植物抗冻蛋白及其高级结构研究进展

抗冻蛋白是一类能控制冰晶生长和抑制冰晶之间发生重结晶的蛋白质．并具有两种明显不同的活性——热滞活性和重结晶抑制活性．植物抗冻蛋白的抗冻特性是低热滞活性和高重结晶抑制活性．植物抗冻蛋白虽然在DNA和氨基酸水平上具有较大的差异-几乎没有同源性，但却拥有相似的高级结构．冰晶结合位点也有一定的相似性．而且都可以通过“表面互补”模型较好地解释它们与冰晶之间的相互作用．全面介绍了植物抗冻蛋白的种类及特性，以及它们的高级结构和冰晶结合位点．     

4种木霉对核盘菌的抑制作用

通过对峙培养以及对木霉菌(Trichoderma spp.)次生代谢物对病原菌的抑菌率进行了测定,结果表明,供试哈茨木霉(T.harzianum)、绿色木霉(T.viride)、康氏木霉(T.koningii),以及一株未知种名的木霉菌株TY(Trichoderma sp.)均对核盘菌(Sclerotinia sclerotiorum (Lib) de Bary)有明显的抑制作用.在对峙培养中,木霉菌菌落与核盘菌的菌落相向生长一定时间后,挑取核盘菌菌落内部的菌丝进行镜检,发现有木霉菌孢子存在,且核盘菌发生菌丝变形,如原生质浓缩,菌丝断裂,或菌丝消解等现象.并发现不同种木霉所产生的次生代谢物的活性不同,对核盘菌的抑制作用也存在显著差异.     

Characterization of an Insoluble and Soluble Form of Melanin Produced by Streptomyces cavourensis SV 21, a Sea Cucumber Associated Bacterium

Melanin is a widely distributed and striking dark-colored pigment produced by countless living organisms. Although a wide range of bioactivities have been recognized, there are still major constraints in using melanin for biotechnological applications such as its fragmentary known chemical structure and its insolubility in inorganic and organic solvents. In this study, a bacterial culture of Streptomyces cavourensis SV 21 produced two distinct forms of melanin: (1) a particulate, insoluble form as well as (2) a rarely observed water-soluble form. The here presented novel, acid-free purification protocol of purified particulate melanin (PPM) and purified dissolved melanin (PDM) represents the basis for an in-depth comparison of their physicochemical and biological properties, which were compared to the traditional acid-based precipitation of melanin (AM) and to a synthetic melanin standard (SM). Our data show that the differences in solubility between PDM and PPM in aqueous solutions may be a result of different adjoining cation species, since the soluble PDM polymer is largely composed of Mg²⁺ ions and the insoluble PPM is dominated by Ca²⁺ ions. Furthermore, AM shared most properties with SM, which is likely attributed to a similar, acid-based production protocol. The here presented gentler approach of purifying melanin facilitates a new perspective of an intact form of soluble and insoluble melanin that is less chemical altered and thus closer to its original biological form.     

Semi-Synthesis,Cytotoxic Evaluation,and Structure—Activity Relationships of Brefeldin A Derivatives with Antileukemia Activity

Brefeldin A (1), a potent cytotoxic natural macrolactone, was produced by the marine fungus Penicillium sp. (HS-N-29) from the medicinal mangrove Acanthus ilicifolius. Series of its ester derivatives 2–16 were designed and semi-synthesized, and their structures were characterized by spectroscopic methods. Their cytotoxic activities were evaluated against human chronic myelogenous leukemia K562 cell line in vitro, and the preliminary structure–activity relationships revealed that the hydroxy group played an important role. Moreover, the monoester derivatives exhibited stronger cytotoxic activity than the diester derivatives. Among them, brefeldin A 7-O-2-chloro-4,5-difluorobenzoate (7) exhibited the strongest inhibitory effect on the proliferation of K562 cells with an IC₅₀ value of 0.84 µM. Further evaluations indicated that 7 induced cell cycle arrest, stimulated cell apoptosis, inhibited phosphorylation of BCR-ABL, and thereby inactivated its downstream AKT signaling pathway. The expression of downstream signaling molecules in the AKT pathway, including mTOR and p70S6K, was also attenuated after 7-treatment in a dose-dependent manner. Furthermore, molecular modeling of 7 docked into 1 binding site of an ARF1–GDP-GEF complex represented well-tolerance. Taken together, 7 had the potential to be served as an effective antileukemia agent or lead compound for further exploration.     

氟节胺抑制枇杷嫁接成苗砧木萌蘖相关机理初探

为寻找更长效、无害的化学控蘖剂以减轻田间管理劳动强度,以枇杷(Eriobotrya japonica)嫁接成苗为材料,喷施有效成分浓度为0.050%,0.071%和0.125%的氟节胺,于不同时期取样观察控蘖效果,并尝试探究相关机理。结果表明,氟节胺减少了叶片单位面积光能的吸收量、抑制了光化学转换,但叶片的热耗散量并无明显变化。进一步测定植株氧化还原状态发现,氟节胺能够快速促进细胞内超氧阴离子(O₂^.-)积累并延缓或抑制H₂O₂的转化,从而诱发氧化胁迫。由此可见,氟节胺可能通过抑制叶片光化学转换和热耗散来破坏新生萌蘖的光合性能,进而抑制其萌发生长,而成熟叶片则通过提升自身抗氧化系统活性耐受氟节胺造成的轻度胁迫。在本试验中,各浓度的氟节胺对控制砧木萌蘖有不同程度的效果,其中浓度为0.125%的氟节胺控蘖效果相对最佳,且基本不会对砧木和嫁接成苗造成持续伤害,其作为化学控蘖剂有一定的应用潜力。     

伐地那非增加EGCG-β-乳球蛋白纳米粒对肝癌细胞的抑制作用

较高浓度的EGCG才能抑制癌细胞的增殖,通过纳米化和EGCG与其他药物的联合使用是提高EGCG生物活性的重要策略。本研究将EGCG和伐地那非（VD）同时包埋于β-乳球蛋白（β-Lg）纳米载体中,制备出EGCG-VD-β-Lg纳米粒（EVβ-NPs）,体外试验证实,EVβ-NPs能提高人肝癌细胞（HepG2细胞）中Caspase-3活性,使HepG2细胞在S期产生明显的阻滞,诱发细胞核分裂,从而导致HepG2细胞凋亡。研究结果表明,将EGCG与微量的VD联合使用,并通过纳米化包埋可以显著提高EGCG的抗癌活性。这一方法在EGCG抗癌制品的开发方面具有潜在的价值。     

High-throughput screening reveals small molecule modulators inhibitory to Acidovorax citrulli

Acidovorax citrulli is a seed-transmitted gram-negative bacterium that can cause substantial economic yield loss in watermelon and melon production worldwide. Four small-molecule libraries containing 4,952 compounds were selected for high-throughput screening against A. citrulli wild-type strain Xu3-14 by evaluation of growth inhibition. One hundred and twenty-seven molecules (2.5% hit rate) were identified as bactericidal or bacteriostatic against A. citrulli at 100 μM. Secondary screens indicated that 27 candidate compounds were more effective against A. citrulli Group II strains than Group I strains (classified using repetitive element PCR). Several compounds were inhibitory to other pathogenic bacteria, including Clavibacter michiganensis subsp. michiganensis and Xanthomonas campestris pv. campestris, but did not affect the growth of plant beneficial bacteria Pseudomonas fluorescens and Bacillus subtilis. More than half of the compounds did not inhibit germination of Arabidopsis or watermelon seeds. The effect of small molecules on A. citrulli seed-to-seedling transmission was evaluated by applying each compound to inoculated watermelon seeds and assessing seedling infection. Nine compounds were chosen for further investigation based on their reduction of percentage seedling infection and compiling scores on their specificity, sensitivity, and phytotoxicity obtained in the secondary screens. The five best compounds were selected (thiamphenicol, nadifloxacin, pipemidic acid, ciclopirox, and zinc pyrithione) for greenhouse tests and were found to effectively reduce the seed-to-seedling transmission of A. citrulli in both artificially and naturally infested seeds. These top five compounds provide a basis for future development of an A. citrulli-specific bactericide.     

硝化抑制剂对枸杞园土壤的作用效果研究

通过室内恒温培养试验,筛选出效果最佳的硝化抑制剂剂型及剂量并应用于枸杞园土壤,研究其对枸杞产量及品质的影响。室内恒温培养试验供试硝化抑制剂为2-氯-6-三氯甲基吡啶(Nitrapyrin)、双氰胺(DCD)和3,4-二甲基吡唑磷酸盐(DMPP),共设17个处理：未添加硝化抑制剂(CK),添加Nitrapyrin(纯氮量的0.1%、0.2%、0.3%、0.4%、0.5%、0.6%),添加DMPP(纯氮量的0.5%、1.0%、1.5%和2.0%),添加DCD(纯氮量的1.0%、2.0%、3.0%、3.5%、4.0%和5.0%)。结果表明：在砂土的培养中,三者硝化抑制效果表现为DMPP≥Nitrapyrin＞DCD;DMPP和Nitrapyrin的硝化抑制率分别为71.90%~75.17%和4.83%~77.28%。但由于DMPP的价格(240~360元·kg^-1)及用量均高于Nitrapyrin(155元·kg^-1),故选择纯氮量0.5%的Nitrapyrin应用于大田试验。田间试验设置4个处理：农民习惯施肥为SF100,SFN100、SFN80及SFN60处理是在SF100处理基础上分别减少0%、40%、60%的枸杞专用肥同时添加纯氮量为0.5%浓度的Nitrapyrin。结果表明：田间试验中施用Nitrapyrin处理的产量较SF100处理分别提高了6.67%,5.80%及3.52%,同时与SF100处理相比,SFN100处理的多糖及蛋白质含量分别提高了16.22%及8.67%。综合经济效益及生态效益,浓度为纯氮量0.5%的Nitrapyrin为最佳处理。在大田试验中施用Nitrapyrin同时减少枸杞专用肥的用量,枸杞产量及效益均有所提高,且蛋白质及多糖含量有显著增加。因此,可初步认为硝化抑制剂的施用对枸杞的种植有“减肥增效”的作用。