不同保存条件下鱼类组织基因组DNA的提取效果分析

对-20℃下冰冻，常温下酒精、甲醛溶液和Bouin氏液等保存条件下的鱼体组织（血液、肌肉、鳞片、鳍条）的DNA提取方法作了比较，并经琼脂糖凝胶电泳、紫外分光光度计检测及RAPD扩增结果验证。结果表明，除鳞片外，新鲜组织、-20℃冰冻组织以常规方法可提取出较高质量的DNA；保存于70％酒精或10％甲醛溶液中的组织，经预处理，亦可得高质量DNA；新鲜、冰冻、70％酒精或短时间10％甲醛固定的鳞片经适当预处理也可提取出较高质量的DNA。     

总状蕨藻多糖提取工艺的研究

采用酶法提取与热水抽提相结合的方法研究了总状蕨藻多糖的提取工艺,结果显示,采用固液质量比1∶10,蛋白酶用量136.5 U/g,于60℃下酶法提取2.5 h后,再将固液质量比调至1∶20,在温度95℃条件下继续进行热水提取2.5 h,多糖得率为6.7%,比单用热水提取法提高了131.0%。     

米曲霉中性蛋白酶特性的研究

研究了米曲霉中性蛋白酶的特性。结果表明,米曲霉中性蛋白酶的最佳提取方法为加入pH 7.2的缓冲液研匀,置恒温(30℃)培养箱中提取30 min,每10 min搅动1次,最后用4层纱布过滤。酶反应最适温度为55℃,最适pH值为7.2。NaCl和EDTA对其有一定的抑制作用。酶的热稳定性好,在45℃的温度下处理24 h后,酶活力仍有45.5%。米曲霉中性蛋白酶与培养物一起保存时,稳定性也较好,在4℃保存一个月后,酶活力几乎无变化。     

牡蛎壳制备活性钙

为充分利用牡蛎壳的钙资源 ,对乙醇在不同浓度、不同烧煮时间、不同配比条件对牡蛎壳粉中钙的活化效果及不同浓度的乳酸对活性钙的提取效果进行了研究。结果表明 :以活化温度 12 0℃ ,m(牡蛎壳粉 ) / g∶V(酒精 ) / m L为 1∶ 5,烧煮时间 3.5h活化效果最好 ;用体积分数为 85%乳酸提取钙的效果最佳。由牡蛎壳制备的乳酸钙含水分 1.9% ,游离酸 1.6% ,乳酸钙 95.1% ,磷 0 .3% ,钙 14.8%。     

简化CTAB法快速微量提取丝状真菌DNA

应用分子标记技术如RAPD、AFLP进行植物病原真菌的群体遗传学、系统演化[1～ 4] 等方面的工作 ,特别是进行群体遗传结构及系统演化研究时需要研究大量样品 ,提取DNA的工作量十分大 ,需要花很多的时间。通常提取植物病原真菌DNA时 ,获取菌丝后需要经低温真空干燥处理 ,利用液氮研磨 ,再用CTAB(十六烷基三乙基溴化铵 )或SDS(十二烷基磺酸钠 )提取[5 ] 。我们在研究水稻纹枯病菌群体遗传结构的过程中发现 ,采取通常程序提取DNA ,过程复杂 ,操作中容易造成交叉污染。基于快速、简洁、经济有效的原则 ,我们将CTAB法进行简化 ,采用多…     

鳙鱼鱼鳔黏多糖的提取及其抗氧化性活性评价

【目的】探讨鳙鱼(Aristichthys nobilis)鱼鳔黏多糖的提取工艺及提取物的化学组成及抗氧化活性。【方法】采用热水浸提法和碱液浸提法提取黏多糖,用GC-MS法测定分析所提取多糖的化学组成和抗氧化活性等性质。【结果】碱液浸提法的最佳工艺条件为料液质量体积比(g/m L)1∶20、碱液质量分数1%、提取温度70℃、提取时间为4 h,在此条件下,黏多糖的提取率为5.58%;热水浸提和碱液浸提得到的鳙鱼鱼鳔黏多糖量分别为5.3%和25.33%;气质联用色谱(GC-MC)测定结果显示,热水提取的黏多糖(WANVM)主要由质量分数13.72%木糖、19.57%甘露糖、16.11%葡萄糖及32.49%半乳糖构成;碱液提取的黏多糖(AANVM)主要由质量分数50%甘露糖和34.87%半乳糖构成。在相同质量分数条件下,WANVM清除羟基自由基、DPPH自由基及总还原力均高于AANVM。【结论】碱液浸提对黏多糖的提取效果优于热水浸提,AANVM、WANVM组分不同,且前者抗氧化活性略高于后者。     

酶解法提取鱼油的工艺参数优化

利用蛋白酶酶解法从黄鳍金枪鱼(Thunnus albacares)加工的下脚料———鱼头中提取鱼油。以鱼油提取率和感官特征为指标,通过正交优化实验设计,获得了胰蛋白酶提取鱼油的最佳酶解工艺参数:酶解温度45℃,酶添加量1.5%,料液质量比1∶1,酶解时间4 h,酶解pH 8。在该条件下,鱼油的提取率为4.34%,理化指标除过氧化值外均达到SC/T3502-2000的粗鱼油二级标准,多不饱和脂肪酸总含量高达38.47%,其中DHA和EPA的含量分别为23.63%和4.84%。     

正交设计法提取江蓠高活性膳食纤维

采用正交设计法 ,研究了江蓠高活性膳食纤维的提取工艺条件 ,筛选出江蓠碱处理和漂白的最佳工艺条件 ,提取出江蓠高活性膳食纤维。结果表明 ,江蓠高活性膳食纤维的产率达 2 8.3% ,颜色较白 ,膳食纤维干基质量分数达 92 .83% ,钙质量分数 1.0 8% ,磷质量分数 0 .0 6 8% ,膨胀力为 9.0 m L / g,持水力为 82 5% ,质量和功能性指标远远超过西方国家麸皮膳食纤维的标准 (膳食纤维干基质量分数 4 7%、膨胀力 4 m L/ g、持水力 4 0 0 % )     

超高压结合酶解法提取鱼油工艺优化

【目的】优化黄鳍金枪鱼(Thunnusalbacares)副产品(鱼头)提取鱼油的工艺参数。【方法】以鱼油提取率为评价指标,采用单因素实验确定最佳超高压处理压强和保压时间,采用Box-Behnken响应面试验设计优化关键酶解工艺参数。【结果】建立鱼油提取率与加酶量、液料质量比、酶解温度和酶解时间之间的回归模型,加酶量、液料质量比、酶解温度和酶解时间对鱼油提取率均有显著影响(P <0.05),影响由高到低依次为加酶量、液料质量比、酶解时间、酶解温度;当超高压处理压强200 MPa、处理时间10 min、加酶量(质量分数)为1.125%、液料质量比为1∶1、酶解温度65℃和酶解时间60min时,根据质量法计算,鱼油提取率达99.51%,与模型预测值之间差异无统计学意义(P> 0.05)。【结论】采用响应面法建立的回归模型较好地描述超高压结合酶解提取鱼油的工艺过程,优化的工艺参数可有效提高鱼油提取率。     

结合Gabor小波和形态学的高分辨率图像树冠提取方法

树冠信息的遥感提取能有效辅助森林参数反演、林分长势监测、树种识别等森林调查活动。随着遥感信息自动化提取的需求不断加强,本文基于高空间分辨率遥感数据,以滁州市皇甫山林场为研究区域,设计了一种结合Gabor小波和形态学的树冠提取方法。该方法首先采用Gabor小波提取出纹理特征,其次结合K-means聚类分析方法,对PCA降维后的纹理特征向量提取出阔叶林区,最后基于形态学理论降低影像噪声,并利用前景后景标记的分水岭方法进行单木树冠提取。经过与人工解译的树冠信息结果对比发现,在郁闭度较高的阔叶林区,该自动化方法提取树冠精度较高,分割准确率A_d为79.59%,F测度达到了79.00%能有效提供精确的单木树冠信息,为林业经济调查技术的发展具有一定的实践意义。     

Quality Assessment of Frozen Solenocera crassicornis Treated with Sodium Metabisulphite by Soaking or Spraying

The present work was carried out to evaluate the safety of shrimp(Solenocera crassicornis)treated with different concentrations of sodium metabisulfite(SMB)by soaking or spraying during frozen storage.Shrimps soaked in higher concentrations of SMB showed higher sensory scores,lower total color differences,and better anti-melanosis effects than shrimps in the control and other treatment groups throughout frozen storage(−18℃).Lower total volatile basic nitrogen and thiobarbituric acid reactive substances and higher salt soluble protein contents were detected in shrimp soaked with high doses of SMB compared with other samples.In addition,lower counts of total aerobic plates and psychrotrophic bacteria were observed in shrimp treated by soaking with higher doses of SMB than those in control shrimp and shrimp treated with other methods during frozen storage(−18℃).However,the SO2 content of 5%SMB-soaked samples exceeded the maximum allowable limit of 100mgkg−1.Overall,the use of 1.5%SMB soaking to treat shrimp results in good antioxidant and antimicrobial effects and,thus,may be suggested to preserve S.crassicornis under frozen conditions.The results of this study present important guidance on the use of SMB to maintain the quality of marine-trawling shrimp from manufacturing to consumption.     

温度、盐度对马氏珠母贝外套膜Hsp70基因表达量的联合影响

设定温度范围16～40°C,盐度范围10～50,采用复合设计和响应曲面分析法在实验室条件下研究不同温度、盐度对马氏珠母贝(P.fucata)外套膜Hsp70基因表达量的综合效应并建立模型。采用方差分析(ANOVA)、决定系数等对实验所得回归方程模型进行显著性及拟合度检验。结果表明,温度的一次、二次效应对马氏珠母贝外套膜Hsp70基因表达量影响显著(p值<0.05);盐度的一次效应对马氏珠母贝外套膜Hsp70基因表达量无显著影响(p值>0.05),二次效应对马氏珠母贝外套膜Hsp70基因表达量影响极显著(p值<0.05);温度、盐度之间的互作效应对马氏珠母贝外套膜Hsp70基因表达量无显著影响(p值>0.05)且温度的效应大于盐度的效应。经响应曲面分析法和优化,得到马氏珠母贝外套膜Hsp70基因表达量在适宜的温度、盐度范围内呈峰值变化,在温度26.85°C、盐度29.39时,马氏珠母贝外套膜Hsp70基因表达量达到最小值0.95,满意度达到99.36%。建立了马氏珠母贝外套膜Hsp70基因表达量模型方程,其决定系数98.69%,矫正决定系数97.38%,预测决定系数91.42%,模型的拟合度极高,可用于预测马氏珠母贝外套膜Hsp70基因表达量。     

基于NDVI阈值法的森林冰冻受灾范围精确提取

快速提取森林冰雪受灾范围,有利于准确掌握森林受灾情况,为此类灾害性气候事件防灾减灾、森林资源管理和生态保护提供科学依据。本文利用2001-2007年NDVI数据,提取灾前植被NDVI参考值和正常波动范围,结合2008年NDVI数据提取冰雪冻灾范围。该方法弥补了基于单一时相的传统方法（NDVI差值法）忽略植被指数正常波动的问题,分像元提取植被NDVI正常波动范围,使提取结果更加客观合理。与传统方法提取结果对比,省级尺度的验证结果相同（即森林受灾率均为34.72%,而实地调查森林受灾率35.3%）,但2种方法在县市行政单元提取的森林受灾率相差较大。NDVI阈值法提取的森林冰雪受灾范围主要分布于湖南省南部地区,北部地区分布相对较少,而传统方法提取结果主要分布于湖南省北部地区,南部地区分布相对较少。根据实地考察资料显示,相比于传统方法,NDVI阈值法提取结果与实际森林冰雪冻灾空间分布信息更接近,精度更高,更适合于区域大尺度提取森林冰雪受灾范围。     

Zooplankton community analysis in the Changjiang River estuary by single-gene-targeted metagenomics

DNA barcoding provides accurate identification of zooplankton species through all life stages. Single-gene-targeted metagenomic analysis based on DNA barcode databases can facilitate longterm monitoring of zooplankton communities. With the help of the available zooplankton databases, the zooplankton community of the Changjiang (Yangtze) River estuary was studied using a single-gene-targeted metagenomic method to estimate the species richness of this community. A total of 856 mitochondrial cytochrome oxidase subunit 1 (cox1) gene sequences were determined. The environmental barcodes were clustered into 70 molecular operational taxonomic units (MOTUs). Forty-two MOTUs matched barcoded marine organisms with more than 90% similarity and were assigned to either the species (similarity>96%) or genus level (similarity<96%). Sibling species could also be distinguished. Many species that were overlooked by morphological methods were identified by molecular methods, especially gelatinous zooplankton and merozooplankton that were likely sampled at different life history phases. Zooplankton community structures differed significantly among all of the samples. The MOTU spatial distributions were influenced by the ecological habits of the corresponding species. In conclusion, single-gene-targeted metagenomic analysis is a useful tool for zooplankton studies, with which specimens from all life history stages can be identified quickly and effectively with a comprehensive database.     

贝类精子质量检测与评价方法的研究Ⅴ:栉江珧精子线粒体活性的荧光检测

采用罗丹明123(Rh123)和碘化丙啶(PI)两种荧光染料对栉江珧(Atrina pectinata)精子线粒体活性进行检测,比较不同染液浓度和染色时间的检测效果,并用该方法检测和比较超低温冷冻保存对线粒体活性的影响。结果显示:Rh123和PI染色的最适染液浓度均为10μg/mL,染色时间控制在10 min之内;染色后具有线粒体活性的精子发出绿色荧光,死精子发出红色荧光,质膜受损但线粒体还未受损的精子发出红绿色荧光,表明Rh123和PI双荧光染色检测栉江珧精子线粒体活性是可行的;精液经冷冻后,线粒体活性为61.9%、质膜完整率56.8%,显著低于新鲜精液组(95.3%、93.5%),表明冷冻对精子结构会造成不同程度的损伤,Rh123和PI双荧光染色法可用来检测栉江珧精子质量。     

Preliminary Analysis of Population Genetic Diversity of Cultivated Laminaria japonica Sporophyte via AFLP Technique

The amplified fragment length polymorphic DNA (AFLP) technique was adopted to estimate the population genetic polymorphism among 30 sporophytes of Laminaria japonica collected from a cultivating farm in Rongcheng,China.Three methods were used for genomic DNA extraction from Laminaria japonica sporophyte and only the products obtained using the improved genomic DNA extraction kit method proved qualified for AFLP analysis.The parameters of the method were optimized.Samples of forty milligrams and the cell lysis time of 120 min were suggested to replace the parameters recommended by the manufacturer.Thirty individuals of Laminaria japonica from the same cultivating site were investigated using one pair of selective primers.A total of 21 loci were obtained and 17 of them were polymorphic.The mean percent age of polymorphic loci of this population was 80.95%.The Nei's gene diversity (H) within this population was 0.3028 and the average Shannon's Information index (I) was 0.4498.A genetic distance matrix among different individuals was constructed as well.Through this study,an applicable AFLP genetic analysis working system for Laminaria japonica sporophyte was established.The results of this research also revealed a high level of genetic diversity within the studied population.     

集成夜间灯光数据与Landsat TM影像的不透水面自动提取方法研究

利用多源遥感数据提取不透水面信息是一个重要的研究方向。针对以往研究中多需要人工选取不透水面样本进行模型训练的问题,本文通过整合夜间灯光遥感与Landsat TM影像中的空间和光谱信息实现了不透水面覆盖范围（Impervious Surface Area,ISA）的自动提取。首先根据夜间灯光的分布来定位ISA聚集的城市区域的位置,分别在城市区域内部和外部自动提取可靠性高的ISA及非ISA样本,然后通过迭代分类提取城市区域的ISA,再以此为样本对城市区域外部进行分类,最后将分类结果整合完成整幅影像的ISA提取流程。应用本方法对美国雪城地区的DMSP/OLS夜间灯光影像上提取了84个城市区域,提取精度大于95%。从中分别选择高ISA密度和低ISA密度的2个城市区域作为ISA提取的测试区,本文方法在城市区域内的ISA提取总体精度与kappa系数分别为88.23%和0.63;在城市区域外部为78.6%和0.54,均优于人工样本选取方法的提取精度,表明该方法能够实现精度稳定且高效的ISA自动提取。     

蕴含地理事件微博客消息的自动识别方法

微博客文本蕴含类型丰富的地理事件信息,能够弥补传统定点监测手段的不足,提高事件应急响应质量。然而,由于大规模标注语料的普遍匮乏,无法利用监督学习过程识别蕴含地理事件信息的微博客文本。为此,本文提出一种蕴含地理事件微博客消息的自动识别方法,通过快速获取的语料资源增强识别效果。该方法利用主题模型具有提取文档中主题集合的优势,通过主题过滤候选语料文本,实现地理事件语料的自动提取。同时,将分布式表达词向量模型引入事件相关性计算过程,借助词向量隐含的语义信息丰富微博客短文本的上下文内容,进一步增强事件消息的识别效果。通过以新浪微博为数据源开展的实验分析表明,本文提出的蕴含地理事件信息微博客消息识别方法,识别来自事件微博话题的消息文本的F-1值可达到71.41%,比经典的基于SVM模型的监督学习方法提高了10.79%。在模拟真实微博环境的500万微博客数据集上的识别准确率达到60%。     

Preliminary analysis of population genetic diversity of cultivated <Emphasis Type="Italic">Laminaria japonica</Emphasis> sporophyte via AFLP technique

The amplified fragment length polymorphic DNA (AFLP) technique was adopted to estimate the population genetic polymorphism among 30 sporophytes of Laminaria japonica collected from a cultivating farm in Rongcheng, China. Three methods were used for genomic DNA extraction from Laminaria japonica sporophyte and only the products obtained using the improved genomic DNA extraction kit method proved qualified for AFLP analysis. The parameters of the method were optimized. Samples of forty milligrams and the cell lysis time of 120 min were suggested to replace the parameters recommended by the manufacturer. Thirty individuals of Laminaria japonica from the same cultivating site were investigated using one pair of selective primers. A total of 21 loci were obtained and 17 of them were polymorphic. The mean percent age of polymorphic loci of this population was 80.95%. The Nei’s gene diversity (H) within this population was 0.3028 and the average Shannon’s Information index (I) was 0.4498. A genetic distance matrix among different individuals was constructed as well. Through this study, an applicable AFLP genetic analysis working system for Laminaria japonica sporophyte was established. The results of this research also revealed a high level of genetic diversity within the studied population.     

基于标记控制分水岭分割方法的高分辨率遥感影像单木树冠提取

树冠是树木的重要组成,可以直接反映树木健康状况。高空间分辨率遥感影像和遥感技术为快速获取详细的树冠信息和实时监测林冠变化提供了有效的途径。因此,基于高空间分辨率遥感影像的单木树冠提取方法研究对现代森林管理具有重要意义。本文以黄河三角洲地区孤岛林场人工刺槐林和旱柳为研究对象,以QuickBird影像为数据源,首先利用面向对象方法实现研究区林地和非林地分类;然后以林地为掩膜,提取出树冠分布范围;在此基础上,分别选取疏林区和密林区为试验区域,通过形态学开闭重建滤波,平滑图像,去除噪声;最后,利用标记控制分水岭分割方法分别对疏林区和密林区进行树冠提取。本文以人工勾绘结果为参考进行精度验证,结果显示疏林区F测度达到87.8%,密林区F测度达到65.5%,表明该提取方法简单易行,精度可靠。