Fucose-Rich Sulfated Polysaccharides from Two Vietnamese Sea Cucumbers Bohadschia argus and Holothuria (Theelothuria) spinifera: Structures and Anticoagulant Activity

Fucosylated chondroitin sulfates (FCSs) FCS-BA and FCS-HS, as well as fucan sulfates (FSs) FS-BA-AT and FS-HS-AT were isolated from the sea cucumbers Bohadschia argus and Holothuria (Theelothuria) spinifera, respectively. Purification of the polysaccharides was carried out by anion-exchange chromatography on DEAE-Sephacel column. Structural characterization of polysaccharides was performed in terms of monosaccharide and sulfate content, as well as using a series of non-destructive NMR spectroscopic methods. Both FCSs were shown to contain a chondroitin core [→3)-β-d-GalNAc-(1→4)-β-d-GlcA-(1→]_n bearing sulfated fucosyl branches at O-3 of every GlcA residue in the chain. These fucosyl residues were different in pattern of sulfation: FCS-BA contained Fuc2S4S, Fuc3S4S and Fuc4S at a ratio of 1:8:2, while FCS-HS contained these residues at a ratio of 2:2:1. Polysaccharides differed also in content of GalNAc4S6S and GalNAc4S units, the ratios being 14:1 for FCS-BA and 4:1 for FCS-HS. Both FCSs demonstrated significant anticoagulant activity in clotting time assay and potentiated inhibition of thrombin, but not of factor Xa. FS-BA-AT was shown to be a regular linear polymer of 4-linked α-L-fucopyranose 3-sulfate, the structure being confirmed by NMR spectra of desulfated polysaccharide. In spite of considerable sulfate content, FS-BA-AT was practically devoid of anticoagulant activity. FS-HS-AT cannot be purified completely from contamination of some FCS. Its structure was tentatively represented as a mixture of chains identical with FS-BA-AT and other chains built up of randomly sulfated alternating 4- and 3-linked α-L-fucopyranose residues.     

毛竹入侵对落叶阔叶林土壤理化性质和胞外酶活性的影响

【目的】研究毛竹（Phyllostachys edulis）入侵亚热带北缘落叶阔叶林对土壤理化性质和土壤胞外酶活性的影响,为探究毛竹入侵条件下森林生态系统养分循环过程与固碳潜力及森林的毛竹入侵治理提供科学依据。【方法】在毛竹入侵样带上,选择落叶阔叶林（麻栎林Quercus acutissima）、竹阔混交林（混交林）和毛竹林3种林分为研究对象,采集其林下地表0~10 cm土壤,进行土壤理化性质和土壤酶活性的测定,包括土壤pH、含水率和有机碳、全氮、NH⁺₄-N、NO^-₃-N、全磷、有效磷含量,以及β-葡糖苷酶（BG）、β-N-乙酰氨基葡萄糖苷酶（NAG）、亮氨酸氨基肽酶（LAP）、酸性磷酸酶（AcP）、酚氧化物酶（POX）、过氧化物酶（PER）活性和酶计量(V_C/N、V_N/P、V_C/P),比较不同林分的土壤指标。【结果】①随着毛竹入侵,毛竹林土壤pH值和含水率显著增大,且与混交林、麻栎林差异显著（P<0.05）;其土壤有机碳、NO^-₃-N和全氮含量显著下降（P<0.05）,NH⁺₄-N、有效磷和全磷含量与混交林、麻栎林差异不显著。②土壤碳、氮和磷循环相关的水解酶（BG、NAG+LAP、AcP）活性均随毛竹入侵呈下降趋势,氧化酶（POX、PER）活性变化趋势与之相反;酶计量分析表明,毛竹林土壤的V_N/P显著低于混交林和麻栎林（P＜0.05）,而V_C/N、V_C/P在不同林分间差异不显著。③酶活性的矢量分析表明,毛竹林土壤微生物的磷限制程度高于混交林和麻栎林。④土壤水解酶活性与土壤有机碳、NO^-₃-N、全氮、有效磷含量或pH有显著相关关系,氧化酶活性与土壤性质相关性总体不显著。【结论】毛竹林取代落叶阔叶林（麻栎林）后,土壤有机碳和养分含量及相关水解酶活性降低,不利于原有落叶阔叶林土壤碳库与养分库的保存。     

丛枝菌根真菌对元宝枫生长及其根系形态的影响

【目的】阐明丛枝菌根（arbuscular mycorrhizal,AM）真菌对元宝枫生长状况及根系形态特性的影响,探讨元宝枫根系形态与其生长发育的关系。【方法】在元宝枫幼苗生长基质中接种摩西斗管囊霉（Funneliformis mosseae）和根内根孢囊霉（Rhizophagus intraradices）2种AM真菌,以不接种AM真菌为对照,通过盆栽试验,培养90 d后,测定元宝枫的生长状况、根系活力及根系形态指标,利用单因素方差分析和多重比较方法,分析接菌与不接菌处理之间的差异,采用envfit ()函数、冗余度分析和变差分解分析元宝枫生长状况与其他指标间的关系。【结果】①接种2种AM真菌后,元宝枫株高、地径、茎干质量和根系活力较对照显著提高,根冠比下降。根内根孢囊霉对元宝枫的促生作用比摩西斗管囊霉显著。②与对照相比,接种2种AM真菌后,元宝枫根表面积和根体积显著增大,粗根根长占总根长的比例增加,细根根长占总根长的比例显著降低。根内根孢囊霉对元宝枫根系形态的影响比摩西斗管囊霉显著。③元宝枫生长状况与其根系活力和根系形态密切相关。【结论】接种AM真菌有助于增强元宝枫的根系活力,改善其根系形态,促使其汲取更多养分,进而提高其生物量。     

6个鲜食葡萄品种不同部位生物活性物质含量及抗氧化活性比较

【目的】 对不同色系鲜食葡萄品种各器官生物活性物质含量和各器官提取物总活性进行比较研究,为鲜食葡萄育种和生物活性物质利用提供有效依据。【方法】 以三种色系鲜食葡萄(各含一个有籽和无籽鲜食葡萄品种)为材料,分别测定果梗、果皮、果肉、葡萄籽中多酚、黄酮、原花青素含量,用自由基清除法评价各器官提取物的总活性,并使用相关系数法分析3种生物活性物质含量和总活性的相关性。【结果】 结果表明,不同色系葡萄品种中,黄绿色系与红色系果梗多酚含量高于紫黑色系品种,无籽品种含量高于有籽品种;黄酮含量以红色系品种含量最高,且无籽高于有籽品种。果皮中多酚、黄酮及原花青素含量与果皮颜色深浅有关,颜色越深含量越高;原花青素含量在黄绿色系与红色系葡萄品种中无籽显著高于有籽葡萄。果肉多酚含量紫黑色系有籽葡萄显著低于其它品种;原花青素含量紫黑色系显著高于红色系品种,黄绿色系及红色系品种有籽显著高于无籽品种。葡萄籽中多酚及原花青素含量变化具有随着果皮颜色加深而显著增加的变化趋势。生物活性物质抗氧化活性在果梗中黄绿色系与红色系品种显著高于紫黑色系品种,无籽品种高于有籽品种;在果皮中紫黑色系品种显著高于黄绿色系与红色系品种;在果肉中黄绿色系有籽品种显著高于无籽品种;多酚、黄酮含量与提取物总的抗氧化活性呈极显著正相关关系(P <0.01)。综上可知,鲜食葡萄果梗中含有丰富的多酚和黄酮物质,且红色系无籽品种显著最高,可将葡萄采果后果梗作为多酚和黄酮物质提取的原材料;鲜食葡萄多酚、黄酮、原花青素等生物活性物质随着果皮颜色加深而加深,可选育果皮颜色深且品质好的品种做鲜食葡萄;葡萄籽中含有丰富的多酚、黄酮、原花青素,可作为生物活性物质提取的原材料;鲜食葡萄提取物总抗氧化活性主要由多酚、黄酮决定。     

Analysis of the gacS-gacA regulatory genes of spontaneous mutants of Pseudomonas fluorescens biocontrol strain PfG32R

Pseudomonas fluorescens strain PfG32R, a potential biocontrol agent against soilborne pathogens, frequently loses its antifungal activity and ability to produce enzymes. To characterize genetically the instability of these bacterial functions, we analyzed gacS and gacA genes of PfG32R and three spontaneous mutants of PfG32R (NR1, NR9, and ASW6), which had lost their ability to produce proteases and antifungal activities. The gacS and gacA sequences of PfG32R had 77%–89% and 78%–87% homology, respectively, with several known gacS and gacA homologues. All three spontaneous mutants were subjected to complementation analysis. Introduction of clones containing an intact gacS of PfG32R and another P. fluorescens strain Pf-5 as well as strain tea632 of P. syringae pv. theae complemented all three mutants restoring protease and antifungal activities, indicating a mutation in gacS. In sequencing analysis, the mutants had a deletion or change in amino acids in the conserved sensor kinase domains of GacS. The three mutants maintained both their antibacterial activity against Ralstonia solanacearum and Clavibacter michiganensis ssp. michiganensis and siderophore production, indicating that they are not controlled by the GacS/GacA system. The sequences reported in this article have been deposited in the DDBJ database under accession numbers AB219364 and AB219365.     

Synthesis and insecticidal activities of pyridine ring derivatives of podophyllotoxin

In an attempt to find the biorational pesticides, we synthesized 12 pyridinyl derivatives of podophyllotoxin (PPT) and 4′-demthylepipodophylltoxin (4′-DMEP) in this study. Their structures and the α/β substitution at C-4 were confirmed by ¹H NMR, IR, MS spectral analyses and elemental analysis. The insecticidal activities were tested against fifth-instar larvae of Pieris rapae and the third-instar larvae of Cullex pipiens pallens at concentrations of 250 and 10 μg ml⁻¹. Four derivatives of PPT, 4.1, 4.2, 4.3 and 4.5, showed higher insecticidal activities against P. rapae than PPT, while three derivatives of PPT, 4.4, 4.5 and 4.6, displayed higher mosquito larvicidal activity than PPT, with LC₅₀ values of 1.66, 3.96 and 1.54 mg l⁻¹, respectively. Interestingly, we also found that the pyridine ring derivatives of PPT showed delayed insecticidal activity, which is different from traditional neurotic insecticides. The results suggest that 4′-OCH₃ in the PPT derivatives is essential to keep the insecticidal activity and the insecticidal activities of pyridine ring derivatives of PPT are higher than that of the derivatives of 4′-DMEP, supporting PPT has the potential to be a lead structure of semi-synthetic insecticides.     

辣椒素的抑菌作用及其对某些保护酶活性的影响

 本文对辣椒素的抑菌作用和对寄主体内3种保护酶活性的影响进行了研究,结果表明3.57μg/mL-50.00μg/mL的辣椒素对辣椒枯萎病菌的菌丝生长都有明显的抑制作用,浓度愈高,抑制作用愈强。20μg/mL辣椒素对另外供试的8种病原菌的菌丝具有强烈的抑制作用,其中对番茄灰霉病菌抑制作用最强。辣椒素处理辣椒后1-6 d内,SOD、POD和CAT酶活性的平均增减比率值均明显高于对照,而且对根部和叶部3种保护酶活性的影响显著不同。     

迷迭香精油对几种植物病原菌的抑菌活性研究

采用生长速率法和抑菌圈法测定了迷迭香精油对甜瓜蔓枯病菌、西瓜炭疽病菌、甜瓜灰霉病菌、甜瓜枯萎病菌、梨树腐烂病菌、柑橘黑腐病菌、油菜菌核病菌、蚕豆轮纹病菌和桃细菌性穿孔病菌的抑菌活性.结果表明:迷迭香精油对8种病原真菌都具有很好的抑菌活性,其EC50分别为843.9914、1 084.2372、483.9457、1 735.5866、1 011.7869、901.2150、356.8803、1 197.3657 μL/L;对桃细菌性穿孔病菌也具有一定的抑菌活性,浓度为100 mL/L时,抑菌圈直径为1.00 cm,最低抑菌浓度为1 000 μL/L.     

香菇纤维素酶和半纤维素酶的热效应

本文测定了温度对香菇胞外纤维素酶和半纤维素酶活性的影响。木屑基质培养90天的香菇胞外游纸纤维素酶（C_1酶）、羧甲基纤维素酶（C_X酶）和半纤维素酶（H_C酶）的最适作用温度分别是55℃、55～60℃和50℃。在50～55℃条件下保温1小时后,C_1酶和H_C酶活性大幅度下降,而C_x酶活性下降较为缓慢,具有较高的热稳定性。本文提出相对活力系数（Relative activity coefficient,RAC）作为表示香菇菌丝生长温度与酶最适作用温度之间关系的一个参数。     

异噁草酮对土壤微生物和土壤酶活性的影响

本试验检测了异噁草酮处理土壤后土壤中微生物群落数量和土壤酶活性变化,目的在于研究异噁草酮对土壤微生态产生的影响。结果表明:土壤中异噁草酮有效成分浓度为200、500μg/kg和700μg/kg时,促进土壤中细菌和真菌数量增长,该数量在处理7d之内就会明显变化,并且此影响随异噁草酮使用浓度的提高而增强,但异噁草酮对放线菌的数量无显著影响。施用异噁草酮后,土壤酶活性反应程度由高到低的顺序为:转化酶多酚氧化酶过氧化氢酶。本研究中异噁草酮施入土壤后,除对多酚氧化酶有明显抑制作用,且该抑制作用在短时间内可恢复外,对土壤微生物数量和土壤酶活性都有促进作用。