一株海绵附生芽孢杆菌的抗硅藻附着活性成分的分离鉴定

为寻找天然抗污损活性化合物,以抗硅藻附着活性为导向,采用有机溶剂萃取、半制备高压液相色谱对分离自海绵的芽孢杆菌UST050418-715代谢产物进行分离,纯化抗硅藻附着活性物质,并利用气相色谱-质谱联用仪、核磁共振波谱分析活性物质结构。从菌株UST050418-715代谢产物中分离得到7种具有抗硅藻附着活性的环二肽类化合物,分别鉴定为:(1)环(L-亮氨酸-反式-8-羟基-L-脯氨酸-)、(2)环(L-缬氨酸-L-脯氨酸)、(3)环(D-脯氨酸-L-亮氨酸)、(4)环(L-脯氨酸-D-亮氨酸)、(5)环(甘氨酸-L-脯氨酸)、(6)环(L-苯丙氨酸-顺式-8-羟基-D-脯氨酸-)、(7)环(L-苯丙氨酸-反式-8-羟基-L-脯氨酸-)。说明海绵附生芽孢杆菌UST050418-715代谢产物中存在大量环二肽类化合物,可以帮助宿主海绵实现对硅藻附着的化学防御。     

New polymorphic psyllipsocids from Burmese amber (Psocodea: Psyllipsocidae)

Annulipsyllipsocus andreneli gen. et sp. nov. and A. inexspectatus gen. et sp. nov., the first records of the Psyllipsocidae from the Cretaceous amber of Myanmar are described, illustrated and their taxonomic position discussed. The new genus is characterized by antennae with 16 segments; flagellomeres with secondary annulations; radial cell closed, five-angled, bulged on its R1 border; triangular pterostigma. Species belonging to Annulipsyllipsocus gen. nov. are polymorphic. The described taxa represent the earliest evidence for polymorphism in Psocodea. A checklist of all known species of Psyllipsocidae is given.     

细基江蓠繁枝变型对水体硝态氮变化的生理响应

通过静态模拟实验,比较研究了6个硝态氮(NO3--N)摩尔浓度水平(0,10,20,40,60,80μmol/L)下培养的细基江蓠繁枝变型(Gracilariatenuistipitatavar.liuiZhangetXia)的丙二醛(MDA)、游离脯氨酸含量以及硝酸还原酶(NR)、超氧化物酶歧化酶(SOD)、过氧化物酶(POD)活性的变化。结果表明,随着水体NO3--N摩尔浓度的逐渐增加,细基江蓠繁枝变型NR、SOD和POD的活性均呈现先增加后下降的显著变化(P<0.05),膜脂过氧化产物MDA含量随水体NO3--N摩尔浓度的变化则呈相反的趋势(P<0.05);游离脯氨酸含量则与水体NO3--N摩尔浓度呈正相关(r=0.9161)。综合分析结果认为,单独以硝态氮为氮源时,20μmol/L的供氮水平能使细基江蓠繁枝变型处于相对最佳的生理状态。     

饲料中添加芽孢杆菌组分对凡纳滨对虾幼虾生长、能量代谢和抗WSSV能力的影响

以凡纳滨对虾(Litopnnaus vannamei)幼虾为实验动物,在其基础饲料中分别添加芽孢杆菌(Bacillus sp.)胞外蛋白粗提物(1.0%,胞外蛋白组)、胞内多糖粗提物(1.0%,胞内多糖组),以基础饲料为对照组饲料,进行为期28d的养殖实验,探讨芽孢杆菌胞外蛋白和胞内多糖粗提物对幼虾生长、能量代谢及抗病能力的影响。研究表明,芽孢杆菌2种粗提物对凡纳滨对虾的生长和成活率影响不显著(P>0.05)。饲喂2种粗提物影响了凡纳滨对虾肝胰腺和肌肉中能量代谢相关酶活力。胞外蛋白组肝胰腺中磷酸果糖激酶(PFK)、乳酸脱氢酶(LDH)活力在养殖实验前期显著高于对照组(P<0.05),而肌肉中PFK、LDH活力显著低于对照组(P<0.05);胞内多糖组肝胰腺中己糖激酶(HK)活力及肌肉中PFK、LDH活力显著低于对照组(P<0.05)。饲喂2种粗提物均显著提高了凡纳滨对虾肌肉中琥珀酸脱氢酶(SDH)活力和肝胰腺中电子传递系统(ETS)活力(P<0.05),胞外蛋白粗提物对肝胰腺中的SDH活力也有显著的提高作用(P<0.05)。饲喂2种粗提物显著降低了凡纳滨对虾肝胰腺及肌肉组织中脂肪酸合酶(FAS)含量(P<0.05)。此外,饲喂2种粗提物均会显著提高凡纳滨对虾肠道蛋白酶活力(P<0.05),而胞外蛋白粗提物使凡纳滨对虾肠道脂肪酶活力显著降低(P<0.05)。白斑综合征病毒(WSSV)感染后,与对照组相比,胞外蛋白组和胞内多糖组凡纳滨对虾半致死时间分别延长了26.19%和7.14%。根据以上结果推测,芽苞杆菌2种粗提物能够影响凡纳滨对虾能量代谢水平并提高其抗WSSV能力,且二者之间密切相关。     

Salinivibrio sp.YH4胞外丝氨酸蛋白酶EYHS耐盐性及生物信息学分析

摘要：目的 探讨菌株Salinivibrio sp.YH4分泌的丝氨酸蛋白酶EYHS的耐盐性及结构特征。方法 明胶底物酶谱法分析EYHS的耐盐性。应用生物信息学手段对EYHS及6种耐盐的S8家族丝氨酸蛋白酶结构特征进行分析。结果 EYHS在4 mol/L的NaCl溶液中仍具有活性,属于耐盐蛋白酶。EYHS及6种S8家族丝氨酸蛋白酶分子表面的loop区等无规则卷曲所占比例较高,α-螺旋与β-片层则主要位于酶分子内部。EYHS分子表面酸性氨基酸含量较高,且具有弱疏水内核。多序列比对发现蛋白酶的催化三联体两侧存在高度保守的基序和保守的极性氨基酸及芳香族氨基酸,并存在多个保守的Gly与Ala。同源模建和表面电荷分布显示,α螺旋和β片层围成了蛋白酶的催化腔,EYHS活性中心包含由Asp32、His65与Ser215组成的催化三联体,且催化位点区域表面静电势为负。结论 上述结构特征可能有助于耐盐丝氨酸蛋白酶EYHS在高盐环境下维持其稳定性和适度柔性,并有助于其催化功能的发挥,为深入研究耐盐丝氨酸蛋白酶的高盐环境适应性提供了一定的理论依据。     

A new pelagic Polyconchoecia Xiang, Chen and Du, 2018 (Ostracoda: Myodocopa: Halocyprididae) from the South China Sea

Halocyprid ostracods are appreciable part of ostracods floating through virtually everywhere in marine environment.In this study,we describe a new species of genus Polyconchoecia Xiang,Chen and Du,2018,tribe Conchoeciini Chavtur and Angel,2011,family Halocyprididae Dana,1853 from the middle of the South China Sea.Polyconchoecia chenii sp.nov.is very close to P.commixtus Xiang,Chen and Du,2018.But it differs from P.commixtus by the distinctions of locations of major glands of carapace and the characteristics of appendages:more posteriorly situated left asymmetric gland of carapace,no right asymmetric gland;segmented frontal organ;the endopod 2 of the first antenna with a very small seta;a-and c-setae of the first antenna with long end joint have long end joint,the b-and d-setae have no end joint,spinose e-seta without end joint;the e-seta of the second antenna is present;teeth side is distinctive;the setal counts of the mandible,maxilla,fifth limb,and sixth limb are individual.The locations of the major glands on carapace and the characteristics of the first antenna can be the key of the new species.This work is the second discovery of the genus Polyconchoecia from the world.     

产新琼四糖的海洋微泡菌属AG1热稳定性β-琼胶酶的过量表达与鉴定

从微泡菌属AG1（Microbulbifer sp. AG1）克隆得到1302 bp大小的琼胶酶基因,该基因编码产物为一个成熟蛋白（413个氨基酸残基）外加一个信号肽（20个残基）。将不含信号肽片段的琼胶酶在E. coli BL21 （DE3）中进行了异源表达和纯化。使用琼脂糖作为底物,该重组琼胶酶的最适反应温度和pH分别为60℃和7.5。该重组酶表现出优良的热稳定性,在50℃和60℃下处理1 h,重组琼胶酶仍能分别保持67%和19%的残余酶活力。除了SDS,重组琼胶酶对于其他测试的抑制剂、去垢剂和尿素变性剂有着较好的抗性。利用薄层色谱和以对硝基苯-α/β-D-吡喃半乳糖苷为底物的酶活力分析结果表明,该重组琼胶酶为β型琼胶酶,它水解琼脂糖的主要终产物为新琼四糖,而且不同聚合度的酶解产物具有抗氧化活性。     

渤海裸甲藻和链状亚历山大藻的麻痹性贝毒毒素分析

目的:分析渤海裸甲藻(Gymnodinium sp.)和链状亚历山大藻(Alexandrium catenella)的麻痹性贝毒毒素,为渤海天津海域的赤潮研究积累基础数据。方法:通过实验室培养裸甲藻和链状亚历山大藻,选取对数生长期、平台生长期的裸甲藻以及平台生长期的链状亚历山大藻,利用高效液相色谱法(HPLC)对这两种微藻进行麻痹性贝毒(PSP)毒素分析。结果:裸甲藻细胞内不含有麻痹性贝毒(PSP);链状亚历山大藻细胞内含有C毒素和GTX1-4毒素,该微藻每个细胞毒素含量约为10.81 fmol/cell。结论:裸甲藻细胞内虽不含有麻痹性贝毒(PSP),但不能排除其含有其它毒素的可能。链状亚历山大藻细胞内含有麻痹性贝毒(PSP),属于有毒微藻,需要对其进行密切监测。     

Hybrids between olive flounder Paralichthys olivaceus and stone flounder Kareius bicoloratus: karyotype, allozyme and RAPD analyses

The hybrid between olive flounder Paralichthys olivaceus and stone flounder Kareius bicoloratus was produced by artificial insemination of olive flounder eggs with stone flounder Sperm. Sinistral and dextral are two types of hybrid progeny after metamorphosis. Karyotypes of both hybrid flounders are the same as those of the two parental species. Of the 22 loci examined from 12 allozymes,12 confirmed hybridization of the paternal and matemal loci in hybrids and no difference was found in allozyme patterns of sinistral and dextral hybrid fishes. RAPD patterns of these specimens were also studied with 38 primers selected from 104 tested. Among them, the PCR products of 30 primers showed hybridization of the paternal and maternal bands. Genetic variation between hybrids and their parental stocks was analyzed by RAPD using 10 of the above 38 primers. The average heterozygosity and genetic distance were calculated. The results suggested that the filial generation could inherit a little more genetic materials from paternal fish than that from maternal fish.     

拮抗香蕉枯萎病菌海洋细菌的筛选和鉴定

从93份中国南部海域近海海泥样品中分离出1800多株海洋细菌,以对峙培养法,测定海洋细菌对香蕉枯萎菌的拮抗作用,再以琼脂划线培养法(agar streak)测定菌株的抑菌活性,结果表明:约55.6%的菌株对香蕉枯萎菌有一定的拮抗作用,其中58株菌株抑制香蕉枯萎菌生长所形成的抑菌带宽度大于10.0mm,抑菌作用较好,复筛后其中13个菌株形成的抑菌带宽在15.0mm以上。经反复转接和对峙培养,筛选出拮抗效果好、抑菌活性稳定的菌株TC-1,初步鉴定为芽孢杆菌属的细菌,TC-1对14种病原微生物都有较好的抑菌效果。