Effects of Ceratocystis fimbriata on phenolics content, PPO and PAL activity in sweet potato

选用对甘薯黑斑病抗性不同的品种南京-92(高抗)和烟台-252(高感)的叶片为材料,研究黑斑病对甘薯叶总酚含量、绿原酸含量、类黄酮含量以及苯丙氨酸解氨酶(PAL)活性和酚氧化酶(PPO)活性的影响.结果表明:在未受黑斑病侵染时,南京-92叶片中类黄酮含量、绿原酸含量、PAL活性显著或极显著高于烟台-252,可以作为选育和鉴定抗黑斑病品种的生理指标,但总酚含量和PPO活性差异不显著.接种后2～8 d内,南京-92叶片内总酚含量和PPO活性增加迅速,与烟台-252达到显著或极显著差异,总酚含量和PPO活性的上升速度快、保持时间长,有利于提高对黑斑病的抵抗能力.     

南方根结线虫侵染对2个不同抗性番茄砧木幼苗苯丙烷类代谢的影响

为明确不同抗性番茄(Lycopersicon esculentum)砧木幼苗苯丙烷类代谢与抗南方根结线虫(Meloidogyne incognita)的关系,以高感砧木‘Ls-89’和高抗砧木‘坂砧2号’为试材,采用盆栽人工接种二龄幼虫的方法,研究番茄砧木幼苗苯丙烷类代谢的变化。结果表明,未接种根结线虫时,‘坂砧2号’的根系PAL活性与‘Ls-89’无显著差异,而根系PPO活性及叶片PAL、PPO活性和总酚、类黄酮、木质素含量却显著高于‘Ls-89’。接种根结线虫后,两砧木根系与叶片的PAL、PPO活性与总酚、类黄酮含量均呈周期性升高,木质素含量也逐渐升高,但‘坂砧2号’的升幅均显著高于‘Ls-89’。在一个侵染周期内,‘坂砧2号’根系PAL、PPO活性与总酚、类黄酮含量的峰值分别比‘Ls-89’高28.45%、71.47%、54.24%、91.71%,而叶片的分别高47.29%、64.00%、23.33%、52.61%。这说明南方根结线虫侵染使砧木幼苗苯丙烷类代谢活性显著增强,且‘坂砧2号’显著高于‘Ls-89’,表明番茄砧木幼苗苯丙烷类代谢与抗南方根结线虫水平密切相关。     

UV-B对不同发育时期离体蓝莓主要果实品质及相关酶活性的影响

该研究以幼果期、白果期、转色期的离体‘北陆’蓝莓果实为试材,设置0(CK)、5、10、15min紫外光辐照处理,24h后取样分析蓝莓果实中可溶性糖、总酚、类黄酮和花青苷含量,以及苯丙氨酸裂解酶(PAL)和查尔酮异构酶(CHI)活性的变化,探究UV-B紫外照射处理对不同发育时期蓝莓主要果实品质及相关酶活的影响。结果显示:(1)对于幼果期蓝莓,5min UV-B处理可显著增加果实内可溶性糖含量;10min UV-B处理果实PAL活性增加效果最为显著;15min UV-B处理对果实总酚和花青苷积累的促进作用最大,但显著降低了类黄酮含量和CHI活性。(2)对于白果期蓝莓,5min UV-B处理显著增加了果实类黄酮含量和CHI活性,10min处理使果实可溶性糖和总酚含量较对照分别增加25%和18%;15min处理对果实花青苷含量和PAL活性影响作用最大。(3)对于转色期蓝莓,各处理除果实可溶性糖及类黄酮含量降低外,其余物质含量均显著增加。(4)UV-B处理并未改变果实发育过程中可溶性糖、总酚、类黄酮和花青苷含量及PAL、CHI酶活性的积累规律。(5)蓝莓果内PAL活性与其可溶性糖、总酚和类黄酮的积累呈极显著正相关关系,而CHI活性仅与其可溶性糖呈极显著正相关。研究表明,UV-B辐照处理促进了幼果期和白果期可溶性糖的积累,也能促进不同发育时期蓝莓果实总酚和花青苷及白果期类黄酮的积累,对蓝莓果实主要品质能够产生积极的影响。     

高浓度臭氧对蒙古栎叶片酚类物质含量和总抗氧化能力的影响

采用开顶箱(OTC)法,研究了高浓度臭氧(80±8 nmol·mol^-1)熏蒸处理对蒙古栎叶片 中总酚、类黄酮、单宁及丙二醛(MDA)含量的影响,使用1,1-二苯基苦基苯肼(DPPH) 法测定了蒙古栎叶片的总抗氧化能力.结果表明:蒙古栎叶片中总酚、类黄酮、缩合单宁和MDA的含量都有所增加,其中总酚和缩合单宁含量分别增加了48.17%和26.77%,差异显著（P<0.05）;类黄酮和MDA含量分别增加了24.66%和5.26%,差异不显著(P＞0.05);蒙古栎叶片总抗氧化能力显著增强(P<0.05),且与叶片中总酚和缩合单宁含量呈显著的正相关关系.     

艳婀珍蝶取食对薇甘菊叶片生理指标的影响

艳婀珍蝶取食后,对薇甘菊叶片的超氧物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)、多酚氧化酶(PPO)的活性,总酚含量,有机自由基(DPPH·)清除能力进行分析。结果表明,取食后3h内所测各量即发生变化,但与对照差异不显著。取食4d中,叶片的SOD、POD活性总体上分别高于对照,CAT活性总体上低于对照,说明在艳婀珍蝶胁迫过程中,SOD和POD所起的作用比CAT大。取食叶SOD、POD活性均在48h时达到最大值,之后下降;CAT活性在24h时达到最大值,之后也迅速下降。取食叶PPO活性动态变化程度较大,表现出3个峰值,分别为对照的1·83,1·92倍和2·17倍;总酚含量表现为先上升后下降的趋势;对DPPH·的清除力一直显著性甚至极显著性低于对照。取食叶的SOD与CAT和POD均呈正相关性,且相关性大于对照;PPO与总酚含量在取食叶与对照中也均呈弱正相关性。实验结果表明,薇甘菊的保护酶对艳婀珍蝶胁迫的应激效应是短暂而有限的,艳婀珍蝶的取食破坏了薇甘菊叶片功能,较大程度的干扰了薇甘菊保护酶系统的防御代谢,薇甘菊的总抗氧化能力降低。薇甘菊也不能通过改变酚类物质含量来抵御艳婀珍蝶的取食,艳婀珍蝶取食对薇甘菊有较明显的控制作用。     

不同灰霉病抗性苹果果实中酚类物质代谢特征

以‘秦冠’、‘富士’、‘金冠’苹果果实为材料,通过对损伤接种灰葡萄孢菌(Botrytis cinerea Pers)后果肉组织酚类代谢主要产物和相关酶活性变化的分析测定,揭示苹果采后酚类物质代谢与灰霉病抗性的关系,为苹果灰霉病抗性鉴定和筛选抗灰霉病苹果资源提供理论指导。结果表明:(1)接种灰葡萄孢菌后,3个苹果品种的果实灰霉病发病率和病斑直径大小均为‘秦冠’‘富士’‘金冠’,而且3个品种间的发病率和病斑直径均差异显著,各品种对灰霉病的抗性由强到弱依次为‘秦冠’‘富士’‘金冠’。(2)抗病品种‘秦冠’果肉组织中类黄酮、木质素含量及苯丙氨酸解氨酶(PAL)、过氧化物酶(POD)、多酚氧化酶(PPO)活性均显著高于感病品种‘富士’和‘金冠’,但总酚含量为‘秦冠’‘金冠’‘富士’,且3品种间总酚含量差异显著。研究表明,抗病苹果品种通过调节果肉内酚类物质代谢,增强次生代谢能力,其中类黄酮和木质素含量的增加强化了果实的抗性反应,进而提高对灰霉病的抗性,但总酚含量与植物抗病性关系不大。     

小菜蛾取食后上海青的保护酶活力变化

上海青遭受小菜蛾取食为害后,0.5小时内叶片的超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)、多酚氧化酶(PPO)和总酚含量发生变化。就一天内的变化趋势而言,受害叶片的SOD、CAT、POD、PPO活力和总酚含量总体上比正常叶片低,但在15：00时,正常叶片的SOD、CAT和POD活力最低时,受害叶片的三种酶活力却明显上升,在11：00时,正常叶片PPO活力为最低值时,受害叶片的PPO活力达最高,这可能是过度胁迫造成的,正常叶片的PPO活力与总酚量负相关,但受害叶片不呈现这种关系,此外,受害植株的未受害叶的保护酶活力也不同程度地发生变化,可见上海青对小菜蛾的取食胁迫后保护酶的反应是系统性的。     

辣椒感染疫病后生化指标的响应研究

采用生理生化分析方法研究了辣椒感染疫病后叶片中几个生化指标的变化。结果表明,染病前后感病品种叶片中可溶性总糖含量持续高于抗病品种;抗病类型品种和感病类型品种的可溶性蛋白含量变化规律均表现为先升高后下降,但接种前其叶片中可溶性蛋白含量两者间无明显差异;抗病类型和感病类型辣椒接种后保护酶活性均升高,而且感病类型的POD和ASP酶活性在接种后120h显著高于抗病类型;高抗类型叶片中PPO活性增加幅度显著大于感病类型,但抗病类型品种PPO活性上升趋势比较平缓。接种后,感病品种PAL活性上升幅度小于高抗品种,接种后96h PAL活性开始逐渐下降。可溶性总糖含量和苯丙氨酸解氨酶可以作为辣椒苗期抗疫病鉴定的生化指标。     

不同颜色果袋对‘云红梨2号’果皮色泽形成的影响

研究了不同颜色果袋对‘云红梨2号’果实着色的影响,比较了不同套袋处理下果皮外观着色、叶绿素、类黄酮、总酚、花色素苷含量以及花色素苷合成相关酶活性的差异.结果表明: 发育期的黑暗处理有利于解袋后梨果皮着色;不同套袋处理中,采前解袋自然光照射下梨果皮中花色素苷含量最高,着色最好,白色纸袋次之.不同套袋处理显著影响果皮中叶绿素、类黄酮、总酚和花色素苷含量,从而影响梨果皮的外观色泽.不同套袋处理的花色素苷合成酶活性差异显著;相关性分析表明,果皮中花色素苷含量与二氢黄酮醇-4-还原酶(DFR)和类黄酮3-O-葡萄糖基转移酶(UFGT)活性呈显著正相关,而与苯丙氨酸解氨酶(PAL)活性相关性不显著.     

菠萝叶片色泽、色素及抗氧化活性的关系

以菠萝22个栽培品种的叶片为实验材料,测定其5种色泽参数（L^*、a^*、b^*、c^*和h^*值）、5种色素（叶绿素、类胡萝卜素、花青苷、类黄酮和总酚）含量及3种抗氧化活性指标（ABTS、DPPH自由基和亚硝酸盐的清除能力）,并进行相关性分析。研究结果显示,色泽参数a^*和h^*值可以作为菠萝叶片指示色泽、主要色素含量和抗氧化活性变化的重要指标;菠萝叶片主要色素组成是叶绿素、类黄酮和总酚,且含有少量的花青苷,几乎不含类胡萝卜素。相关性分析结果显示,菠萝叶片类黄酮和总酚含量均与3种抗氧化活性指标极显著正相关,而叶绿素含量与其它指标相关性未达到显著水平,类黄酮和总酚是菠萝叶片抗氧化活性的主要功效成分。     

Role of Phenylalanine Ammonia Lyase and Polyphenol Oxidase in Host Resistance to Bacterial Wilt of Tomato

Plants respond to bacterial pathogen attack by activating various defence responses, which are associated with the accumulation of several factors like defence-related enzymes and inhibitors which serve to prevent pathogen infection. The present study focused on the role of the defence-related enzymes phenylalanine ammonia lyase (PAL) and polyphenol oxidase (PPO) in imparting resistance to tomato against bacterial wilt pathogen Ralstonia solanacearum . The temporal pattern of induction of these enzymes showed maximum activity at 12 h and 15 h for PAL and PPO, respectively, after the pathogen inoculation (hpi) in resistant cultivars. Twenty different tomato cultivars were analyzed for PAL, PPO and total phenol content following pathogen inoculation. The enzyme activities and total phenol content increased significantly (P < 0.05) in resistant cultivars upon pathogen inoculation. The increase in enzyme activities and total phenol content were not significant in susceptible and highly susceptible cultivars. The role of PAL and PPO in imparting resistance to tomato against bacterial wilt disease is discussed.     

马铃薯不同品种感染早疫病菌后防御酶活性变化

通过测定抗性不同的马铃薯品种接种和未接种情况下叶片内防御酶活性,研究马铃薯品种对早疫病的抗性机制。结果表明,在接种处理后,各品种植株体内苯丙氨酸解氨酶（PAL）、过氧化物酶（POD）、多酚氧化酶（PPO）和过氧化氢酶（CAT）活性均提高,抗病品种酶活性增幅高于感病品种,说明上述4种酶与品种抗性有一定的关联。     

Phenylalanine-ammoniaque lyase et composes phenoliques au cours de la maturation de la cerise

L-Phenylalanine-ammonia lyase (PAL) activity and phenolic content were investigated during growth and maturation of sweet cherry fruit on the tree. Phenolic acid, flavonoid and tannin contents, were studied in relation to PAL activity over two years. Concomitant variations in PAL activity and in flavonoid or phenolic acid concentrations were found during growth, suggesting that PAL is the major element controlling the synthesis of phenolic compounds in sweet cherry. During ripening, the increase in PAL activity did not appear to be directly related to anthocyanin synthesis. The concentrations of neochlorogenic acid and narcissin measured over the same period were also related to PAL activity.     

低温与GA_3诱导菠萝黑心病的生理机制

菠萝黑心病是PPO催化氧化酚类物质形成褐色产物所致。低温或GA_3处理提高了PPO活性及其底物——儿茶酚、绿原酸和咖啡酸的含量,也导致了PAL活性增加;低温还使乙烯释放率增大。这些变化均有利于黑心病的发生和发展。     

绿盲蝽取食与机械损伤对棉花叶片内防御性酶活性的影响

为探明绿盲蝽Apolygus lucorum (Mayer-Dür)取食和机械损伤对不同抗性棉花叶片内主要防御酶活性的影响以及防御酶与棉花抗绿盲蝽性的关系,以棉花3个不同抗性品系为材料,室内条件下测定绿盲蝽取食和机械损伤处理后棉叶中苯丙氨酸解氨酶(PAL)、过氧化物酶(POD)和多酚氧化酶(PPO)的活性.结果表明:对...     

The effects of infection by Phytophthora infestans on the control of phenylpropanoid metabolism in wounded potato tissue

During the first 24 h of in vitro incubation of excised potato tuber (Solanum tuberosum L.) discs, the appearance of phenylalanine ammonia-lyase (PAL; EC 3.4.1.5) and the accumulation of chlorogenic acid are both stimulated by infection with Phytophthora infestans (Mont.) de Bary. Whereas in control tissue the level of PAL reached a stable plateau value after 40 h, in infected tissue it subsequently rose again, in one experiment, as the fungal mycelium developed. In the infected but not the control tissue, the level of chlorogenic acid subsequently fell to about to about 20% of its maximum after 50 h. The time courses of increases in cinnamic acid 4-hydroxylase (CA4H; EC 1.14.13.11; 0–60 h) and of caffeic acid acid o-methyltransferase (COMT; EC 2.1.1.42; 0–160 h) are not altered by fungal infection. If the discs are restored to the tuber environment immediately after excision, by placing them inside a host tuber, the activity of PAL as well as those of CA4H and COMT remained at the constant low endogenous level for at least 60 h, irrespective of whether the discs had first been inoculated with P. infestans. The increase in PAL may not be an obligatory feature of the P. infestans/potato compatible interaction but dependent on an underlying wound response. The experiments provide further evidence that PAL is the rate limiting step of chlorogenic acid biosynthesis in potato tuber discs.Abbreviations PAL phenylalanine ammonia-lyase - CA4H cinnamic acid 4-hydroxylase - COMT caffeic acid o-methyltransferase - CGA chlrogenic acid (5-o-caffeoylquinic acid) - gfwt gram fresh weight     

Transgenic sweet potato expressing thionin from barley gives resistance to black rot disease caused by Ceratocystis fimbriata in leaves and storage roots

Black rot of sweet potato caused by pathogenic fungus Ceratocystis fimbriata severely deteriorates both growth of plants and post-harvest storage. Antimicrobial peptides from various organisms have broad range activities of killing bacteria, mycobacteria, and fungi. Plant thionin peptide exhibited anti-fungal activity against C. fimbriata. A gene for barley α-hordothionin (αHT) was placed downstream of a strong constitutive promoter of E12Ω or the promoter of a sweet potato gene for β-amylase of storage roots, and introduced into sweet potato commercial cultivar Kokei No. 14. Transgenic E12Ω:αHT plants showed high-level expression of αHT mRNA in both leaves and storage roots. Transgenic β-Amy:αHT plants showed sucrose-inducible expression of αHT mRNA in leaves, in addition to expression in storage roots. Leaves of E12Ω:αHT plants exhibited reduced yellowing upon infection by C. fimbriata compared to leaves of non-transgenic Kokei No. 14, although the level of resistance was weaker than resistance cultivar Tamayutaka. Storage roots of both E12Ω:αHT and β-Amy:αHT plants exhibited reduced lesion areas around the site inoculated with C. fimbriata spores compared to Kokei No. 14, and some of the transgenic lines showed resistance level similar to Tamayutaka. Growth of plants and production of storage roots of these transgenic plants were not significantly different from non-transgenic plants. These results highlight the usefulness of transgenic sweet potato expressing antimicrobial peptide to reduce damages of sweet potato from the black rot disease and to reduce the use of agricultural chemicals.     

Responses of phenolic acid and defensive enzyme activities to mechanical damage in Artemisia frigida

Aims The study aims at understanding the effects of feed intake and trample damage on the phenolic acid formation and antioxidant enzyme activities in Artemisia frigida, and elucidating the adaptive mechanisms in A. frigida to grazing in secondary metabolites and their related enzyme activities.
Methods We analyzed the phenolic acid content and the activities of polyphenol oxidase (PPO), phenylalanine ammonia-lyase (PAL) and protective enzymes in leaves and roots in A. frigida under three levels (light, moderate, and heavy) of manipulative grazing condition. The measurements of the 9 phenolic acid contents started after 6 h of the mechanical damage of the plants by using the high performance liquid chromatography (HPLC), and the enzyme activities in leaves and roots were measured by a spectrophotometry method.
Important findings The light damage treatment induced productions of PPO, PAL and significantly (p < 0.05) increased antioxidant enzyme activities in the leaves and roots of A. frigida. The contents of PPO, PAL and antioxidant enzymes increased with increasing intensity of mechanical damage. Compared to the control, the content of free caffeic, syringic, ferulic and cinnamic acid in the leaves A. frigida were significantly elevated (p < 0.05) by 150.4%, 93.5%, 154.4% and 121.7%, respectively. They were significantly (p < 0.05) positively correlated with PAL activity in the moderate damage treatment. The content of free chlorogenic acid and catechol decreased by 91.1%, and 69.3%, respectively, compared with the control they had a negative correlation with PPO activity in the heavy damage treatment. The contents of gallic and protocatechuic acids increased (p < 0.05) by 280.6% and 215.7%, respectively, in the heavy damage treatment. With increasing intensity of mechanical damage, the content of 9 free phenolic acids significantly increased in roots but the increasing range was less than the one in leaves. Mechanical damage induced an increasing trend in the total amount of free and bounded phenolic acids in the leaves but a decreasing trend in the total amount of bounded phenolic acids in the roots of A. frigida. The results indicated that mechanical damage could firstly induce an increase of antioxidant enzymes and key enzymes in phenolic metabolism in A. frigida, leading to the accumulation of antioxidant substances of phenolic acids, further regulate the biosynthesis of lignins, quinones and tannins, and then enhance the resistance to mechanical damage and improved the tolerance of A. frigida to grazing.     

Antisense-overexpression of the MsCOMT gene induces changes in lignin and total phenol contents in transgenic tobacco plants

Initially, we isolated the caffeic acid O-methyltransferase (COMT) gene from Miscanthus sinensis (accession number HM062766.1). Next, we produced transgenic tobacco plants with down-regulated COMT gene expression to study its control of total phenol and lignin content and to perform morphological analysis. These transgenic plants were found to have reduced PAL and ascorbate peroxidases expression, which are related to the phenylpropanoid pathway and antioxidant activity. The MsCOMT-down-regulated plants had decreased total lignin in the leaves and stem compared with control plants. Reduced flavonol concentrations were confirmed in MsCOMT-down-regulated transgenic plants. We also observed a morphological difference, with reduced plant cell number in transgenic plants harboring antisense MsCOMT. The transgenic tobacco plants with down-regulated COMT gene expression demonstrate that COMT plays a crucial role related to controlling lignin and phenol content in plants. Also, COMT activity may be related to flavonoid production in the plant lignin pathway.