Molecular cloning, expression of heat shock protein 60 (HSP60) gene in Marsupenaeus japonicus and the association of its polymorphisms with thermal tolerance

日本囊对虾(Marsupenaeusjaponicus)是我国重要经济养殖虾类，但其在南方地区存在着高温度夏死亡率较高的养殖难题。因此，急需了解日本囊对虾的耐热性状及耐温的分子机制，为此，我们开展了日本囊对虾MjHSP60基因的克隆、表达分析及其多态性与耐热性状相关性的研究，获得了如下主要结果： 1.MjHSP60基因克隆 在日本囊对虾MjHSP60cDNA序列基础上克隆到了该基因DNA全序列。日本囊对虾MjHSP60基因全长4055bp，结构与拟穴青蟹一致，均由8个外显子及7个内含子构成，且对应的外显子长度极其相近，序列相似度达94%，而内含子长度存在较大差异。 2.MjHSP60mR...Marsupenaeus japonicus is an important commercial species in China, however, high temperature has been a negative factor for M. japonicus farming in summer. Therefore, it’s important to understand the thermal resistance and mechanism of heat resistance of M. japonicus. In the present study, a HSP60 gene from M. japonicus was cloned, the expression of MjHSP60 mRNA during early developmental stages ...学位：理学硕士院系专业：海洋与地球学院_海洋生物学学号：2232012115132

The physico-chemical properties of the algicidal pigment deinoxanthin and the development of algicidal agent

塔玛亚历山大藻(Alexandrium tamarense)是一种有毒甲藻,常引发赤潮,对海洋环境及人类健康造成极大的威胁.前期研究发现,耐辐射球菌属的一株细菌Y35能够通过分泌杀藻化合物Deinoxanthin达到控制塔玛亚历山大藻生长的效果.为了加强Deinoxanthin的应用效果,对其在不同条件下的稳定性进行了研究.结果表明,杀藻化合物Deinoxanthin有较高的热稳定性,且在中性和碱性条件下稳定,在光照和紫外线辐射下易分解.而通过壳聚糖和海藻酸钠对Deinoxanthin进行包埋固定化,从而构建缓释微球提高了Deinoxanthin的稳定性.杀藻制剂能够高效地表现出杀藻效果,杀藻制剂的研发为杀藻细菌色素应用于藻华治理奠定了基础.Alexandrium tamarense is a toxic dinoflagellate,capable of forming harmful algal blooms( HABs),and poses a substantial threat to both the marine environment and human health. Our previous research found Deinococcus sp. Y35 could secret algicidal pigment deinoxanthin which showed efficient algicidal effect on A. tamarense,resulting in controlling the growth of A. tamarense. In order to enhance the algicidal effect of deinoxanthin,the stability of deinoxanthin under different conditions was investigated. The results show that the thermal stability of deinoxanthin was high and remained stable under neutral and alkaline conditions,yet was prone to decompose in the light and ultraviolet radiation. The stability of deinoxanthin was improved after immobilization by chitosan and sodium alginate. The algicidal agent could exhibit high algicidal effect on A. tamarense,laying the foundation for the application in the control of HABs.国家自然科学基金(No.41576109,31400318);; 河南师范大学博士科研启动基金(No.5101049170160)~

Modified-Prior PLDA Based Speaker Recognition System

为减弱注册语音与测试语音时长不一致对说话人识别性能的负面影响,提出一个概率修正PldA建模方法.根据语音时长自适应改变传统PldA模型中I-VECTOr的概率分布函数,提高PldA对每个说话人每段语音的时长表征能力,以增强说话人类别的区分度.为验证基于概率修正PldA模型的有效性,进行了nIST SrE10 COrECOrE测试集在3种不同时长的评测实验,以及nIST 2014 I-VECTOr MACHInE lEArnIng CHAllEngE测试任务.结果表明,相较于传统的PldA训练模型,通过语音时长的约束提高了说话人识别性能.To reduce the negative impact on the performance of speaker recognition systems due to the duration mismatch between enrollment utterance and test utterance,a modified-prior PLDA method is proposed.The probability distribution function of i-vector was modified by incorporating the covariance matrix with duration of each utterance of each speaker during the PLDA training,which further improved the discriminant capability of speaker classification.To evaluate the robustness of the proposed modified-prior PLDA method,extensive experiments were performed on NIST SRE10 core-core task(female part)in duration mismatch conditions and NIST 2014 i-vector machine learning challenge.Experimental results demonstrated that the duration-based modified-prior PLDA method achieved better performance compared with the traditional PLDA.国家自然科学基金资助项目(61105026

基于电子鼻的慢性胃炎气滞证患者常见病位的口腔呼气气味图谱研究

目的:运用电子鼻探讨慢性胃炎气滞证患者常见病位的口腔呼气气味图谱特征。方法:采用证素辨证的方法,筛选出397例慢性胃炎气滞证患者并判断病位证素,同时运用基于阵列式气体传感器技术的医用电子鼻(EN0l1103-A)采集其口腔呼气的气味图谱,选择气味图谱响应曲线的振幅、斜率作为图谱特征参数,借助分类器算法对慢性胃炎气滞证与非气滞证的口腔呼气气味图谱特征进行模式识别,比较慢性胃炎气滞证患者常见病位的口腔呼气气味图谱特征。结果:慢性胃炎气滞证的主要病位证素分布是胃(91.18%)、脾(38.29%)、肝(23.68%);主要病位为胃、脾胃、肝脾胃、肝胃;采用分类器算法对慢性胃炎气滞证的气味图谱进行模式识别时,运用随机森林算法,对气滞证的准确率可以达到65.85%;病位胃组、脾胃组的气味图谱响应曲线B、C、D、E、F、I、J的振幅均显著低于病位肝脾胃组(P<0.01);病位胃组、脾胃组的气味图谱响应曲线C、D、E、F、I的斜率均显著低于病位肝脾胃组(P<0.01),脾胃组曲线A的斜率低于病位肝脾胃组(P<0.05)。结论:运用电子鼻结合模式识别方法可初步判断慢性胃炎气滞证及其不同病位间的口腔呼气气味差异。国家自然科学基金项目(No.81373552);;福建省自然科学基金项目(No.2014J01362);;福建省中医药科研项目(No.wzpw201313);;福建省教育厅A类项目(No.JA14212);;载人航天领域预先研究项目(No.020104)~

高效溶藻放线菌BS01发酵培养基及发酵条件优化

从漳江口红树林区采集的沉积物样品中分离到1株放线菌菌株BS01 Brevibacterium sp.,其胞外活性产物对塔玛亚历山大藻Alexandrium tamarense具有明显的溶藻作用。采用单因素及均匀设计,通过摇瓶培养对BS01产溶藻活性物质的发酵培养基及发酵条件进行优化。结果表明,在可溶性淀粉为碳源、硝酸钠为氮源、装液量为40%、起始pH值为7.5、培养温度为28℃、转速为150r.min?1、振荡培养时间为48h的条件时,BS01发酵产物的杀藻活性最强。通过均匀设计进行最佳发酵培养基及培养条件优化的结果为:可溶性淀粉为20g.L?1,硝酸钠为0.5g.L?1,pH为7.7,温度为27.2℃。研究结果为杀藻活性物质高效提取及杀藻机制研究奠定了基础

Optimization of Fermentation Conditions of Marine Algicidal Bacterium Microbulbifer sp. BS03 by Uniform Design

采用均匀设计法设计和二次多项式逐步回归分析,对一株高效杀塔玛亚历山大藻微泡菌bS03(MICrObulbIfEr SP.)产杀藻活性物质的发酵培养条件进行优化.通过单因素实验筛选出碳源、氮源、PH、培养时间和接种量为显著影响因子,并对5个显著影响因子采用u15(155)水平对培养基进行优化.结果表明bS03最适发酵培养条件为:蔗糖8 g/l,蛋白胨10.50 g/l,初始PH值7.5,培养时间32 H,接种量3.00%.验证试验结果显示,在此条件下该菌发酵液的干重为4.725 g/l,较优化前增加了31.35%,ld50为0.768%,较优化前降低了25.14%.研究结果为杀藻活性物质以及杀藻机理的研究奠定了理论基础.The optimal parameters of fermentation conditions of marine bacterium Microbulbifer sp.BS03 producing algicidal substances against Alexandrium tamarense were obtained by using Uniform Design and quadric polynomial regression methods.Five main factors were achieved through single factor experiments,that is,carbon source,nitrogen source,pH,incubation time and inoculum size.The U15(155) design result showed the optimum conditions of experiment were as follows: sugar 8 g/L,peptone 10.5 g/L,initial pH 7.5,incubation time 32 h,and inoculum size of 3.00%.According to the verification experiment,under the optimal conditions,the dry biomass was 4.725 g/L which was increased by 31.35% and LD50was 0.768% which was decreased by 25.14% compared with those of the basic fermentation conditions.This outcome will help us to separate active substances from complex components in medium and thus to reveal the mechanism of algicidal activity in the future.国家自然科学基金项目(Nos.40930847;31070442);福建省自然科学基金项目(Nos.2012J01150;2010J01223);福建省教育厅项目(No.JA10232);莆田市科技计划项目[No.2011S09(4)]资助;theOpenFundoftheKeyLaboratoryoftheMinistryofEducationforCoastalandWetlandEcosystemsofXiamenUniversity(CWel0902)---

RNA甲醛琼脂糖凝胶电泳的优化及探讨

目的:优化RNA甲醛琼脂糖凝胶电泳条件。方法:对实验器材进行彻底的RNase灭活处理;在原有电泳过程的基础上,增加预电泳、缓冲液液面的处理和制胶过程中未加入溴化乙锭等优化过程。结果:优化后的电泳不仅能验证总RNA的完整性,而且能鉴定RNA的分子大小。结论:优化的RNA甲醛琼脂糖凝胶电泳法操作简单、省时、快速,RNA条带清晰、定位准确、无弥散及非特异条带

Functional groups of benthic macrofauna in the 14th Yong intertidal zone of Nansha,Guangzhou

大型底栖动物根据食性可分为浮游生物食者(planktophagous,Pl)、植食者(phytophagous,Ph)、肉食者(carnivorous,C)、杂食者(omnivorous,O)和碎屑食者(detritivorous,D)五个功能群。本文根据2007–2008年度和2013–2014年度在茳芏(Cyperus malaccensis)、海桑(Sonneratia caseolaris)2种生境调查获得的各4个季度的大型底栖动物数据,分析了广州南沙十四涌潮间带大型底栖动物功能群的生境差异、季节变化和年际变化。2007–2008年度采集到26种大型底栖动物,低于2013–2014年度的36种。无论是2007–2008年度还是2013–2014年度,茳芏、海桑生境的大型底栖动物均以植食者的栖息密度和生物量最高,碎屑食者的栖息密度和生物量最低。生境比较得出,茳芏生境大型底栖动物浮游生物食者(Pl)的丰富度指数(d)、均匀度指数(J)和多样性指数(H')均高于海桑生境。年度比较得出,在茳芏和海桑生境,2013–2014年度浮游生物食者的丰富度指数、均匀度指数和多样性指数均高于2007–2008年度,这是因为2013–2014年度采集到红树蚬(Gelonia coaxans)和彩虹明樱蛤(Morerlla iridescens)等,而2007–2008年度没有采集到。Benthic macrofauna were classified into 5 functional groups,i.e.planktophagous(Pl),phytophagous(Ph),carnivorous(C),omnivorous(O) and detritivorous(D).The biotope difference,seasonal and annual variations of functional group of benthic macrofauna in the intertidal zone of the 14 th Yong of Nansha,Guangzhou were analyzed in this study according to the four seasonal data of benthic macrofauna collected in two biotopes(Cyperus malaccensis and Sonneratia caseolaris) in 2007–2008 and 2013–2014.A total of 26 species were recorded in 2007–2008,which was lower than 36 species in 2013–2014.Both in 2007–2008 and 2013–2014,phytophagous group showed the highest value while detritivorous group shared the lowest one in density and biomass.Habitat comparison showed that the abundance index(d),evenness index(J) and diversity index(H') of planktophagous group in Cyperus malaccensis habitat were all higher than those in Sonneratia caseolaris habitat.Annual comparison showed that the abundance index,evenness index and diversity index of planktophagous group in 2013–2014 were all higher than those in 2007 –2008 both in Cyperus malaccensis and Sonneratia caseolaris habitats because the planktophagous such as Gelonia coaxans and Morerlla iridescens and so on were collected in 2013–2014 but they were not found in 2007–2008.国家自然科学基金(41176089和41376113

溶藻细菌BS03(Microbulbifer sp.)对塔玛亚历山大藻生长及抗氧化系统的影响

【目的】研究溶藻细菌BS03(Microbulbifer sp.)胁迫下塔玛亚历山大藻细胞光合作用、抗氧化酶系统和半胱氨酸蛋白酶3(Caspase-3)变化,探讨溶藻细菌BS03对塔玛亚历山大藻的溶藻机制。【方法】通过0.5%、1.0%、1.5%、2.0%不同终浓度BS03上清液处理藻细胞后12、24、36、48h取样,测定溶藻过程藻细胞光合色素、叶绿素荧光效率、抗氧化酶系统、Caspase酶活性变化。【结果】(1)BS03上清液处理藻细胞后,藻细胞叶绿素a含量和叶绿素荧光Fv/Fm比值随BS03上清液处理时间延长和浓度的增加呈逐渐下降趋势;低浓度处理组藻细胞类胡萝卜素含量上升到一峰值,高于对照组后逐渐回落,而高浓度处理组类胡萝素含量呈下降趋势,低于对照组;(2)藻细胞抗氧化酶保护系统(SOD和CAT)活性随着BS03上清液处理浓度增加而升高,但随着处理时间的延长呈现先上升后下降趋势。藻细胞膜脂过氧化产物MDA积累量随着BS03上清液处理时间延长和处理浓度的增加而显著提高;(3)处理组藻细胞Caspase-3活性显著高于对照组,呈现出类似程序性死亡特征。【结论】BS03的抑藻机理可能是通过抑制藻细胞光合作用,降低抗氧化酶活性、加大膜脂过氧化起到对塔玛亚历山大藻的溶解作用,并呈现出类程序性死亡特征

理想声源辐射声场的数值分析

通过声场的特性和规律分析可以识别和定位噪声源,对噪声控制以及声源的设计提供参考。数值方法是求解有源声场的重要工具,复杂的声辐射一般可分解为简单的声辐射叠加。研究了单极子、偶极子、活塞等几种理想声源辐射声场的解析解,并用有限元法计算数值解,得到相应的辐射声场,包括声压、速度、指向性等量,有限元法得到的数值结果与解析解吻合;利用有限元法计算了点声源的线性阵列与平面阵列等典型的叠加声场。对各种声源的特性和辐射声场的规律以及在工程领域中的应用进行了归纳。国家自然科学基金项目(51505261);;山东省自然科学基金项目(ZR2015AM013