青香蕉和黄熟香蕉果皮感病过程中5种酶活性的变化

以同一品种的未后熟的青香蕉和已黄熟的香蕉为材料,测定了接种炭疽病菌5d内果皮PPO、POD、SOD、PAL和LOX活性的变化,探讨与香蕉抗病性相关的酶类。结果表明,不论接种与否,青香蕉果皮中的PPO、POD和SOD的活性明显低于黄熟香蕉,而PAL和LOX活性高于黄熟香蕉;接种对PPO、POD和LOX活性影响不大,但引起SOD和PAL活性下降。青香蕉果皮中较低的PPO、POD和SOD的活性有利于活性氧和酚类物质的积累而产生过敏性反应,较高的PAL和LOX活性则利于苯丙烷类代谢和脂质过氧化,有利于抗病物质的形成。PAL和LOX是香蕉抗病性形成的关键酶。     

杧果采前喷施茉莉酸甲酯对其抗病性和采后品质的影响

 以‘台农1号’杧果（Mangifera indica L.‘Tainong 1’）为试材,研究了采前喷施茉莉酸甲酯（MeJA）处理对杧果抗病性和采后品质的影响及其相关机理。结果表明,与对照果实相比,50 μmol · L^-1 MeJA采前处理显著降低了采收时的病果率和贮藏期的病情指数,抑制了接种炭疽病菌果实的病斑直径;有利于杧果贮藏品质的保持,提高了果肉中维生素C含量,延缓了可溶性糖含量的升高和可滴定酸含量的降低;同时,采前MeJA处理可以提高杧果果皮中苯丙氨酸解氨酶（PAL）、过氧化物酶（POD）、多酚氧化酶（PPO）和β–1,3–葡聚糖酶（GLU）等防御酶的活性,提高贮藏早期的过氧化氢（H₂O₂）水平,抑制贮藏后期H₂O₂和MDA含量的积累。这些结果表明,采前MeJA处理提高杧果抗病性和保持果实品质与激活杧果的防御系统及降低膜脂过氧化程度有关。     

激发子物质处理富士苹果果实后抗轮纹病病菌侵染的研究

富士苹果果实经硝普钠(SNP)、茉莉酸甲酯(MeJA)、水杨酸(SA)和氯化钙处理后再接种轮纹病菌,各个处理均能不同程度地防止轮纹病菌的侵染和扩展,其中MeJA和SNP处理效果最明显,烂果率和烂果面积分别比对照减少26.7%、26.7%和41.6%、36.2%。另外,还对各处理果实的PAL、SOD、POD和PPO活性变化进行了分析,结果表明,接种轮纹病菌的72h内,4个处理的PAL和PPO活性均表现出先升后降的趋势,而SOD和POD活性变化除SA处理一直保持平稳波动上升外,其余3个处理也表现为先升后降趋势;各处理的PAL和POD峰值出现的时间均早于SOD和PPO。PAL、POD和PPO活性峰值和平均值均以MeJA处理最高,SOD活性峰值和平均值SNP处理最高。     

香蕉枯萎病菌4号生理小种毒素解毒剂的筛选及对香蕉防御酶活性的影响

从15种候选解毒剂中筛选出对香蕉枯萎病菌(Fusarium oxysporum f.sp.cubense)4号生理小种(简称Foc 4)毒素解毒效果最好的2种解毒剂——硫酸锌和井冈霉素。在2 g.L-1浓度下,硫酸锌和井冈霉素对Foc 4毒素的钝化率分别为76.67%和71.33%,而在5 g.L-1浓度下,它们的钝化率分别为90.33%和80.00%。探索了毒素—硫酸锌和毒素—井冈霉素处理体系在2 g.L-1和5 g.L-1浓度下对香蕉苗5种防御酶(PAL、POD、PPO、SOD和CAT)活性的影响。结果表明,在这个处理体系中,香蕉苗PAL和PPO酶活高峰出现的时间(PAL 36 h、PPO 36 h和48 h)较毒素单独处理(PAL 48 h、PPO 60 h)早;SOD和CAT酶活高峰出现的时间与对照基本一致;而POD酶活高峰出现的时间较复杂。总体上,这个处理体系的酶活高于毒素单独处理和无菌水对照。"毒素—硫酸锌处理体系"在PAL、SOD整体酶活上高于"毒素—井冈霉素处理体系",而后者在其他3种酶活上普遍高于前者。     

钙对茉莉酸甲酯诱导番茄抗灰霉病及防御酶活性的调控作用

以番茄品种L402幼苗为试材,通过外源施钙和缺钙处理,探讨钙对茉莉酸甲酯（MeJA）诱导番茄抗灰霉病的作用及其生理机制。结果表明：（1）外源MeJA显著诱导番茄抗灰霉病|外源钙离子进一步增强MeJA诱导的抗病程度|而钙螯合剂EGTA和质膜钙通道抑制剂LaCl3则不同程度抑制MeJA诱导番茄的抗病性。（2）MeJA单独处理后2～5 d内PAL、PPO和POD活性显著高于对照,外源钙离子进一步增强MeJA诱导的上述防御酶活性,两种缺钙处理则不同程度降低防御酶活性。（3）外源施钙及不同缺钙处理,对MeJA诱导的CAT和SOD活性没有规律性影响。由此认为,钙增强MeJA诱导番茄抗灰霉病的程度,可能与其对MeJA诱导番茄叶片中PAL、PPO和POD等防御酶活性的调控有关。     

促生菌CH1诱导黄瓜对猝倒病抗性的研究

 采用分根法研究了植物生长促生菌CH1诱导黄瓜对猝倒病的抗性以及黄瓜植株经CH1处理后根部与叶部组织的超氧化物歧化酶( SOD) 、过氧化物酶( POD) 、过氧化氢酶(CAT) 、多酚氧化酶( PPO) 和苯丙氨酸解氨酶( PAL) 的活性变化。结果表明, 促生菌CH1提高了黄瓜的成苗率, 处理后的28 d, 猝倒病相对防效高达78.64% , 推测促生菌CH1使黄瓜产生的诱导抗性是其防病的主要原因。黄瓜经CH1处理并同时接种腐霉菌后, 处理根部与未处理叶部组织中SOD、POD、CAT、PPO和PAL活性都明显高于其它处理, 其根部这些酶的活性分别在处理后3、3、5、5和5 d时出现高峰, 叶部酶活性变化基本与根部的相吻合。表明这些酶活性的提高与CH1诱导的黄瓜对猝倒病抗性可能有一定的相关性, 而且CH1对这些酶活性的诱导具有系统性。     

苯并噻二唑处理提高采后沙糖橘对指状青霉菌的抗性

对沙糖橘(Citrus reticulata Blancdo‘Shiyueju’)果实进行苯并噻二唑(S-methyl benzo thiadiazole-7-carbothioate,BTH)渗透处理后接种指状青霉菌孢子,12h后即可明显诱导果皮中抗病相关蛋白CR-CT1和CR-BG3基因表达并在贮藏期间维持较高水平;不同浓度BTH处理果实的发病率和病斑面积均比对照低,其中以0.025g·L-1BTH处理的效果最好;BTH处理提高了果皮几丁质酶(CT)、β–1,3–葡聚糖酶(GUN)、苯丙氨酸解氨酶(PAL)、多酚氧化酶(PPO)、过氧化物酶(POD)活性和H2O2含量,表明BTH诱导的沙糖橘果实抗病性与果实CR-CT1和CR-BG3基因的表达增强,CT、GUN、PAL、PPO和POD等相关酶活性和H2O2含量提高密切相关。     

BTH诱导甜瓜愈伤组织活性氧代谢和抗性酶活性的信号途径

通过对含有苯并噻二唑(BTH)、咖啡酸(CA)或氯化钴(CC)等不同MS培养上甜瓜愈伤组织的研究,测定了超氧阴离子(O2■)产生速率、过氧化氢(H2O2)含量及抗性相关酶活性的变化,探讨了BTH诱导的甜瓜ROS代谢和抗性酶活性及其相关的信号途径。结果表明,CA可以消除BTH对甜瓜愈伤组织O2■增加和H2O2积累的诱导作用;CA或CC能够部分抵消BTH对SOD增强的诱导作用;CC对BTH诱导的POD增强具有抑制效应,CA+CC可完全消除BTH对POD的诱导作用;CA和CC均能促进BTH对PAL增强的诱导作用,且具有协同效应。BTH对甜瓜O2■升高、H2O2积累和CAT降低的诱导主要通过SA途径实现,对POD的影响主要通过ET途径实现,对SOD升高的诱导通过SA和ET 2条途径实现,而对PAL活性的影响通过ET和SA以外的途径实现。     

Bioassay method for testing Fusarium wilt disease tolerance in transgenic banana

Fusarium wilt caused by the fungus, Fusarium oxysporum f.sp. cubense is a major factor limiting commercial banana production in Malaysia. Field evaluation of banana plants for Fusarium wilt disease has not been effective as the infection period is slow and other variables such as distribution of inoculum concentration is difficult to control. Therefore, a study was carried out to develop a semi-qualitative bioassay for early and rapid susceptibility tests against Fusarium wilt disease in banana plantlets. The induction of sporulation and germination of Foc race 1 (VCG 01217) were firstly determined. Then, the effect of inoculum density of fungal spores on the magnitude of infection in Pisang Rastali (AAB) plantlets was carried out. The concentration of hydrogen peroxide (H₂O₂) and other enzyme activities such as phenylalanine ammonia lyase (PAL), chitinase, glucanase, peroxidase (PER) and polyphenol oxidase (PPO) in infected roots of banana plants were determined to relate to the levels of tolerance or susceptibility to Fusarium wilt disease. Using similar conditions, transgenic Pisang Rastali (AAB) was tested to investigate the effectiveness of this bioassay. Hydrogen peroxide and phenylalanine ammonia lyase is the most sensitive compound and enzyme detectable after inoculated with Fusarium spores. The chitinase, β-1,3-glucanases, peroxidase and polyphenol oxidase enzymes activity of transgenic plants were higher than untransformed plantlets. The results obtained indicate that the use of these biochemical parameters has the potential to predict the levels of tolerance or susceptibility to Fusarium wilt disease in other banana cultivars.     

Induction of resistance in Yali pear (Pyrus bretschneideri Rehd.) fruit against postharvest diseases by acibenzolar-S-methyl sprays on trees during fruit growth

The objective of this work was to evaluate how disease resistance in harvested fruit of Yali pears (Pyrus bretschneideri Rehd.) may be affected by acibenzolar-S-methyl (ASM) sprays on trees. Results showed that disease incidence and lesion diameter in mature pears from trees sprayed with ASM for three times during growth and inoculated with Penicillium expansum or Alternaria alternata after harvest were 27.9 and 42.7%, or 29.1 and 23.4% lower, respectively, than in control fruit 17 days after inoculation. Mature fruit from ASM-treated trees exhibited higher activities of defense enzymes including phenylalanine ammonia-lyase (PAL), chitinase and β-1,3-glucanase than in control at harvest. In young pears, activities of peroxidase, polyphenoloxidase, PAL, chitinase and β-1,3-glucanase were significantly enhanced by ASM after first spray on trees. The ASM spray also significantly increased H₂O₂ level and glutathione reductase activity, but reduced activities of catalase and ascorbate peroxidase in young pears. The study indicated that enhancement of disease resistance in harvested Yali pear fruit could be the result of multiple effects of several factors related to plant defenses induced by ASM sprays on trees during fruit growth. Application of ASM in the field holds great promise for controlling postharvest diseases of the fruit.     

壳寡糖诱导和白粉菌侵染的辣椒叶片防御酶系活性研究

以遵义朝天椒为供试材料,用壳寡糖喷雾处理辣椒叶片后再接种辣椒白粉菌,系统地研究了壳寡糖对辣椒防御酶活性变化的影响。结果表明:壳寡糖喷雾处理辣椒后,辣椒叶片中多酚氧化酶(PPO)、过氧化物酶(POD)、苯丙氨酸解氨酶(PAL)活性较对照都有明显增加,其中PPO和PAL峰值提前(处理后接种)。试验证明,壳寡糖处理后,随着酶活性增强,辣椒的系统抗病性得到表达,二者有一定的相关性。     

钙素对SA诱导番茄幼苗抗灰霉病的调控作用

 为探索钙对水杨酸（SA）诱导番茄（Solanum lycopersicum）幼苗抗灰霉病的作用,采用‘L402’番茄品种,通过分别喷施8 mmol • L^-1 CaCl₂和5 mmol • L^-1 EGTA后再喷施2 mmol • L^-1 SA,3 d后接种灰霉病菌孢子的方法,研究了不同处理对番茄幼苗灰霉病病情指数、活性氧积累、主要防御酶活性及其基因表达的影响。结果表明：接种灰霉病菌孢子5 d后,SA处理的番茄幼苗病情指数比对照降低37.27%,Ca + SA处理较SA处理进一步降低18%;接种1 d和2 d后,叶片中产生速率和H₂O₂含量分别出现应激高峰,且处理间存在差异,与对照相比Ca + SA处理分别提高33.05%和29.31%,EGTA + SA处理分别降低32.62%和46.34%;叶片中抗病相关酶活性和基因表达在接种病菌后也出现应激高峰,其中Ca + SA处理显著提高了PAL、几丁质酶、β–1,3–葡聚糖酶活性,EGTA处理及EGTA + SA处理显著降低了PAL、几丁质酶、β–1,3–葡聚糖酶活性。上述结果表明,Ca²⁺在SA诱导番茄抗灰霉病中具有正调控作用,而且这种作用与PAL、几丁质酶、β–1,3–葡聚糖酶活性及其基因表达密切相关。     

轮纹病菌对不同抗性苹果品种防御酶的影响

实验研究了3个苹果品种被轮纹病菌(Botryosphaeriaberengrianaf.piricola)侵染后苹果枝干轮纹病防御酶系的变化。结果表明,在受病菌侵染后,苯丙氨酸解氨酶(PAL)、超氧化物歧化酶(SOD)、过氧化物酶(POD)和多酚氧化酶(PPO)均表现为先升后降的趋势,且抗病品种的峰值出现较晚。抗病品种鸡冠的PAL、SOD和POD活性变化峰值和均值要高于国光和富士,而PPO活性变化的峰值和均值则低于国光和富士。     

Improving disease resistance in peach fruit during storage using benzo-(1,2,3)-thiodiazole-7-carbothioic acid S-methyl ester (BTH)

Summary

The compound benzo-(1, 2, 3)-thiodiazole-7-carbothioic acid S-methyl ester (BTH) effectively induces a systemic acquired resistance (SAR) response in plants and increases plant resistance to disease. To learn how BTH may affect post-harvest quality and disease resistance in peach, (Prunus persica L. cv. ‘Jiubao’) peach fruit were treated with 200 mg l^–1 BTH for 5 min immediately after harvest, then incubated for 60 h at 20°C, 85–95% RH. Sixty h after treatment, fruit were inoculated with spore suspensions of Penicillium expansum at 1.2 10⁴ CFU ml^–1. The incidence of disease and lesion areas on peach fruit were significantly reduced by BTH-treatment (P < 0.05). In addition, chitinase and -1,3-glucanase activities in peach fruit were significantly enhanced by BTH-treatment (P < 0.05). Moreover, the total levels of phenolic compounds and chlorogenic acid, and the lignin content of BTH-treated fruit were significant higher than in control fruit (P < 0.05). The effects of BTH on fruit quality were also analysed. Soluble solids content, soluble pectin, titratable acidity and ascorbic acid levels in peach fruit were not significantly affected by treatment with BTH. Our results suggest that the application of BTH may control post-harvest diseases in peach fruit during commercial production.     

Control of black spot disease caused by Alternaria alternata on jujube (Ziziphus jujuba Mill. cv. Dongzao) using HarpinXoo protein

Summary

Currently, no adequate control measures exist for the fungal disease, black spot (Alternaria alternata (Fr.) Keissler) on ‘Dongzao’ jujube (Ziziphus jujuba Mill. cv. Dongzao). Black spot is an important factor limiting jujube fruit yield under certain conditions. In this study, three concentrations (10, 20, or 30 µg ml^–1) of the Harpin_Xoo protein, or water (negative control), were sprayed onto ‘Dongzao’ jujube trees to control black spot disease. Following Harpin_Xoo protein treatment, its effects on the control of black spot disease were determined, and the levels of a defence-related substance [i.e., salicylic acid (SA)], and the activities of peroxidase (POD), polyphenol oxidase (PPO), and phenylalanine ammonia lyase (PAL) were measured. The incidence of black spot disease was significantly decreased, compared with water-treated control, fruit by ≤ 88.4%. After 2 d, SA levels, and POD, PPO, and PAL activities in Harpin_Xoo-treated fruit were clearly higher than those in control fruit. After fruit had been stored for 45 d at 0ºC, SA levels, and POD and PPO activities showed no differences between the Harpin_Xoo treatments and the controls, but PAL activities in Harpin_Xoo-treated fruit were lower than those in the controls. The most appropriate concentration of Harpin_Xoo protein to control black spot disease was 20 µg a.i. ml^–1. These results suggest that the Harpin_Xoo protein induced systemic acquired resistance in ‘Dongzao’ jujube trees and may offer a worthwhile strategy for disease control.     

诱抗处理对甜瓜叶片酚类物质代谢的影响

以甜瓜抗白粉病品种‘银帝’和感白粉病品种‘卡拉克赛’为材料,研究诱导抗性处理对其叶片酚类物质代谢的影响及其与抗病性的关系。结果表明：苯丙噻重氮（BTH）和水杨酸（SA）处理以及接种白粉菌均可显著诱导甜瓜叶片苯丙氨酸解氨酶（PAL）、过氧化物酶（POD）、多酚氧化酶（PPO）活性和绿原酸、总酚、类黄酮含量升高,但纳米硅（SiO2）诱导无效。经诱导和接菌后,甜瓜抗病品种叶片PAL、POD、PPO活性和总酚、绿原酸、类黄酮含量高于感病品种。BTH和SA诱导甜瓜抗病性增强的原因之一是提高了叶片次生代谢相关酶的活性,增加了叶组织次生代谢物质的含量。     

茉莉酸甲酯诱导辣椒抗青枯病与活性氧代谢的关系

为探究茉莉酸甲酯(Methyl Jasmonate,Me JA)诱导辣椒抗青枯病效应与活性氧代谢相关酶的关系,以辣椒易感青枯病品种‘粤红1号’和抗青枯病品种‘辛香8号’幼苗为试验材料,在0.1mmol·L-1 Me JA喷雾处理后12、24、48、72和96 h接种青枯劳尔氏菌(Ralstonia solanacearum),对其进行青枯病病情指数与活性氧代谢相关酶——超氧化酶歧化酶(Superoxide Dismutase,SOD)、过氧化氢酶(Catalase,CAT)、过氧化物酶(Peroxidase,POD)和抗坏血酸过氧化物酶(Ascorbate Peroxidase,APX)活性以及丙二醛(Malondialdehyde,MDA)含量测定及相互关系的分析。结果表明:‘粤红1号’和‘辛香8号’的病情指数随喷Me JA处理时间的推移表现为先降后升,但都低于对照;而诱导效果则相反,Me JA处理对两个辣椒品种幼苗抗性的最适诱导时间均为接种前48 h。接种后0~96 h,Me JA喷雾处理的辣椒幼苗叶片SOD、CAT、POD和APX酶活性均显示先升后降的趋势,且明显高于对照,接种后24~48 h达到最高,而MDA含量则明显低于对照。因此,Me JA可诱导辣椒幼苗抗青枯病,其实现途径可能与提高活性氧代谢相关酶活性和缓解膜脂过氧化有关。     

钾、钙、硼、硅对茄子抗黄萎病效果的影响

以茄子黄萎病为研究对象,采用田间施用钾（K）、钙（Ca）、硼（B）、硅（Si）的方法,探讨4种元素对接种黄萎病菌后茄子幼苗的发病率、植株生长状况及防御酶活性的影响。结果表明,与对照相比,施用4种元素的各处理均降低了茄子黄萎病的发病率和病情指数,低浓度的K、Ca、B、Si促进了茄子幼苗株高、茎粗、根长、全株鲜质量、根系活力及POD、PPO、PAL活性,而后随着浓度增大,促进作用减弱。其中以B 50 mg·L^-1处理抗病性最好,其发病率比对照低46.66个百分点,各生长指标显著高于对照,根系活力比对照提高286 %;POD、PPO、PAL活性分别比对照提高20 %、78 %、87 %。说明B对诱导茄子黄萎病抗性有显著效果,可应用到生产。     

甘蓝链格孢菌侵染对白菜保护酶活性和H2O2代谢的影响

 以白菜抗黑斑病苏州青自交系和感黑斑病短白梗自交系为材料,研究其接种甘蓝链格孢菌后保护酶活性及H₂O₂代谢的变化。结果显示,抗感病材料的多酚氧化酶（PPO）、超氧化物歧化酶（SOD）和过氧化氢酶（CAT）与接种前相比均升高,且抗病材料的PPO、SOD始终高于感病材料,CAT活性低于感病材料;接种后抗病材料苯丙氨酸解氨酶（PAL）活性升高,而感病材料的PAL活性降低;抗感病材料接种后过氧化物酶（POD）、H₂O₂均呈上升趋势,且抗病材料的H₂O₂能很快降至较低水平。由此表明：甘蓝链格孢菌侵染白菜后,抗病材料与感病材料相比,能生成较多的PAL和PPO抵抗病原菌的侵染,并能短时迸发H₂O₂刺激抗氧化酶活性升高,使植物体内H₂O₂处于相对较低的水平,降低活性氧伤害。     

白粉菌侵染过程中不同抗性辣椒品种叶片3种酶活性变化

以辣椒4个不同抗性品种为材料,研究成株期感染白粉菌后,叶片组织内苯丙氨酸解氨酶（PAL）、β-1,3-葡聚糖酶和几丁质酶的活性变化。结果表明,4个材料接种白粉病病原菌后3种酶活性均有所提高。受白粉菌侵染后,抗病品种的PAL和β-1,3-葡聚糖酶活性明显高于感病品种。几丁质酶的活性变化与辣椒对白粉病的抗性无关。