不同坡位沙棘光合日变化及其主要环境因子

沙棘是我国干旱半干旱地区重要的生态经济树种,被广泛应用于黄土高原造林活动中。为探明不同坡位沙棘光合特征及影响沙棘光合作用的主要环境因子,在沙棘生长旺盛期分坡位测定光合特性及环境因子日变化进程,并采用通径分析法分析各环境因子对沙棘净光合速率的直接和间接影响。结果表明：1、各环境因子之间存在相互作用,光合有效辐射是空气温度变化的主要驱动因子,而空气温度变化引起的空气密度变化对空气相对湿度和CO2浓度有显著影响。在上、中和下坡位环境变化可以分别解释85.6%、86.0%和 55.22%净光合速率的变异。2、下坡位沙棘净光合速率最小,日均值为9.38μmol/m2/s,上、中坡位分别为14.22μmol/m2/s,15.94μmol/m2/s。3、10:00之前中上坡位沙棘水分利用效率明显高于下坡位,但3坡位沙棘水分利用效率日均值无明显差异。4、上坡位净光合速率主要受到光合有效辐射、空气相对湿度的影响;中坡位光合有效辐射仍为主要环境因子,此外空气温度和空气湿度之间的相互作用对净光合速率也有一定影响,为主要限制因子;下坡位影响沙棘光合速率主导因素为光合有效辐射。5、沙棘在10:00之前光合有效辐射和空气湿度较高而空气温度较低的环境下,净光合速率和水分利用效率最高。作为阳生物种,沙棘需要足够的光照维持生长,而以往研究表明适度水分胁迫可以提高沙棘光合过程中对水的利用效率。本研究所选流域年降水量535mm,中上坡位水分胁迫并非十分严重。因此,种植于中上坡位的沙棘即可接受足够光照（特别是10:00以前的光照）进行光合保证生长速率,又能在光合过程中保持较高水分利用效率,符合干旱半干旱地区生态经济发展的需求。     

宁海地区香鱼弧菌病病原菌鉴定

摘要：【目的】香鱼弧菌病对中国沿海地区的香鱼养殖业造成了巨大的危害,然而,病原不明导致了防治上的许多问题。本文鉴定了引起宁海地区香鱼爆发性弧菌病的病原。【方法】采用TCBS平板分离优势菌;采用回归感染试验确认病原菌,采用改进的寇氏法计算LD50;采用形态学观察、生理生化特征测定、细菌特异性引物PCR扩增检测及细菌16S rRNA和金属蛋白酶（MP）基因序列分析鉴定细菌;采用药敏实验测定它对部分抗生素的敏感性。【结果】分离并鉴定优势菌株ayu-H080701为宁海地区香鱼弧菌病的病原菌,它对香鱼的半致死量为1.2×104 CFU。形态学观察和生理生化特征测定表明,ayu-H080701与鳗利斯顿氏菌最为接近。PCR扩增检测表明,细菌16S rRNA 基因通用引物和鳗利斯顿氏菌MP基因特异引物均能扩增到预期大小的特异性条带。ayu-H080701与鳗利斯顿氏菌16S rRNA基因核苷酸序列同源性最高,为99.4%～99.5%,与同属的海弧菌和美人鱼发光杆菌分别为94.3%和91.9%;ayu-H080701与鳗利斯顿氏菌MP氨基酸序列同源性高达97.6%～98.8 %,与其它弧菌科成员则低于75.6 %,系统进化树分析也揭示ayu-H080701与鳗利斯顿氏菌进化相关性最高。【结论】引起宁海地区香鱼弧菌病的菌株ayu-H080701被鉴定为鳗利斯顿氏菌。     

洋底深部裂褶菌在不同环境条件的生长行为

[目的] 二甲基巯基丙酸内盐（dimethylsulfoniopropionate,DMSP）及其裂解产物二甲基硫（dimethyl sulfide,DMS）在海洋硫循环中发挥重要作用。目前关于DMSP降解细菌的分布已有部分报道,但其合成细菌的研究才刚刚起步。本文拟研究中国东海水体DMSP合成与降解菌及基因的水平和垂直分布（1000 m水深）差异,分析其对环境梯度变化的响应。[方法] 利用流式细胞仪计数海水样品中微微型浮游生物的数量,通过荧光定量PCR和高通量测序手段定量测定DMSP合成基因（dsyB和mmtN）及物种、DMSP降解基因（dddP和dmdA）及物种的丰度,分析其在东海海域水平及垂直方向上的分布差异。[结果] 在垂直方向上,聚球藻、原绿球藻、微微型真核生物和异养细菌丰度随着水深的增加而先增后减,最大值位于30-50 m附近。表层（4 m左右）水体的DMSP合成及降解基因丰度最高,DMSP合成菌（如Alteromonas、Phaeobacter和Pelagibaca等）丰度也最高;随着水深增加,表层以下水体中DMSP合成及降解基因和物种丰度先增加后降低,峰值均出现在100-150 m;100 m以下,DMSP降解基因丰度迅速下降,而合成基因丰度下降程度较低,而且接近底层（>500 m）时出现随水深逐渐增加的趋势。水平方向二者变化规律不明显。浅层水体（≤100 m）和深层水体（>100 m）细菌群落结构存在显著差异,前者拥有较高比例的黄杆菌纲、放线菌纲和蓝细菌纲细菌,后者α变形菌纲细菌丰度较高。[结论] 100 m及以浅和100 m以深的浮游细菌群落结构存在显著差异。表层水体中DMSP合成和降解细菌的丰度最高,100-150 m水体次之,但100-1022 m介导的DMSP合成和降解细菌丰度的变化趋势有较大差别。     

贝莱斯芽孢杆菌CC0955发酵培养基优化及其泡腾颗粒的研制

【背景】植物病害的生物防治及生防产品的开发一直是植物保护研究的重点方向,但现有的生防产品多为可湿性粉剂和水剂,存在剂型单一、货架期短及运输和使用不便等问题。【目的】优化贝莱斯芽孢杆菌CC0955菌株发酵培养基,并利用其开发出一种容易使用和保存的新生防产品——泡腾颗粒。【方法】采用Plackett-Burman设计、中心组合设计和响应面分析等方法,优化了贝莱斯芽孢杆菌CC0955的发酵培养基成分;采用L₉（3³）正交设计,以溶液pH和活菌数为指标,优化了贝莱斯芽孢杆菌CC0955泡腾颗粒配比,并评价了其物理性质和抑菌效果。【结果】贝莱斯芽孢杆菌CC0955最优发酵培养基成分为（g/L）:蛋白胨12.00,酵母粉1.00,葡萄糖15.00,MgSO₄·7H₂O 0.40,K₂HPO₄ 0.05。用此培养基发酵48h,发酵液对立枯丝核菌的抑制率达到89.78%。泡腾颗粒最佳配比为:碱酸摩尔比为2.00,白炭黑1.50 g,黄腐酸钾0.03 g。泡腾颗粒的平均崩解...     

重组酶介导等温核酸扩增方法快速检测土壤沙门氏菌

【背景】沙门氏菌(Salmonella)是一种可以引起人畜患病的致病菌,也是最主要的食源性细菌之一。土壤中的沙门氏菌可通过蔬菜等植物进入人体,引发食物中毒。但由于土壤性质及其他微生物的干扰,如何快速甄别土壤是否受到沙门氏菌的污染仍是一个难题。【目的】建立一种快速、灵敏检测土壤沙门氏菌的实时重组酶介导等温核酸扩增(Real-Time Recombinase Aided Amplification,RT-RAA)方法。【方法】针对沙门氏菌invA基因序列设计特异性引物和探针,构建含有invA基因待检片段的重组质粒,评价RT-RAA方法的灵敏度;分别以肠炎沙门氏菌、大肠杆菌、福氏志贺氏菌和金黄色葡萄球菌的基因组DNA为模板,评价RT-RAA方法的特异性;RT-RAA方法用于番茄、生姜土壤中沙门氏菌的检测,同时用平板培养法进行验证。【结果】RT-RAA方法可用于重组质粒中invA基因片段的检测,在39℃条件下,20 min内即可获得检测结果,最低检测质粒拷贝数为10拷贝/反应,而且与大肠杆菌、福氏志贺氏菌和金黄色葡萄球菌无交叉反应。土壤样品DNA的RT-RAA检测结果显示,供试番茄土已被沙门氏菌污染,而生姜土则没有,与平板培养结果一致。【结论】RT-RAA方法具有灵敏度高和特异性强的特点,可用于土壤沙门氏菌污染的快速检测。     

Immunogenicity and Efficacy of Intramuscular Replication-Defective and Subunit Vaccines against Herpes Simplex Virus Type 2 in the Mouse Genital Model

Herpes simplex virus type 2 (HSV-2) is a sexually transmitted virus that is highly prevalent worldwide, causing a range of symptoms that result in significant healthcare costs and human suffering. ACAM529 is a replication-defective vaccine candidate prepared by growing the previously described dl5-29 on a cell line appropriate for GMP manufacturing. This vaccine, when administered subcutaneously, was previously shown to protect mice from a lethal vaginal HSV-2 challenge and to afford better protection than adjuvanted glycoprotein D (gD) in guinea pigs. Here we show that ACAM529 given via the intramuscular route affords significantly greater immunogenicity and protection in comparison with subcutaneous administration in the mouse vaginal HSV-2 challenge model. Further, we describe a side-by-side comparison of intramuscular ACAM529 with a gD vaccine across a range of challenge virus doses. While differences in protection against death are not significant, ACAM529 protects significantly better against mucosal infection, reducing peak challenge virus shedding at the highest challenge dose by over 500-fold versus 5-fold for gD. Over 27% (11/40) of ACAM529-immunized animals were protected from viral shedding while 2.5% (1/40) were protected by the gD vaccine. Similarly, 35% (7/20) of mice vaccinated with ACAM529 were protected from infection of their dorsal root ganglia while none of the gD-vaccinated mice were protected. These results indicate that measuring infection of the vaginal mucosa and of dorsal root ganglia over a range of challenge doses is more sensitive than evaluating survival at a single challenge dose as a means of directly comparing vaccine efficacy in the mouse vaginal challenge model. The data also support further investigation of ACAM529 for prophylaxis in human subjects.     

Evaluation of a botulinum fragment C-based ELISA for measuring the humoral immune response in primates.

An enzyme-linked immunosorbent assay (ELISA) using botulinum neurotoxin serotype B recombinant fragment C (rBoNTB(HC)) was developed to measure specific humoral immune responses of monkeys vaccinated with a vaccine consisting of rBoNTB(HC). Several fundamental parameters for a bioassay were evaluated. The evaluation results demonstrated that using BoNTB(HC) as the capture antigen led to a specific and sensitive ELISA for botulinum type B antibody with excellent precision, accuracy, and linearity. There was a good correlation (r=0.91) between ELISA titers and neutralization bioassay titers. Experimental results suggested that the ELISA could be useful for detecting botulinum type B antibody levels and may supplement mouse neutralization bioassays during planned clinical manufacturing and clinical trials of rBoNTB(HC) vaccine.     

A proteomic view of <Emphasis Type="Italic">Caenorhabditis elegans</Emphasis> caused by short-term hypoxic stress

Background  

The nematode Caenorhabditis elegans is both sensitive and tolerant to hypoxic stress, particularly when the evolutionarily conserved hypoxia response pathway HIF-1/EGL-9/VHL is involved. Hypoxia-induced changes in the expression of a number of genes have been analyzed using whole genome microarrays in C. elegans, but the changes at the protein level in response to hypoxic stress still remain unclear.     

Effects of a GTP-insensitive mutation of glutamate dehydrogenase on insulin secretion in transgenic mice

Glutamate dehydrogenase (GDH) plays an important role in insulin secretion as evidenced in children by gain of function mutations of this enzyme that cause a hyperinsulinism-hyperammonemia syndrome (GDH-HI) and sensitize beta-cells to leucine stimulation. GDH transgenic mice were generated to express the human GDH-HI H454Y mutation and human wild-type GDH in islets driven by the rat insulin promoter. H454Y transgene expression was confirmed by increased GDH enzyme activity in islets and decreased sensitivity to GTP inhibition. The H454Y GDH transgenic mice had hypoglycemia with normal growth rates. H454Y GDH transgenic islets were more sensitive to leucine- and glutamine-stimulated insulin secretion but had decreased response to glucose stimulation. The fluxes via GDH and glutaminase were measured by tracing 15N flux from [2-15N]glutamine. The H454Y transgene in islets had higher insulin secretion in response to glutamine alone and had 2-fold greater GDH flux. High glucose inhibited both glutaminase and GDH flux, and leucine could not override this inhibition. 15NH4Cl tracing studies showed 15N was not incorporated into glutamate in either H454Y transgenic or normal islets. In conclusion, we generated a GDH-HI disease mouse model that has a hypoglycemia phenotype and confirmed that the mutation of H454Y is disease causing. Stimulation of insulin release by the H454Y GDH mutation or by leucine activation is associated with increased oxidative deamination of glutamate via GDH. This study suggests that GDH functions predominantly in the direction of glutamate oxidation rather than glutamate synthesis in mouse islets and that this flux is tightly controlled by glucose.     

叶绿体转基因植物——一种新型生物反应器

叶绿体转基因植物作为生物反应器, 具有外源蛋白表达量高和环境安全性好等优点, 近年来呈现出诱人的发展前景。本文综述了叶绿体基因工程的优越性, 并重点介绍了叶绿体转基因植物作为生物反应器在生产疫苗、药用蛋白及生物可降解塑料等物质方面的最新研究进展。