226 resultados para 188-1165C


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The glutathione peroxidases are essential enzymes of the cellular antioxidant defence system. In the present study, the full-length cDNA sequence encoding an extracellular glutathione peroxidase (designated CfGPx3) was isolated from Zhikong scallop Chlamys farreri. The complete cDNA was of 1194 bp, containing a 5' untranslated region (UTR) of 50 bp, a 3' UTR of 490 bp and an open reading frame (ORF) of 654 bp encoding a polypeptide of 217 amino acids. CfGPx3 possessed all the conserved features critical for the fundamental structure and function of glutathione peroxidase, such as the selenocysteine encoded by stop codon UGA, the GPx signature motif ((96)LGVPCNQFI(103)) and the active site motif ((WNFEKF184)-W-179). The high similarity of CfGPx3 with GPx from other organisms indicated that CfGPx3 should be a new member of the glutathione peroxidase family. By fluorescent quantitative real-time PCR, the CfGPx3 mRNA was universally detected in the tissues of haemocytes, gill, gonad, muscle and hepatopancreas with the highest expression in hepatopancreas. After scallops were challenged by Listonella anguillarum, the expression level of CfGPx3 transcript in haemocytes was significantly up-regulated (P<0.05) at 8 h post challenge. These results suggested that CfGPx3 was potentially involved in the immune response of scallops and perhaps contributed to the protective effects against oxidative stress. (C) 2010 Elsevier Inc. All rights reserved.

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Although the deep-sea sediments harbor diverse and novel bacteria with important ecological and environmental functions, a comprehensive view of their community characteristics is still lacking, considering the vast area and volume of the deep-sea sedimentary environments. Sediment bacteria vertical distribution and community structure were studied of the E272 site in the East Pacific Ocean with the molecular methods of 16S rRNA gene T-RFLP (terminal restriction fragment length polymorphism) and clone library analyses. Layered distribution of the bacterial assemblages was detected by both methods, indicating that the shallow sediments (40 cm in depth) harbored a diverse and distinct bacterial composition with fine-scale spatial heterogeneity. Substantial bacterial diversity was detected and nine major bacterial lineages were obtained, including Acidobacteria, Actinobacteria, Bacteroidetes, Chloroflexi, Nitrospirae, Planctomycetes, Proteobacteria, and the candidate divisions OP8 and TM6. Three subdivisions of the Proteobacteria presented in our libraries, including the alpha-, gamma- and delta-Proteobacteria. Most of our sequences have low similarity with known bacterial 16S rRNA genes, indicating that these sequences may represent as-yet-uncultivated novel bacteria. Most of our sequences were related to the GenBank nearest neighboring sequences retrieved from marine sediments, especially from deep-sea methane seep, gas hydrate or mud volcano environments. Several sequences were related to the sequences recovered from the deep-sea hydrothermal vent or basalt glasses-bearing sediments, indicating that our deep-sea sampling site might be influenced to certain degree by the nearby hydrothermal field of the East Pacific Rise at 13A degrees N.

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Up to now, accurate determination of the growth age and hiatuses of the Co-rich crust is still a difficult work, which constrains the researches on the genesis, growth process, controlling factors, regional tectonics, paleo-oceanographic background, etc. of the Co-rich crust. This paper describes our work in determining the initial growth age of the Co-rich crust to be of the late Cretaceous Campanian Stage (about 75-80 Ma), by selecting the Co-rich crust with clear multi-layer structures in a central Pacific seamount for layer-by-layer sample analysis and using a number of chronological methods, such as Co flux dating, dating by correlation with Os-187/Os-188 evolution curves of seawater, and stratigraphic division by calcareous nannofossils. We have also discovered growth hiatuses with different time intervals in the early Paleocene, middle Eocene, late Eocene and early-middle Miocene, respectively. These results have provided an important age background for further researches on the Co-rich crust growth process and the paleo-oceanographic environment evolution thereby revealed in the said region.

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趋磁细菌(Magnetotactic bacteria)的研究是国际微生物学研究热点之一。趋磁细菌体内含有纳米单磁畴的氧化铁/硫化铁(Fe3O4或Fe3S4)晶体,称为磁小体。由于趋磁细菌营养条件要求苛刻,在环境中需要微好氧条件,且营养类型属于化能自养,使得培养趋磁细菌时常遇到问题。 本研究首先通过正交试验优化趋磁细菌AMB-1菌株培养条件,在培养条件铁源为奎尼酸铁0.02 mmol/L,装瓶量75% ,pH值6.7,温度25 ℃时,AMB-1 OD600达到0.440(1.166×109 cells/ml)。同时运用磁收集传代法,使带有磁小体的AMB-1细胞比例占95%以上(Cmag值稳定在1.9-2.0)。 在AMB-1具有较好的生物量,同时又具有较好的含磁小体细胞比例后,研究磁小体的变化过程。通过透射电镜观察磁小体变化过程,发现培养24 h细菌体内已有较小晶体形成(平均27 nm,n=188)且沿长轴分布;48 h晶体长大(平均43 nm,n=203)且形成分段链沿长轴排列;72 h晶体进一步成熟(平均50 nm,n=191)仍以分段链沿长轴排列;随后细菌逐渐衰亡磁小体变小,168 h可见部分自溶细菌中仍有磁小体链(平均37 nm,n=186);192 h细菌自溶磁小体链(平均33 nm,n=184)分散到环境中。 通过透射电镜在细胞水平上研究趋磁细菌细胞分裂时发现,磁小体在细菌分裂时采用两种分离方式:一种为磁小体分配到两个子细胞;另一种为磁小体只分配到一个子细胞。无磁小体的子细胞,在随后的生长过程又分为两种情况:一种为细胞逐渐产生磁小体,另一种为不再产生磁小体。这种现象的发现,解释了随着传代次数的增多,细菌磁性有所下降的原因(Cmag值降低)。 在对趋磁细菌磁小体合成机制的研究中,常使用基因敲除的办法获得缺陷型,并与野生型对比进行研究。但是,利用基因敲除获得缺陷型不仅操作繁琐并且所得缺陷型不稳定。本研究利用特殊的磁富集传代法,先将带有磁小体的菌体收集并连续传代,筛选获得了高磁菌株;利用这种方法,收集不含磁小体的菌体并连续传代,筛选获得了无磁菌株。 趋磁细菌磁小体在医疗、环保等领域具有广阔应用价值,但是目前由于趋磁细菌难以大规模培养,并且磁小体纯化存在成本高等原因,将磁小体真正实际应用尚有一段距离。通过研究磁小体在趋磁细菌中的变化过程发现,AMB-1菌株在培养192 h后自溶,并且磁小体随着细胞的破碎释放到环境中去。

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从山东黄岛海水养殖场分离到一株弧菌V134,从中克隆得到琼胶酶基因agaV,并将其在大肠杆菌中表达,纯化得到重组的琼胶酶AgaV。酶活分析发现该琼胶酶的最适温度在40℃左右,对pH比较敏感,pH 7.0时具有最高的琼胶裂解活性。对AgaV进行了两种应用性探索:(1)利用AgaV从琼脂糖凝胶中回收DNA,回收效率可达90%以上;(2)利用agaV作为报告基因构建了捕获分泌序列的载体pBU,并用其从革兰氏阳性细菌(G+菌)和革兰氏阴性细菌(G-菌)中筛选出了一系列分泌蛋白。将利用pBU从一株哈维氏弧菌T4中筛选出的6个分泌蛋白分别进行基因克隆、蛋白表达纯化和牙鲆免疫实验,发现其中一个蛋白,命名为DegQVh,具有免疫保护效应,其免疫保护率(RPS)可达64%。为了提高DegQVh的免疫保护效应,将AgaV的分泌结构域与DegQVh融合,构成融合抗原AgaV-DegQVh。利用大肠杆菌作为载体菌构建了AgaV-DegQVh融合抗原递呈系统,用其作为疫苗进行免疫,发现其RPS可达到95%。酶活分析表明DegQVh在50℃、pH 8.0时具有最高的活性。突变分析表明83位的组氨酸、113位的天冬氨酸和188位的丝氨酸以及两个PDZ结构域是DegQVh活性所必需的。表达分析发现degQVh表达受温度和细胞浓度调控,并且其上游有一个受E调控的启动子。进一步的分析发现DegQVh能够与大肠杆菌的DegP功能互补。

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过量氮和磷引起的海水富营养化引起了一系列的生态环境效应,探讨二者的耦合关系对阐明全球海洋生态环境变化机制意义重大。本论文首次探讨了过量氮和磷引起的富营养化对海水无机碳体系的影响,通过室内系统模拟实验,构筑了过量氮和磷与无机碳体系变化的定量耦合关系,在此基础上获得了中国近海典型海域特别是南黄海近十年氮和磷演变引起的无机碳源汇格局变化,获得了以下一系列新的结果和认识: 1、不同营养盐(NO3-、NH4+、NO2-和PO43-)对水体溶解无机碳体系均有重要影响,且影响程度有较大的差异。 在模拟实验中,当NO3-<188 μmol/L,NH4+<126 μmol/L,NO2-<39.5 μmol/L时均可明显提高水体的固碳能力,相应水体中的DIC、HCO3-和 pCO2均较空白对照组时有一定程度的下降。当N浓度达到一定值后,其固碳能力减弱,DIC、HCO3-和pCO2出现相反变化。当NH4+>126 μmol/L和NO2->39.5 μmol/L时pCO2增加,向大气释放CO2。在PO43-添加组中,当浓度小于19.5μmol/L时DIC、pCO2较空白对照组下降,水体表现为大气CO2的汇,其固碳能力高于N组。在孔石莼添加组中,各营养盐组水体的固碳能力均高于空白对照组,相应水体中DIC、HCO3-和pCO2的下降明显。其中,同一营养盐浓度水平下,NH4+组中各无机碳组分的变化幅度明显高于NO3-和NO2-组,PO43-组的无机碳体系变化幅度低于N组。其中,当NO3-、NH4+和NO2-浓度分别在71 μmol/L,49.7 μmol/L和11.7 μmol/L时,pCO2下降明显,水体表现为大气CO2的强汇。对无机碳各参数的变化量与营养盐进行多元回归分析,得到以下统计关系: ⊿DIC=-0.937(⊿PO43-)-0.34(⊿NO3-)-0.46(⊿NH4+)+0.11(⊿NO2-)(R2=0.69, n=30,Sig.<0.05) ⊿HCO3-=-1.357(⊿PO43-)-0.35(⊿NO3-)-0.57(⊿NH4+)-0.013(⊿NO2-)(R2=0.76, n=32, Sig.<0.05) ⊿CO32-=0.344(⊿PO43-)+0.16(⊿NO3-)+0.18(⊿NH4+)+0.076(⊿NO2-)(R2=0.69, n=32, Sig.<0.05) ⊿pCO2=-1.321(⊿PO43-)-0.12(⊿NO3-)-0.31(⊿NH4+)-0.032(⊿NO2-)(R2=0.84, n=35, Sig.<0.01) 2.氮、磷对海水无机碳体系的源汇格局影响主要受控于水体浮游藻类对营养盐的利用。 模拟实验各种情况下Chl-a的浓度均有不同程度的变化。在N组中,当NH4+<126 μmol/L,随着NH4+浓度的增加,Chl-a浓度增加,而高浓度(NH4+>126 μmol/L)的实验组中,Chl-a则随浓度的增加而下降。当NO2-和PO43-的浓度分别大于39.5 μmol/L和19.5 μmol/L时,水体中Chl-a随营养盐浓度的增加而出现下降趋势,不同的是NO3-添加组在实验浓度范围内藻类均出现了不同程度的增长,这可能与浮游藻类对过量营养盐的耐受性不同有关。在低浓度时,PO43-组pCO2的下降最为明显,这与 PO43-组Chl-a的含量较高相一致,其次为NH4+,NO3-高于NO2-,可见在实验水体中PO43-对于藻类新陈代谢作用影响明显,从而作用到无机碳上的影响也就最为明显。对各实验水体⊿Chl-a与无机碳各组分作相关性分析发现,⊿Chl-a与⊿pCO2的相关性最为明显(R2=0.75,p<0.0001),这可能是因为浮游藻类光合作用和分解作用消耗或生成的首先是海水中的游离CO2(即pCO2)。在孔石莼组中,由于大藻对过量营养盐的耐受性较强,因此各组中藻类干重均有一定程度的增长。藻类干重的增加以NH4+最为明显,其次为NO3-和NO2-,这可能与其对N盐的优先吸收顺序NH4+>NO3->NO2-有关。加PO43-组中藻类的干重增加量低于NH4+和NO3-组,这与大藻的生理特性有关。对藻类干重增加量⊿m与⊿DIC作相关性发现,二者相关性明显(R2=0.64,p<0.01),这是由于孔石莼光合作用同时吸收HCO3-和CO2为碳源有关。 3、近年来,中国近海由于海水过量氮磷引起的富营养化,可使大部分海域海水无机碳汇强度增加 应用室内模拟获得的无机碳与氮、磷之间的定量耦合关系,估算了过量氮、磷引起的富营养化条件下,中国近海水体无机碳源汇的变化情况。结果表明,近十年来,由于氮、磷浓度的增加而导致渤海、南黄海、北黄海和东海每年从大气中多吸收0.197×106t C、0.302×106t C、0.039×106t C和2.233×106t C,东中国近海营养要素的含量的变化及组成比例的时间性和地区性差异是造成各海区碳通量差异的重要因素;对2006年南黄海水文、化学、生物与无机碳体系之间的相互关系进行综合分析,可知水体无机碳体系与营养盐水平密切相关。

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采用乙酸地衣红染色技术(Acetic orcein staining technique)较系统地研究了长牡蛎 Crassostrea gigas (Thunberg)三倍体产生的卵子在受精且第一极体释放受抑制后的减数分裂过及染色体的分离行为以阐明可存活四倍体的产生体的机制。用浓度为0.5 mg/L的细胞松驰素B (CB)处理受卵以抑制其第一极体的释放。在观察到个别受精卵出现第一极体时开始CB处理,持续至对照组中50%的受精卵出现的第一极体。对处理组和对照组的受精卵从受精后隔5分钟取样一次,用卡诺氏液(Carnoy's fixative, 冰醋酸和甲醇按1:3的体积比充分混合)固家样品。采用0.5%的乙酸-地衣红染料进行受精卵的染色,而后压片观察受精卵染色体行为。长牡蛎三倍体产生的卵子,中期I同源染色体构型呈现单价体(Univalents),二价体(Bivalents)、三价体(Trivalents)以及大于三价体的多价体(Multivalents)混合出更的特征。在第一极体释放受抑制的受精卵的第二次减数分裂过程中,可确认四种染色体分离类型:三极分离(Tripolar segregation) (54.5%)、联合二极分离(United bipolar segregation) (12%)、独立二级分离(Incomplete united bipolar segregation)(4%)。其余卵子的染色体分离行为(23%)不规律,呈现不同程度的紊乱,但总体看来介于上述四种分离类型之间。此外,某些特定的独立二级也可能是四位体形成的最主要的细胞遗传学体制。此外,某些特定的独立二极分离也可能产生四倍体。轻细胞体驰素B 处理的受精卵的减数分裂过程具有显著的不同步性,表现在三个方面:第一,在两个重复组之间,即两个雌体之间,存在第二次减数分裂的时间进程的不同步性;第二,同一个雌体产生的卵子之间的发育速度不同步性,表现为不同的卵子进入第一次减数分裂的时间不同;第三,同一卵子内的染色体之间,其行为有时存在的不同步性。另外,探讨了中心类在支配第二减数分裂时各种染色体分离行为的可能机制。以长牡蛎二倍体与近江牡蛎二倍体的染交作为对照,探讨了能长牡蛎四倍体与近江牡蛎二倍体杂并诱导异源三倍体的可行性。长牡蛎Crassostrea gigas (Thunberg)四倍体和二倍体与近江牡蛎Crassostrea rivularis (Gould)二倍体的杂交以及相应的对照组共进行了三批重复实验,杂交实验采用高密度的精子。研究结果表明,自交组平均受精率依次为94%(GG)、77% (RR),88% (G/GG)和85% (GG/G)。双方差分析(ANOVA)表明,各自交组之间受精率没有显著差异(F=3.118, P=0.132)。在杂交组,直接授精后180分钟,尚未观察到受精迹象,因而无法估计受精率。授精后48小时的孵化率各组之间差异很大,并经双方差分析(ANOVA)表明存在显著性差异,(F=3.188, P=0.018)。其中GGR和RGG组的孵化率相近似,产生的幼虫数量明显少于对照组。在四种类型的杂交实验中,二倍体C. gigas (雌体) * 二倍体 C. rivularis (雌体)(GR)早最成功的。虽基GR组幼虫的生长率低于对照组,但其存活率接近于对照组。长牡蛎四倍体与近江牡蛎二倍体杂交组(GR),在授精后两天的孵化率较低,但幼虫的生长状况与对照组接近。另外两个杂交组,即近江牡蛎二倍体与长牡蛎四倍体(RGG),二倍体近江牡蛎江与二倍体长牡蛎(RG),授精后两天的孵化率很低,幼虫生长得缓慢。三个重复组的GR杂交组和一个重复组的GGR杂交组获得稚贝。聚合酶链式反应/限制性酶切片段长度的多态性(PCR/PFLP)检支分析结果证实这些稚贝均是杂交种;流式细胞术分析结果证明GGR获得的稚贝是三倍体,从而证明获得了长牡蛎与近江牡蛎的异源三倍体。有迹象表明三倍体与二倍体杂交种之间(GGR对GR)存在生长上的差异。首先,GGR的眼点幼虫大约比GR组早出现5-7天即仅次于对照组GG,G/GG,和GG/G;第二,尽管仅获得少量GGR幼贝,这些幼贝在授精后90天的大小显著大于GR组的个体。在RGG和RG组中,幼虫没能存活到眼点幼点阶段。细胞学检查结果表明,杂交组的绝大多数卵子发育停滞在第一次减数分裂中期(Metaphase I),这一过程至少持续到授精后180分钟。仅有2%的GGR 组的卵子在授精后180分钟进入第一次减数分裂后期)(Anaphase I). 而在此时期,GR,RGG和RG组的卵子中,仍只观察到10第二价体(Bivalents).

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High molecular weight dissolved organic matter (HMW-DOM, > 1000 Da) represents a major fraction (> 30%) of dissolved organic carbon (DOC) in the ocean and thus plays an important role in the global biogeochemical cycling of carbon and many other elements. Its organic sources and formation mechanisms, however, are still not well understood especially in estuarine and coastal regions where multiple natural and anthropogenic sources contribute to total HMW-DOM. In this paper we report our measurements of natural radiocarbon (C-14) abundances and stable carbon isotope (C-13) compositions of the major biochemical compound classes: amino acids, carbohydrates and lipids separated from eight HMW-DOM samples collected from five US estuaries as part of our on-going study of sources, distribution and transport of chromophoric dissolved organic matter (CDOM) in estuarine and coastal waters. Distinct differences in both C-14 and C-13 values were found among the bulk HMW-DOM samples as well as the individual compound classes. Radiocarbon ages of the major compound classes varied by as much as 27,000 years in a single sample. The calculated average radiocarbon ages of the compound fractions of HMW-DOM indicate that the total lipid fraction is very "old", while the acid-insoluble fraction is slightly younger. Total amino acid and carbohydrate fractions, however, have relatively modern apparent C-14 ages. The significant variability in C-14 ages among the compound classes indicates not only multiple organic carbon sources but also different formation and turnover pathways controlling the cycling of different biochemical components of HMW-DOM in estuarine and coastal waters. (c) 2006 Elsevier Ltd. All rights reserved.

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High-speed counter-current chromatography (HSCCC) technique in semi-preparative scale has been successfully applied to the separation of bioactive flavonoid compounds, liquiritigenin and isoliquiritigenin in one step from the crude extract of Glycyrrhiza uralensis Risch. The HSCCC was performed using a two-phase solvent system composed of n-hexane-ethyl acetate-methanol-acetonitrile-water (2:2:1:0.6:2, v/v). Yields of liquiritigenin (98.9% purity) and isoliquiritigenin (98.3% purity) obtained were 0.52% and 0.32%. Chemical structures of the purified liquiritigenin and isoliquiritigenin were identified by electrospray ionization-MS (ESI-MS) and NMR analysis. (c) 2005 Published by Elsevier B.V.

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柴达木盆地是青海省主要的商品粮基地,由于长期以来粗放耕作,造成春小麦田以野燕麦、藜等杂草为主的农田杂草大面积危害,严重影响春小麦产量和品质。通过化学除草剂单剂和复配剂筛选,结合化学除草剂施用方法的改进,配合农艺措施,达到综合控制该地区麦田杂草的目的。

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针对我国目前草原生态建设中存在的一些理论和实践问题,如草原退化的面积和程度,造成退化原因的作用和权重等不确定性问题;载畜量及其估测的理论基础问题;“围封转移”和“春季休牧”措施的理论基础以及草原生态系统多功能等问题.认为这些问题将严重困扰着我国草业能否健康、和谐的可持续发展,因此在研究我国草业发展战略时,必须面对并迫切需要解决.

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生物多样性是测度生态系统内物种组成、结构多样性和复杂化程度的客观指标,是生态系统内生物群落对生物和非生物环境综合作用的外在反映.生物多样性研究已经成为当今植物生态学研究的热点之一[2,3,6,7,1],对于具体的植物群落来说,大的气候条件相对一致,群落生境的差异可能是形成物种多样性的主要原因,而土壤因子可能是一个重要的环境因子,因此,研究物种多样性与土壤环境的关系有重要的意义,国内外都对此做了大量的研究[3,5,10,17].利用数学方法研究鸟岛地区物种多样性与土壤环境的关系,一方面可以进一步了解这一片新生土地的特点,增加高海拔地区的资料,另一方面可以为保护鸟岛这一青海湖物种多样性关键地区提供理论支持.

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按照青藏公路建设和整修的不同阶段,利用样方调查植被空间分布变化,得出高寒草原植被的自然演替进程遵循以下规律。在工程结束2年,8年,26年后,群落植被覆盖度和生物多样性指标分别达到原生植被的2%~4%,6%一23%;32%~54%,46%~50%:95%以上和100%左右.青藏公路沿线高寒草原植被的人为破坏影响是明显的,植被的自然恢复需要20年左右的时间.工程建设破坏面积大于1500m~2,植被难以恢复,土壤沙化和水土流失,影响周边地区生态环境质量.因此,在青藏铁路工程建设中首先应当减少对地形地貌的破坏,其次应当重视对地表土壤的保护并辅助人工植被恢复措施,促进植被的自然恢复.

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通过野外控制实验,研究了高寒矮篙草草甸不同植物类群、群落对模拟增减降水条件下的响应。结果表明,不同植物类群(禾草类、莎草类)增加降水20%,地上生物量分别比对照提高103.63、77.12g/m^2。在植物生长期(6月),增加降水20%及40%,植物群落物种多样性指数(H)分别比对照提高0.188和0.735;而均匀度指数(J)在增加降水40%时,提高了0.086。生长期(7月)增加降水20%,物种多样性指数(H)和均匀度指数(J)分别提高0.409和0.07。当降水增加20%时,植物群落中禾草类的重要值较对照提高了0.92。

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通过野外控制实验,研究了高寒矮嵩草草甸群落植物多样性、初级生产力对模拟降雨条件的响应.结果表明:(1)在植物生长期(6月),增加降雨20%、增加降雨40%,植物群落物种多样性指数(H)和均匀度指数(J)分别比对照提高了0.188和0.011、0.735和0.076,生长期(7月)增加降雨20%物种H和J提高了0.409和0.07;(2)禾草类:增加降雨20%处理的地上生物量与对照相比没有明显的显著性差异(P>0.05),增加降雨40%处理的地上生物量与对照相比差异显著(P<0.05),说明过多增加降雨会抑制禾草的生长发育.杂类草:减少降雨50%处理的地上生物量与对照相比差异显著(P<0.05),其地上生物量对减少降雨的反映比较敏感.莎草类:其地上生物量对增加和减少降雨都没有显著变化;(3)0~10 cm和0~30 cm土层地下生物量均在增加降雨20%时最高,地下生物量的总量也在增加降雨20%时最高;(4)矮嵩草草甸地下生物量与地上生物量、总生物量的比值接近于生长季末时最大,且在模拟增加降雨20%的水平时,7、8、9月份地下和地上生物量较其它处理组高.