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1.
An (AC)n-microsatellite-enriched library for Larimichthys crocea was constructed in this study. Primers for fifty simple sequence repeat (SSR) loci were synthesized and genotyped on 30 L. crocea individuals from Guanjingyang wild population (WP) in Fujian Province and 38 individuals from Ningbo cultured population (CP) in Zhejiang Province. Only 21 loci were successfully amplified and polymorphic in two populations. In WP, the observed heterozygosity (H_O ) ranged from 0.233 to 0.900 and the expected heterozygosity (H_E ) ranged from 0.326 to 0.893, with an average of 7.8 alleles/locus, the polymorphism information content (PIC) ranged from 0.283 to 0.866 (mean 0.731). In CP, the H O ranged from 0.189 to 0.892 and the H E ranged from 0.333 to 0.800, with an average of 4.4 alleles/locus. The probability test showed significant departures from HWE in 9 and 2 of the 21 loci in WP and in CP, respectively. Deficiency of heterozygotes at four loci showed the presence of null alleles (P <0.01). The PIC of 20 microsatellite loci in WP were greater than 0.50. Overall, these novel markers are potentially useful for the study of population genetics, construction of genetic linkage and quantitative trait loci maps in large yellow croaker.  相似文献   

2.
夏季高温是限制龙须菜(Gracilariopsis lemaneiformis)生长和发育的一个主要问题。本研究以野生龙须菜作为材料,经过28℃的热锻炼预处理72 h,再分别置于32、36、40℃高温胁迫24 h,并测定了高温胁迫条件下龙须菜的丙二醛(MDA)、过氧化氢(H2O2)和可溶性蛋白质的含量,以及超氧化物歧化酶(SOD)活性和抗坏血酸(AsA)水平等生理指标,研究了龙须菜对高温胁迫的响应。结果表明:热锻炼能维持藻体较低的MDA含量,降低H2O2的产生速率,缓解藻体的膜脂过氧化伤害;同时热锻炼缓解了可溶性蛋白质的降解,增加了SOD酶的比活力,缓解了AsA的下降速率,说明热锻炼能缓解高温胁迫对藻体的伤害,有效地提高了龙须菜对高温胁迫响应的能力。  相似文献   

3.
王菁  牛从从  王辉  郇丽 《海洋科学》2016,40(1):64-67
浒苔(Ulva prolifera)是我国黄海海域绿潮暴发的主要成因种,其孢子囊的形成和孢子的释放是浒苔大量、快速增殖的基础。本文以浒苔为研究材料,比较分析了还原剂(DTT)和氧化剂(H_2O_2)对直径为1 mm的圆形藻片从营养细胞形成孢子囊过程的影响。研究结果表明,DTT对浒苔孢子囊的形成具有明显的抑制作用,用H2O2处理的浒苔藻片与对照组相比没有显著性的差异,因此浒苔细胞处于还原状态时不利于孢子囊的形成。研究结果为探究浒苔孢子形成的调控机制提供了科学依据。  相似文献   

4.
以哈维弧菌(Vibrio harveyi)、鲍类疱疹病毒(Abalone herpesvirus,Ab HV)悬液对杂色鲍(Haliotis diversicolor)注射刺激,研究其血淋巴中可溶性总蛋白浓度,免疫相关酶超氧化物歧化酶(Superoxide Dismutase,SOD)、酸性磷酸酶(Acid Phosphatase,ACP)、碱性磷酸酶(Alkaline Phosphatase,AKP)活性的变化,杂色鲍外套膜、鳃、肝胰腺、腹足中血蓝蛋白(Hemocyanin,Hc)基因Hc1、Hc2相对表达量的变化以及感染后杂色鲍无细胞血浆的抑菌性。结果显示:(1)杂色鲍血淋巴中可溶性总蛋白浓度变化显著,呈现出先升高后降低的趋势。(2)无细胞血浆中的SOD、ACP、AKP活性与对照组相比变化显著,哈维弧菌注射组的SOD活性在感染后第6小时显著升高;Ab HV注射组SOD活性与对照组比较在注射后的各个时相均呈下降趋势,从注射后6 h各个时相的SOD活性与对照组相比均差异显著(P0.05)。哈维弧菌注射组ACP活性在注射后的第48小时显著升高;Ab HV注射组ACP活性在注射后的第12小时显著升高。哈维弧菌注射组AKP活性在注射后48 h显著升高;AbHV注射组AKP活性在注射后第12小时显著升高。(3)杂色鲍外套膜、鳃、肝胰腺以及腹足中血蓝蛋白两个基因Hc1、Hc2的相对表达量均在注射后的不同时间点出现升高。(4)杂色鲍无细胞血淋巴对哈维弧菌、创伤弧菌和溶珊瑚弧菌这三种贝类病原均表现出了不同程度的抑菌性增强的作用。这些结果表明细菌和病毒的刺激引起了杂色鲍免疫相关因子的变化,使得血淋巴的抑菌性增强。本研究为今后进一步研究杂色鲍的非特异性免疫以及在生产实践中应用免疫技术防治病害提供参考。  相似文献   

5.
谷胱甘肽过氧化物酶(Glutathione peroxidase,GPx)是生物体内重要的抗氧化酶,能防止过氧化氢对生物体的氧化应激。该研究利用RACE技术获得了斑节对虾(Penaeus monodon)GPx3a(PmGPx3a)的全长c DNA序列,进行了相关生物信息学分析。作者利用荧光定量PCR方法研究了PmGPx3a在斑节对虾不同组织的表达情况。探究了PmGPx3a在不同胁迫条件下(盐度、重金属和细菌)的表达情况。结果表明PmGPx3a c DNA全长1135 bp,其中开放阅读框(ORF)长651 bp,预测编码216个氨基酸。PmGPx3a推导的氨基酸序列与其他动物的GPx3a氨基酸序列具有高度一致性。实时定量PCR结果显示,在高低盐胁迫下,PmGPx3a在肝胰腺中相对表达量都为上升的(p0.05)。在铜、锌、铬胁迫中,鳃中的PmGPx3a的相对表达量总体呈现下降趋势,在肝胰腺中呈现上升趋势。在哈维弧菌(Vibrio harveyi)刺激下,PmGPx3a在血淋巴中的相对表达量总体呈现上升趋势,24 h后表达量最大,显著高于对照组2.2倍(P0.05)。以上研究结果表明,PmGPx3a基因参与了斑节对虾对环境胁迫和氧化应激的适应性反应。  相似文献   

6.
构建了文蛤铁蛋白的重组表达质粒 pGEX-4T-1-MmeFer, 在大肠杆菌 Escherichia coli 中获得了重组蛋白, 经酶解和质谱鉴定该蛋白为重组文蛤铁蛋白(rGST-MmeFer), 切除 GST 标签的重组文蛤铁蛋白(rMmeFer)具有氧化和吸收铁离子的功能。利用 rGST-MmeFer 制备了兔抗文蛤铁蛋白的多克隆抗体, 进而利用 Western blot 的方法对文蛤铁蛋白的组织表达分布进行了研究。结果表明, 铁蛋白在文蛤各组织中均有分布, 其中消化腺中表达量最高, 足中表达量最低。此外, 利用实时定量PCR 分析了文蛤消化腺、外套膜和鳃组织中铁蛋白 mRNA 的表达, 发现铁蛋白 mRNA 在外套膜中表达量最高, 消化腺次之, 鳃中表达量最低, 差异达到显著水平(P<0.05)。上述结果为进一步研究铁蛋白参与贝类贝壳形成的分子机制奠定了基础。  相似文献   

7.
Solid-state voltammetric (micro)electrodes have been used in a variety of environments to study biogeochemical processes. Here we show the wealth of information that has been obtained in the study of sediments, microbial mats, cultures and the water column including hydrothermal vents. Voltammetric analyzers have been developed to function with operator guidance and in unattended mode for temporal studies with an in situ electrochemical analyzer (ISEA). The electrodes can detect the presence (or absence) of a host of redox species and trace metals simultaneously. The multi-species capacity of the voltammetric electrode can be used to examine complex heterogeneous environments such as the root zone of salt marsh sediments. The data obtained with these systems clearly show that O2 and Mn2+ profiles in marine sedimentary porewaters and in microbial biofilms on metal surfaces rarely overlap indicating that O2 is not a direct oxidant for Mn2+. This lack of overlap was suggested originally by Joris Gieskes' group. In waters emanating from hydrothermal vents, Fe2+, H2S and soluble molecular FeS clusters (FeSaq) are detected indicating that the reactants for the pyrite formation reaction are H2S and soluble molecular FeS clusters. Using the ISEA with electrodes at fixed positions, data collected continuously over three days near a Riftia pachyptila tubeworm field generally show that O2 and H2S anti-correlate and that H2S and temperature generally correlate. Unlike sedimentary environments, the data clearly show that Riftia live in areas where both O2 and H2S co-exist so that its endosymbiont bacteria can perform chemosynthesis. However, physical mixing of diffuse flow vent waters with oceanic bottom waters above or to the side of the tubeworm field can dampen these correlations or even reverse them. Voltammetry is a powerful technique because it provides chemical speciation data (e.g.; oxidation state and different elemental compounds/ions) as well as quantitative data. Because (micro)organisms occupy environmental niches due to the system's chemistry, it is necessary to know chemical speciation. Voltammetric methods allow us to study how chemistry drives biology and how biology can affect chemistry for its own benefit.  相似文献   

8.
改性粘土治理赤潮生物的主要原理是絮凝作用,但水体中未被去除的微藻对改性粘土有怎样的生理生化响应尚不清楚。本文以小球藻(Chlorella vulgaris)为模式生物,考察了改性粘土絮凝后水体中残余小球藻生理生化性质的变化情况。研究发现,经改性粘土或原土絮凝后,残余小球藻的生长受到了明显抑制,其丙二醛(malondialdehyde,MDA)含量迅速上升,改性粘土组的MDA含量高于原土组。除了原土组过氧化氢酶(catalase,CAT)活性相比对照组没有明显变化外,改性粘土组和原土组超氧化物歧化酶(superoxide dismutase,SOD)、CAT、抗坏血酸过氧化物酶(ascorbate peroxidase,APX)和谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-PX)等酶活,均呈现先上升后下降、并稳定在较高水平的变化趋势。原土组各酶活普遍低于改性粘土组,但GSH-PX活性则相反。结果表明,改性粘土不仅能高效去除水体中的小球藻,而且能够刺激残余的藻细胞积累较多的O2-·和H2O2,从而对藻细胞造成过氧化伤害,抑制未被去除部分海洋微藻的正常生长。  相似文献   

9.
利用可口革囊星虫重组铁蛋白富集Pb2+、Fe3+、Cd2+、Cr3+、Cu2+、Mn2+、Sn2+和Zn2+等8 种重金属离子后, 通过透射显微镜和傅里叶红外光谱测定蛋白的超微结构变化和内部基团特性。以 天然的马脾铁蛋白为对照, 观察重组铁蛋白与天然铁蛋白的差别。结果显示, 重组铁蛋白富集Pb2+ 和其它7 种重金属离子后蛋白体积大小与重金属离子种类直接相关。重组铁蛋白相对天然铁蛋白来 说在富集不同种类重金属离子后, 除了非特异性酰胺特征峰和蛋白特征峰外, 还出现一些专门富集 某些重金属离子的特异基团, 如:-CH3、-CH2-、-CH-、-NH-、-SO3-、-COOH、-S=S-和-C-S-C 等。 通过细胞实验进一步验证了铁结合蛋白能够通过富集重金属离子Pb2+来保护小鼠成骨前体细胞 MRC3T3-E1, 使细胞的死亡率明显降低(P<0.05).  相似文献   

10.
采用体外实验筛选益生菌结合16SrDNA序列分析法,从124株不具有溶血作用的皱纹盘鲍(Haliotis discus hannai)肠道细菌中筛选得到5株潜在益生菌并进行了分子鉴定,进一步对5株潜在益生菌进行了安全性实验及体内饲喂实验。结果表明,2株具有拮抗哈维弧菌和灿烂弧菌能力的潜在益生菌分别被鉴定为Shewanella sp.(WA64)和Shewanella sp.(WA65),一株产海藻酸酶和淀粉酶的潜在益生菌被鉴定为Vibrio sp.(WA51),产蛋白酶潜在益生菌被鉴定为Bacillus sp.(FA12),产琼脂酶潜在益生菌被鉴定为Tamlana sp.(FA86);安全性实验表明5株潜在益生菌在107cfu/ml下对皱纹盘鲍没有明显的毒害作用;通过体内饲喂实验发现,潜在益生菌WA64、WA65的复合作用能够显著提高幼鲍增重率和存活率(P<0.05),并在生产条件下能够明显降低幼鲍的死亡量。经抗生素敏感性实验,WA64菌株对15种抗生素均敏感或中度敏感,WA65菌株仅对庆大霉素和链霉素2种抗生素产生耐药。  相似文献   

11.
It is widely thought that redox-active metals in cells such as iron or copper catalyze the reduction of hydrogen peroxide to toxic hydroxyl radicals or their equivalent. However, this has not been directly demonstrated in vivo. To probe this requirement, the freshwater microorganism Euglena gracilis was used. Its intracellular iron content can be modulated by the concentration of iron in the defined growth medium without effect on the proliferation rate of the cells. E. gracilis contains two large storage pools of cytosolic iron which can be monitored to assess cellular iron status. The toxicity of H2O2 in E. gracilis was inversely related to the amount of iron in the extracellular medium. At high levels of external iron, the metal carried out the Fenton reaction with H2O2 outside the cell, producing hydroxyl radicals as detected by electron-spin resonance spin-trapping experiments. This reaction reduced the amount of H2O2 that could diffuse into cells to cause toxicity. When cells with different intracellular iron content were placed in iron-deficient media, the the toxicity of H2O2, measured by inhibition of proliferation, was directly related to the concentration of internal iron. Using cells deficient in iron, this oxidant did not inhibit proliferation at low concentration but was somewhat effective at higher concentrations. Although the iron chelating agents, 1,10-phenanthroline and desferrioxamine, also depressed cytosolic iron, they were cytotoxic to E. gracilis and so could not be used unambiguously to examine the role of intracellular iron in the toxic effects of hydrogen peroxide.  相似文献   

12.
Although there has been an increase in our understanding of the shell-boring polydorids that infest abalone Haliotis midae in South Africa, abalone from a limited number of farms, and wild populations from east of Cape Agulhas only, have been examined. To gain further knowledge and a more complete understanding of the local distribution of polydorids, we examined up to 30 abalone from each of 14 farms in the Northern, Western and Eastern Cape provinces, and five wild sites in the Western Cape, west of Cape Agulhas. Farm and wild communities were significantly different from each other (ANOSIM, r = 0.632, p < 0.002) and Bray–Curtis cluster analysis showed that most farms clustered separately from wild sites. Boccardia proboscidea was present on all but one of the infested farms, whereas some were also infested by Dipolydora capensis. By contrast, D. capensis was present at all wild sites sampled, whereas B. proboscidea was absent from these sites. Polydora hoplura, a common shell-boring pest, was present at only two farms and two wild sites. There appeared to be some exchange of polydorids among farms and between farms and wild abalone. The farm on the East Coast did not cluster with any other sites, indicating a unique composition of polydorids on that coast. No new, potentially problematic, species was encountered, although four species showed an increase in their known distribution.  相似文献   

13.
鲍是重要的海珍品,羊鲍(Haliotisovina)是主要生活在热带岩岸潮间带大型经济鲍类底栖动物。为了研究南海岛礁羊鲍资源及其栖息环境,我们在宣德群岛的永兴岛和七连屿,永乐群岛的羚羊礁、甘泉岛和晋卿岛进行了羊鲍资源分布及其栖息水环境研究。按照《海岛调查技术规程》中潮间带岩岸大型底栖生物调查方法,对羊鲍进行取样调查。按照《海洋调查规范第4部分:海水化学要素调查》(GB T 12763.4-2007),对羊鲍栖息地水环境的总无机氮、活性磷酸盐、悬浮物和叶绿素a等海水化学要素进行分析。结果表明,羊鲍在永乐群岛和宣德群岛都有分布,这些岛礁按永兴岛、七连屿、羚羊礁、甘泉岛和晋卿岛的顺序,分布密度依次增加,个头大小依次增大,总体来说羊鲍资源量小,有的岛礁海区几乎绝迹,亟需开展生态增殖和资源养护研究工作。水质调查表明,除永兴岛站位的悬浮物指标因填海作业未达标外,各站位的水质指标均达一类水标准且具有一定稳定性。因此,尽管南海岛礁受人类活动的影响,但该海域仍适合羊鲍栖息,是羊鲍资源养护、底播生态增殖和生态养护的适宜场所,符合建立海洋保护区的国家水质标准和环境条件。  相似文献   

14.
采用微卫星DNA分子标记方法对中国南部沿海多鳞鱚(Sillago sihama)广西北海、广东湛江、汕尾及海南三亚4个野生地理群体进行了遗传多样性及遗传分化等研究。结果表明:(1)用于研究的34对微卫星引物在4个群体中具有丰富的多态性,有效等位基因数介于1.71—20.46之间,多态信息含量PIC介于0.40—0.95之间。4个群体总的平均期望杂合度(He)为0.598,观测杂合度(Ho)为0.572,SY群体的杂合度最大(He=0.606,Ho=0.623),SW群体的最小(He=0.586,Ho=0.508)。(2)群体间F-统计量分析表明,群体遗传分化处于高度分化水平(Fst平均值为0.306);基于奈氏遗传距离(DA)构建的NJ和UPGMA聚类树均显示地理位置相邻的群体聚在一起。  相似文献   

15.
海水中的高溶解氧浓度、低温和UV辐射导致了氧胁迫是北极海洋细菌面临的主要胁迫因素之一。本文对北极细菌Pseudoalteromonas sp.A2对H2O2导致的氧胁迫的应答特征进行了转录组测序和基因差异表达的比较分析,以期发现与氧胁迫相关的关键功能基因。研究表明,与对照组相比,1 mmol/L的H2O2可导致菌株A2转录组的很大变化,包括109个基因的显著上调与174个基因的显著下调。COG分析表明,在功能已知的基因中,与转录后修饰、蛋白质转换和分子伴侣相关的基因大部分显著上调,而与氨基酸运输和代谢等相关基因则大部分显著下调。值得指出的是,有18个与鞭毛相关的基因和4个与热激蛋白相关的基因显著上调;同时,有9个与细胞色素和细胞色素氧化酶相关的基因和5个与TonB依赖受体相关的基因显著下调。在GO分析表明具有抗氧化活性的18个基因中,只有1个基因的表达显著上调,而有2个显著下调。简言之,RNA-Seq表明,除了传统的抗氧化基因和应激蛋白外,鞭毛相关基因和功能未知基因在菌株Pseudoalteromonas sp.A2的氧胁迫适应性中起着重要作用。该转录组分析和氧胁迫相关基因的发现有助于了解北极细菌的氧胁迫适应机制。  相似文献   

16.
采用实验生态学方法研究了不同盐度梯度(10、15、20、25、30和35)和规格体重[S组(1.55±0.37) g, M组(3.92±0.74) g, L组(9.08±1.38) g]对横带髭鲷(Hapalogenys mucronatus)幼鱼耗氧率(RO)和排氨率(RN)的影响,以及氧氮比(O:N)对横带髭鲷幼鱼能源物质的分析。结果表明,当盐度恒定时,横带髭鲷幼鱼的耗氧率和排氨率均随体重(W)的增加而下降,体重与单位体重的耗氧率之间呈一元三次函数关系,体重与单位体重的排氨率之间呈幂函数关系;盐度和体重对横带髭鲷幼鱼的耗氧率、排氨率均有显著影响(P<0.05),但盐度和体重的交互作用对耗氧率、排氨率影响均不显著(P>0.05)。随着盐度的升高,三种规格横带髭鲷幼鱼耗氧率和排氨率均呈先升高后降低的趋势,耗氧率在盐度20时出现相对最大值,而排氨率则在盐度25时出现相对最大值。三种规格横带髭鲷幼鱼O︰N值均随盐度的上升呈波动趋势, O:N值波动范围在5.035~7.533之间,其总平均值为5.916;同一规格不同盐度间O︰N值差异不...  相似文献   

17.
过氧化氢酶(CAT)和硒谷胱甘肽过氧化物酶(Se-GPx)在抗氧化防御体系中扮演重要的角色,在清除多余的活性氧自由基(ROS)防止机体氧化损伤方面具有重要的作用.本文利用RACE-PCR方法首次从脊尾白虾肝胰腺组织中克隆获得了CAT cDNA全长序列,该基因全长2649 bp,包括5′非编码区(UTR) 78 bp、开放阅读框(ORF)1554 bp和3′非编码区(UTR) 1017 bp,共编码517个氨基酸,预测分子量为58.46 kDa,理论等电点为6.64,利用PROSITE tools预测CAT基因的功能位点,发现该氨基酸序列包括酶活性中心序列“FDRERIPERVVHAKGAG”,亚铁血红素结合信号序列“RLFSYPDTH”和 3个催化位点残基His71,Asn144和Tyr354.同源性分析表明,脊尾白虾CAT氨基酸序列与其他物种的相似性为68%-92%.荧光定量PCR分析结果表明,CAT基因在肝胰腺、血细胞、心脏、肌肉、卵巢、鳃、胃和眼柄均有表达,其中在肝胰腺中表达量最高.低盐胁迫后脊尾白虾鳃和肝胰腺中CAT和GPx基因分别在48 h和6 h达到最大值,且与对照组差异显著(P< 0.05),表明CAT和GPx基因参与了低盐胁迫反应,并表现出一定时间效应关系.综合分析结果指出脊尾白虾CAT基因是CAT家族主要成员之一,可能在低盐环境胁迫过程中具有重要的作用.  相似文献   

18.
秦松  王希华  童顺  曾呈奎 《海洋与湖沼》1995,26(S1):126-129
于1993年1月 - 1993年4月,为了制备遗传性纯一的基因工程受体,用酶法从海带雌配子体单性生殖系分离雌性单倍体原生质体。从皱纹盘鲍消化腺及消化器官提取出鲍酶,在25°C下、1%(WV)的鲍酶与2%(WV)的纤维素酶(Onozuka R-10)合用能有效解离单细胞和原生质体。海带雌性原生质体经2d培养再生了细胞壁。  相似文献   

19.
于2006年2月、5月、8月和11月四个季度月采用呼吸瓶法现场研究了胶州湾菲律宾蛤仔的呼吸排泄作用。结果表明,胶州湾菲律宾蛤仔呼吸排泄速率具有明显的季节变化趋势,春、夏季高于秋、冬季。不同生物学规格的蛤仔呼吸排泄速率具有明显的差异,单位个体耗氧率、排氨率和排磷率均随着个体大小的增加而增加,而单位体重的耗氧率、排氨率和排...  相似文献   

20.
利用缢蛏(Sinonovacula constricta)重组铁蛋白富集Fe~(3+)和Mn~(2+)制备重组Fe-铁蛋白和Mn-铁蛋白,通过扫描电镜、X射线能量色散能谱仪(EDS)和MALDI TOF/TOF质谱系统测定蛋白的表面形貌变化、金属元素的能量变化和肽段分子量。利用综合物性测量系统(PPMS)测定蛋白纳米颗粒在室温300K,外加磁场3T下的磁学性质变化。结果显示,重组Fe-铁蛋白和Mn-铁蛋白与空白相比,表面形貌发生明显变化,Fe-铁蛋白仍为小球状,Mn-铁蛋白聚集体呈片层花球状,Cd-铁蛋白聚集体呈小圆球状,Mn-铁蛋白富集Cd~(2+)后呈片层花瓣散落状。Fe-铁蛋白和Mn-铁蛋白分别检测出相应金属元素且都有其特征能量态。两种重组蛋白的肽谱图与空白组相比,除铁蛋白保守肽段外还出现各自的特征肽段,推测与铁蛋白对Fe~(3+)和Mn~(2+)的富集功能密切相关。Fe-铁蛋白和Mn-铁蛋白纳米颗粒磁滞回线形状与铁蛋白空白组基本相同,呈顺磁性特征,磁性强度随Fe~(3+)和Mn~(2+)富集量的增加而增大。通过比较Fe-铁蛋白和Mn-铁蛋白与空白组在富集Hg~(2+)、As O43–和Cd~(2+)三种重金属离子方面能力的差异,发现Fe-铁蛋白对Hg~(2+)、AsO_4~(3–)和Cd~(2+)三种重金属的富集能力是空白组的2.4倍、1.7倍和3.7倍。Mn-铁蛋白对Hg~(2+)、AsO_4~(3–)和Cd~(2+)三种重金属离子在相同条件下的富集能力也有明显提高,分别为铁蛋白空白组的1.8倍、3.0倍和4.6倍。本研究结果为Fe-铁蛋白和Mn-铁蛋白在重金属污染治理方面的应用提供了数据参考。  相似文献   

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