The salicylic acid-dependent defence pathway is effective against different pathogens in tomato and tobacco

The role of salicylic acid (SA) was investigated in basal defence and induced resistance to powdery mildew ( Oidium neolycopersici ) and grey mould ( Botrytis cinerea ) in tomato ( Lycopersicon esculentum ) and tobacco ( Nicotiana tabacum ). A comparison of NahG transgenic tomato and tobacco (unable to accumulate SA) to their respective wild types revealed that in tomato, SA was not involved in basal defence against O. neolycopersici but NahG tobacco showed an enhanced susceptibility to O. neolycopersici infection, the effect becoming more obvious as the plants grew older. In contrast, SA played no role in the basal defence of tobacco against B. cinerea , but seemed to contribute to basal defence of tomato against B. cinerea. Activation of the SA-dependent defence pathway via benzothiadiazole (BTH) resulted in induced resistance against O. neolycopersici in tobacco but not in tomato. Microscopic analysis revealed that BTH treatment could prevent penetration of the Oidium germ tube through tobacco leaves, whereas penetration was successful on tomato leaves irrespective of BTH treatment. In contrast, soil or leaf treatment with BTH induced resistance against B. cinerea in tomato but not in tobacco. It is concluded that the SA-dependent defence pathway is effective against different pathogens in tomato and tobacco.     

Oidium neolycopersici: intraspecific variability inferred from amplified fragment length polymorphism analysis and relationship with closely related powdery mildew fungi infecting various plant species

Previous works indicated a considerable variation in the pathogenicity, virulence, and host range of Oidium neolycopersici isolates causing tomato powdery mildew epidemics in many parts of the world. In this study, rDNA internal transcribed spacer (ITS) sequences, and amplified fragment length polymorphism (AFLP) patterns were analyzed in 17 O. neolycopersici samples collected in Europe, North America, and Japan, including those which overcame some of the tomato major resistance genes. The ITS sequences were identical in all 10 samples tested and were also identical to ITS sequences of eight previously studied O. neolycopersici specimens. The AFLP analysis revealed a high genetic diversity in O. neolycopersici and indicated that all 17 samples represented different genotypes. This might suggest the existence of either a yet unrevealed sexual reproduction or other genetic mechanisms that maintain a high genetic variability in O. neolycopersici. No clear correlation was found between the virulence and the AFLP patterns of the O. neolycopersici isolates studied. The relationship between O. neolycopersici and powdery mildew anamorphs infecting Aquilegia vulgaris, Chelidonium majus, Passiflora caerulea, and Sedum alboroseum was also investigated. These anamorphs are morphologically indistinguishable from and phylogenetically closely related to O. neolycopersici. The cross-inoculation tests and the analyses of ITS sequences and AFLP patterns jointly indicated that the powdery mildew anamorphs collected from the above mentioned plant species all represent distinct, but closely related species according to the phylogenetic species recognition. All these species were pathogenic only to their original host plant species, except O. neolycopersici which infected S. alboroseum, tobacco, petunia, and Arabidopsis thaliana, in addition to tomato, in cross-inoculation tests. This is the first genome-wide study that investigates the relationships among powdery mildews that are closely related based on ITS sequences and morphology. The results indicate that morphologically indistinguishable powdery mildews that differed in only one to five single nucleotide positions in their ITS region are to be considered as different taxa with distinct host ranges.     

Basal tomato defences to Botrytis cinerea include abscisic acid-dependent callose formation

In Arabidopsis, abscisic acid (ABA) application can induce resistance by priming for callose deposition; this resistance is impaired in ABA-deficient and -insensitive mutants. In tomato, ABA-deficiency causes resistance to Botrytis cinerea. Here, we show that callose deposition after B. cinerea inoculation is weaker in the ABA-deficient sitiens tomato mutant compared to the wild type (WT). Inhibition of callose synthesis did not affect resistance in sitiens, but caused additional susceptibility in WT. These findings indicate that callose deposition is not part of sitiens defence responses that are effective in blocking B. cinerea and suggest that callose deposition only contributes to WT basal resistance. Furthermore, also in tomato callose formation is at least in part ABA-dependent. However, it seems that in contrast to Arabidopsis, basal ABA levels in tomato are sufficiently high to prime for callose deposition.     

干旱胁迫对苗期木薯内源激素含量的影响

以SC124、KU50、C4和SC8四个木薯品种为试验材料,运用酶联免疫吸附测定法,研究了盆栽条件下干旱胁迫对苗期木薯根系与叶片中内源激素脱落酸（ABA）、吲哚乙酸（IAA）、赤霉素（GA）和玉米素核苷（ZR）的含量以及四种激素间平衡关系的影响。结果表明：随着干旱程度的加强,四种激素的含量在不同木薯品种内呈现不同的变化趋势。干旱至第10天时,ABA含量在苗期木薯叶片内增加,在根系内含量也增加（KU50除外）;IAA在叶片中含量下降,根系内含量因品种不同,变化趋势相差明显,其中SC124和KU50中IAA含量显著下降,而在SC8和C4中含量上升;GA含量在四种品种的根系中均下降,叶片GA含量在C4和KU50中略有下降而在SC124和SC8中上升;ZR含量在四种品种的根系中均下降,叶片中ZR含量在C4中上升,SC8中下降,而在KU50和SC124中无明显变化。IAA/ABA ,ZR/ABA在根系中比值下降,SC124降幅最大,SC8降幅最小,KU50、C4降幅居中,说明IAA/ABA ,ZR/ABA与木薯的抗旱能力具有一定相关性。该研究结果解释了苗期木薯根系和叶片内源激素对干旱胁迫的适应性反应,为筛选不同木薯抗旱品种提供了理论依据。     

Ethylene-insensitive tobacco shows differentially altered susceptibility to different pathogens

ABSTRACT Transgenic tobacco plants (Tetr) expressing the mutant etr1-1 gene from Arabidopsis thaliana are insensitive to ethylene and develop symptoms of wilting and stem rot when grown in nonautoclaved soil. Several isolates of Fusarium, Thielaviopsis, and Pythium were recovered from stems of diseased Tetr plants. Inoculation with each of these isolates of 6-week-old plants growing in autoclaved soil caused disease in Tetr plants but not in nontransformed plants. Also, when 2-week-old seedlings were used, nontransformed tobacco appeared nonsusceptible to the Fusarium isolates, whereas Tetr seedlings did develop disease. Tetr seedlings were not susceptible to several nonhost Fusarium isolates. In contrast to results with Fusarium isolates, inoculation of 2-week-old seedlings with a Thielaviopsis isolate resulted in equal symptom development of nontransformed and Tetr tobacco. In order to explore the potential range of pathogens to which Tetr tobacco plants display enhanced susceptibility, the pathogenicity of several root and leaf pathogens was tested. Tetr plants were more susceptible to the necrotrophic fungi Botrytis cinerea and Cercospora nicotianae and the bacterium Erwinia carotovora, but only marginally more to the bacterium Ralstonia solanacearum. In contrast, the biotrophic fungus Oidium neolycopersici, the oomycete Peronospora tabacina, and Tobacco mosaic virus caused similar or less severe symptoms on Tetr plants than on nontransformed plants. Total peroxidase activity of Tetr plants was lower than that of nontransformed plants, suggesting a role for peroxidases in resistance against necrotrophic microorganisms. A comparable range of pathogens was examined on Arabidopsis and its ethylene-insensitive mutants etr1-1 and ein2-1. With the exception of one Fusarium isolate, ethylene insensitivity increased susceptibility of Arabidopsis plants to a similar spectrum of necrotizing pathogens as in tobacco. Thus, both ethylene-insensitive tobacco and Arabidopsis plants appear to be impaired in their resistance to necrotrophic pathogens.     

盐胁迫对苗期向日葵内源激素含量的影响

对水培20 d的苗期向日葵进行浓度为0、50、100 mmol·L-1以及时间为10、20、30 h的盐胁迫处理,运用酶联免疫吸附法(ELISA)分别测定了各组向日葵的根部、茎秆和叶片中内源激素的浓度变化。结果表明:随着盐浓度的增加以及胁迫时间的延长,向日葵根、茎、叶中四种激素呈现不同的变化趋势。IAA,ABA和ZR三种激素总体表现为先增后降的趋势,盐浓度越大,植物激素上升或下降的幅度越大,变化速度也越快。GA没有显著变化且波动在±2.0 ng·g-1范围内。而根、茎、叶中IAA/ABA,GA/ABA,(ZR+IAA+GA)/ABA的比值总体呈下降趋势,表明了盐胁迫对生长的抑制。盐胁迫下向日葵叶片和茎秆中这几种激素的变化更为明显,而根部的变化则较为缓慢。     

Evaluation of systemic fungicides for use in the glasshouse industry

The primary evaluation of ten fungicides for the control of cucumber powdery mildew (Sphaerotheca fuliginea), tomato leaf mould (Cladosporium fulvum) and tomato grey mould (Botrytis cinerea) indicated that nine of them were systemically active against one or more of the pathogens when applied as soil drenches. Benomyl, thiophanate, thiophanate-methyl, thiabendazole and Cela W52A showed activity against all three diseases, but dimethirimol, triarimol and dodemorph were more specific and mainly active against cucumber powdery mildew. An in vitro bioassay generally indicated that fungitoxicity was not translocated from sprayed to unsprayed leaves, and though toxicity to B. cinerea, following soil drenches of benomyl, thiophanate and thiophanate-methyl, was detected by bioassay in leaf sap and calyces it was not detected in fruit. Furthermore, extended evaluation of benomyl for the control of grey mould on cropping tomotoes showed that in the presence of abundant B. cinerea inoculum, benomyl soil drench treatment effectively controlled flower and leaf infection but not fruit ghost-spotting.     

Long‐lasting β‐aminobutyric acid‐induced resistance protects tomato fruit against Botrytis cinerea

Minimizing losses to pests and diseases is essential for producing sufficient food to feed the world's rapidly growing population. The necrotrophic fungus Botrytis cinerea triggers devastating pre‐ and post‐harvest yield losses in tomato (Solanum lycopersicum). Current control methods are based on the pre‐harvest use of fungicides, which are limited by strict legislation. This investigation tested whether induction of resistance by β‐aminobutyric acid (BABA) at different developmental stages provides an alternative strategy to protect post‐harvest tomato fruit against B. cinerea. Soil‐drenching plants with BABA once fruit had already formed had no impact on tomato susceptibility to B. cinerea. However, BABA application to seedlings significantly reduced post‐harvest infection of fruit. This resistance response was not associated with a yield reduction; however, there was a delay in fruit ripening. Untargeted metabolomics revealed differences between fruit from water‐ and BABA‐treated plants, demonstrating that BABA triggered a defence‐associated metabolomics profile that was long lasting. Targeted analysis of defence hormones suggested a role of abscisic acid (ABA) in the resistance phenotype. Post‐harvest application of ABA to the fruit of water‐treated plants induced susceptibility to B. cinerea. This phenotype was absent from the ABA‐exposed fruit of BABA‐treated plants, suggesting a complex role of ABA in BABA‐induced resistance. A final targeted metabolomic analysis detected trace residues of BABA accumulated in the red fruit. Overall, it was demonstrated that BABA induces post‐harvest resistance in tomato fruit against B. cinerea with no penalties in yield.     

Spatial and temporal variation of reactive oxygen species and antioxidant enzymes in o-hydroxyethylorutin-treated tomato leaves inoculated with Botrytis cinerea

Pretreatment of detached tomato leaves with o -hydroxyethylorutin reduced the percentage leaf area affected, and delayed the appearance of necrosis, following inoculation with conidial suspensions in droplets of the grey mould fungus Botrytis cinerea . o -Hydroxyethylorutin delayed, but did not inhibit, in vitro germination of conidia, although overall percentage germination was reduced compared with water controls. Both the reactive oxygen species (ROS) – superoxide anions and hydrogen peroxide – increased twice as much in o -hydroxyethylorutin-treated leaf tissue 2 and 6 h postinoculation with B. cinerea conidia in tissues under inoculation drops, as well as in surrounding tissues, whereas in plants not pretreated with the compound ROS generation was noticed later, and only in tissues under inoculation drops. Compared with these compounds, changes in the levels of hydroxyl radicals, lipid peroxidation and the activity of the enzymes superoxide dismutase, guaiacol peroxidase, ascorbate peroxidase and catalase were largely unchanged. In stimulating ROS in inoculated tomato tissue, B. cinerea appeared to be affected directly pre- and postinfection, but indirect effects increasing host resistance cannot be excluded.     

干旱胁迫及复水对豌豆叶片内源激素含量的影响

采用盆栽试验方法, 研究了干旱胁迫及复水对豌豆叶片脱落酸(ABA)、吲哚乙酸(IAA)、赤霉素(GA)和玉米素(ZT)等4种内源激素含量的影响.结果表明: 苗期轻度和重度干旱胁迫分别导致叶片ABA含量增加1.03%和1.10%,IAA含量增加16.44%和61.99%,GA含量降低1.44%和4.79%,ZT含量降低2.10%和3.16%;始花期轻度和重度干旱胁迫分别导致叶片ABA含量增加1.60%和1.90%,IAA含量增加14.42%和27.75%,GA含量降低1.17%和1.79%,ZT含量降低16.65%和19.83%;灌浆期轻度和重度干旱胁迫分别导致叶片ABA含量增加0.83%和1.63%,IAA含量增加12.25%和29.02%,GA含量降低0.87%和1.76%,ZT含量降低7.89%和13.23%.同时对各种内源激素比例产生影响.复水可对豌豆叶片内源激素含量产生补偿效应, 其补偿量决定于豌豆生育时期、干旱胁迫强度和历时、复水历时.复水可促进豌豆叶片内源激素比例发生变化.     

干旱胁迫对不同根型苜蓿内源激素含量的影响

为探究不同根型苜蓿在不同生长年限的抗旱性与内源激素间的关系,采用高效液相色谱法测定根茎型、直根型、根蘖型苜蓿播种当年及生长第2年在干旱胁迫后地上与地下部内源激素吲哚乙酸（IAA）、赤霉素（GA₃）、玉米素（ZT）和脱落酸（ABA）含量。结果表明,随干旱胁迫加剧,播种当年及生长第2年,根茎型、直根型和根蘖型苜蓿地上、地下部IAA和ZT含量显著降低,相比CK,IAA的年均降幅分别为50.09%、36.52%、49.15%和58.17%、53.42%、56.62%,ZT的年均降幅分别为38.11%、52.57%、36.18%和43.86%、63.94%、54.17%。ABA含量显著升高,随生长年限延长增幅降低,种植当年及生长第2年的增幅分别处于62.50%～358.82%和19.70%～85.19%之间;而GA₃含量在播种当年显著下降,生长第2年中、重度胁迫下达到峰值。相比CK,播种当年地上、地下部GA₃/ABA、IAA/ABA、（IAA+GA₃+ZT）/ABA比值及干质量随胁迫加剧呈下降趋势;生长第2年...     

冠菌素诱导水稻幼苗抗旱性的生理效应

以水稻日本晴为材料,在正常生长条件和用质量分数为17.5%的聚乙二醇(PEG)模拟干旱条件下,分析了0.1 μmoL/L冠菌素(coronatine,COR)处理对水稻幼苗生物量的积累、叶片中抗氧化酶[超氧化物岐化酶(SOD)、过氧化氢酶(CAT)及过氧化物酶(POD) ]活性以及渗透调节物质(脯氨酸和可溶性糖)、可溶性蛋白、内源激素[脱落酸(ABA)、吲哚-3-乙酸(IAA)、茉莉酸甲酯(Me-JA)、玉米素核苷(ZR)和赤霉素(GA4)]含量的影响。结果表明:在正常条件下,COR处理对幼苗生物量的积累、抗氧化酶、渗透调节物质和可溶性蛋白的含量均无显著影响,但可增加其体内ZR和GA4 2种激素的含量;在模拟干旱胁迫条件下,COR处理可促进幼苗生物量的积累,维持较高的抗氧化酶活性,促进叶片中脯氨酸、可溶性糖和可溶性蛋白的积累,调节植物体内渗透势,维持[JP2]细胞膨压,同时促进ABA和ZR含量大量累积。研究结果表明,COR通过增强水稻叶片中抗氧化酶活性、调节体内渗透调节物质以及内源激素的含量,可改善水稻幼苗在干旱胁迫条件下的耐旱能力。     

五种脱落酸功能类似物对提高小麦抗盐胁迫的效应和机理

土壤盐渍化是降低作物产量的非生物胁迫因素之一,对小麦种子出苗和苗期生长造成严重影响。本研究以小麦品种“鲁原502”为材料,采用室内水培法探讨了5种脱落酸(ABA)功能类似物——萘酮戊酸(natenpac,NA),quinabctin (QB)及其3种结构类似物tetrafluoroquinabctin (TFQB)、indabactin (IQB)和tetrafluorothioquinabctin (TFTQB)对提高其幼苗抗盐性的影响。结果表明:苗期喷施上述5种化合物均能显著促进在300 mmol/L氯化钠胁迫下小麦幼苗的生长,其中喷施TFQB、TFTQB和NA的效果最好;NA和TFTQB能够通过增加根长、根表面积和根体积而显著促进盐胁迫下小麦幼苗的根系生长,从而增强小麦幼苗根系的吸水能力;5种ABA功能类似物对盐胁迫下小麦幼苗的光合效应、抗氧化酶活性、细胞膜受损状况、渗透调节物质积累等均有显著改善,其中NA处理可显著减轻PSⅡ最大光能转换效率 (Fv/Fm) 下降的幅度,维持胞间二氧化碳(CO₂)浓度的功能优于其他4种化合物;除IQB外,其他4种化合物均能有效降低丙二醛(MDA)含量、显著降低内源生长素(IAA)含量并提高内源ABA含量,其中NA在启动ABA信号途径以提高抗盐胁迫方面比其他4种化合物更有效。本研究结果对揭示ABA功能类似物的作用机制、提高小麦耐盐性具有重要的理论意义和潜在的应用价值。     

植物干旱胁迫中的ABA研究

抗旱机埋的研究不断深入，ABA生理功能的认识不断深入，工厂规模化生产ABA的技术取得突破。在以上几重背景下对植物干旱胁迫中ABA的相关研究作了较详细和全面的综述，主要包括ABA在植物体内的分布与积累，干旱胁迫下植物根、茎、叶中ABA的反应ABA对干旱胁迫下植物的作用，ABA与基因的干旱胁迫诱导，ABA与干旱胁迫的信号转导等。     

Infection of Commercial Hybrid Primula Seed by Botrytis cinerea and Latent Disease Spread Through the Plants

ABSTRACT Botrytis cinerea occurred commonly on cultivated Primula xpolyantha seed. The fungus was mostly on the outside of the seed but sometimes was present within the seed. The fungus frequently caused disease at maturity in plants grown from the seed, demonstrated by growing plants in a filtered airflow, isolated from other possible sources of infection. Young, commercially produced P. xpolyantha plants frequently had symptomless B. cinerea infections spread throughout the plants for up to 3 months, with symptoms appearing only at flowering. Single genetic individuals of B. cinerea, as determined by DNA fingerprinting, often were dispersed widely throughout an apparently healthy plant. Plants could, however, contain more than one isolate.     

ALA对NaCl 胁迫下不同品种番茄植株光合作用、保护酶活性及果实产量的影响

以耐盐性不同的番茄品种金鹏超冠(耐盐性较强)和中杂9号(耐盐性较弱)为材料,研究了叶面喷施5-氨基乙酰丙酸(ALA)对150 mmol/L,250 mmol/L NaCl胁迫下番茄光合作用、保护酶系统及产量的影响。结果表明,ALA能使NaCl胁迫下番茄叶片的叶绿素含量、净光合速率(Pn)、蒸腾速率(Tr)、气孔导度(Gs)升高,胞间二氧化碳浓度(Ci)降低;ALA处理显著提高了高盐胁迫下的SOD、POD和CAT活性,且降低了丙二醛含量,尤其是中杂9号更为显著,ALA处理可能是通过促进保护酶系统活性,保护细胞的光合性能,促进光合作用,从而促进番茄的生长发育,提高产量,缓解盐胁迫对番茄的抑制作用。     

Significance of leaf infection by Botrytis cinerea in stem rotting of tomatoes grown in non-heated greenhouses

The most serious symptom of Botrytis cinerea in tomatoes grown in greenhouses is stem rotting. Lesions on the stem may result from direct infection or from progression of the rot along infected leaves, until infection approaches the stem. In a set of experiments conducted in commercial greenhouses, an experimental greenhouse and growth chambers, the significance of the two types of stem infections was studied. In non-heated greenhouses most of the stem lesions originated from progression of the pathogen along infected petioles. The rate at which B. cinerea had progressed on infected petioles was 0.3–0.5 cm/day, an average of ca. 6 weeks was needed for a leaf infection to approach the stem. Application of Trichoderma harzianum T39 extended this time by 1–2 weeks and application of chemical fungicides by 3 weeks. Influence of the environment on the progression of B. cinerea along infected petioles was then determined. Within range of 5–30 °C, the higher the temperature, the more rapid was the rate of disease progression. The fungus progressed more rapidly on tomato petioles incubated at high vapour pressure deficit (VPD) rather than at low VPD. The source-sink relationship of the plant governed the rate of B. cinerea progression along the petioles as well: it was more rapid when the source was restricted (by shading) and slower when the sink was restricted (by removal of flowers and small fruits). The possibility that sanitation of infected leaflets would reduce the incidence of stem rotting was examined in two experiments. In plots not treated with a fungicide, the sanitation treatment substantially decreased the incidence of stem lesions and this treatment was as effective as weekly application of chemical fungicides.     

灰葡萄孢拮抗细菌的筛选

采集扬州、淮阴两地１６种常见植物根部土壤、根和叶片样本１８２份,分离纯化后得到细菌６３２株,抑菌圈法测定对灰葡萄孢有拮抗性的５８株,拮抗性稳定的１３株。其中拮抗性最强的Ｙ２－１１－１菌株为芽孢杆菌,抑菌圈直径达２９．７ｍｍ。接种试验表明,Ｙ２－１１－１以番茄叶片和果实灰霉病的防效分别达６８．７％和７５．０％。部分菌株对其他重要植物病原真菌也有一定的拮抗作用。     

Virus-induced changes in the response of faba bean to infection by Botrytis

Prior infection of faba bean with the viruses bean yellow mosaic and bean leaf roll increased host susceptibility to subsequent infection by Botrytis fabae and B. cinerea. Cell necrosis beneath inoculum droplets, rate and extent of lesion spread and sporulation of B. fabae were all increased on detached leaves from virus-infected compared with healthy plants. Changes were most marked in young leaves showing conspicuous symptoms of systemic virus infection and in plants virus-infected for at least 2-4 weeks. Localized lesions produced by B. cinerea or a low concentration of B. fabae conidia (10³ spores/ ml) showed increased cell necrosis but were not transformed into aggressive, spreading lesions on virus-infected leaves.