品种间大豆脂肪氧化酶活性差异及β-胡萝卜素漂白性研究

为了解东北地区大豆品种间大豆脂肪氧化酶(LOX)活性差异规律,整理适于东北地区种植的大豆种质资源307份(6份半野生品种,17份地方品种及4份国外品种,其余为栽培品种),统一种植,秋季收获后,测定各品种成熟籽粒脂肪氧化酶总活性。分析品种间脂肪氧化酶总活性分布规律、大豆脂肪氧化酶总活性与β-胡萝卜素漂白性之间关系。结果表明,307份大豆品种间脂肪氧化酶总活性差异极显著,平均值为21.68 U·mL~(-1),最小值为2.51 U·mL~(-1),最大值为42.38 U·mL~(-1)。根据脂肪氧化酶总活性差异,通过聚类分析将307份大豆品种分为3类。低活性品种占总量43.65%,脂肪氧化酶总活性平均值为15.25 U·mL~(-1);中等活性品种占总量43.97%,脂肪氧化酶活性平均值为24.64 U·mL~(-1);高活性品种占总量12.38%,脂肪氧化酶总活性平均值为33.87 U·mL~(-1)。脂肪氧化酶总活性与油分含量呈正相关,与蛋白含量呈负相关,与漂白性呈正相关。     

Increased abundance of patatins,lipoxygenase and miraculins in a thaxtomin A-habituated potato Russet Burbank somaclone with enhanced resistance to common scab

Potato common scab caused by the actinobacterium Streptomyces scabiei is characterized by the formation of corky lesions on tubers that reduce their marketability. Management of common scab is very complex and often ineffective under various environmental conditions. Using potato varieties that are more resistant to common scab remains one of the most efficient strategies to control this disease. However, very little is known about the factors associated with resistance to common scab. Somaclone RB9 regenerated from thaxtomin A-habituated potato Russet Burbank calli produced tubers more resistant to common scab than the original variety. Comparison of the RB9 tuber proteome with that of Russet Burbank using label-free quantitative proteomic analysis revealed changes in the accumulation of defence-related proteins from the patatin and lipoxygenase (LOX) families, which are involved in the metabolism of lipids, and of two miraculins of the Kunitz-type protease inhibitors family. The implication of LOX during common scab infection was studied using synchronized minitubers developed from leaf-bud cuttings. S. scabiei infection stimulated the accumulation of LOX in both Russet Burbank and RB9 minitubers, but this accumulation was intensified in RB9 minitubers. Infection also increased LOX activity in Russet Burbank and RB9 minitubers. However, LOX activity measured in noninfected RB9 minitubers was similar to that of infected Russet Burbank minitubers, indicating endogenous activation of LOX activity in RB9 minitubers. We discuss how increased LOX abundance and activity in the somaclone RB9 may contribute to improving tuber defence against common scab.     

水杨酸对采后桃果实脂氧合酶及相关指标的影响

水杨酸 (salicylicacid ,SA)是一种植物体内产生的简单酚类化合物 ,广泛存在于高等植物中。已发现 ,外源施用SA可诱导某些植物开花和产热 ,诱导烟草和黄瓜等植物对病毒、真菌及细菌等病害的抗性 ,Raskin( 1 992 )提出SA可以被看作是一种新的植物内源激素 ,其作用和机制受到人们的关注。外源SA处理对园艺产品采后的作用研究还较少 ,仍有许多问题尚待探讨。本试验以 2个不同品种的桃果实为试材 ,研究了SA处理后果肉组织脂氧合酶活性以及其他相关指标的变化 ,并测定了果肉的硬度 ,为进一步探讨桃果实采后成熟衰老机制 ,改进现有贮藏方式提…     

紫外分光光度法测定啤酒大麦籽粒脂氧合酶活力的研究

为了明确影响啤酒大麦籽粒脂氧合酶(LOX-1)活力的参数,以甘啤4号啤酒大麦籽粒为研究对象,采用紫外分光光度法探究底物浓度、提取缓冲液p H、反应体系缓冲液p H、提取时间、粗酶加入量∶底物加入量等对LOX-1活力的影响。结果表明,LOX-1活力随底物浓度的增加表现为先增加后减小的趋势,当底物浓度达到0.30mmol·L~(-1)时,LOX-1活力显著高于其它处理;随提取缓冲液p H的增加,LOX-1活力表现为双峰变化趋势,且在p H值5.0处,酶活力为9.85U·g~(-1),显著高于其它处理;粗酶提取时间为30min时,LOX-1活力显著高于其它处理,低于或者高于30min,酶活力均呈现降低趋势;LOX-1活力随反应体系p H的增加,表现为双峰变化趋势,当p H值为6.4时,酶活力最大,且与其它处理间存在显著差异;在其它条件不变,粗酶加入量为50μL的情况下,增加底物加入量,LOX-1活力表现为先增加后减小的趋势,且底物加入量为200μL时,酶活力显著高于其它处理。最终确定紫外分光光度法的测定参数:粗酶提取时间为30min,底物浓度为0.25mmol·L~(-1),提取液和反应体系缓冲液分别为p H值5.0醋酸盐缓冲液与p H值6.4磷酸盐缓冲液,粗酶加入酶量∶底物加入量为1∶4。研究结果为紫外分光光度法在啤酒大麦籽粒LOX-1活力测定中的应用提供了参考。     

大豆脂氧酶的热失活动力学与其二级结构的圆二色谱表现

应用大豆脂氧酶的变温圆二色谱图研究了大豆脂氧酶(LOX)的热失活与二级结构变化过程.结果表明:在50一65℃条件下,10 min内,LOX的酶活随热处理时间延长而下降,其热失活过程遵循一级动力学,活化能Ea值为217 kJ.mol-1.在相同的热处理条件下,LOX中二级结构a-螺旋和β-折叠的相对含量均在20%左右浮动...     

四川不同产区烟叶脂氧合酶活性及基因表达水平的差异

为烟叶香气物质形成机制的研究提供参考,以云烟87和红花大金元2个烤烟品种为试验材料,在四川6个产区采用田间试验研究烤烟旺长期、现蕾期和成熟期烟叶的脂氧合酶活性、基因表达量差异及其相关性。结果表明:在四川6个产区2个烤烟品种不同生育期的脂氧合酶活性及其基因相对表达量存在差异,但变化趋势相同,并多在现蕾期达最高,且各产区云烟87和红花大金元烤烟不同生育期烟叶的LOX酶活性差异多达显著水平。在旺长期和现蕾期云烟87烟叶LOX基因的相对表达量与LOX酶活性呈极显著正相关,相关系数分别为0.96和0.97;在成熟期呈不显著正相关,相关系数为0.14。在旺长期、现蕾期和成熟期红花大金元烟叶的LOX基因相对表达量与LOX酶活性均呈极显著正相关,相关系数分别为0.98、0.95和0.97。烟叶脂氧合酶活性与其基因相对表达量呈正相关关系,且在旺长期、现蕾期和成熟期其相关性均达到极显著水平。     

Prokaryotic Expression, Purification and Characterization of a Novel Rice Seed Lipoxygenase Gene OsLOX1

Lipoxygenase (LOX, EC1.13.11.12) is a key enzyme during the degradation of lipids in animals and even plants, and also the first key enzyme responsible for the biosynthesis of jasmonate. To purify and characterize the OsLOX1 gene from rice seeds, the entire coding region of the OsLOX1 gene was inserted into an expression vector pET30a(+) and transformed into Escherichia coli BL21 (DE3). Expression of the fusion protein was successfully induced by isopropyl-β-D- thiogalactopyranoside (IPTG) and the purified ...     

Role of LOX3 Gene in Alleviating Adverse Effects of Drought and Pathogens in Rice

Lipoxygenase 3 (LOX3) is a major component of the LOX isozymes in mature rice seeds. To investigate the role of LOX3 gene under stresses, a plant expression vector containing antisense cDNA of LOX3 was constructed. Rice varieties Wuyunjing 7 and Kasalath were transformed by the Agrobacterium-mediated method and transgenic rice plants were generated. PCR and Southern blot results showed that the antisense LOX3 gene was integrated into the rice genome. Analyses of embryo LOX3 deletion and semi-quantitative RT-PCR confirmed the antisense suppression of LOX3 gene in transgenic plants. The T2 antisense plants of LOX3 were sensitive to drought stress, rice blast and bacterial blight compared with non-transgenic plants. These results suggest that the LOX3 gene might function in response to stresses.     

水稻种子脂氧合酶基因OsLOX1的原核表达、纯化及鉴定

 脂氧合酶是动植物体内催化脂质降解的关键酶，也是茉莉酸合成途径的第一个关键酶。以先前克隆到的水稻种子脂氧合酶基因OsLOX1全长cDNA为模板，用含有特异酶切位点的P1、P2为引物，通过PCR方法将它构建到大肠杆菌表达载体pET30a(+)上并转化到大肠杆菌菌株BL21（DE3）中，获得相应的重组工程菌。经过20℃条件下的IPTG诱导，OsLOX1重组蛋白在BL21（DE3）菌株中得到表达，经生化特性分析，发现该重组蛋白具有LOX催化活性，其最适pH和温度分别为4.8和30℃。该重组子可进一步应用于体外生产茉莉酸和研究植物种子LOX结构与功能的关系。