膜脂过氧化与水稻对稻瘟病抗性的关系

喷雾接种稻瘟菌后测定了水稻幼苗叶片内丙二醛含量及相对电导率的变化。结果表明：表现非亲和反应的水稻H8R叶片中丙二醛含量及相对电导率在接种后18 h开始增加,均于30 h和60 h形成一小一大的峰值,这正是水稻叶片过敏性坏死斑出现到形成时;而在亲和反应的H8S叶片中二者均只在接种后48～72 h间有一个相对升高期,其增幅均低于非亲和反应。同时证明,接种后30 h,H8R叶片中亚麻酸和亚油酸含量明显降低,而在亲和反应中基本不变。非亲和反应的水稻叶片中脂氧合酶活性在接种后18 h开始上升,在24 h和72 h分别出现两个峰值;亲和反应中该酶活性在互作后期（60～72 h）略有增加。这些结果说明脂氧合酶活性变化与膜脂过氧化表现明显的时间动态相关,膜脂过氧化引起的电解质渗漏是过敏性反应的主要生理机制之一。     

Hydroperoxides of fatty acids induce programmed cell death in tomato protoplasts

Arachidonic acid (AA) is a fatty acid elicitor abundant in the glycerolipids of the late blight pathogen Phytophthora infestans and related Oomycete species. Lipoxygenases (LOX), which catalyze the addition of molecular oxygen to the 1 or 5 position of a cis, cis -1,4- Z, Z -pentadiene system in polyunsaturated fatty acids, is induced in host plants such as tomato and potato during infection by P. infestans. Here it is reported that AA, the LOX metabolites of AA, 5- and 15-hydroperoxyeicosatetraenioc acid (5- and 15-HpETE), and the LOX metabolite of linoleic acid, 9-hydroperoxyoctadecadienoic acid (9-HpODE), are potent inducers of programmed cell death (PCD) in tomato protoplasts. 5- and 15-HpETE increased DNA fragmentation as detected by t erminal deoxynucleotidyl transferase-mediated d U TP-X n ick e nd l abeling (TUNEL) and increased DNA laddering as visualized by ligation-mediated PCR. Background levels of DNA laddering were decreased in intensity by Zn²⁺ and increased by Ca²⁺, effects that are consistent with the reported action of these cations on PCD-associated endonucleases in other systems. H₂O₂, methyl jasmonate, and linoleic and linolenic acids were not toxic to tomato protoplasts at concentrations up to 350 μ , and lipid peroxides (LD₁₀₀ = 80 μ ) were far more potent inducers of death than free AA within this same concentration range. These results indicate the potential of fatty acid peroxides and LOX-related metabolism to engage an apoptotic type of PCD in higher plant cells.     

Genotypic lipoxygenase variation in soybean seeds and response to nitrogen nutrition

Decreasing the lipoxygenase content of soybean (Glycine max (L.) Merr.) seed is a prerequisite for its increased use in human foods. These investigations aim to analyse the variation of lipoxygenase contents in relation to the genotype or the nitrogen nutrition of the plant. Four genotypes from a divergent selection for N₂ fixation and the recurrent parent, Weber, were grown in pots, and two Bradyrhizobium japonicum strains (G.49 and SMGS1) were used. Dinitrogen fixation was estimated in situ using the acetylene–reduction assay. The lipoxygenase activity was determined polarographically on seeds taken from plants grown under controlled conditions in the presence or absence of symbiotic N₂ fixation. The large genotypic variation observed for the lipoxygenase contents was enhanced by high N₂ fixation. The variation range of lipoxygenase contents, which was obtained through the combination of the genotypic variation and the N₂ fixation activity, was relatively large with a ratio of 2 on a seed dry–weight basis and a ratio of 2.6 when the total proteins were considered. Decreased lipoxygenase content and increased total protein content can thus be considered in parallel in a genetic context, together with more efficient N₂ fixation.     

罗非鱼鳃组织中脂肪氧合酶的性质研究

试验结果表明脂肪氧合酶最适反应温度为30℃,最适pH为10.0和4.0。在pH10.0的条件下,最适底物浓度为2.5×10-4mol/L,Km值为0.073mmol/L,Vmax值为3.08×104U/mgprotein.min。EDTA对该酶有较强的抑制作用。而在pH4.0的条件下,最适底物浓度为5×10-4mol/L,BHA、BHT、TBHQ等抗氧化剂有较强的抑制作用。     

莲原花青素在油脂体系中的抗氧化作用

【目的】研究莲原花青素在油脂体系中的抗氧化作用,为开发新型健康的天然食品抗氧化剂提供理论依据。【方法】通过添加不同浓度的原花青素以及将原花青素和卵磷脂等抗氧化剂复配,研究原花青素在油酸和各类动植物油脂中的抗氧化作用。采用亚油酸氧化体系表征原花青素对脂肪氧合酶活性的抑制。【结果】原花青素对油酸和动植物油脂均显示不同程度的抗氧化效果,对试验所用的油脂抗氧化效果依次为:茶籽油猪油棉籽油豆油,其中茶籽油的氧化诱导期延长了2.2倍;并且卵磷脂和维生素E对原花青素有很好的增效作用,尤其与维生素E的复配,能将油酸的诱导期延长14.7倍,猪油的诱导期延长3.6倍。在抑制脂肪氧合酶活性方面,原花青素与茶多酚有着相同的效果。【结论】原花青素对油脂体系有着优良的抗氧化作用,对油脂氧化抑制效果显著。     

小麦TaLox-B等位变异对脂肪氧化酶活性和面粉色泽的影响

【目的】小麦籽粒脂肪氧化酶(Lox)与面粉色泽密切相关,研究TaLox-B位点上的不同等位变异类型对Lox活性和面粉色泽的影响,为面粉色泽的改良和相关的品质育种工作提供参考依据。【方法】选用122份河南小麦品种(系)为试验材料,用紫外分光光度计和色差仪分别测定其Lox活性和面粉色泽,并利用控制脂肪氧化酶活性位于4BS上的TaLox-B1、TaLox-B2和TaLox-B3位点上的功能标记Lox16、Lox18和Lox-B23对参试材料的Lox基因型进行鉴定。【结果】不同品种间Lox活性及其色泽性状差异达到极显著水平。基因型鉴定结果表明,参试材料的TaLox-B1位点存在TaLox-B1a和TaLox-B1b两种等位变异,所占比例分别为63.9%和39.1%,TaLox-B2位点存在TaLox-B2a和TaLox-B2b两种等位变异,所占比例分别为57.4%和42.6%,TaLox-B3位点也存在TaLox-B3a和TaLox-B3b两种等位变异,所占比例分别为41.8%和48.2%。分析其基因型组合发现,参试材料中共有6种基因型组合类型,依次为TaLox-B1a/TaLox-B2a/TaLox-B3a、TaLox-B1a/TaLox-B2a/TaLox-B3b、TaLox-B1a/TaLox-B2b/TaLox-B3b、TaLox-B1b/TaLox-B2a/TaLox-B3a、TaLox-B1b/TaLox-B2a/TaLox-B3b和TaLox-B1b/TaLox-B2b/TaLox-B3b,所占比例分别为41.8%、15.6%、6.6%、28.7%、5.7%和1.6%。分析不同TaLox-B位点基因与Lox活性及红度(a*值)、黄度(b*值)、亮度(L*值)、白度(Wht值)等面粉色泽性状的关系表明,单基因等位变异对Lox活性和面粉色泽的影响不同,3个基因等位变异a*值差异均不显著,对Lox活性的效应,TaLox-B2a高于TaLox-B2b(P0.05),TaLox-B3a高于TaLox-B3b(P0.01),TaLox-B2a基因型的Wht值低于TaLox-B2b基因型(P0.01),TaLox-B3a基因型的Wht值也低于TaLox-B3b基因型(P0.05)。说明TaLox-B2和TaLox-B3对Lox活性和面粉色泽具有重要影响。进一步分析发现,拥有TaLox-B1a/TaLox-B2a/TaLox-B3a基因型组合的小麦品种(系)的Lox活性和b*值最高,a*值和Wht值最低,而TaLox-B1b/TaLox-B2b/TaLox-B3b基因型组合的小麦品种(系)的L*值、a*值和Wht值最高,其Lox活性和b*值最低。【结论】河南小麦中所发现的6种不同Lox基因型组合中,拥有TaLox-B1a/TaLox-B2a/TaLox-B3a基因型组合的小麦品种(系)的Lox活性相对较高,Wht值相对较低(P0.05),而拥有TaLox-B1b/TaLox-B2b/TaLox-B3b基因型组合的小麦品种(系)的Lox活性相对较低,Wht值相对较高(P0.05)。Lox活性的遗传控制在面粉色泽品质改良进程中发挥着关键作用。     

Pepper 9- and 13-lipoxygenase genes are differentially activated by two tobamoviruses and by hormone treatments

Two novel pepper 13-lipoxygenase (LOX) genes were cloned and their expressions were compared with those of three 9-LOX genes in pepper leaves inoculated with two different tobamoviruses. Obuda pepper virus (ObPV) inoculation led to a massive induction of pathogenesis-related genes and to the development of hypersensitive reaction (incompatible interaction), while Pepper mild mottle virus (PMMoV) inoculation resulted in a compatible interaction. Both virus infections markedly activated the expression of the two novel 13-LOXs. The magnitudes of induction of 13-LOXs did not differ substantially between the ObPV- and PMMoV-inoculated leaves. The induction of three 9-LOXs was markedly more robust and rapid in ObPV-inoculated leaves than in PMMoV-inoculated ones. LOXs were very differentially activated in pepper leaves treated with defense hormones. A large number of hormone-related cis-regulatory elements were identified in the promoter regions of LOXs. ObPV inoculation resulted also in the substantial up-regulation of an omega-6-fatty acid desaturase gene. Our results suggest that 9-LOX-dependent pathways are more probably involved in the suppression of virus replication than 13-LOX-dependent plant responses.     

Differential responses of Brachypodium distachyon genotypes to insect and fungal pathogens

To increase understanding of the interactions between Brachypodium distachyon (purple false brome) and its pathogens, six diploid and two hexaploid Plant Introductions (PI) lines were assessed for their resistance/susceptibility to nine economically important fungal pathogens and two species of insect pests affecting closely related grass species. Naturally occurring variation in resistance was found, with two lines being the most resistant and one line being the most susceptible to most of the insects and pathogens tested. Evidence was found for differential activation of key genes in pathogen defense response pathways between susceptible and resistant lines.     

黄淮麦区部分小麦种质脂肪氧化酶活性分析及等位基因检测

【目的】小麦中的脂肪氧化酶（LOX）是影响小麦面粉颜色和其储藏特性的主要因素之一,对中国黄淮麦区的306份小麦种质资源进行脂肪氧化酶活性分析及等位基因检测,为小麦品质育种提供理论参考。【方法】利用紫外分光光度计和酶标仪测定参试样品的脂肪氧化酶（LOX）活性,并利用控制脂肪氧化酶活性的位于1AL上的QLpx.caas-1AL位点紧密连锁的分子标记Xwmc312以及位于4BS上的TaLOX-B1位点的功能标记LOX16和LOX18,采用特异引物的PCR扩增技术以及琼脂糖凝胶电泳和聚丙烯酰胺凝胶电泳分离技术进行参试材料的LOX基因型鉴定。【结果】表型测定结果表明,参试的黄淮麦区小麦品种（系）中脂肪氧化酶活性的平均值为65.73 AU·min^-1·g^-1,标准差为13.54,变幅为27.09-99.55 AU·min^-1·g^-1,变异系数为20.6%。参试材料中小于40 AU·min^-1·g^-1和大于90 AU·min^-1·g^-1的小麦品种（系）分别为7个和8个,可作为对当前主栽小麦品种进行脂肪氧化酶活性改良的重要资源材料。基因型鉴定结果表明,参试材料的QLpx.caas-1AL位点存在3种等位变异类型,Xwmc312₂₂₇、Xwmc312₂₃₅和Xwmc312₂₄₇所占比例分别为30.4%、18.9%和50.6%。参试材料的TaLOX-B1位点存在TaLOX-B1a和TaLOX-B1b两种变异类型,所占比例分别为 28.7%和71.2%。分析其基因型组合发现,参试材料中共有6种基因型组合类型,依次为TaLOX-B1a/Xwmc312₂₂₇、TaLOX-B1a/Xwmc312₂₃₅、TaLOX-B1a/Xwmc312₂₄₇、TaLOX-B1b/Xwmc312₂₂₇、TaLOX-B1b/Xwmc312₂₃₅和TaLOX-B1b/Xwmc312₂₄₇,所占比例分别为7.2%、9.5%、12.1%、23.2%、9.5%和38.6%。分析不同LOX基因型与脂肪氧化酶活性的关系表明,1AL位点上拥有Xwmc312₂₃₅基因型的小麦品种脂肪氧化酶活性显著高于其他两种基因型（P＜0.05）,4BS位点拥有TaLOX-B1a基因型的小麦品种脂肪氧化酶活性显著高于拥有TaLOX-B1b基因型的小麦品种（P＜0.05）。进一步分析不同LOX基因型组合与脂肪氧化酶活性的关系表明,拥有TaLOX-B1a/Xwmc312₂₃₅基因型组合的小麦品种脂肪氧化酶活性（76.80 AU·min^-1·g^-1）显著高于其他5种基因型组合,而拥有TaLOX-B1b/Xwmc312₂₂₇基因型组合的小麦品种脂肪氧化酶活性显著低于其他5种基因型组合,仅为62.44 AU·min^-1·g^-1。【结论】中国黄淮麦区的小麦品种脂肪氧化酶活性绝大多数处于中间类型,拥有极低（小于40 AU·min^-1·g^-1）或极高（大于90 AU·min^-1·g^-1）脂肪氧化酶活性类型的小麦品种所占比例较低。黄淮麦区所发现的6种不同LOX基因型组合中,拥有TaLOX-B1a/Xwmc312₂₃₅基因型组合的小麦品种脂肪氧化酶活性相对较高（P＜0.05）,而拥有TaLOX-B1b/Xwmc312₂₂₇基因型组合的小麦品种脂肪氧化酶活性相对较低（P＜0.05）。     

龙眼采后果肉脂氧合酶活性和膜脂脂肪酸组分的变化

以‘福眼’龙眼果实为试验材料,分析果实采后贮藏过程中脂氧合酶(LOX)活性和细胞膜脂脂肪酸组分的变化,探讨其与果实成熟衰老的关系。结果表明,龙眼果肉脂肪酸组分主要有豆蔻酸(14:0)、棕榈酸(16:0)、硬脂酸(18:0)、油酸(18:1)和亚油酸(18:2),未检出亚麻酸。随着龙眼采后贮藏时间的延长,不饱和脂肪酸(油酸、亚油酸)和饱和脂肪酸(豆蔻酸、棕榈酸、硬脂酸)的含量变化不明显。果皮中未检出脂肪酸。果肉中的LOX活性伴随着膜脂过氧化作用的加强而逐渐增强,但由于LOX的活性较低,所以上升的幅度相对来说比较小,推测LOX可能不是导致果实衰老腐烂的关键酶。