稻瘟菌诱导性稻叶脂氧合酶的进一步纯化及抗体制备   总被引：1，自引：1，他引：1  

杨依军  秦引萍 《植物病理学报》1997,27(3):243-249

 采用DEAE-Toyopearl离子交换、Buty l-Toyopear l疏水层析、CM-Toyopearl离子交换、Sephacryl S100凝胶过滤和FPLCMono Q等步骤,从非亲和性稻瘟菌侵染稻叶中纯化了两种诱导性脂氧合酶,即CM-Loxl和CM-Lox2。SDS-PAGE检测结果表明:CM-Lox1和CM-Lox2为单链多肽,它们的分子量分别为98 kd和102 kd。利用微量免疫法制备了CM-Lox1和CM-Lox2的抗体,制备的抗体效价达到1:1000倍,能检测0.5~2.0 ng的抗原。免疫印迹杂交证明CM-Lox1和CM-Lox2之间存在密切的血清学关系。此外,Anti-CM-Lox1和Anti-CM-Lox2与水稻中另一稻瘟菌诱导性脂氧合酶RLL以及发芽种子出现的RL-2也存在交叉反应。  相似文献   

Cloning and Sequence Analysis of Lipoxygenase Gene cDNA from Cucumber Fruit （Cucumis sativus L.）     

Lipoxygenases are nonheme-iron-containing dioxygenases that catalyze the hydroperoxidation of unsatrated fatty acids containing a cis  cis-  -pentadiene structure producing hydroperoxy acids with conjugated dienes.…… 《分子植物育种》2007,5(2):188-189

Lipoxygenases are nonheme-iron-containing dioxygenases that catalyze the hydroperoxidation of unsatrated fatty acids containing a cis, cis-1,4-pentadiene structure producing hydroperoxy acids with conjugated dienes. LOX activity has been found in a wide range of plants. Typical substrates for LOX in plants are linoleic acid and linilenic acid fatty acids. The function of various LOXs in plants is unknown, but their participation in all stages of plant growth and development has been suggested （Hildebrand, 1989; Siedow, 1991）. Some of the physiological processes in whicn lipoxygenses have been implicated include wounding （Saravitz and Siedow, 1996）, pathogen attack （Melan et al., 1993）, seed germination （Kato et al., 1992）, fruit ripening （Ferrie et al., 1994）, plant senescence （Paliyath and Droillard, 1992）. The study on the role of lipoxygenase in ripening and senescence fruit focused on tomato and strawberry. Cloning LOX gene of cucumber fruit will make us further understand the molecularaction mode of this enzyme during fxuit ripening and senescence. In this paper we isolated the partial nucleotide sequences of cucumber fiuit lipoxygenase gene and discuss the characterization of it.  相似文献   

PG和LOX对采后番茄果实软化及细胞超微结构的影响   总被引：17，自引：2，他引：15  

生吉萍  罗云波  申琳 《园艺学报》2000,27(4):276-280

以番茄 (Lycopersiconesculentum)品种‘丽春’为试验材料研究了果实采后多聚半乳糖醛酸酶 (PG)和脂氧合酶 (LOX)的活性变化与果实硬度的关系 ,并观察了细胞超微结构的变化。结果表明 ,随着果实采后呼吸强度和乙烯释放量不断增加 ,PG和LOX活性迅速增加 ,而果实硬度逐渐下降。呼吸强度和LOX活性高峰出现在发白期 (BR) ,而乙烯释放量高峰延迟至转色期 (TU)。PG活性随着果实的成熟逐渐增加 ,而LOX活性在发白期之后保持较高水平。分别用外源LOX和LOX加底物处理番茄组织切片 ,发现组织细胞的衰老进程加速 ,进而导致细胞膜瓦解、胞壁纤维松弛以及细胞器空胞化。PG和LOX都与果实的后熟软化有关 ,LOX与衰老的启动有关  相似文献   

104份甘肃小麦品种脂肪氧化酶和多酚氧化酶活性基因等位变异的检测   总被引：3，自引：0，他引：3  

曹东  张雪婷  王世红  王东霞  赖勇  孟亚雄  王化俊  杨文雄 《麦类作物学报》2014,34(4):467-473

面粉和面制品的色泽是评价小麦品质的重要指标。小麦脂肪氧化酶(LOX)和多酚氧化酶(PPO)活性对面粉白度及面制品的色泽具有重要影响。为给甘肃省小麦品质育种提供参考依据,以104份甘肃省育成的小麦品种为材料,利用功能标记LOX16、LOX18、PPO18、PPO16和PPO29检测TaLox-B1、PpoA1及Ppo-D1位点的等位变异,分析甘肃小麦品种资源中LOX和PPO活性基因的组成和分布特点。结果表明,在甘肃小麦中,等位变异TaLox-B1a和TaLox-B1b的频率分别为22.12%和77.88%;其中,甘肃冬小麦品种高LOX活性等位变异TaLox-B1a的分布频率(30.56%)高于春小麦(3.13%)。等位变异Ppo-A1a、PpoA1b、Ppo-D1a和Ppo-D1b的频率分别为49.04%、50.96%、50.96%和49.04%;两个PPO基因的等位变异组合Ppo-A1a/Ppo-D1b、Ppo-A1a/Ppo-D1a、Ppo-A1b/Ppo-D1b和Ppo-A1b/Ppo-D1a的分布频率依次为28.85%、20.19%、20.19%和30.77%。说明在甘肃小麦品种中,低LOX活性等位变异(TaLox-B1b)品种比例较高;低PPO活性等位变异(Ppo-A1b、Ppo-D1a)品种分布比例略高于高PPO活性类型;其中,32份小麦品种在TaLox-B1、Ppo-A1和Ppo-D1三个位点同时含有高LOX活性和低PPO活性的等位变异。  相似文献   

果蔬成熟和衰老中的重要酶-脂氧合酶   总被引：2，自引：0，他引：2  

生吉萍  申琳  罗云波 《果树学报》1999,16(1):2

脂氧合酶是广泛存在于植物体中的一种酶,已有研究表明,它可能参与植物生长、发育、成熟、衰老和防御过程的调控。近年来,随着对果蔬采后生理研究的深入,越来越发现脂氧合酶与果蔬采后的变化有关,它调节某些物质的合成,参与乙烯的代谢途径,并与果蔬的抗性、风味、颜色、硬度等都有密切的关系。综述了脂氧合酶在采后的研究进展,提出了该酶可能是控制采后生理变化的关键酶,并提出寻找脂氧合酶特异性抑制剂,用反义基因技术控制该酶的生物合成,培育耐贮新品种的设想  相似文献   

猕猴桃成熟果实中脂氧合酶基因的克隆   总被引：11，自引：0，他引：11  

陈昆松 Ross.  GS 《园艺学报》1998,25(3):230-235

采用ＰＣＲ扩增方法,从成熟猕猴桃果实组织中克隆到一个８２４ｂｐ的脂氧合酶（ＬＯＸ）基因片段（ｐＫＬＣ１）,该片段含有２７５个氨基酸组成的开放阅读框架,它与拟南芥菜、黄瓜、豌豆、土豆、水稻、大豆、烟草和番茄的核苷酸同源性为５９．０％～７４．６％,氨基酸同源性为６３．１％～７６．３％。Ｓｏｕｔｈｅｒｎ杂交表明,ｐＫＬＣ１片段为一低拷贝数基因家族所编码。  相似文献   

Molecular simulations probing Kushecarpin A as a new lipoxygenase inhibitor     

Nisar M  Kaleem WA  Khan I  Adhikari A  Khan N  Shah MR  Khan IA  Qayum M  Samiullah  Ismail M  Aman A 《Fitoterapia》2011,82(7):1008-1011

Zizyphus oxyphylla Edgew is used in Pakistan as a folk medicinal remedy for inflammatory conditions, pains especially rheumatic pain, fevers, allergy and diabetes. The aim of the current study was to scientifically validate the folk use of Z. oxyphylla Edgew by using the isolated compound in vitro and in vivo levels. Kushecarpin A was isolated from ethyl acetate fraction of the plant crude extract. Molecular docking simulations predicted Kushecarpin A as a potential new lipoxygenase (LOX) inhibitor. Kushecarpin A showed significant lipoxygenase inhibition (IC50: 7.2 ± 0.02 μM) thus validated computational prediction. It also exhibited significant and highly significant inhibition (p < 0.05 and p < 0.01) of carrageenan-induced hind paw oedema at the doses of 5, 10 and 20 mg/kg. Kushecarpin A seems to be a potentially new anti-inflammatory compound responsible for anti-inflammatory activities of Z. oxyphylla Edgew. In vitro and in vivo anti-inflammatory inflammatory activities were found in good agreement with the folk medicinal use of Z. oxyphylla Edgew in inflammatory disorders.  相似文献   

等电聚焦凝胶电泳检测5个大豆杂交组合中脂肪氧化酶的缺失突变     

闫彩霞  马峙英  张彩英  秦君  高雪峰  张孟臣 《安徽农业科学》2008,36(16):6691-6694

[目的]为大豆的分子标记辅助育种提供理论依据。[方法]应用改进的等电聚焦凝胶电泳(IEF-PAGE)技术,对由不同脂肪氧化酶(Loxs)缺失类型亲本配置的5个大豆杂交组合F2代进行逐粒检测,鉴定其Loxs缺失类型。[结果]杂交组合0129和0124为一类,其F2代有4种表现型(-Lox2-、Lox1Lox2、-Lox2Lox3和-Lox1Lox2Lox3);0139和0155为一类,F2代有6种表现型,分别为-Lox2、-Lox3、-Lox2Lox3、N(正常)、H(Lox2杂合)和H-Lox3(Lox2杂合且Lox3缺失);0134单独为一类,F2代仅3种表现型,分别为-Lox2、N和H。Lx3/Lx1和lx3/lx1为显隐性等位基因,Lx2/lx2为共显性等位基因。[结论]杂合基因型在蛋白水平上的特异表达为筛选优异的种质资源提供了一种辅助手段。  相似文献   

桃果实采后脂氧合酶活性和膜脂脂肪酸组分的变化   总被引：16，自引：0，他引：16  

吴敏  陈昆松  吴平  张上隆 《园艺学报》2001,28(3):218-222

 以‘湖景蜜露’桃果实为试材, 分析了果实后熟软化进程中的乙烯释放、脂氧合酶(LOX) 活性和细胞膜脂脂肪酸组分的变化, 探讨其与果实软化的关系。结果表明, 细胞脂脂肪酸组分主要有棕榈酸(16∶0) 、硬脂酸(18∶0) 、油酸(18∶1) 、亚油酸(18∶2) 和亚麻酸(18∶3) 。在果实采后20 ℃后熟过程中, 三种不饱和脂肪酸组分比例变化较大(尤其是亚油酸和亚麻酸) , 而两种饱和脂肪酸的变化很小。在LOX 自我活化之前, 亚麻酸可能是LOX催化反应的主要底物。果实组织的后熟软化与LOX参与的细胞膜脂过氧化作用有关, 乙烯跃变可能是桃果实成熟启动后的伴随因子, 参与后熟软化进程。  相似文献   

Expression of a lipoxygenase encoding gene (BoLOX1) during postharvest senescence of broccoli     

María Eugenia Gomez-LobatoPedro M. Civello  Gustavo A. Martínez 《Postharvest Biology and Technology》2012,64(1):146-153

Lipoxygenases (LOX) belong to a large family of plant enzymes that catalyze the hydroperoxidation of polyunsaturated fatty acids. Most of them are expressed during senescence and contribute to membrane deterioration and biosynthesis of jasmonic acid, a known senescence enhancer. In this work, we cloned a fragment of a gene encoding a LOX from broccoli (BoLOX1). The analysis of the sequence revealed that BoLOX1 is closely related to other LOX from higher plants. Furthermore, we analyzed the expression of BoLOX1 and detected a larger increase during postharvest senescence. A slight increase of total lipoxygenase activity was also found during senescence. In other sets of experiments, broccoli heads were treated with plant hormones, such as cytokinin and ethylene, as a way to assess the effect of such compounds on the expression of BoLOX1. Cytokinin treatment delayed the increase of BoLOX1 expression and lipoxygenase activity whereas ethylene accelerated both processes. Also, several postharvest treatments were applied in order to delay senescence in broccoli florets and to evaluate their effects on BoLOX1 expression. Samples treated with modified atmosphere, hot air, UV-C or white light showed a delay in chlorophyll degradation and degreening. In most cases, the treatments also delayed the increase of BoLOX1 expression, reaffirming the relationship between the expression of this gene and senescence. However, treatments like modified atmospheres and visible light markedly increased lipoxygenase activity, which suggests a lack of correlation between BoLOX1 expression and lipoxygenase activity.  相似文献