的理论基础。     

QuEChERS-超高效液相色谱-串联质谱技术同时测定大蒜中10种农药残留

建立了快速、准确测定大蒜中10种农药多残留的QuEChERS-超高效液相色谱-串联质谱检测技术。样品经酸化乙腈提取,无水硫酸镁(500 mg)除水后,用N-丙基乙二胺(PSA,500 mg)和十八烷基键合硅胶(C₁₈,500 mg)净化,以乙腈和0.1%甲酸水溶液为流动相进行梯度洗脱,Waters C₁₈色谱柱分离,采用正离子多反应监测(MRM)模式检测,基质匹配标准溶液外标法定量。10种农药的检出限在0.020~0.800 μg·kg^-1,定量限在0.067~2.670 μg·kg^-1。线性范围内相关系数均大于0.99。加标回收率在73.4%~109.0%,相对标准偏差在1.2%~9.8%。结果表明,该方法简便、快速、灵敏度高、重现性好,可同时测定大蒜中10种农药。     

Integrated Evaluation and the Physiological and Biochemical Responses of Semi-Wild Cotton under Complex Salt-Alkali Stress

[Objective] The genetic diversity of semi-wild cotton was abundant, so more new elite resistant genes could be found and applied to improve the resistance of cotton. The aim of this study was to primarily study on the regulation of response to salt-alkali stress, explore the method of integrated evaluation and characterize the resistance of semi-wild cotton. [Method] Four semi-wild and two cultivated Gossypium hirsutum accessions were evaluated for phonotypical, physiological and biochemical traits under alkali-salt versus normal conditions in hydroponic solutions. The alkali-salt tolerance was determined based on principal component analysis, cluster analysis, correlation analysis, grey rational analysis and analysis of variance. [Result] On the basis of the overall results, the alkali-salt stress tolerance of the six accessions was ranked as: Marie-galante85 > Latifolium32 > CRI16 > CRI12 > Latifolium40 > Latifolium130. Roots were found to be the most important responsive systems to complex alkali-salt stress. At seedling growth stage, the active scavenging system played a crucial role in response to alkali-salt stress. In addition, the salt tolerant and salt sensitive accessions showed different response tends towards leaf peroxidase, root ascorbate peroxidase and root catalase within 48 h, suggesting the accessions have different levels of response to salt stress. [Conclusion]Our study identified the most alkali-salt tolerant accessions and provided basic concept of complex alkali-salt tolerance mechanism within cotton accessions. And, our study provided a simple and rapid, highly accurate and precise method for evaluating salt resistance of cotton, and proved that the balance of ROS system play an important role in response to salt-alkali stress. Hence, mining of salt tolerant genes from these accessions may facilitate the development of novel salt tolerant variety.     

中华绒螯蟹微孢子虫研究进展

微孢子虫是专性细胞内寄生的真核生物,有着广泛的宿主。中华绒螯蟹微孢子虫可以引起中华绒螯蟹微孢子虫病,因此而受到重视。本文简要综述了中华绒螯蟹微孢子虫的生物学特性,检测方法以及中华绒螯蟹微孢子虫病的研究现状,以期为有效防治河蟹微孢子虫病提供参考。     

加勒帕戈斯群岛野生棉第二次考察简报

在2015年考察的基础上，于2016年9月21日-10月2日再次赴厄瓜多尔的加勒帕戈斯群岛，重点对Santa Cruz、Isabela和San Cristobal三个岛上的达尔文氏棉和克劳茨基棉的自然分布进行调查。观察到28个野生棉自然居群，遗传多样性丰富；特别是发现了达尔文氏棉新类型，具有珍贵的保存和研究价值，充实和丰富了我国棉花种质资源基因库。同时分析了达尔文氏棉的地理分布、形态特征和生态适应性，并对其后续研究和利用方向进行讨论和展望。     

花前氮亏缺对水稻叶片氮代谢酶活性的影响

以籼型杂交稻中浙优1号和籼粳型杂交稻甬优12为材料,试验分析了花前不同时期氮亏缺处理对水稻叶片氮代谢酶活性的影响。结果表明,花前氮亏缺导致植株上3叶氮浓度、NR、GS、GOGAT、GOT和GPT酶活性大幅下降,GDH活性显著增长,且各叶位对花前氮亏缺敏感度总体上表现为剑叶倒2叶倒3叶;破口期亏氮对籼型杂交稻中浙优1号上3叶氮同化酶活性的影响远小于减数分裂期处理,与之相比,籼粳杂交稻甬优12对破口期亏氮胁迫仍较敏感,表明中浙优1号植株氮代谢酶的亏氮敏感性由减数分裂期至破口期逐步下降,对土壤速效氮的需求同步降低,甬优12则对土壤供氮存在更高需求。     

Replacement of Fishmeal by a Mixture of Soybean Meal and Chlorella Meal in Practical Diets for Juvenile Crucian Carp,Carassius auratus

A growth trial was conducted to evaluate the effect of a mixture of soybean meal and Chlorella meal (SCM) as a dietary fishmeal (FM) substitute on growth performance, apparent digestibility coefficients (ADCs), digestive enzymatic activities, and histology of juvenile crucian carp, Carassius auratus. Five isonitrogenous and isoenergetic diets were formulated to replace 0 (SCM0), 25 (SCM25), 50 (SCM50), 75 (SCM75), and 100% (SCM100) of protein from FM with SCM, respectively. The diets were fed to triplicate groups of juvenile crucian carp for 6 wk. Weight gain, specific growth rate, feed intake, protein efficiency ratio, and intestinal digestive enzymatic activities (amylase, trypsin, and lipase) tended to decline with increasing FM replacement levels (P > 0.05). Dietary SCM substitution significantly influenced dry matter content in muscle, and crude protein and lipid contents in liver (P < 0.05). ADCs for dry matter, protein, lipid, energy, and most amino acids showed no significant differences between the control and SCM25 group, but tended to decline with replacement levels over 25%. Higher SCM substitution (50–100%) caused karyopyknosis and necrosis in liver, but intestinal histology did not show noticeable pathological changes. The present study indicated that FM could be replaced by 25% of SCM, without significant adverse growth performance, feed utilization, and histology of crucian carp.     

蜗牛消化酶提取河蟹微孢子虫基因组反应条件的优化

PCR是目前能够特异地检测河蟹微孢子虫的最灵敏的方法。河蟹微孢子虫孢子壁坚厚不易破碎,对许多化学物质有很强的抵抗性,常规的核酸抽提方法效果不够理想。本文通过对比PCR的结果,研究蜗牛消化酶对DNA抽提效果的影响,确定了酶作用的最佳反应条件,提高了PCR检测河蟹微孢子虫的灵敏性。结果显示:蜗牛消化酶提取微孢子虫DNA的酶促反应温度为37℃,底物浓度为20 mg,酶的使用浓度为酶/底物重量比400 mg/g,酶促反应时间为4 h。实验表明,使用蜗牛消化酶处理样品,有助于更加灵敏地检测出微孢子虫。该研究结果将有助于提高河蟹微孢子虫病的检验检测能力,对河蟹微孢子虫的检疫和防治具有积极意义。     

Identification,characterization and full-length sequence analysis of a novel endornavirus in common sunflower (Helianthus annuus L.)

To identify the possible quarantine viruses in seven common sunflower varieties imported from the United States of America and the Netherlands, we tested total RNAs extracted from the leaf tissues using next-generation sequencing of small RNAs. After analysis of small RNA sequencing data, no any quarantine virus was found, but a double-stranded RNA(dsRNA) molecule showing typical genomic features of endornavirus was detected in two varieties, X3939 and SH1108. Full-length sequence and phylogenetic analysis showed that it is a novel endornavirus, temporarily named as Helianthus annuus alphaendornavirus(HaEV). Its full genome corresponds to a 14 662-bp dsRNA segment, including a 21-nt 5′ untranslated region(UTR), 3' UTR ending with the unique sequence CCCCCCCC and lacking a poly(A) tail. An open reading frame(ORF) that encodes a deduced 4 867 amino acids(aa) polyprotein with three domains: RdRP, Hel and UGT(UDP-glycosyltransferase). HaEV mainly distributed in the cytoplasm but less in the nucleus of leaf cells by fluorescence in situ hybridization(FISH) experiment. This virus has a high seed infection rate in the five varieties, X3907, X3939, A231, SH1108 and SR1320. To our knowledge, this is the first report about the virus of the family Endornaviridae in the common sunflower.