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。     

降温方法对不同采收期鸭梨采后果心褐变和膜脂组分的影响

为了探讨缓慢降温抑制采后鸭梨果实褐变的机理,研究了不同降温方法对不同采收期鸭梨果心膜脂脂肪酸组分、含量、膜相变温度、脂氧合酶（LOX）活性以及果心褐变的影响。结果表明：鸭梨果心含有月桂酸、豆蔻酸、棕榈酸、棕榈油酸、珠光酸、硬脂酸、油酸、亚油酸、亚麻酸,其中含量较多的是亚油酸、棕榈酸和油酸。缓慢降温提高了早采鸭梨果心的亚麻酸和亚油酸相对含量及不饱和脂肪酸与饱和脂肪酸比值（U/S）,降低了果心的膜相变温度,明显抑制了果心LOX活性和褐变。推迟采收提高了鸭梨果心的U/S值,降低了膜相变温度,但由于采收期晚,果实衰老较快,后期果心LOX活性快速升高,膜脂过氧化严重,导致晚采果更容易褐变。总之,适当早采结合缓慢降温可以提高鸭梨果心膜脂不饱和脂肪酸含量和U/S值,抑制果心褐变。     

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。     

苹果果实发育成熟软化过程中脂氧合酶活性变化及采后乙烯调控

以富士、金冠和嘎拉苹果果实为试材,研究了脂氧合酶（LOX）在果实生长发育至采后软化过程中的变化规律及采后乙烯调控对其活性的影响。结果表明：在果实发育及后熟过程中,富士、金冠和嘎拉3个品种果实的LOX活性呈相似的变化规律,在幼果期均出现较高活性高峰,之后下降,至成熟后果实LOX活性又开始提高。品种间比较结果显示,虽然富士果实LOX在幼果期出现较高峰值,但之后下降迅速直至采收初期,而此间金冠和嘎拉果实LOX活性及增加速率均高于富士,说明LOX活性表达状况可能是衡量果实贮藏特性的一个重要因素。采后乙烯调控后,乙烯抑制剂1-MCP能极显著地抑制苹果果实LOX活性,在贮藏期间其活性基本恒定,而乙烯利对其活性虽有促进作用,但不如1-MCP的作用显著,因品种贮藏性不同而有所差异,其确切机理有待进一步探讨。     

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。     

荔枝果实采后脂氧合酶活性的变化

研究了采后冷藏、外源ABA、亚精胺(Spd)、1-甲基环丙烯(1-MCP)、热处理及酸处理对荔枝果实脂氧合酶(LOX)活性变化的影响。荔枝果实组织中LOX活性测定反应系统的最适pH是6．5。与3℃冷藏相比,0℃下糯米糍果皮LOX活性升高较快,丙二醛(MDA)含量积累加快,在21d时分别高出34．9％和25．6％。0℃下桂味果皮LOX活性先升后降,于14d达到高峰。冷害促进了果皮LOX活性增加,但0℃下糯米糍果肉的LOX活性和MDA含量均低于3℃下果肉。外源ABA、Spd、1-MCP处理不同程度地降低了果皮LOX活性,减轻了果实冷害的发生程度。推测由LOX主导的膜脂过氧化作用引起的膜结构破坏在荔枝果实衰老和冷害褐变中可能起重要作用。46．2～80．0℃的热水处理诱导LOX活性的升高,适当浓度的酸则显著抑制该酶的活性。     

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.     

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.     

甘蔗脂氧合酶基因ScLOX1的克隆与表达分析

LOX属于脂氧合酶超家族(lipoxygenase superfamily),是脂肪氧化途径的重要因子,广泛参与植物生长发育的调节和对外界刺激的抵御。本研究基于甘蔗(Saccharumspp.)转录组数据库,通过RT-PCR技术,首次从新台糖22号(ROC22)蔗芽中克隆获得ScLOX1基因(GenBank登录号为MK106188)的cDNA全长序列。生物信息学分析显示,ScLOX1基因cDNA序列长度为2813 bp,开放读码框全长2664 bp,编码887个氨基酸,其编码蛋白的理论等电点为6.23,不稳定系数为39.77,亲水性平均值为-0.437,无信号肽和跨膜结构,但含有PLATLH2和Lipoxygenase活性位点,与高粱(Sorghum bicolor) LOX (XP002466613.1)的氨基酸序列相似性高达95.96%。预测ScLOX1基因的编码蛋白为酸性稳定亲水性非分泌蛋白,属于type I类非传统9-LOX。qPT-PCR分析结果显示,ScLOX1基因在蔗芽组织中特异性表达。接种甘蔗黑穗病菌(Sporisorium scitamineum)后, ScLOX1基因的表达量在抗病品种崖城05-179中短暂上升,但在感病品种ROC22中显著下降。分别对瞬时表达ScLOX1基因的本氏烟(Nicotiana benthamiana)植株叶片接种烟草茄病镰刀菌蓝色变种(Fusarium solani var. coeruleum)和青枯菌(Ralstonia solanacearum),表型观察、3,3’-二氨基联苯胺(3,3’-diaminobenzidine,DAB)染色和烟草免疫相关基因的表达情况分析显示,ScLOX1基因的过表达能够增强本氏烟对烟草茄病镰刀菌蓝色变种的防御,但对烟草青枯菌的作用与对照相比无明显差异。研究还发现,ScLOX1基因的表达受茉莉酸甲酯和水杨酸抑制下调,但受脱落酸、氯化钠和聚乙二醇诱导上调。以上结果为深入研究甘蔗ScLOX1基因的功能提供参考资料。