外源ATP对盐胁迫下油菜种子萌发及幼苗生长的影响

以‘陇油8号’油菜为试验材料,研究了不同浓度(1、10、25、50、100μmol·L~(-1))外源ATP预处理对盐胁迫下油菜种子萌发及幼苗生物量、H_2O_2和·OH含量、抗氧化酶活性、膜损伤程度、渗透调节物质及相关基因表达的影响。结果表明,与对照(不用ATP或NaCl处理)相比,用不同浓度的外源ATP处理,可不同程度地提高油菜种子的发芽率和发芽势,25μmol·L~(-1)的ATP处理表现较为显著,发芽率和发芽势分别提高了26.9%和17.2%。25μmol·L~(-1)的ATP+NaCl处理相比较于单独NaCl胁迫处理,油菜种子的发芽率和发芽势分别提高了13.7%和15.0%,油菜幼苗的生物量(根长、株高、鲜重)、2种抗氧化酶活性(CAT、POD)、可溶性糖和脯氨酸含量及相关基因(MAPK3和MAPK6、SOS1和NHX1)的表达量均显著增加,其中根长、株高、鲜重分别增加了23.6%、27.3%、28.6%,CAT、POD活性、可溶性糖和脯氨酸含量分别增加了10.3%、27.0%、15.9%、41.0%;而H_2O_2和·OH含量、丙二醛(MDA)含量及相对电导率分别显著降低了13.6%、6.3%、27.6%、20.5%。以上结果表明,外源ATP浸种能够促进NaCl胁迫处理下油菜的种子萌发,显著提高幼苗的生物量、抗氧化能力及相关基因的表达,降低其膜损伤,增强油菜幼苗的耐盐性。     

Vibrio harveyi (VirB11) recombinant vaccine development against vibriosis in orange‐spotted grouper (Epinephelus coioides)

Despite significant improvements in aquaculture to compensate wild catch, disease organisms have thrived in limiting its national and global potential. Using antibiotics, in a bid to remedy the havoc, has given rise to complications, attracting attention to disease prevention by immune enhancement against diseases. Grouper production has been inhibited for the threats of bacterial infection, particularly of Vibrio origin. Considering the rise in vibriosis cases, improved vaccines are necessary; moreover, recombinant vaccines, the choice for trial in the present experiment have been effective and more specific in improving immunity. The current work deals with grouper immune system enhancement with a recombinant vaccine developed from VirB11 gene in Vibrio harveyi. VirB11 was cloned in V. harveyi for recombinant vaccine development against vibriosis in orange‐spotted grouper (Epinephelus coioides). As indicated by the results, recombinant VirB11 protein showed effectiveness in conferring protection against vibriosis with observable specific antibody response in enzyme‐linked immunosorbent assay (ELISA) analysis; a significant increase (p < 0.05) in antibody levels was observed after a week and after 8 weeks post‐vaccination. From the weeks post‐vaccination, log2 (antibody titres) in the sera of vaccinated groups reached a peak of 14.2 at week 5 in the vaccinated group in comparison with a peak of approximately 5 and 2 in adjuvant and PBS controls. As indicated by the challenge results, 90% relative survival was observed in vaccinated group and 13% relative survival in control group I (adjuvant control). The cumulative performance of protein concludes VirB11 commendable for recombinant vaccine development.     

Protective immune responses of recombinant outer membrane proteins OmpF and OmpK of Aeromonas hydrophila in European eel (Anguilla anguilla)

Outer membrane proteins (Omps) of Gram‐negative bacteria have been proven to be efficient subunit vaccines against bacteriosis. In this study, OmpF and OmpK of Aeromonas hydrophila were expressed, and their immune protective effects in European eel (Anguilla anguilla) were evaluated. The genomic DNA of A. hydrophila 322A was used as a template, and two kinds of prokaryotic expression plasmids, pET‐32a‐OmpF and pET‐32a‐OmpK, were constructed. Recombinant OmpF protein (r‐OmpF) and r‐OmpK were purified and were proven to have antigenicity by Western‐blot analysis. r‐OmpF and r‐OmpK were used as immunogens to immunize European eel by intraperitoneal injection. The mRNA expression of 6 immune‐related genes (IgM, IL‐10, IRF3, IRF7, LysG4 and HexB) in the liver tissues of eels at 1 hr, 3 hr, 6 hr, 12 hr, 24 hr, 72 hr and 10 days postimmunization was analysed by real‐time PCR. At 30 dpi, the serum antibody response was measured by ELISA. Fish were attacked at 15 dpi by live 322A to assess the protective immunity of r‐OmpF and r‐OmpK. All the six tested genes responded to r‐OmpF or r‐OmpK vaccination at varying degrees. The serum antibody titre of r‐OmpF‐ and r‐OmpK‐immunized groups was 1:1,600 and 1:3,200 respectively. In addition, r‐OmpF gave 35.5% of the relative immune protection rate to European eels, while r‐OmpK gave 70.0%. By analysing the protective immunity and the regulatory role in the immune‐related gene expression of the two recombinant proteins that were studied, it was found that r‐OmpK was a potential vaccine candidate against A. hydrophila.     

狂犬病病毒HEP-Flury M基因重排后在小鼠神经母细胞瘤细胞中的表型分析

【目的】探究狂犬病病毒HEP-Flury M基因重排对基因转录和蛋白表达的影响,揭示病毒在小鼠神经母细胞瘤(NA)细胞中的表型变化与M基因重排的相关性。【方法】通过荧光定量PCR、Western blot以及病毒在NA细胞中的生长和扩散试验,对亲本毒株rHEP-Flury和M基因重排毒株M2、M4在NA细胞中的基因转录、表达、生长和扩散进行比较。【结果】狂犬病病毒结构基因的转录和表达主要受病毒基因组RNA合成的影响,但是在一个完整转录过程中单个结构基因的转录比例与其所在位置相关,M基因重排病毒的Leader RNA (LeRNA)和L mRNA的转录比例显著高于亲本毒株rHEP-Flury。M基因重排病毒在NA细胞中的生长和扩散都劣于亲本毒株rHEPFlury。【结论】狂犬病病毒亲本毒株rHEP-Flury具有狂犬病病毒原始的基因组顺序,在NA细胞中的生长和扩散都明显优于M基因重排病毒。结构基因在基因组中的位置主要决定其在一次转录过程中的转录比例,进而影响病毒在NA细胞中的生长和扩散。     

Using vibrational molecular spectroscopy to detect moist heating induced carbohydrates structure changes in cool-climate adapted barley grain

There is limited research to study how moist heating affects internal structure of barley grain on a molecular basis. The objectives of this study were to use vibrational molecular spectroscopy: 1) to determine the moist heating induced changes of barley carbohydrate (CHO) structure on a molecular basis, 2) to study the effects of moist heating on CHO chemical profiles, Cornell Net Carbohydrate and Protein System (CNCPS) subfractions, in situ rumen degradation, and predicted intestinal carbohydrate supply of barley grain; and 3) to reveal the association between molecular structure spectral features and CHO related metabolic characteristics. Barley samples (CDC cowboy) were collected from Kernen Crop Research Farm (Saskatoon, Canada) during two consecutive years. Half of each sample was kept as raw barley and the other half underwent moist heating (autoclaving at 120 °C for 60 min). The molecular spectroscopy (attenuated total reflectance-fourier transform infrared, ATR-FTIR) was used to detect the barley CHO related molecular structure spectral features. Moist heating did not affect carbohydrate related chemical profiles and CNCPS subfractions but it decreased rumen degradable carbohydrate. Rumen undegradable and intestinal digestion of CHO subfractions were not affected by moist heating. The advanced vibrational molecular spectroscopy can be used to detect carbohydrate molecular spectral features. Nutrient utilization prediction using molecular spectral characteristics is warranted and further investigation is encouraged.     

间作木薯对橡胶树根际土壤真菌群落结构的影响

研究间作木薯对橡胶树根际土壤真菌群落多样性变化规律,为判断胶园间作木薯的可行性提供理论支撑。以橡胶与木薯间作为处理,橡胶单作为对照,采用传统方法和高通量测序技术分析橡胶树根际土壤理化性状及真菌群落结构。结果表明：（1）间作模式下橡胶树根际土壤的碱解氮、速效磷、速效钾、有机质含量和真菌Shannon指数显著降低,真菌Simpson指数显著提高,pH和Chao1指数虽有降低,但差异未达到显著水平;（2）门分类水平上,2个处理的橡胶树根际土壤真菌群落结构组成相近,共有的3大优势菌门均为Ascomycota、Zygomycota和Basidiomycota。间作木薯降低了橡胶树根际土壤中Zygomycota和Basidiomycota的相对丰度,但提高了Ascomycota相对丰度,达到91.54%;（3）属分类水平上,2个处理中相对丰度比例前10的优势菌属占所有检测出菌属的61.30%~65.59%,间作木薯提高了橡胶根际土壤中Phyllosticta、Gibberella和Lecythophora的相对丰度;（4）土壤碱解N、速效P和速效K含量对橡胶树根际土壤中真菌群落α-多样性产生影响,土壤真菌多样性变化与土壤有机质含量有关。因此,间作木薯增加了植物对土壤矿质营养元素的吸收,导致橡胶树根际土壤肥力下降,进一步降低了橡胶树根际土壤真菌的群落多样性,同时增加了病原真菌的丰度。     

硝酸钠和乙酸铵配施对黄果柑幼苗光合特性的影响

利用Li–6400便携式光合测定系统，测定了硝酸钠与乙酸铵11个不同配比处理的黄果柑幼苗在不同季节的光合能力。结果表明：1) 硝酸钠与乙酸铵配比处理黄果柑幼苗的光合能力均表现为夏季最强，冬季最弱，秋季强于春季；叶绿素a、叶绿素b和总叶绿素含量均表现为夏季最高，冬季最低，秋季高于春季。2) 硝酸钠和乙酸铵配比(质量比)为8∶2时，黄果柑幼苗叶片的净光合速率、蒸腾速率、水分利用率和气孔导度以及叶绿素a、叶绿素b和总叶绿素含量均高于其他处理，单施硝酸钠或乙酸铵处理都低。综合比较分析，硝酸钠和乙酸铵配比为8∶2时，黄果柑幼苗的光合能力最强。     

罗氏沼虾低温有水保活试验

通过对保活温度(12、15、17、20、23、25 ℃)，虾与水质量比(1∶6、1∶8、1∶10、1∶12、1∶15)和暂养时间(0、12、24、36、48 h) 3个因素的单因素试验和正交试验，确定罗氏沼虾低温保活的最佳工艺条件，并检测低温保活过程中的水质指标及肌肉、血液、肝脏等虾体组织指标的变化，探究影响罗氏沼虾的保活机理。结果表明：罗氏沼虾的临界温度为9 ℃；单因素试验中，最佳保活温度为20 ℃，最佳虾与水质量比为1∶10，最佳暂养时间为24 h；正交试验中，最优工艺为保活温度20 ℃、虾与水质量比1∶12、暂养时间24 h，保活24 h内存活率可达100%；以正交优化后的最优工艺条件进行罗氏沼虾的低温保活试验，水样中氨氮质量浓度和菌落总数显著上升(P＜0.05)，但溶解氧质量浓度、pH均仍在适宜罗氏沼虾存活范围内变化，肌肉糖原质量分数、pH和三磷酸腺苷(ATP)质量分数下降，乳酸浓度显著上升(P＜0.05)，血液谷丙转氨酶(ALT)和谷草转氨酶(AST)活力显著上升(P＜0.05)，肝脏糖原质量分数显著下降(P＜0.05)，总超氧化物歧化酶(T–SOD)活力先降后升，而丙二醛(MDA)含量则先升后降。说明水质指标的变化会影响虾体肌肉、血液及肝脏各组织正常的生理代谢，从而影响其保活时间和存活率。     

Analysis of sinking death using video images of the swimming performance of Pacific Bluefin tuna (Thunnus orientalis) larvae

In this study, we aimed to clarify the mechanism of sinking death during the larval stage of Pacific Bluefin tuna Thunnus orientalis by investigating the effects of swimming performance on sinking death, using a behavioral approach. Swimming performance was examined 3–9 days after hatching (DAH) under day and night light conditions in cuboid experimental tanks. Swimming behavior variables such as swimming speed and swimming angle were measured under both light conditions. Larvae in the daytime experiment and larvae with inflated swim bladders at night were distributed on the surface layer of the water column. In contrast, larvae with uninflated swim bladders at night were frequently observed swimming vertically or sinking to the bottom of the tank. Larvae with inflated swim bladders at night were always distributed beneath the surface until the next morning (survival rates were 100 %). However, larvae with uninflated swim bladders at night swam upward repeatedly and later sank to the bottom of the tank (survival rates were 60 % and 38 % at 5 and 9 DAH, respectively). Larvae with uninflated swim bladders were not always able to maintain their swimming depth by swimming until the next morning. Additionally, their swimming speed and vertical swimming frequency (ratio) depended on the illumination and swim bladder conditions. Our findings show that larvae with uninflated swim bladders at night were associated with a higher risk of sinking death. The swimming energy capacity of Pacific Bluefin tuna larvae, which indicates the total amount of the energy that enables individuals to swim throughout the night without feeding, was found to be linked to sinking death.