BTH处理对采后草莓果实还原势和抗病性的调控作用

为明确苯丙噻唑硫代乙酸甲酯(BTH)处理抑制草莓果实采后灰霉病的相关机理,本研究分析了BTH对果实在20 ℃贮藏期间还原势及抗病性的调控作用。结果显示,单一BTH处理在贮藏1 d后可有效诱导草莓果实磷酸戊糖代谢途径关键酶G6PDH(葡萄糖-6-磷酸脱氢酶)、6PGDH(6-磷酸葡萄糖酸脱氢酶)活性以及AsA-GSH(抗坏血酸-谷胱甘肽循环)循环关键酶MDHAR(单脱氢抗坏血酸还原酶)、DHAR(脱氢抗坏血酸还原酶)、GR(谷胱甘肽还原酶)和APX(抗坏血酸过氧化物酶)活性的上升,促进NADPH(还原型烟酰胺腺嘌呤二核苷酸磷酸)、GSH(还原型谷胱甘肽)和AsA(抗坏血酸)等还原性物质含量的上升,并抑制NADP+(烟酰胺腺嘌呤二核苷磷酸)、GSSG(氧化型谷胱甘肽)和DHA(脱氢抗坏血酸)等氧化产物的合成,从而提高果实内部还原势;伴随着还原势的升高,果实中FaPR1、FaPR5、FaCHI-1和Faβglu等PRs基因表达丰度出现明显的上升趋势,同时果实由Botrytis cinerea导致的灰霉病发病率显著(p<0.05)降低。另一方面,经BTH处理的果实在接种病原菌B. cinerea后,其果实内PPP(磷酸戊糖)途径和AsA-GSH循环关键酶活性以及PRs基因表达丰度在整个贮藏期间出现较单一BTH处理更为显著的提高(p<0.05),同时果实发病率和病斑直径也显著下降(p<0.05)。这些结果说明,0.1 mmol/L BTH可通过诱导Priming抗性,使草莓果实在受到B. cinerea侵染时产生强烈抗病性;同时,BTH处理可提高PPP途径和AsA-GSH循环关键酶活性,促进果实中还原性物质积累,从而为PRs基因表达提供高还原势的有利环境。     

β-氨基丁酸处理对采后桃果实还原势的影响及抗病性的诱导作用

采用10 mmol/L β-氨基丁酸（β-aminobutyric acid,BABA）和病原菌Rhizopus stolonifer处理采后‘白凤’水蜜桃,以此研究BABA处理诱导水蜜桃防卫反应的模式,并从还原势变化角度分析相关诱导抗性机理。结果显示：经10 mmol/L BABA处理的桃果实在20 ℃下贮藏2 d后,伴随着果实的发病,其还原性内源信号分子NO含量以及PpNPR1-like、PpCHI和PpGNS基因表达丰度均明显高于对照组;而BABA＋病原菌处理组PpNPR1-like、PpCHI和PpGNS表达丰度在贮藏前3 d整体上显著高于BABA处理和病原菌接种果实（P＜0.05）,在整个贮藏期间3 种基因的表达丰度均处于较高水平。此外,与对照组相比,BABA处理和BABA＋病原菌接种均可显著诱导NO的积累,并同时提升桃果实中磷酸戊糖途径关键酶葡萄糖-6-磷酸脱氢酶和6-磷酸葡萄糖酸脱氢酶活力,促使果实中还原型辅酶II（nicotinamide adenine dinucleotide phosphate,NADPH）和还原型谷胱甘肽大量生成,并降低NADP＋和氧化型谷胱甘肽含量,因而有效提高果实组织还原势。通过这些结果可推测,10 mmol/L BABA可通过诱导Priming反应的方式赋予桃果实在受病原菌侵染时更强的抗病性,从而抑制采后软腐病的发生;同时,BABA处理可通过诱导还原性信号分子积累并提高磷酸戊糖途径关键酶活力,进而提升桃果实还原势、活化相关转录因子,以此诱导PRs基因的表达。     

2,4-表油菜素内酯处理对桃果实软腐病及能量代谢的影响

研究了2,4-表油菜素内酯（2,4-epibrassionolide,EBR）处理对‘白凤’桃果实采后软腐病发生及能量代谢的影响。桃果实先用不同浓度（0、1、5、10 μmol/L）EBR溶液浸泡10 min后接种匍枝根霉,后在（20±1）℃下贮藏60 h。结果表明：5 μmol/L的EBR处理能最显著抑制桃果实病斑直径和发病率的上升（P＜0.05）,同时保持桃果实较好的贮藏品质。与仅接种匍枝根霉的对照组相比,EBR处理能提高Ca2＋-ATPase、H＋-ATPase、琥珀酸脱氢酶和线粒体细胞色素c氧化酶活力,延缓三磷酸腺苷和二磷酸腺苷含量的下降,抑制一磷酸腺苷含量的上升,从而维持较高的能荷水平;相关性分析显示,对照组（r＝－0.939,P＜0.01）和EBR处理组（r＝－0.962,P＜0.01）桃果实的发病率和能荷水平均呈显著负相关。因此,EBR处理抑制桃果实采后软腐病的发生与其维持较高的能荷水平有关。     

2,4-表油菜素内酯对葡萄果实采后灰霉病的抑制作用机理

为研究2,4-表油菜素内酯（2,4-epibrassionolide,EBR）处理对‘巨峰’葡萄果实采后灰霉病的抑制效果及作用机制,将葡萄果实经5 μmol/L EBR溶液处理12 h后刺伤接种灰葡萄孢（Botrytis cinerea）病原菌,然后置于25 ℃下贮藏60 h。结果显示：EBR处理有效抑制了葡萄采后灰霉病的发生和病斑直径的扩展,与单一接种病原菌和EBR处理相比,先用EBR处理后接种B. cinerea葡萄果实展现出更强、更快的抗病反应,表现为几丁质酶、β-1,3-葡聚糖酶和苯丙氨酸解氨酶等抗病相关酶活力及总酚含量显著提高（P＜0.05）,H2O2积累及超氧化物歧化酶、过氧化氢酶、抗坏血酸过氧化物酶活力提高,同时VvCHI、VvGNS和VvPAL-like抗病基因表达显著增加（P＜0.05）;体外实验发现,EBR处理可显著抑制B. cinerea孢子的萌发,但对芽管伸长长度没有显著影响。结论：EBR处理主要是通过敏化反应机制诱导提高葡萄果实的抗病性,从而减轻灰霉病的发生。     

重组葡萄糖脱氢酶酶学性质研究

葡萄糖脱氢酶可用于高纯度低聚果糖和葡萄糖酸钙的生产，一种重组葡萄糖脱氢酶表达、纯化后，对该酶酶学性质进行了研究。最适反应pH值8．4，最适反应温度37℃，在温度低于45℃条件下，酶活力稳定性好。在最适温度和pH条件下，以葡萄糖为底物时葡萄糖脱氢酶的米氏常数Km=20．09mmol／L，以烟酰胺腺嘌呤二核苷酸磷酸（NADP）为底物时的米氏常数Km=0．10mmol／L。     

Trichothecium roseum菌丝体细胞壁提取物对苹果青霉病的控制效果及其诱导抗病机理

以‘红富士’苹果为试材,通过粉红单端孢（Trichothecium roseum）菌丝体细胞壁提取物处理后24 h损伤接种扩展青霉（Penicillium expansum）,研究其对苹果果实青霉病的抑制效果,并通过生化方法进一步揭示了T. roseum菌丝体细胞壁提取物的部分诱导抗病机理。结果表明,T. roseum菌丝体细胞壁提取物处理能有效抑制损伤接种P. expansum苹果青霉病的扩展,其中以150 mg/mL T. roseum菌丝体细胞壁提取物处理效果最好,处理后第5天病害程度相比于对照组降低了29.5%。进一步研究表明,150 mg/mL T. roseum菌丝体细胞壁提取物处理显著提高了贮藏期间果实组织中多酚氧化酶、过氧化物酶、苯丙氨酸解氨酶、4-香豆酰-辅酶A连接酶、肉桂酸-4-羟化酶和肉桂醇脱氢酶的活力（P＜0.05）,同时提高了果实组织中总酚、类黄酮和木质素的含量。进一步推断出T. roseum菌丝体细胞壁提取物可通过促进苯丙烷代谢、提高抗性相关酶的活力和抗性物质的积累而增加果实的抗病性。     

罗非鱼红色肉在冷藏期间的肉色稳定性

为研究罗非鱼红色肉冷藏期间的肉色的稳定性,该文测定了不同包装方式贮藏的红色肉的色差值、pH值、肌红蛋白含量、高铁肌红蛋白还原酶活力、线粒体膜通透性、琥珀酸脱氢酶及乳酸脱氢酶活性、烟酰胺腺嘌呤二核苷酸含量。结果表明,冷藏期间红色肉的L值无明显变化,托盘包装红色肉红度值不断降低,真空包装红色肉红度值先升高后无明显变化。托盘包装红色肉中高铁肌红蛋白含量显著增加,而在真空包装中无明显变化。2种包装方式下氧合肌红蛋白含量、高铁肌红蛋白还原酶活力、线粒体膜通透性、琥珀酸脱氢酶活性、乳酸脱氢酶活性、烟酰胺腺嘌呤二核苷酸含量不断降低,但真空包装均高于托盘包装,证明无氧条件冷藏能够更好地维持肌肉内肌红蛋白的氧化还原过程,从而提高肉色稳定性。通过各因素相关性分析,红度值变化与高铁肌红蛋白含量呈极显著负相关,而与氧合肌红蛋白含量、高铁肌红蛋白还原酶活性、乳酸脱氢酶活性、琥珀酸脱氢酶活性、烟酰胺腺嘌呤二核苷酸含量呈现极显著正相关,说明线粒体活性降低,线粒体内乳酸和琥珀酸分解能力降低,电子传递链产生的烟酰胺腺嘌呤二核苷酸(nicotinamide adenine dinucleotide,NADH)含量减少,使...     

高氧动态气调对双孢蘑菇细胞壁代谢及木质化进程的影响

研究高氧动态气调（high-oxygen dynamic controlled atmosphere,HO-DCA）贮藏过程中双孢蘑菇的几 丁质、纤维素和木质素含量,以及细胞壁代谢和木质素合成相关酶活力的变化规律。结果表明：在贮藏后期, HO-DCA处理组的双孢蘑菇几丁质含量显著高于静态气调组与对照组（P＜0.05）,几丁质酶活力显著低于对照 组（P＜0.05）,说明HO-DCA处理有效抑制了细胞壁中几丁质的降解。HO-DCA处理显著抑制了纤维素酶活力 （P＜0.05）,维持了较高的纤维素含量,保持了较高的组织抗压力。双孢蘑菇在其贮藏后期,菌柄木质素含量 急剧升高,木质化程度不断加深,菌柄木质化程度大于菌盖。4-香豆酸-辅酶A连接酶（4-coumarate: coenzyme A ligase,4CL）和肉桂醇脱氢酶（cinnamyl alcohol dehydrogenase,CAD）活力与木质素含量呈显著正相关, 而苯丙氨酸解氨酶活力与木质素含量相关性不显著。HO-DCA处理组的双孢蘑菇木质素含量显著低于对照组 （P＜0.05）,说明HO-DCA处理通过抑制双孢蘑菇4CL和CAD的活力抑制了木质素的积累,延缓了其木质化进 程。研究明确高氧动态气调对其细胞壁代谢及木质化进程的影响,可为有效控制双孢蘑菇质地劣变提供理论依据。     

鹿茸菇多糖对小鼠抗疲劳能力的影响

为研究鹿茸菇多糖对小鼠抗疲劳能力的影响,将小鼠随机分为5个实验组（共50只,每组10只）,分别为鹿茸菇多糖低剂量组[100 mg/（kg·d）]、中剂量组[200 mg/（kg·d）]、高剂量组[400 mg/（kg·d）]、阳性对照组[红景天200 mg/（kg·d）]和空白对照组（蒸馏水）,连续灌胃30 d,鹿茸菇多糖低、中、高剂量组分别与空白对照组和阳性对照组比较,评价小鼠抗疲劳能力相关指标。结果表明：鹿茸菇多糖低、中、高剂量组小鼠负重游泳力竭时间比空白对照组显著增加（P＜0.05）,力竭运动后鹿茸菇多糖各剂量组血清葡萄糖、肌糖原和肝糖原含量显著增加（P＜0.05）,血清中乳酸脱氢酶活性均显著升高（P＜0.05）、乳酸和尿素氮含量均显著降低（P＜0.05）,肝组织中谷胱甘肽过氧化物酶、超氧化物歧化酶和过氧化氢酶的活性均显著升高（P＜0.05）、丙二醛含量显著降低（P＜0.05）、钠钾三磷酸腺嘌呤核苷酸酶和钙镁三磷酸腺嘌呤核苷酸酶活性显著升高（P＜0.05）;鹿茸菇多糖各剂量组与阳性对照组比较,各项指标没有显著差异。研究表明鹿茸菇多糖具有改善运动性疲劳的作用。     

粉红单端孢侵染和苯并噻重氮诱抗对甜瓜脂氧合酶代谢及特征香气的影响

C6和C9醇醛类挥发性物质是甜瓜果实的特征香气化合物,主要来源于不饱和脂肪酸的脂氧合酶 （lipoxygenase,LOX）代谢途径。本研究以‘玉金香’甜瓜（Cucumis melo L. cv. Yujinxiang）为材料,分析甜瓜 采后粉红单端孢（Trichothecium roseum）侵染和苯并噻重氮（benzothiadizaole,BTH）诱抗对其LOX代谢酶活力和 C6、C9醇醛类挥发性物质释放的影响,探讨各因子之间的相关性。结果表明：粉红单端孢侵染可增加对照组（蒸 馏水处理）和BTH处理组果实的LOX代谢酶活力,且峰值出现时间较未侵染果实提前;对于BTH处理组,果实果 皮和果肉的LOX、醇脱氢酶活力峰值分别比对照组高3.49%、3.93%和5.13%、7.21%,而醇酰基转移酶活力受到抑 制,抑制率分别为19.03%和14.79%。粉红单端孢侵染可促进C6和C9醇醛类挥发性物质的释放,释放量与LOX代谢 酶活力呈正相关;而BTH处理组的C6和C9醇醛类挥发性物质释放量低于对照组,释放量与LOX酶活力无明显相关 性。诱抗剂BTH可提高甜瓜果实采后的抗病性,并抑制挥发性物质的释放。     

采前喷施胺鲜酯对采后龙眼果实贮藏期间果皮能量代谢的影响

本实验研究采前喷施含量为10 mg/kg的胺鲜酯(diethyl aminoethyl hexanoate,DA-6)对采后'福眼'龙眼果实(Dimocarpus longan Lour.cv.Fuyan)贮藏期间果皮能量代谢的影响.贮藏期间取样测定龙眼果实的果皮腺苷三磷酸(adenosine triphosphate...     

Discovery of systematic responses and potential biomarkers induced by ochratoxin A using metabolomics

Ochratoxin A (OTA) is known to be nephrotoxic and hepatotoxic in rodents when exposed orally. To understand the systematic responses to OTA exposure, GC-MS- and ¹H-NMR-based metabolomic techniques together with histopathological assessments were applied to analyse the urine and plasma of OTA-exposed rats. It was found that OTA exposure caused significant elevation of amino acids (alanine, glycine, leucine etc.), pentose (ribose, glucitol, xylitol etc.) and nucleic acid metabolites (pseudouridine, adenosine, uridine). Moreover, myo-inositol, trimethylamine-oxide (TMAO), pseudouridine and leucine were identified as potential biomarkers for OTA toxicity. The primary pathways included the pentose phosphate pathway (PPP), the Krebs cycle (TCA), the creatine pathway and gluconeogenesis. The activated PPP was attributed to the high requirements for nicotinamide adenine dinucleotide phosphate (NADPH), which is involved in OTA metabolism through cytochrome P450. The elevated gluconeogenesis and TCA suggest that energy metabolism was involved. The up-regulated synthesis of creatinine reveals the elevated catabolism of proteins. These findings provide an overview of systematic responses to OTA exposure and metabolomic insight into the toxicological mechanism of OTA.     

Trichothecium roseum蛋白激发子诱导杏果抗黑斑病的作用机理

研究非亲和菌株粉红单端孢（Trichothecium roseum）菌体蛋白激发子对杏果实抗黑斑病的诱导效果及其作用机理。以‘兰州大接杏’为试材,通过处理后不同时间损伤接种杏果黑斑病菌（Alternaria alternata）,研究T. roseum蛋白激发子对杏果实黑斑病抗病性的诱导及其时效性,同时进一步研究最佳处理质量浓度对杏果实组织活性氧及抗性相关酶的影响。结果表明,T. roseum蛋白激发子处理可显著抑制损伤接种杏果病斑的扩展（P＜0.05）,其中14.4 μg/mL蛋白激发子的控制效果最好,且诱导处理72 h时杏果产生最大抗性。T. roseum蛋白激发子处理后杏果组织O2－·的生成速率和H2O2含量分别在48 h和72 h出现明显的峰值,同时处理的果实组织超氧化物歧化酶、过氧化物酶、多酚氧化酶、β-1,3-葡聚糖酶和几丁质酶活性均有所增强。T. roseum菌体蛋白激发子处理能显著增强杏果组织的抗性,其可能是通过激发活性氧爆发和提高杏果实抗病性相关酶的活性来实现的。可见T. roseum蛋白激发子可以作为一种有潜力的诱抗剂应用于采后病害的控制。     

Gene Cloning of an alcohol dehydrogenase from thermophilic alkane-degrading Bacillus thermoleovorans B23

The gene encoding an alcohol dehydrogenase (Bt-ADH) was cloned from a newly isolated thermophilic alkane-degrading Bacillus thermoleovorans, strain B23. The gene conferred 1-tetradecanol dehydrogenase activity on Escherichia coli cells. Bt-ADH is composed of 249 amino acid residues and the calculated molecular mass is 27,196 Da. A tyrosine residue in the active site and a glycine-rich sequence (GGXXGI/LG) constituting probable nicotinamide adenine dinucleotide (NAD+) or nicotinamide adenine dinucleotide phosphate (NADP+) binding site were completely conserved in the Bt-ADH sequence at positions 155 and 11, respectively. A phylogenetic analysis of Bt-ADH suggested that the enzyme belongs to the zinc-independent ADH Group II. Its highest similarity (48% identical) was to a hypothetical oxidoreductase from a hyperthermophile, Thermotoga maritima.     

Hexose shunt dehydrogenase activity in leukocytes isolated from bovine milk

A spectrophotometric method for measuring small changes in light absorption at specific wavelengths from highly turbid suspensions of biological material (dual-beam spectrophotometry) has been used to measure glucose-6-phosphate oxidation and nicotinamide adenine dinucleotide phosphate (oxidized) reduction catalyzed by lysates of milk leukocytes. This reduction was correlated .84 with cellular protein concentration and, therefore, with cell concentration.     

n-Hexanol Formation from n-Hexanal by Enzyme Action in Soybean Extracts

n-Hexanal in soybean homogenates decreased during incubation at alkaline pH. When soybean extracts were dialyzed and then incubated with n-hexanal and cofactors [nicotinamide adenine dinucleotide (NAD⁺), the reduced form (NADH), nicotinamide adenine dinucleotide phosphate (NADP+), and the reduced form (NADPH)], NADH and NADPH stimulated the enzymatic reduction of n-hexanal with NADH being more effective than NADPH. When undialyzed preparations were incubated, all the cofactors stimulated enzymatic activity, with NADH being the most effective. The reaction product in all the incubation mixtures was n-hexanol; n-hexanal was reduced to n-hexanol stoichiometrically. It was suggested that alcohol dehydrogenase was responsible for the decrease of n-hexanal in soybean homogenate at alkaline pH.     

沙枣胶/枯茗醛复合涂膜对番茄采后红粉病的抑制作用及机制

红粉病是番茄果实主要的病害之一,采前采后均可发生,给番茄生产和贮运造成了较大损失。本实验研究枯茗醛对番茄红粉病病原真菌粉红单端孢（Trichothecium roseum Pers. Link）的体外抑制活性,并制备沙枣胶与枯茗醛复合涂膜‘千禧’樱桃番茄,果实经损伤接种后探究该涂膜对番茄红粉病的抑制作用及其对番茄果实抗性的影响。结果显示,枯茗醛在离体条件下能有效抑制T. roseum孢子萌发和菌丝生长,在0.1 μL/mL枯茗醛作用下,对菌落生长的抑制率达80%以上。在体内条件下,沙枣胶与枯茗醛复合涂膜能明显抑制果实上T. roseum菌体生长,延缓病斑扩展并降低番茄果实接种发病率,在贮藏第9天,质量浓度30 mg/mL沙枣胶与体积分数0.2%枯茗醛复合涂膜组病斑直径和发病率分别为对照组的36.7%和69.2%。复合涂膜可显著提高番茄果实接种前期H2O2含量,并显著提高果实贮藏期间过氧化物酶、苯丙氨酸解氨酶和4-香豆酸辅酶A连接酶的活力,促进果实中总酚和类黄酮的积累。沙枣胶与枯茗醛复合涂膜能够有效抑制番茄采后红粉病的发生与发展,其作用机制不仅与枯茗醛抑制病原菌生长有关,还与诱导番茄果实增强抗性有关。     

Membranes of mammary gland. XI. Marker enzyme distribution profiles for membranous components from bovine mammary gland.

Enzyme distribution profiles of clarified bovine mammary homogenates separated by equilibrium centrifugation on linear sucrose gradients suggested that several of the commonly utilized marker enzymes for rat liver are also valid markers for mammary cellular components. These marker enzymes include: Succinate dehydrogenase (mitochondria), nicotinamide adenine dinucleotide phosphate cytochrome c reductase and, to a lesser extent, retenone insensitive nicotinamide adenine dinucleotide cytochrome c reductase (endoplasmic reticulum), galactosyl transferase (Golgi apparatus), 5'-nucleotidase (plasma membranes), uric acid oxidase (microbodies), and acid phosphatase (lysosomes). Rotenone sensitive nicotinamide adenine dinucleotide cytochrome c reductase and sodium, potassium, magnesium-stimulated adenosine triphosphatase were widely distributed among subcellular fractions and are not valid marker enzymes. The boyant densities determined for the above fractions should aid in design of methods to obtain enriched sources of these components for analysis.