A new method to detect Lonsdalea quercina in infected plant tissues by real‐time PCR

Presymptomatic and accurate diagnoses of pathogens are essential for disease prediction and the timely application of bactericide. The bacterium Lonsdalea quercina (=Brenneria quercina) has been reported as the causal agent of drippy nut and bark canker disease on oak in California (US) and Europe. In recent years, it is also found on Populus × euramericana trees in Henan province of China. This bacterium causes longitudinal cankers of a few centimetres in size on the bark surface of the upper trunk. In this study, we developed two species‐specific PCR assays using primer pairs LqfF/LqfR and LqgF/LqgR for the rapid and accurate detection of the pathogenic bacteria in diseased plant tissues. The results show that the LqfF/LqfR primers amplified only a single PCR band of approximately 382 bp and the LqgF/LqgR primers yielded a PCR product of approximately 286 bp. The two primers were successfully adapted to real‐time PCR based on SYBR Green I used with the ABI 7500 system. The detection limit of the reaction was 0.1 pg genomic DNA per 20 μl PCR reaction volumes. The pathogen was mainly detected in the phloem of cankers as well as in the exudates of diseased trees, but was not found in the xylem or leaves. The size of pathogen in distribution was larger than the lesion. The results demonstrate that real‐time PCR assays can be used to detect the pathogen by extracting DNA directly from infected plant tissues. This method is a rapid, reliable method for the presymptomatic and accurate detection of L. quercina, providing a useful insight into epidemiological studies.     

Insecticidal benzoylphenylurea-S-carbamate: a new propesticide with two effects of both benzoylphenylureas and carbamates

New propesticides with two effects of both benzoylphenylureas and carbamates were designed and synthesized by the key intermediate N-chlorosulfenyl-N-methylcarbamate, which was prepared for the first time. These benzoylphenylurea-S-carbamates were identified by 1H NMR spectroscopy and elemental analyses. The bioactivities of the new compounds were evaluated. These benzoylphenylurea-S-carbamates exhibited excellent larvicidal activities against Oriental armyworm, some of which were good as compared to the parent benzoylphenylureas. Toxicity assays indicated that these benzoylphenylurea-S-carbamates had knockdown activities of carbamates at higher concentrations and insect growth regulator activities of benzoylphenylureas at lower concentrations. We found that the title compounds exhibited good systemic larvicidal activities against Oriental armyworm, which were especially advantageous when combating sucking pests. Some of these title compounds can kill aphids and mosquitoes as well.     

基于EEM-PARAFAC分析冻融作用对高寒泥炭湿地土壤溶解性有机质的影响

  目的  全球气候变暖加速了土壤冻融格局的改变,而冻融作用又对土壤溶解性有机质（DOM）产生深刻的影响。  方法  以高寒泥炭湿地土壤为研究对象,采用室内模拟冻融的方式开展了两种冻融条件下（日冻融:?3 ℃/5 ℃,12 h/12 h,30次;深度冻融:?24 ℃/5 ℃,20 d/10 d,1次）的试验研究,基于紫外?可见吸收光谱（UV-vis）和三维荧光光谱结合平行因子（EEM-PARAFAC）分析探究土壤DOM数量和质量的变化。  结果  （1）经过30 d培养后,两种冻融处理均显著提高了高寒泥炭湿地土壤溶解性有机碳（DOC）的含量,日冻融和深度冻融条件下土壤样品的DOC含量分别为183.26和187.06 mg/kg,相比于对照组分别增加了18.80%和21.26%（P < 0.05）。（2）不同冻融条件下的土壤DOM的紫外?可见吸收曲线线型差异较小,无明显特征吸收峰,且显示出吸光度随波长呈指数衰减的趋势,但经冻融处理的样品出现了明显的红移现象。（3）两种冻融处理均使得土壤DOM的腐殖化指数（HIX）值（从4.42增加至11.73和17.10）、254 nm处的单位比色皿光程下的紫外吸收值（SUVA₂₅₄）（从1.35 mg/（L·cm）增加至1.37和1.40 mg/（L·cm））以及A₂₅₃/A₂₀₃值（0.38增加至0.40和0.43）增大,波长275 ~ 295 nm处光谱斜率系数减小（从13.34 μm?1减小至13.23和12.91 μm?1）。（4）EEM-PARAFAC鉴别出的荧光组分有类富里酸物质、类腐殖酸物质、溶解性微生物代谢产物和类蛋白物质,冻融改变了土壤DOM的组成以及各组分的贡献率。对照组土壤DOM组分C1、C2和C3的贡献率分别为37%、32%和31%;日冻融条件下,组分C1、C2和C3的贡献率分别为41%、34%和 25%;深度冻融条件下,组分C1、C2和C3的贡献率分别为44%、35%和 21%。  结论  冻融改变了土壤DOM的数量和质量,表现为DOC含量的增加和腐殖化程度的提高。      

不同热预处理方式对酸豆乳品质特性的影响

为研究热预处理方式对酸豆乳品质特性的影响.采用持续加热(Continuous heating soymilk,CHS)和热磨浆(Heat soymilk,HS)2种工艺制备豆乳,温度115℃下灭菌10,15,20 min后发酵、后熟得到酸豆乳.在感官评价的基础上,进一步研究发酵期间和后熟酸豆乳的pH、酸度、活菌数、持水性、粘度、色度和质构特性的变化.结果表明:CHS处理酸豆乳凝乳均一稳定、紧密不分层、无乳清析出、酸甜可口、发酵香味浓郁、无豆腥味、酸度适中,总体接受度好;理化指标、微生物指标和质构指标表明,CHS处理酸豆乳发酵过程中,在1～6h内的pH显著低于HS处理,到9 h时2种酸豆乳的pH趋于一致;后熟8 d内CHS和HS酸豆乳的pH变化不显著;酸度变化与pH变化趋势一致;CHS处理酸豆乳持水性和粘度均低于HS处理;发酵6 h时,2种豆乳活菌数基本保持在7.2×107～7.9×107 CFU/mL,之后基本保持不变;后熟过程中不同处理方式的活菌数基本无明显变化,CHS处理的黏性和粘结性优于HS处理.说明磨浆时热预处理工艺直接影响酸豆乳品质特性,持续加热磨浆显著提高酸豆乳的品质.     

大弹涂鱼血细胞发生的研究

通过对大鲜涂鱼血液、肾脏、脾脏组织涂片的观察，发现其肾脏、脾脏内白细胞的发育大致经过了3个阶段，即原始阶段、幼稚阶段、成熟阶段。造血母细胞从造血器官释放入外周血中有一成熟过程。着重描述了大弹涂鱼的红细胞、嗜中性粒细胞、淋巴细胞和单核细胞发生过程中各阶段细胞的形态特征。     

Production, disinfection and evaluation for aquaculture applications of rotifer resting eggs from Bohai Bay, P.R. of China

The dynamics of resting egg production of the rotifer Brachionus plicatilis originating from a wild population in the Tanggu Saltworks (P.R. China) was investigated. In the natural environment as well as in semi-controlled rearing conditions an increased resting egg production was noticed with declining food availability. Processed resting eggs had a hatching efficiency of 3 x 10 6 rotifers per gramme irrespective of their origin. Hatching started 22 h after the initiation of incubation and was completed after 36 h. Rotifers obtained from resting eggs could be further cultured on Culture Selco® and enriched with Super Selco®. The fatty acid profile of these rotifers were not divergent from reference rotifers (originating from the Laboratory of Aquaculture and Artemia Reference Centre) demonstrating that this strain was not catabolizing essential fatty acids and could be used for enrichment purposes. Storage of resting eggs at 4°C resulted in a 50% lower hatching after 1 year but remained stable during the next 2 years. The resting eggs used for storage could easily be disinfected without affecting their hatching characteristics. These results indicate that this material could be used as inocula for mass cultures of live food for commercial hatcheries.     

刺参生殖周期的研究

本文根据大连沿海刺参生殖腺的活体观察、分期特征以及生殖腺指数、生殖腺主分枝的最大直径和消化管重量的季节变化，研究了刺参的产卵习性及产卵盛期。通过对生殖腺组织学的研究，论述了刺参的性腺发育阶段及其生殖周期。     

Effect of dietary vitamin C supplementation on the oxygen consumption, ammonia-N excretion and Na/K ATPase of Macrobrachium nipponense exposed to ambient ammonia

The 96-h LC₅₀ of ammonia-N and the effects of dietary vitamin C on oxygen consumption, ammonia-N excretion and Na⁺/K⁺ ATPase activity of Macrobrachium nipponense exposed to ambient ammonia were investigated. The results showed that the 96-h LC₅₀ of ammonia-N was 36.6 mg l⁻¹ for the freshwater prawn, M. nipponense, at pH 8.0. When prawns were exposed to high ambient ammonia-N concentrations, the oxygen consumption rate increased and ammonia excretion decreased. Dietary vitamin C supplementation led to higher oxygen consumption and lower ammonia excretion. Na⁺/K⁺ ATPase activity increased with increased ambient ammonia-N exposure in the range of 0–18.3 mg l⁻¹, and then was reduced at ambient ammonia-N 36.6 mg l⁻¹. Na⁺/K⁺ ATPase activities of prawns fed a vitamin C-supplemented diet were significantly lower than those of prawns fed a diet which was not supplemented with vitamin C.     

降胆固醇能力强的耐酸耐胆盐乳酸菌的筛选及鉴定

对从发酵酸菜中分离的乳酸菌,进行降胆固醇能力强的乳酸菌的筛选和鉴定,并对其耐酸耐胆盐能力进行测定。从分离的乳酸菌中,筛选到一株胆固醇降解率达到52.21%的乳酸菌M16。M16菌株具有较强的耐酸耐胆盐能力,在p H 2环境中30℃培养2 h后活细胞数仍然可以达到4.5×106 CFU·m L-1,在1.0%胆盐的环境中培养4 h后活细胞数仍然可以达到1.0×106 CFU·m L-1。通过菌体形态、菌落特征和生理生化实验,将M16乳酸菌鉴定为乳杆菌属的乳酸菌。     

蓝粒小麦4Ag染色体的显微分离与鉴定

本研究采用显微分离技术从蓝粒小麦(Triticum aestivum L.(2n=6x=42)×Thinopyrum ponticum Liu &Wang(2n=10x=70))根尖细胞中期分裂相中分离出具有4Ag染色体形态特征的单条染色体,对分离后的细胞进行原位杂交(FISH),结果显示细胞中所剩的和显微分离到的单条染色体均为4Ag染色体.利用Sau3A接头介导的PCR方法对分离出的单条4Ag染色体进行体外扩增,扩增的DNA片段大小约为200～2 000 bp,主要集中在250～750 bp之间.以DIG-dUTP标记的蓝粒基因组DNA为探针,与该扩增产物进行Southern杂交,结果表明显微分离出的染色体得到了有效的扩增.利用该分离染色体的PCR扩增产物为探针,对蓝粒小麦进行原位杂交,证明显微分离的染色体体外扩增片段确实来源于4Ag染色体.本研究拓宽了染色体显微分离的范围,为构建4Ag染色体文库和克隆位于该染色体上的重要农艺性状基因提供了新途径.