青霉菌Snef1216发酵液诱导番茄抗南方根结线虫研究

 通过Snef1216发酵液处理番茄根系和包衣种子,研究番茄抗南方根结线虫的组织病理学变化和作用方式。结果显示:处理番茄根系且接种15 d,应答根系内根结线虫的数量减少47.5%,且巨细胞出现空泡;包衣番茄种子,接种4、8和12 d,根系内2龄幼虫分别减少54.1%、3.4%和41.7%,接种10、20和30 d,根系内2龄、3龄、4龄幼虫和雌虫的数量也均比对照组番茄明显减少。表明Snef1216发酵液诱导了番茄对南方根结线虫的抗性。     

华南地区主要番茄品种对南方根结线虫的抗性评价

通过人工接种方式, 对华南地区番茄主要栽培品种对优势根结线虫——南方根结线虫（Meloidogyne incognita）的抗性进行了评价。结果表明,广东栽培品种年丰、红宝石、新星、福安、粤红玉、穗丰、金丰、益丰、丰顺、大丰顺、满丰和法国品种SAINT PIERRE均高度感病; 而法国品种 PIERSOL VFN高度抗病,根结率为0。根据SAS8.0多重比较分析表明,在F0.01的水平上, 品种PIERSOL VFN与其余品种之间对根结线虫的抗性均存在显著性差异,而其他12个品种间对根结线虫的抗性差异不显著。     

南方根结线虫生防菌X-20的鉴定及其作用机制研究

南方根结线虫是设施蔬菜上的重要植物病原物,但目前缺乏安全高效的生物防治措施。本研究从根结线虫罹病土壤中分离到一株生防菌X-20,其发酵液对南方根结线虫二龄幼虫校正死亡率达100%。通过形态学、生理生化特征和分子生物学方法将菌株X-20鉴定为荧光假单胞菌Pseudomonas fluorescens。菌株X-20发酵液处理南方根结线虫时,当处理浓度1 mg/mL,虫体出现扭曲、僵直变化,内部组织分解;处理1 h后,虫体波动频率降幅47.26%、头部摆动频率波动性变化、虫体弯曲频率成S型变化;处理24 h后,二龄幼虫的体长减少4.68%,口针增加7.64%,透明尾长增加83.90%;J2虫体体液出现外渗,且电导率随着处理浓度的增大和时间延长而增加;发酵液甲醇提取物1mg/mL处理虫体,6h后糖含量比对照增加39.10%,甘油含量比对照高出66.67%,可溶性蛋白含量降低55.73%。由此可见,荧光假单胞菌X-20菌株代谢产物通过破坏根结线虫体壁及虫体内部结构,引起虫体液渗漏,扰乱虫体内糖类、脂类、蛋白类代谢,造成虫体死亡。     

黄瓜幼苗内生真菌诱导植株对南方根结线虫的抗性

在2007-2008年的一项温室测验中,10株黄瓜幼苗内生真菌表现出了对南方根结线虫较好的防效,为了阐明其作用方式,用裂根试验设计研究了10个菌株诱导黄瓜产生根结线虫抗性的能力.同一植株的根系被分为彼此隔离的挑战根系和应答根系,镰刀菌Fusarium spp.菌株Fu234和Fu654、毛壳菌Chaetomium sp.菌株Ch1001、叶点霉Phyllosticta sp.菌株Ph511在挑战根系中接种后,应答根系中的线虫入侵总数显著降低,较对照分别降低了42.4％、35.7％、38.4％、23.6％(第1次测定)和63.6％、45.2％、51.0％、37.0％(重复测定),这4个菌株诱导了黄瓜对根结线虫入侵的抗性.同时,Fu234、Fu654、Ch1001和拟青霉Paecilomyces sp.菌株Pa972接种后应答根系中的卵与雌虫的比值显著减少,相比对照其比值分别降低了24.3％、37.7％、48.3％、21.2％(第1次测定)和21.6％、39.8％、30.0％、46.2％(重复测定),这4个菌株诱导了黄瓜对根结线虫繁殖的抗性.     

生姜对南方根结线虫侵染的生理生化反应

 本试验就生姜感染南方根结线虫后几种生理生化指标的变化进行了研究.结果表明,接种南方根结线虫后,生姜体内各种生理生化指标主要在初次侵染前期和二次侵染前期表现出一定的反应,其余时间则基本丧失这种反应,且生姜不同部位对南方根结线虫侵染的反应程度也有很大差异,以叶片的反应最为剧烈,根系次之,根茎反应最弱.结合南方根结线虫的生活周期,本文提出了生姜对南方根结线虫的生理生化反应主要表现在侵染初期,对再次侵染则部分丧失敏感性的论点.     

南方根结线虫侵染对黄瓜植株生长和抗氧化系统的影响

为探明野生黄瓜对南方根结线虫的抗性机制,采用温室盆栽苗期人工接种技术,研究了线虫侵染对黄瓜抗感品种植株生长和抗氧化系统的影响。南方根结线虫的侵染显著降低了感病品种津春4号的株高、叶面积、地上部鲜重与干重及根系活力,并显著增加其根系鲜重与干重,而对抗病品种西印度瓜的各项生长指标均未造成明显影响。西印度瓜中超氧阴离子(O.2-)、过氧化氢(H2O2)、电导率和丙二醛(MDA)含量增加幅度均小于津春4号,超氧化物歧化酶(SOD)活性降低,过氧化物酶(POD)、苯丙氨酸解氨酶(PAL)活性增加幅度均显著高于津春4号。研究表明西印度瓜遭受的氧化胁迫较小,其抗氧化系统有效清除了氧化损伤。     

不同杀线虫剂对南方根结线虫的田间防效

为评价新型杀线虫剂三氟吡啶胺防治南方根结线虫的效果,设置了该药剂与作为对照的3种常见杀线虫剂灌根或拌土穴施处理,比较各处理防治西瓜田南方根结线虫的效果。在防控南方根结线虫导致的死棵方面,对照药剂氟吡菌酰胺、阿维菌素、噻唑膦处理区西瓜死棵率为14.70%～44.85%,清水对照区为46.97%,而三氟吡啶胺处理区死棵率仅5.61%;在根结防效方面,三氟吡啶胺处理为70.43%,显著优于药剂对照;在虫口防效方面,三氟吡啶胺处理为66.72%,为各处理中最高。建议将三氟吡啶胺推广应用于南方根结线虫的田间防治。     

长枝木霉菌株TL16防治南方根结线虫的作用机理

为探明长枝木霉Trichoderma longibrachiatum菌株TL16防治南方根结线虫Meloidogyne incognita的作用机理,采用原生质体转化法获得绿色荧光蛋白(green fluorescent protein,GFP)标记菌株GFP-TL16,通过测定菌株TL16和GFP-TL16对南方根结线虫卵和2龄幼虫(2nd-stage juvenile,J2)的寄生与致死作用,其发酵液对卵孵化的抑制作用和对J2的致死作用,以及菌株GFP-TL16在黄瓜根系的定殖情况和菌株TL16对番茄根结形成的抑制作用来综合分析其作用机理。结果显示:菌株TL16菌丝对南方根结线虫卵无寄生作用,处理19 d后卵降解率为26.33%,致死作用较低;菌株TL16分生孢子悬浮液处理南方根结线虫J2后72 h的致死率为1.65%,且无寄生作用。菌株TL16发酵液处理南方根结线虫J2后48 h的校正死亡率为10.71%,处理卵15 d后对卵孵化的相对抑制率为77.11%。菌株GFP-TL16可定殖于黄瓜根系中,经菌株TL16处理后接种南方根结线虫J2,番茄根结减退率为55.88%。表明长枝木霉菌株TL16可通过抑制根结线虫卵孵化和诱导番茄产生抗病性来防治根结线虫病。     

Cf-2/Rcr3pim番茄品系对南方根结线虫的抗性测定

Cf-2/Rcr3~(pim)基因型番茄不仅能够抵御番茄叶霉菌的侵染,而且对马铃薯金线虫的寄生也有一定的抑制效果。为挖掘根结线虫的新抗性资源,本研究采用室内人工接种法测定了Cf-0/Rcr3~(pim)、Cf-2/Rcr3-3和Cf-2/Rcr3~(pim)基因型番茄品系对南方根结线虫的抗感性。抗性评价结果显示,Cf-0/Rcr3~(pim)品系对南方根结线虫表现高感,Cf-2/Rcr3-3品系为中感,而Cf-2/Rcr3~(pim)品系则为感病。与Cf-0/Rcr3~(pim)和Cf-2/Rcr3-3基因型相比,Cf-2/Rcr3~(pim)基因型番茄品系虽然对南方根结线虫侵染的敏感性略低,但是不能阻止线虫在根系上的大量繁殖,不适于根结线虫的防控应用。     

厚壁孢子轮枝菌防治南方根结线虫研究初报

从山东、江苏大豆孢囊线虫和番上的南方根给线虫卵中筛选分离到厚壁孢子轮枝菌。琼脂平板法测试这种食线虫真菌有寄生南方根结线虫卵的能力，发现一菌株的卵寄生率达９０．８％，高于同时分离到的淡紫似青霉。试验还表明不同来源的菌株，卵寄生半差异很大。温室盆钵试验６０天结果，０．１％的菌剂用量即可明显降低番茄上的南方根结线虫幼虫群体数量和根结指数，卵寄生率可达５３．７％。该菌在根围土壤和根表面的残留量分别是３．７×１０￣５个菌落／克土壤和６．３×１０￣５个菌落／克根。菌剂处埋对番茄植株的影响，１％的用量，地上部鲜重平均比对照重３．３克，每克根长比对照增长近１倍。各处理的番茄根和叶片均未出现任何病斑。     

干旱胁迫下桑树活性氧信号传导及转录组分析

对干旱胁迫处理后的桑树幼苗(‘湖桑32’和‘德果1号’)进行ROS信号组织定位和叶的转录组测序，旨在从细胞和分子角度探究干旱胁迫后ROS信号在桑树组织细胞中的分布规律及与之相关的代谢通路和基因调控表达变化。结果表明:在正常情况下桑叶组织细胞中会产生少量的■，而干旱胁迫处理后桑叶组织细胞中ROS积累增多，推测ROS作为一种动态信号分子，会向毗邻细胞或远端细胞进行信号转导，进而与多种信号组分共同完成系统响应表达以调控桑树的生长发育及抗逆性。转录组数据表明:重度干旱胁迫处理后‘湖桑32’有5677个差异表达基因显著多于‘德果1号’(5575)，且差异显著基因(DEGs)主要富集在半乳糖代谢、淀粉和蔗糖代谢、果糖和甘露糖代谢、碳代谢等通路中，推测‘湖桑32’的抗旱性与渗透调节物质的合成有关。同时，挑选11个基因进行qRT-PCR验证，其中11个DEGs的表达趋势与对应的RNA-Seq数据一致。研究也发现调控MKK4/5和WRKY22表达的相关基因LOC21384958和LOC21396104，在弱抗旱桑‘德果1号’中分别上调表达0.72(P<0.05)和2.16(P<0.05)，而...     

Application of energy-filtering electron microscopy (EFEM) for analysis of hydrogen peroxide and lignin in epidermal walls of cucumber leaves triggered by acibenzolar-S-methyl treatment prior to inoculation with Colletotrichum orbiculare

Hydrogen peroxide (H₂O₂) and lignin induced by acibenzolar-S-methyl (ASM) during the expression of systemic acquired resistance (SAR) of cucumber plants against the fungus Colletotrichum orbiculare was analysed with electron microscopy and energy-filtering electron microscopy (EFEM). ASM pretreatment consistently led to the blockage of appressorial penetration pegs into leaves at early stages after inoculation, whereas the fungus was able to grow into the vascular tissue after distilled water pretreatment. The blocked pegs, seen frequently at epidermal pectin layers in leaves of ASM-treated plants, were surrounded by abundant electron-dense amorphous material and dots. These were identified as lignin, based on their appearance and high reactivity with KMnO₄. Lignin first appeared as an amorphous material at pectin layers, and then as dots in some areas of the material. The results showed that ASM caused SAR in leaves via faster formation of lignin within 1 day after inoculation. After CeCl₃ treatment to detect H₂O₂, H₂O₂-reactive products (cerium perhydroxides) were seen near ASM-induced lignin formation sites in pectin layers. EFEM analysis showed that Ce and O were located at the same sites as the lignin dots, suggesting that lignin was associated with sources of H₂O₂ generation. The results indicate that the faster H₂O₂ generation and lignification induced by ASM were the potential causes of SAR.     

l‐DOPA inhibited the root growth of lettuce by inducing reactive oxygen species generation

l ‐3,4‐dihydroxyphenylalanine (l ‐DOPA) is a bioactive natural compound that inhibits the growth of several plant and weed species. However, its mode of action is not yet well understood in plants. The present study was conducted to explore the mechanism of growth inhibition by l ‐DOPA in lettuce (Lactuca sativa L.). The results revealed that l ‐DOPA induced cell death and inhibited the root growth of lettuce. Moreover, 0.1 mmol L^–1 l ‐DOPA enhanced the polyphenol oxidase activity, production of reactive oxygen species ( and H₂O₂) and lipid peroxidation in the lettuce roots. The results suggest that l ‐DOPA‐induced growth inhibition in lettuce roots might be related to oxidative stress.     

Characterization of reactive oxygen species-involved oxidative damage in Hapalosiphon species crude extract-treated wheat and onion roots

Crude extract-induced oxidative damage using the cyanobacterium, Hapalosiphon sp., was investigated in wheat seedlings ( Triticum aestivum L. cv. Norin 61) and onion seedlings ( Allium cepa L. cv. Raputa II). The analysis of root cell viability or cell death using Evans blue uptake showed that the root-tip cells of wheat and onion lost viability after 24 h and 48 h treatment with 3 g dry weight (DW) L⁻¹ of the crude extract, respectively. Lipid peroxidation was induced in the roots of both species and the shoots of onion, whereas no increase in lipid peroxide formation was observed in the wheat shoots. In onion, the degree of random DNA fragmentation increased with the increasing concentration of the extract and laddering of the DNA was observed with 6 g DW L⁻¹ of the extract, but no apparent DNA ladder formation occurred in the wheat. Pretreatment for 1 h with the NADPH oxidase inhibitors, diphenyleneidonium or imidazole, reduced the crude extract-induced root cell death in both species. From the results, we suggest that the Hapalosiphon sp. crude extract might enhance reactive oxygen species (ROS) production, which causes membrane lipid damage and fragmentation of the DNA of plant cells, resulting in cell death and growth inhibition. The crude extract-mediated phytotoxic damage might be caused by ROS, triggered by NADPH oxidase.     

莲子草假隔链格孢对空心莲子草活性氧代谢的影响

为了明确活性氧(ROS)在莲子草假隔链格孢Nimbya alternantherae侵染空心莲子草过程中的作用,采用氯化硝基四氮唑蓝(NBT)光化学还原法、二氨基联苯胺(DAB)组织染色法和分光光度法测定了莲子草假隔链格孢SF-193侵染空心莲子草后超氧阴离子(O2-)产生速率、过氧化氢(H2O2)含量、抗氧化酶体系(超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和过氧化物酶(POD))活性和丙二醛(MDA)含量.结果显示,接种莲子草假隔链格孢SF-193的叶片经NBT和DAB染色后检测到ROS的大量累积.叶片内的O2-产生速率和H202含量随接种时间显著升高,接种后48 h均达到峰值,分别为2.33 nmol·min-1·g-1FW和11.78 μmol·g-1FW,是对照的2.1倍和2.5倍;SOD、CAT和POD活性均在接种后8h达到峰值,分别为277.75、23.47和444.92 U·min-1·g-1FW,之后逐渐下降;MDA含量随接种时间持续上升,72h时达到峰值,为5.20μmol·g-1 FW.表明莲子草假隔链格孢侵染空心莲子草打破了ROS产生与清除之间的平衡,导致植物细胞内的ROS迅速上升,对叶片细胞造成严重破坏.     

外源NO对渗透胁迫下黑麦草种子萌发及活性氧代谢的影响

以黑麦草为试材,研究了一氧化氮(NO)供体硝普钠(SNP)对聚乙二醇6000渗透胁迫下种子萌发、幼苗叶片活性氧代谢和渗透溶质积累的影响,探讨外源NO对渗透胁迫下牧草生理响应的调节作用.结果表明:100μmol/L SNP显著提高了渗透胁迫下黑麦草种子的发芽率、发芽指数和活力指数,缓解了植株鲜重和干重的下降幅度,提高了幼苗叶片SOD、POD、APX、质膜H+-ATPase活性及谷胱甘肽、可溶性糖、游离氨基酸、脯氨酸、K+、Na+含量和植株含水量,降低了O2-.产生速率及H2O2、丙二醛和可溶性蛋白含量.表明外源NO能够促进渗透胁迫下黑麦草种子的萌发,对渗透胁迫引起的黑麦草幼苗膜脂过氧化具有缓解作用.     

Generation of reactive oxygen species by a novel anthraquinone derivative in the cyanobacterium Planktothrix perornata (Skuja)

A water-soluble anthraquinone derivative (2-[methylamino-N-(1′-methylethyl)]-9,10-anthraquinone monophosphate), previously found to be selectively toxic towards Planktothrix perornata at submicromolar concentrations, was studied to determine its toxic mode of action towards this cyanobacterium. Chlorophyll fluorescence was monitored as an indicator of photosynthetic efficiency, and measurement of reactive oxygen species (ROS) was performed using the ROS-sensitive probe 2′,7′-dichlorodihydrofluorescein diacetate. The effects of the herbicide paraquat (a ROS generator) as well as ascorbate and α-tocopherol (ROS scavengers) on ROS formation by P. perornata were studied. Also, the effects of different concentrations of ascorbate, α-tocopherol, and the herbicide diuron on reducing the toxicity of the water-soluble anthraquinone derivative towards P. perornata were determined. Our results indicate that the water-soluble anthraquinone derivative does not inhibit photosynthetic electron transport directly, but does generate ROS at levels that may cause toxicity towards P. perornata.     

小麦-条锈菌互作过程中活性氧及保护酶系的变化研究

 对条锈菌与小麦不同互作体系中组织学特征、活性氧产生及其相关酶系的变化进行了研究。组织学观察表明:非亲和组合相对于亲和组合存在明显差异,表现为菌丝生长受抑,吸器母细胞和吸器形成减少,寄主细胞在接种后18h左右出现过敏性坏死。生化测定结果表明:在非亲和组合中,O₂^-的产生速率和H₂O₂的含量均高于亲和组合,且O₂^-产生速率在接种后12h达到一个峰值,H₂O₂的含量在接种后20和72h出现2个高峰。而在亲和组合中,O₂^-产生速率低于对照或与对照相似,H₂O₂的含量虽然高于对照,但却普遍低于非亲和组合;SOD在亲和组合中的活性总体上要高于非亲和组合;接种24h后,CAT在2种组合中的活性均高于对照,在接种后36和48h时,亲和组合中的CAT活性高于非亲和组合,而在接种60h后又开始低于非亲和组合;POD活性在接种24h后均明显升高,但亲和组合中POD活性增幅大;MDA在非亲和组合中于接种后72h含量明显上升。结果表明,亲和与非亲和组合中条锈菌扩展、活性氧的产生及相关酶活变化都存在明显差异,这些差异与小麦抗锈性的表达可能有密切联系。     

猕猴桃品种中糖分及木质素含量 与抗溃疡病的关系

以安徽省猕猴桃主栽品种金魁、早鲜、魁蜜和金丰为研究对象，于展叶孕蕾期分别取发病的枝条、叶片，以未发病的健株相应组织为对照，采用生理生化的方法，分析枝条、叶片中木质素和可溶性糖的含量变化以及与抗溃疡病的关系。结果表明：抗病品种金魁健株枝条、叶片中可溶性糖及木质素含量显著高于感病品种金丰。自然发病后，抗感病品种枝条、叶片中可溶性糖含量都降低，感病品种降低更多，金魁叶片可溶性糖含量下降4．20％，金丰叶片可溶性糖含量下降55.35％；木质素含量都升高，且抗病品种金魁叶片中的木质素含量比感病品种金丰高得多，其变化率分别为7．17％、3．01％，枝条中的木质素含量变化率分别为110．39％、68．98％，其差异达到显著水平。相关分析表明，枝条中木质素、可溶性糖含量与品种发病率也呈负相关，γ分别为-0．9583和-0．9282；叶片中木质素、可溶性糖含量与品种发病率呈负相关，γ分别为-0．8099和-0、8266。从而说明枝条、叶片中可溶性糖及木质素含量与品种抗性呈正相关。抗感品种淀粉含量无明显规律性变化，与品种抗性关系不大。