抗褐变剂对阿月浑子外植体褐变以及几种酶活性的影响

为了探明阿月浑子组织培养过程中的褐变机制,以阿月浑子组培丛生芽为外植体,在其继代培养基(1/2DKW)中分别添加抗褐变剂PVP和Cys,分析组培苗叶片、茎段和愈伤组织的总酚含量以及PPO、POD、PAL、C4H酶活性的变化,研究其与褐变的关系。结果表明,在一个继代培养周期内,愈伤组织中PPO活性呈增加趋势,而叶片和茎中PPO活性呈"V"字形变化。POD活性随继代时间增长而逐渐升高,继代28 d后开始下降,愈伤组织中POD活性变化范围较叶片和茎段中要高一个数量级。PAL活性总体呈下降趋势,而C4H酶活性在继代前期上升,继代14 d或21 d后下降。结合相关性分析可知,阿月浑子各组织的总酚含量与褐变率呈显著正相关,并且POD酶在促褐变反应中发挥主要作用。在实验中发现,PVP与Cys虽然对POD、PPO、CH4酶活性也有一定影响,但其主要是通过抑制PAL的酶活性,显著减少了组培苗和愈伤组织的酚类物质含量,从而降低褐变率。     

儿茶素对蝴蝶兰叶片离体培养褐变发生的影响

用0.3 g/L儿茶素处理蝴蝶兰叶片外植体,在MS+3 mg/L 6-BA(pH 5.8)培养基上培养4 d,外植体褐变率高于对照227.6%.用香草醛-盐酸溶液染色反应法发现鞣质分布在维管束和细胞间隙.叶片剪切的外植体(培养0 d)鞣质含量较高,培养4 d降低,8 d后再升高.儿茶素处理的外植体鞣质含量明显高于对照.在培养期间,儿茶素处理的外植体PPO、POD和PAL活性高于对照外植体,其中POD活性在第8天达到最大值,PPO和PAL活性在培养第12天达到高峰.     

白及原球茎褐变过程中总酚及相关酶活性的变化

以不同褐变程度的白及原球茎为试材,研究了白及原球茎褐变度与总酚含量、多酚氧化酶(PPO)、过氧化物酶(POD)、苯丙氨酸解氨酶(PAL)活性的关系.结果表明:在一定褐变度范围内,随褐变程度的增加,总酚含量升高,超过一定范围,总酚含量随褐变程度增加而降低;PPO与PAL活性变化趋势一致,在褐变前期与褐变度呈正相关,后期活性随褐变度增加而降低;POD活性随褐变度增加而略有降低.     

鲜切生菜贮藏期间产品品质变化及褐变发生规律

为了研究鲜切生菜贮藏期间品质变化和褐变发生的规律及其内在机理,以新鲜圆生菜为试材,经切分处理,置于4℃下贮藏,观测贮藏期间鲜切生菜外观品质、主要营养成分以及与褐变相关的褐变度,苯丙氨酸解氨酶(PAL)、多酚氧化酶(PPO)、过氧化物酶(POD)活性变化规律。结果表明:随着贮藏时间的延长,经过切分处理的鲜切生菜的含水量逐步降低,失重率增大,后期腐烂明显;主要营养成分叶绿素、还原型维生素C(VC)、可溶性蛋白质含量均出现不同程度的下降;褐变度呈现上升趋势,组织中的PAL活性和总酚含量先升高后逐步下降;PPO活性在整个贮藏时期内呈现出下降趋势,POD活性在贮藏的后期迅速升高。综合分析认为,切分损伤能够诱使PAL活性提高,并促进酶促褐变底物酚类物质的大量积累,通过PPO和POD协同作用,加剧褐变发生;鲜切生菜随贮藏时间延长,其综合品质下降,建议最佳食用期0~3d,货架期0~8d。     

黄金梨CA贮藏中果心褐变与相关理化因子的关系

文章研究了不同气体成分对黄金梨果心褐变指数的影响,分析了果心酚类物质、丙二醛(MDA)含量以及多酚氧化酶(PPO)、过氧化物酶(POD)、过氧化氢酶(CAT)活性与果心褐变的关系。结果表明,MDA含量、PPO活性与果心褐变指数呈正相关性,CAT活性与果心褐变指数呈负相关性。较高CO_2和O_2条件促进了果心酚类物质、MDA含量以及PPO活性升高和POD、CAT活性下降,果心褐变发生较早、程度较重。本试验在较低CO_2(1%)和O_2(5%)条件下可使黄金梨贮藏180d以上。     

不同品种菊芋总酚含量、PPO活性、POD活性及其与褐变的关系

对5个菊芋品种的总酚含量、PPO活性、POD活性与褐变强度进行了测定。结果表明：不同菊芋品种的总酚含量、PPO活性、POD活性与褐变强度差异显著,其中3个北方品种的各项指标均高于2个南方品种。相关性分析显示,4个外形呈不规则瘤形品种的PPO活性与褐变强度之间呈显著正相关,相关系数r=0.982。褐变主成分分析表明：PPO活性、POD活性和总酚含量这3个变量共同影响菊芋的褐变。     

不同气体配比对丰水梨采后贮藏期间褐变相关理化因子的研究

以丰水梨为试材,经过5组不同的气体配比处理,分析低温(0±0.5)℃条件下贮藏过程中果实呼吸速率、乙烯释放量、相对电导率、丙二醛(MDA)含量、果心褐变指数、多酚氧化酶(PPO)活性、过氧化物酶(POD)活性的变化。结果表明:丰水梨是呼吸跃变型果实,适宜的气体配比可以降低果实褐变指数的上升,推迟果实PPO、POD活性高峰的出现时间,抑制酚类物质降解,减少MDA生成量,延缓果实褐变发生时间,其中以5%O2+1%CO2处理组果实抑制褐变效果最好。果皮中的酚类物质含量和PPO活性最高,其次是果心,最后是果肉,而POD活性依次为:果皮果肉果心,但整个贮藏过程中,果皮与果肉均没有发生褐变。     

不同光照强度对蝴蝶兰组培中外植体褐化的影响

本试验研究了不同光照强度培养下,蝴蝶兰初代培养过程中外植体内多酚氧化酶(PPO)活性、总酚含量以及褐化百分率的变化规律.结果表明:1 000Lx和3 000Lx光强培养可降低褐化率,2000Lx光强培养的外植体褐化严重且PPO活性高于其他两处理,差异显著;三处理间总酚含量差异不显著.二者相关性分析表明,PPO活性与外植体中总酚含量呈中等相关.     

不同丝瓜品种褐变相关基因的表达分析

以7种不同丝瓜品种为试验材料,对采后丝瓜果实进行褐变鉴定,测定多酚氧化酶(PPO)活性、过氧化物酶(POD)活性及相关基因的表达量。结果表明:"苏丝3号""苏丝6号""苏丝7号"属于抗褐变品种;"苏丝4号""苏丝5号""苏丝10号"属于耐褐变品种;"苏丝8号"属于严重褐变。从褐变酶POD和PPO变化来看,"苏丝8号"POD活性高达456.7 U·mg-1,抗褐变品种"苏丝3号""苏丝6号""苏丝7号"POD活性均在250 U·mg-1。"苏丝8号"PPO活性为68.2 U·mg-1,"苏丝3号"PPO活性最小,低至25.2 U·mg-1。从褐变基因的变化来看,"苏丝8号"在LcPOD家族基因表达中均高于对照"苏丝3号",其中LcPOD3基因表达量高达5.23,是对照的5倍以上,其它的丝瓜品种LcPOD3基因变化不显著。褐变品种"苏丝8号"在LcPPO基因家族中上调显著,表明"苏丝8号"果实褐变时促进LcPPO基因家族的表达,从而转录翻译成PPO加快果实的褐变。与对照相比,丝瓜褐变基因LCWRKY在"苏丝8号"中上调显著,表明其参与了丝瓜果实采后褐变。其它丝瓜品种LCWRKY上调不显著。     

柑橘衰退病毒侵染对寄主植物同工酶及酸性病程相关蛋白表达的影响

以柑橘衰退病毒(Citrus tristeza virus,CTV)不同致病力株系接种寄主植物,比较分析了寄主植物体内过氧化物酶(peroxidase,POD)和多酚氧化酶(polyphenol oxidase,PPO)活性变化特点。结果表明,该病毒侵染对这2种酶活性的影响因寄主品种和CTV株系而异,CTV强毒和弱毒株系侵染均导致墨西哥莱檬植株中POD活性明显降低;在甜橙上,CTV强毒株系接种植株中POD活性则显著性提高,而弱毒株系接种对POD的活性无明显影响;在对该病毒具有较高抗性的枳壳中,接种CTV不同株系对POD活性均无明显影响。而在接种强毒株系N25的墨西哥莱檬中PPO活性有所增强,在枳壳上则表现为接种植株的PPO活性降低,在甜橙上PPO活性无明显的变化。POD、PPO和酯酶(esterase,EST)同工酶酶谱分析的结果显示,CTV弱毒株系接种枳壳后诱导产生了2条新的EST同工酶谱带,而在其他CTV接种植株上均未诱导表达新的POD、PPO及EST酶谱。酸性病程相关蛋白分析结果显示,CTV强毒株系在甜橙上诱导产生有6条明显的酸性病程相关蛋白条带。     

蝴蝶兰外植体酚类物质和活性氧代谢与组培褐变的关系

（1Institute of Vegetables and Flowers,Chinese Academy of Agricultural Sciences,Beijing 100081,China;2College of Horticulture and Landscape Architecture,Southwest University,Chongqing 400716,China）     

梨果实发育过程中石细胞团及几种相关酶活性变化的研究

以梨品种西子绿、菊水及砀山酥梨为试材,对不同发育时期梨果实石细胞团的密度、大小及几种相关酶的活性变化进行了研究。结果表明石细胞团的分布密度在幼果期较高,以后随着果实的发育和膨大,密度逐渐减小,接近成熟前1个月左右基本稳定;石细胞团的直径随果实的生长而逐渐变大;苯丙氨酸解氨酶在果实发育初期表现出较高的活性,随着果实的生长逐渐降低,花后75d后保持低水平不变;过氧化物酶活性高峰出现稍迟于苯丙氨酸解氨酶,高峰后呈现出下降的趋势;多酚氧化酶活性波动较大,总的变化趋势逐渐下降。品种间苯丙氨酸解氨酶、过氧化物酶活性与石细胞团的形成之间表现为正相关。     

一氧化氮熏蒸处理对双孢蘑菇采后褐变及衰老的影响

以双孢蘑菇为试材,研究了在(10±0.5)℃条件下,不同浓度一氧化氮(NO) (30、60、90 μL/L)气体熏蒸处理对双孢蘑菇贮藏过程中感官品质、褐变度、多酚氧化酶(PPO)和过氧化物酶(POD)活性的影响.结果表明:外源NO熏蒸处理显著抑制了PPO活性的升高(8.90％～33.13％),减轻贮藏期的褐变程度.提高了贮藏后期POD活性(12.28％～76.27％),延缓了双孢蘑菇的衰老,有效延长了货架期(2 d).其中,NO浓度为60 μL/L时效果最好.     

茭白肉质茎膨大过程中3种保护酶的活性变化

以双季茭白浙茭3号、浙茭5号为试验材料,选择不同生长阶段的肉质茎,分别测定多酚氧化酶(PPO)、过氧化氢酶(CAT)、过氧化物酶(POD)的活性。试验结果表明,在茭白肉质茎的膨大过程中,PPO、CAT的活性呈膨大前期下降,后期上升的趋势。PPO的活性在生长后期有明显的上升。POD的活性总体呈下降趋势,生长后期活性趋于稳定。浙茭3号、浙茭5号3种保护酶的变化趋势一致,但品种间不同酶的活性有差异。     

鸭梨果心褐变过程中膜质过氧化及细胞内膜微粘度的变化

采用急降温和缓降温两种方法处理鸭梨果实，并对冷藏中果心褐变指数、保护酶活性、膜质过氧化水平和细胞内膜微粘度进行测定。结果表明：急降温处理的果实约３０天果心出现褐变症状，６０天后褐变指数达到１００％，而缓降温处理的果实在贮藏中无褐变产生。在６０天的贮藏过程中，急降温处理的果实果心组织的超氧化物歧化酶（ＳＯＤ）、过氧化物酶（ＰＯＤ）、过氧化氢酶（ＣＡＴ）活性均呈下降趋势，丙二醛（ＭＤＡ）水平和细胞内膜微粘度则显著升高。缓降温果实果心组织中ＳＯＤ、ＰＯＤ活性呈上升趋势，ＣＡＴ活性虽呈下降趋势，但下降水平明显低于急降温处理。ＭＤＡ水平及细胞内膜微粘度的上升率明显低于急降温处理组。实验结果说明：急降温处理导致果心细胞膜质过氧化作用加剧，同时造成细胞内膜流动性下降，从而造成细胞膜系统组分及功能发生变化，导致细胞中区域化分布被打破，造成果实组织褐变。     

Variations in contents of browning substrates and activities of some related enzymes during litchi fruit development

Pericarp browning is a major factor to limit storage and transportation of postharvest litchi fruit. Experiments were conducted firstly to determine variations in contents of browning substrate (−)-epicatechin in pericarp tissues of litchi fruit during development. Changes in the activities of polyphenoloxidase (PPO), peroxidase (POD) and phenylalanine ammonialyase (PAL) in association with the synthesis or oxidation of (−)-epicatechin were also examined. (−)-Epicatechin content tended to decrease during fruit development. PAL activity had a high level at the initial stage of fruit development, then decreased rapidly, and finally increased slightly, which was in agreement with the variation in (−)-epicatechin content. The higher contents of total phenolics and (−)-epicatechin of fruit of ‘Nuomici’ than ‘Feizixiao’ were also related to the higher PAL activity. PPO had a similar change to PAL in activity, but POD activity slowly decreased at the early fruit development and then increased markedly. It is suggested that (−)-epicatechin content might be largely regulated by PAL activity. This study could help to understand the variations in the content of the browning substrate and the activities of some substrate-related enzymes of litchi fruit during development and then control better pericarp browning of harvested litchi fruit by the application of appropriate preharvest treatments.     

Inhibition of powdery mildew development and activation of antioxidant enzymes by induction of oxidative stress with foliar application of a mixture of riboflavin and methionine in cucumber

Powdery mildew (Sphaerotheca fuliginea Pollacci) infection in cucumber was significantly reduced by foliar application of a mixture of riboflavin and methionine (RM). The effects of fungicidal activity on leaves applied with RM were detected through restriction of progress of colonies and disease severity compared with control plants. The initial response to foliar application of RM was abrupt generation of hydrogen peroxide in the leaves of cucumber plants. Activities of antioxidant enzymes such as SOD and POD were abruptly increased by foliar application of RM. However, activities of antioxidant enzymes in control plants were increased with disease development 9 d after pathogen inoculation. Cucumber leaves have six major SOD isoforms. When plants were foliar-applied with RM, densities of three SOD isozyme bands at SOD-1, SOD-2, and SOD-3 were increased 3 d after foliar application. Leaves of cucumber plants have three major POD isozyme bands. Densities of three POD isozyme bands were increased 3 d after foliar application with RM. Four major PPO isozyme bands were determined in cucumber leaves. Though the overall banding patterns of PPO in control and RM-applied plants were similar, the band profiles in leaves applied with RM were characterized by high densities of the three major isoforms. Activities of PPO in leaves applied with RM increased rapidly during the 3 d after foliar application, and then remained relatively constant for 15 d. Although activities of PPO in the leaves of control plants also abruptly increased after 9 d, it was lower than those of RM-applied plants during the whole time. The difference in lignin content between control and RM-applied plants was detected 9 d after foliar application; it was high in leaves applied with RM.     

阿月浑子外植体褐变抑制方法

外植体褐变是阿月浑子组培的主要障碍之一。对阿月浑子外植体母树年龄、遮光、外植体枝条部位与采集季节、组培条件、添加剂以及多酚氧化酶活性等与外植体褐变的关系进行了研究,结果表明,阿月浑子外植体的褐变率夏季最高、秋季其次、春季最低,枝条中部最低,而枝条上端最高,1年生与7年生树褐变率差异不大,与母树是否遮光关系不明显,在培养基中添加抑制剂PVP能减轻褐变,而添加维生素C、Na2S2O3与活性碳的效果不显著,低光和低温对控制外植体褐变有显著效果,但从低温下转入常温后褐变率高于一直在常温下培养的,阿月浑子褐变率与多酚氧化酶活性关系不显著,可能主要决定于外植体多酚含量,但需要进一步实验数据支持。