红海榄根际土壤来源的青霉属真菌XGH2321及其抑菌活性

本研究根据形态学和ITS序列分析结果,将一株分离自海南东寨港红树林保护区红海榄根际土壤的菌株XGH2321鉴定为青霉属的一种真菌.通过对培养基中的碳源、氮源和盐度的优化,确定了适合该真菌菌株分泌抑菌活性物质的改良查氏培养基(4%玉米浆,0.3%NaNO3,0.05%KC1,0.1%K2HPO4,0.05%MgSO4,pH 7.4,9%盐度).将活化后的菌株XGH2321接种到该培养基中,按28℃、160r/min振荡培养7d,获得的发酵液水萃取物对金黄色葡萄球菌、藤黄八叠球菌和枯草芽孢杆菌的生长具有明显的抑制作用(MIC为400μg/mL),而发酵液的乙酸乙酯萃取物对上述微生物的生长也具有一定的抑制作用(MIC为800μg/mL);同时,上述2种萃取物对植物病原菌立枯丝核菌的生长也具有明显的抑制作用,MIC分别为200μg/mL和400μg/mL.     

木榄内生真菌菌株ZD6及其代谢产物的抑菌活性

根据形态学特征和ITS序列分析结果,将一株从木榄茎中分离到的内生真菌菌株ZD6鉴定为桔青霉Penicillium citrinum Thom.,该菌株在优化后的发酵培养基(1%麦芽糖,2%甘露醇,1%谷氨酸钠,0.5%蛋白胨,0.15%酵母膏,200g/L土豆汁,pH6.5)中28℃、160r/min振荡培养7d后的发酵液具有明显的抑菌活性。利用柱层析及重结晶等方法从该发酵液的乙酸乙酯和正丁醇提取物中分离得到4个化合物:大黄素、环(丙氨酸-甘氨酸)、赤藓醇和甘露醇,其中的大黄素和赤藓醇为首次在桔青霉中发现,它们对枯草芽孢杆菌Bacillus subtilis的生长具有明显的抑制作用,最低抑菌浓度(MIC)分别为25μg/mL和50μg/mL;大黄素对绿脓杆菌Pseudomonas aeruginosa的生长也具有一定的抑制作用,MIC为100μg/mL。     

微量液基稀释法测定中药活性成分的体外抗曲霉菌活性*

通过测定中药活性成分肉桂醛和柠檬醛对常见深部条件致病性真菌黄曲霉、烟曲霉的抗菌活性,为建立中药抗曲霉菌药敏试验标准提供参考依据。参照美国国家临床试验标准化委员会(National Committee for Clinical Laboratory Standards,NCCLS)提出的标准,用微量液基稀释法分别测定肉桂醛和柠檬醛对黄曲霉、烟曲霉的抗菌活性。肉桂醛对黄曲霉、烟曲霉最低抑菌浓度(Minimal inhibitory concentration,MIC)分别为：0.100μg/mL,0.050μg/mL,柠檬醛对黄曲霉、烟曲霉的MIC分别为：2．600μg/mL、0．650μg/mL。中药活性成分肉桂醛和柠檬醛具有高效抗曲霉作用。该研究可为制定中药抗曲霉作用评价标准提供参考依据。     

青蒿内生真菌Alternaria sp.(PQH12)提取物体外细胞毒活性及抗氧化活性的研究

本文分别采用环己烷、正丁醇、乙酸乙酯、丙酮、甲醇5种极性不同的溶剂分别提取青蒿内生真菌Alternaria sp.(PQH12)的菌丝体。采用MTT法分别测定不同极性溶剂提取物对3种肿瘤细胞株(HL-60、MCF-7和COLO205)的生长抑制率,采用DPPH自由基清除法和超氧阴离子O-·2清除法评价了青蒿内生真菌Alternaria sp.(PQH12)不同极性溶剂提取物的抗氧化活性。结果显示,正丁醇和乙酸乙酯提取物均表现出对肿瘤细胞生长的抑制作用;浓度为20μg/mL时,正丁醇提取物对HL-60细胞的生长抑制率为80.56%,乙酸乙酯提物对MCF-7和COLO205细胞的生长抑制率分别为54.47%和83.09%;在两种抗氧化活性评价体系中,甲醇提取物的抗氧化活性均明显优于其他溶剂的提取物,其IC50值分别为0.20 mg/mL(DPPH)和0.19 mg/mL(O-·2)。该结果表明,青蒿内生真菌Alternaria sp.(PQH12)具有较好的细胞毒和抗氧化潜力,具有一定的开发利用价值,可以作为进一步实验研究的对象。     

一株来自于伊犁贝母的内生尖孢镰孢菌Y1的抑菌活性初步研究

从新疆巩留县伊犁贝母的新鲜鳞茎中分离到一株具有分泌抑菌活性物质的内生尖孢镰孢菌Fusarium oxysporumY1,该菌在7种不同培养基上生长时显示出不同的菌落生长特征,而且只在沙氏培养基中生长时才具有分泌抑菌活性物质的能力。抑菌活性筛选结果表明:由该菌及其发酵液制备的发酵液浸膏、菌体裂解液浸膏以及经进一步纯化后获得的乙酸乙酯浸膏和正丁醇浸膏均具有明显的抑菌活性,其中以发酵液的乙酸乙酯浸膏和菌体裂解液的正丁醇浸膏活性最强,它们对金黄色葡萄球菌Staphylococcus aureus、表皮葡萄球菌Staphylococcus epidermidis、枯草芽孢杆菌Bacillus subtilis、藤黄八叠球菌Sarcina lutea和大肠杆菌Escherichia coli的最低抑菌浓度均小于25μg/mL。     

湘西虎杖抑菌活性成分提取研究

对湘西虎杖抑菌活性物提取条件进行研究,通过抑菌实验得到最佳提取条件为90%乙醇、60℃、3 h,该条件下的提取物对细菌生长的抑制活性最强,但对酵母菌无明显的效果,对金黄色葡萄球菌、大肠杆菌、痢疾杆菌的最小抑菌浓度（MIC）为0.25 g/mL～0.5 g/mL.提取物经85℃、100℃,15 min或pH 2～8处理后仍具有较强的抑菌活性.     

多孔板-MTT比色法评价植物和微生物代谢产物的抗真菌活性

多孔板-MTF比色法测定植物和微生物代谢产物对真菌抑制活性的步骤为:在多孔板的每孔中依次加入浓度为105孢子/mL的供试真菌孢子悬液90μL,不同浓度的药液10 μL.25℃暗培养48 h,然后每孔中加入8mg/mL的MTT溶液10μL,继续培养10 h后,离心去上清,加入DMS0 150 μL,振荡30 min,离心后上清液在510nm测定吸光值.采用上述条件测定了白屈菜红碱对稻瘟病菌和西瓜枯萎病菌的MIC值分别为80和1.5μg/mL,IC50值分别为21.99和0.78 μg/mL;Diepoxinζ对稻瘟病菌的MIC和IC50值分别为200和96.21 μg/mL.多孔板-MTT比色法为快速有效地筛选和评价植物和微生物抗真菌活性成分创造了条件.     

浙贝母内生真菌Nectria sp. JZ6分泌抑菌活性代谢产物的初步研究

Nectria sp.JZ6是从浙贝母新鲜鳞茎中首次分离到的一株内生真菌,其发酵液具有抑菌活性。为建立该真菌稳定分泌抑菌活性成分的发酵体系,本研究利用单因素实验和正交实验确定了发酵培养基的配方并初步优化了发酵条件。结果表明,将活化后的菌种接种于改良查氏培养基(8%葡萄糖,0.5%蛋白胨,0.05%KCl,0.1%K2HPO4,0.15%MgSO4,pH6.5)中,28℃、150r/min振荡培养6d后获得的发酵液抑菌活性比优化前提高了26%。该发酵液经100℃水浴30min不失活,在pH1-5时抑菌活性相对稳定,其后逐渐减弱,并在pH9.0及以上时丧失。Nectriasp.JZ6发酵液的乙酸乙酯浸膏抑菌活性最强,对金黄色葡萄球菌、藤黄八叠球菌、枯草芽孢杆菌、绿脓杆菌和大肠杆菌的最低抑菌浓度(MIC)分别为0.625、0.625、1.25、1.25和1.25mg/mL。     

川芎内生真菌Cladosporium sp.次级代谢产物及抗菌活性研究

为了增加川芎用途,本研究从川芎中分离鉴别出了一株内生真菌Cladosporium sp. IS384,并首次对其次级代谢产物进行研究。用硅胶柱层析、制备薄层层析和重结晶等方法,从该菌发酵液的乙酸乙酯萃取相中首次分离获得4个单体化合物,运用现代波谱技术以及文献数据对照,鉴定其结构分别为cladospolide B(1),iso-cla-dospolide B(2),麦角甾醇(3),水苏碱(4)。对提取部位及单体化合物进行抗菌实验,发现乙酸乙酯部位对粪链球菌ATCC 29212的MIC值达到3. 91μg/mL,化合物1对ATCC 29212的MIC为0. 31μg/mL,化合物2对ATCC12228的MIC值为0. 63μg/mL。本文通过对川芎内生真菌的研究发现了具有一定抗菌活性的物质,为后续川芎内生真菌研究提供很好的理论依据。     

迷迭香酸对几种植物病原真菌的抗菌活性*

研究了迷迭香酸对不同植物病原真菌菌丝生长和孢子萌发的抑制活性。试验结果表明,迷迭香酸对供试的8种植物病原真菌菌丝生长均有抑制作用,其中对番茄灰霉病菌、芒果灰斑病菌、柑桔青霉和梨黑斑病菌抑制作用较强,EC50分别为615．04μg/mL、698．23μg/mL、714．50μg/mL和809．10μg/mL;对杉木猝倒病菌和苹果树腐烂病菌抑制作用次之,EC50分别为1039．92μg／mL和1044．72μg/mL;对松枯梢病菌和种实霉烂病菌的抑制作用较弱,EC50分别为1256．90μg/mL和1270．87μg/mL。迷迭香酸对供试的6种植物病原真菌孢子萌发也有明显的抑制作用,EC50大致在400～700μg/mL范围,其中对梨黑斑病菌孢子萌发抑制作用最强,EC50为395．37μg/mL。     

Stable soil pH of thai mangroves in dry and rainy seasons and its relation to zonal distribution of mangroves

Soil characteristics of mangrove forests at Amphur Laemngob, Trat Province, east Thailand were investigated in both dry andrainy seasons. Sonneratia alba, Avicennia alba andRhizophora mucronata mainly grew as seaward mangroves.Rhizophora apiculata, Ceriops tagal, Excoecaria agallocha andBruguiera gymnorrhiza grew as meso mangroves in most cases.Lumnitzera racemosa was found as a landward mangrove. The salinity of the soil in which these 8 species grew was significantly higher in the dry season than in the rainy season. Soil pH did not change irrespective of the season. Each species seemed to grow at inherent soil pH. Bruguiera gymnorrhiza, Sonneratia alba andAvicennia marina were found both in Japan and Thailand. The soil pH of each mangrove species was similar in Japan and Thailand. The soil salinity of each mangrove species in Japan was similar to that in Thailand in the dry season. It was suggested that soil pH and salinity in the dry season are the important factors governing the zonal distribution of mangroves.     

Soil Characteristics and Vegetation Structure in a Heavily Deteriorated Mangrove Forest in the Caribbean Coast of Colombia

Mass mortality and biomass of mangrove trees are related to soil factors in the Caribbean Coast of Colombia. Soil properties measured were particle size distribution, bulk density, organic matter, total nitrogen, total phosphorus, pH, redox potential, salinity, and extractable nitrogen (NO₂, NO₃, NH₄) and phosphorus (PO₄). Sampling was performed at three locations along two 50 m transects at each site. Soil nutrient concentrations of either exchangeable nitrogen or phosphorus were comparable to other reported values. Redox potentials (E_h) were variable probably due to microtopographic conditions and to the amount of water and organic matter present in the soil. Hydrogen potentials (pH) are within the range reported as normal for mangrove soils. Locations with highest biomass had an average soil salinity of 35 with a range of 20 to 53. Sites with dead or dwarfed vegetation had an average soil salinity of 74 and minimum and maximum values observed were 52 and 100 respectively. Statistical analysis exhibited temporal and spatial differences in soil salinity and suggest that this parameter is most correlated to the development and distribution of mangrove vegetation in the area indicating that basal area and biomass volume are inversely correlated with soil salinity.     

Aerobic Degradation of 3-Methylindole by Pseudomonas aeruginosa Gs Isolated from Mangrove Sediment

3-Methylindole (3MI), an N-heterocyclic aromatic compound also called skatole, is associated with animal waste and industrial processing. A pure culture of bacterium capable of using 3MI as the sole source of carbon and energy was isolated from mangrove sediment using an enrichment technique and identified as Pseudomonas aeruginosa Gs based on 16S rDNA sequence. Microbial degradation of 3MI was studied in batch culture experiments for several factors, including initial substrate concentrations, pH, and salinity. The optimum pH and salinity was 7.0 and 5‰, respectively. Degradation of 3MI by P. aeruginosa Gs was quantified by reversed-phase high-performance liquid chromatography. Two metabolites of 3MI degradation were detected and proposed to be indoline-3-carboxylic acid and indoline-3-ol based on data obtained from HPLC/MS. Our results suggest that 3MI can be rapidly degraded by indigenous microorganisms found in mangrove sediment.     

Effects of Salinity Fluctuation on Photosynthetic Gas Exchange and Plant Growth of The Red Mangrove (Rhizophora mangle L.)

The red mangrove (Rhizophora mangle L.) in southern Floridaoccurs frequently in two distinct growth forms, tall and scrubplants, with the scrub form usually found in coastal inlandareas having a higher fluctuation of soil water salinity. Inthe present study, effects of constant and fluctuating salinitieson leaf gas exchange and plant growth of red mangrove seedlingswere investigated under greenhouse conditions. Both constantand fluctuating salinity treatments significantly affected leafgas exchange and plant growth of red mangrove seedlings. Seedlingssubjected to the fluctuating salinity with the mean of both100 and 250 mol m^–3 NaCl showed significantly lower photosynthesisand plant growth than those subjected to the corresponding constantsalinity with the same mean. The photosynthetic and growth ratesof the seedlings under these fluctuating treatments were aslow as, or even lower than those expected if they were growingunder the high constant salinity of their respective fluctuationtreatments. Seedlings subjected to the fluctuating salinitywith the mean of 500 mol m^–3 NaCl, however, demonstratedslightly higher CO₂ assimilation rate and stomatal conductance,but the same plant growth rates as those under the constant500 mol m^–3 NaCl treatment. These results suggest that,in general, fluctuating salinity has significant negative effectson photosynthesis and plant growth relative to constant salinitywith the same mean. If this finding can be applicable to fieldsituations, the low photosynthesis and plant growth observedpreviously in several scrub mangrove forests probably can beattributed in part to the salinity fluctuation of soil waterin these mangrove forests. Key words: Fluctuating salinity, photosynthesis, growth, growth forms, mangroves     

Soil salinity and pH in Japanese mangrove forests and growth of cultivated mangrove plants in different soil conditions

Soil conditions of mangrove forests in southern Japan were found to correlate largely with zonal distributions of the species.Kandelia candel grew in soils with low salinity and low pH,Avicennia marina, Rhizophora stylosa andSonneratia alba in soils with high salinity and high pH, andBruguiera gymnorrhiza in soil with a wide range of pH but limited range of salinity.Lumnitzera racemosa colonized soil with a wide range of pH and medium salinity. Seedlings ofKandelia candel, Bruguiera gymnorrhiza andRhizophora stylosa were planted in soils with differing salinity and pH. Optimum seedling growth ofKandelia, Bruguiera andRhizophora occurred when plants were cultivated in soils similar to those of their natural habitats, suggesting that growth of mangrove species and their zonal distributions were regulated by salinity and soil pH.     

Elution of dissolved Fe from mangrove soil by tannin solution

To reveal the role of tannins in mangroves, tannins in mangrove leaves and the Fe eluted from mangrove soil by adding tannin solutions of different salinity levels was investigated. Leaves of six mangrove and 16 non-mangrove species, and samples of a mangrove floor, Andosol and dark red soil were collected. Results were: (1) Increasing tannic acid concentration to ~50 mM, increased the Fe eluted from mangrove soil to ~20 μgg^?1. (2) When a 100 mM tannic acid solution was added, the Fe eluted from mangrove soil was 5.5 times higher than dark red soil. (3) Although elution of Fe from mangrove soil was higher than in Andosol one day after submersion in a 10 mM tannic acid solution, the difference was stable after 2 days. (4) The elution of Fe from all soils significantly decreased with increasing salinity of a 10 mM tannic acid solution. However, the amount from mangrove soil was 6.1 times higher than dark red soil even with 35 ‰ salinity. (5) The tannin content in the mangrove leaves was 99 ± 16 mgg^?1 and non-mangrove leaves was 76 ± 19 mgg^?1. (6) The Fe eluted from mangrove soil had a positive correlation with the tannin concentrations in the added leaf solution. Tannins in mangrove species promote the elution of Fe from mangrove floor soil even in saline water. Fe complexes were formed when mangrove soil was mixed with leaf tannins suggesting that Fe produced by tannins in mangrove leaves growing in land/sea interfaces likely plays a direct role in marine ecosystems.     

珠江河口湿地沉积物硝化作用强度及影响因素研究

采用现场调查和实验室模拟的方法,研究了珠江河口红树林、芦苇和光滩表层湿地沉积物氨氧化细菌(AOB)的分布、硝化强度及其主要影响因素。研究结果表明:三种不同生态环境中AOB数量差别较大。温度、pH、盐度和氨氮的浓度对沉积物硝化作用均有影响。其中盐度和pH对沉积物硝化强度影响最为显著,较高的盐度和较低的pH对硝化过程有明显的抑制作用。偏碱性环境下(pH=8.5)红树林、芦苇和光滩的硝化强度分别是偏酸性环境(pH=6.5)时2.5倍。最适宜的硝化温度在28℃左右,氨氮对氨氧化细菌的生长有促进作用,在含氨氮浓度较高的培养液中硝化强度较强。大型植物根系对硝化作用有明显的促进作用,对河口湿地氮循环具有重要的意义。     

The effect of atmospheric carbon dioxide concentrations on the performance of the mangrove Avicennia germinans over a range of salinities

By increasing water use efficiency and carbon assimilation, increasing atmospheric CO₂ concentrations could potentially improve plant productivity and growth at high salinities. To assess the effect of elevated CO₂ on the salinity response of a woody halophyte, we grew seedlings of the mangrove Avicennia germinans under a combination of five salinity treatments [from 5 to 65 parts per thousand (ppt)] and three CO₂ concentrations (280, 400 and 800 ppm). We measured survivorship, growth rate, photosynthetic gas exchange, root architecture and foliar nutrient and ion concentrations. The salinity optima for growth shifted higher with increasing concentrations of CO₂, from 0 ppt at 280 ppm to 35 ppt at 800 ppm. At optimal salinity conditions, carbon assimilation rates were significantly higher under elevated CO₂ concentrations. However, at salinities above the salinity optima, salinity had an expected negative effect on mangrove growth and carbon assimilation, which was not alleviated by elevated CO₂, despite a significant improvement in photosynthetic water use efficiency. This is likely due to non‐stomatal limitations to growth at high salinities, as indicated by our measurements of foliar ion concentrations that show a displacement of K⁺ by Na⁺ at elevated salinities that is not affected by CO₂. The observed shift in the optimal salinity for growth with increasing CO₂ concentrations changes the fundamental niche of this species and could have significant effects on future mangrove distribution patterns and interspecific interactions.     

三种红树植物对盐胁迫的生理适应

无瓣海桑（Sa）、海桑（Sc）、红海榄（Rs）都属于乔木红树植物。这3种红树植物对盐度的敏感程度存在着差异,因此对不同标准的盐度的适应性也大不相同。通过对这3种红树植物用不同的盐度的水3个月的处理,发现Sa和Sc叶片的净光合作用速率、气孔导度、蒸腾速率都随着盐度的增加而降低。Sa,Sc,Rs叶片中的可溶性总糖含量随着盐度的升高整体上有上升趋势。Sa和Sc茎、叶中丙二醛（MDA）含量在低盐度时（〈10）略有降低,随着盐度升高,MDA含量急速升高,而Rs茎、叶中MDA只是在盐度超过40时才会有明显增长,3种红树植物根部的MDA含量变化都不明显。Rs可以依靠超氧化物歧化酶（SOD）来消除活性氧自由基,而红树植物Sa和Sc的耐盐性稍差,SOD对膜的保护能力不强。根据实验结果,可以得出对3种红树植物对盐度的适应范围,这将为指导中国南海海岸线上的红树造林计划提供依据。     

产木聚糖酶的沿海红树林真菌筛选及其培养与酶活测定条件优化

从香港海岸红树林分离到的 77株真菌中有 34株可产生木聚糖酶 ,从中选出CY2 80 9(Staganosporasp .)、CY4 786和CY5 0 4 0等 3菌株与已知陆生产酶菌株HU5 0 4 8(Aspergillusawamori)进行产木聚糖酶的比较研究。根据培养液中菌丝生物量、木聚糖酶活力和木糖等价还原糖含量等指标的测定 ,菌株CY4 786在起始pH 7 8的木聚糖 酵母膏 海盐液体培养基中 2 5℃下震荡 (10 0r min)培养 7d产酶最佳 ;粗酶液在 5 0℃和pH 4 6的优化条件下进行测定 ,木聚糖酶活力达到 1 0 7× 10 4 U mL。结果表明 ,红树林真菌起着半纤维素降解者的作用 ,沿海红树林环境中存在着可资利用的木聚糖酶产生菌。作者讨论了利用发酵液中木糖等价还原糖含量的动态变化作为快速筛选产木聚糖酶菌株的指标的可能性