番茄资源对叶霉病的抗性鉴定和SSR标记遗传变异分析

 对62份番茄种质资源进行了抗番茄叶霉病鉴定, 共筛选出抗病材料17份, 其中对全部生理小种(1.2.3、1.2、2.3、1.2.3.4、1.2.4)均表现抗病或免疫的材料9份;抗3个生理小种的材料4份;抗2个生理小种的材料2份;抗4个生理小种的材料2份。并应用SSR标记技术对62份种质资源进行了遗传多样性分析, 50对SSR引物中筛选出11对多态性明显、条带清晰、反应稳定的引物, 共扩增出94条谱带, 平均每个引物扩增出8.55条带, 多态性条带比例达63.94%, 遗传相似系数在0~1之间, 表现出丰富的遗传多态性, 可将番茄野生种和栽培种分开, 反映了种间的遗传差异。     

番茄与叶霉菌互作的分子机理

 番茄和叶霉菌(Cladosporium fulvum Cooke)系统是研究植物和病原物互作分子机理的模式系统。4个叶霉菌无毒基因已被克隆。这些无毒基因编码含偶数个半胱氨酸的小分子量蛋白,在叶霉菌毒性菌株中的存在与否及存在方式各不相同。Avr4具有几丁质结合活性;而Avr9可能在叶霉菌氮素代谢中起调节作用。7个具有功能的番茄抗叶霉菌Cf基因已被克隆。它们与其同源基因一起以基因簇形式存在于复合基因座中,其成员称为Hcr (homologues of C. fulvum resistance gene Cf)基因。Cf蛋白定位于细胞质膜,但主体在膜外,主要结构域为富含亮氨酸重复(LRR)和跨膜结构域。LRR重复单元数目以及N端序列决定了Cf蛋白对Avr的识别特异性。Cf对Avr的识别遵守"保卫"假说。在Cf-2对Avr2的识别系统中,蛋白酶Rcr3为"被保卫"蛋白。Cf识别Avr后迅速激活下游信号传导和防卫反应,包括过敏性反应的产生和氧化迸发,以及K⁺离子通道、各类蛋白激酶、SGT1、硫氧还蛋白及磷脂酸途径的活化。温度、湿度和光照等环境条件显著影响Cf介导的过敏性反应和抗病性。不同Cf对Avr的识别机理及其下游信号传导途径有显著差别。     

Allelic test proves genes Cf4 and Cf8 for resistance to Cladosporium fulvum (Fulvia fulva) on tomato to be undistinguishable

In an allelic test it was proven that the genes Cf₄ and Cf₈ for resistance toC. fulvum in tomato are undistinguishable, confirming a recent suggestion that Cf₈ does nott provide a novel source for resistance toC. fulvum.Samenvatting In een allelie-toets werd aangetoond dat de genen Cf₄ en Cf₈ voor resistentie tegenC. fulvum van tomaat niet te onderscheiden zijn. Hiermee werd een recente suggestie bevestigd, dat Cf₈ geen nieuwe resistentie tegenC. fulvum verschaft.     

Accumulation of pathogenesis-related proteins in the epidermis of tomato leaves infected by Cladosporium fulvum

Upon infection byCladosporium fulvum, tomato plants start to produce pathogenesis-related (PR) proteins. The PR proteins 1,3-β-glucanase, chitinase, and PR-1b accumulated near the stomata in the lower epidermis ofC. fulvum-inoculated tomato leaves as could be determined by immunolocalization with polyclonal antibodies. However, no differences in accumulation of PR proteins between a compatible and an incompatible interaction were found. Results obtained from enzyme activity measurements of 1,3-β-glucanase and chitinase on similar leaf material as used for the immunolocalization did not fully reflect the immunolocalization data. The antibodies possibly detect only the extracellular but not the intracellular enzymes. The accumulation of PR proteins near the stomata might be part of a general defence response of plants against pathogens and potential pathogens.     

Race 2.5.9, a new race of Cladosporium fulvum (fulvia fulva) and sources of resistance in tomato

Samenvatting Cladosporium fulvum staat bekend om zijn vermogen zich snel aan te passen aan nieuw geïntroduceerde resistentiegenen. Die laatste jaren bleven tomaterassen met C5 resistentie in de praktijk resistent. Deze rassen zijn zelfs resistent tegen het complexe fysio 2.4.5 door het bezit van Cf9 of een ander resistentie-gen. Een fysio 2.4.5.9 trad weliswaar op bij proeven op het IVT (Wageningen), maar werd nooit in de praktijk gevonden.In 1985 werd in Frankrijk het ras Rianto F1 in een praktijkteelt aangetast. Een isolaat vanC. fulvum van dit C5-resistente ras bleek fysio 2.5.9 te zijn. Een deel van de thans bij onderzoek gebruikte resistentiebronnen, zoals Bulgaria no. 1, Japan 74S en Japan 76S, is resistent voor dit nieuwe fysio.Het optreden van dit nieuwe fysio onderstreept de wenselijkheid ten minste twee nieuwe resistentiegenen in nieuwe rassen te gebruiken ter verbetering van de duurzaamheid van de resistentie.     

Race 2.5, a new race of Cladosporium fulvum (Fulvia fulva) on tomato

Samenvatting VanCladosporium fulvum, de veroorzaker van bladvlekkenziekte van tomaat, zijn talrijke fysio's bekend. Van tomaterassen met resistentiegen Cf5 werd, zowel bij materiaal uit Bulgarije als uit Frankrijk, een nieuw fysio, 2.5, geïsoleerd. Dit bezit naast het noodzakelijke virulentiegen 5 ook het overbodige gen 2. Dit is wéér een voorbeeld van afwezigheid van stabilising selection.     

The effect of hydrogen peroxide on the viability of tomato cells and of the fungal pathogen Cladosporium fulvum

An oxidative burst was previously demonstrated to be induced in tomato plants by race specific elicitors of the fungal pathogen Cladosporium fulvum . The in planta levels of H₂O₂estimated to occur during elicitor treatment, were compared with the levels required to show toxicity to host cells and to the fungal pathogen. Injection of Cf-9 tomato leaves with 100 m m H₂O₂caused an insignificant degree of necrosis and 1m H₂O₂was required to cause complete leaf necrosis comparable to that induced by the AVR9 elicitor. Assays with Cf-5 tomato cell suspensions confirmed the low toxicity of H₂O₂to tomato cells but, as expected, the addition of Fe²⁺with H₂O₂(or with intercellular fluids containing AVR5 elicitor) enhanced cell death as determined by the Evans Blue assay. Germination and germ tube growth of conidia of C. fulvum were significantly retarded by 4–5 m m H₂O₂, and at higher concentrations, death of germ tubes was observed (ED₅₀=22 m m), as determined by the fluorescein diacetate assay. The addition of Fe²⁺with H₂O₂had little effect on fungal growth or viability in vitro . These results suggest that the amount of H₂O₂accumulating during an elicitor-induced response in leaves may be sufficient to affect fungal colonization but not to affect viability of host cells unless the Fe²⁺status in the apoplast is in some way altered by the elicitor to facilitate OH^.production via the Fenton reaction.     

A light and scanning-electron microscopic study of infection of tomato plants by virulent and avirulent races of Cladosporium fulvum

Infection of tomato plants byCladosporium fulvum Cooke was studied using light and scanning-electron microscopy. Races 1.2.3 and 4 ofCladosporium fulvum were used, whereas tomato cultivars, carrying the Cf2 gene (susceptible to race 1.2.3 and immune to race 4) and the Cf4 gene (immune to race 1.2.3 and susceptible to race 4) served as differentials. No differences were observed in growth between compatible and incompatible combinations during germination, subsequent formation of runner hyphae and stomatal penetration. Runner hyphae did not show directional growth towards stomata. Penetration usually occurred on the third or fourth day after inoculation. In compatible combinations the fungus grew intercellularly, often in close contact with spongy mesophyll cells. Under optimal conditions it did not cause visible damage to plant cells during early stages of infection. Under suboptimal conditions in winter, the host cells often reacted with callose deposition, but growth of the fungus did not appear to be inhibited. Ten to twelve days after inoculation conidiophores emerged through the stomata and produced conidia. In incompatible combinations fungal growth was arrested one to two days after penetration and confined to stomata and surrounding cells. Very soon the host cells, in contact with the fungus, deposited extensive amounts of callose. Later these cells turned brown and collapsed. At the surface of the host cells, contacted by fungal hyphae, abundant extracellular material could be observed by scanning-electron microscopy. Removing the epidermis of leaves before inoculation delayed the resistant response. On stripped leaves the rate of fungal growth was equal for both interactions up to ten days after inoculation, but the incompatible combination lacked sporulation.     

Characterization of elicitor activities of apoplastic fluids isolated from tomato lines infected with new races of Cladosporium fulvum

Apoplastic fluids were isolated from the near-isogenic lines Cf2, Cf4 and Cf5 of the tomato (Lycopersicon esculentum Mill.) cultivar ‘Moneymaker’ infected with a set of newCladosporium fulvum races 2.5, 2.4.11, 2.5.9, 2.4.5.11, 2.4.9.11 and 2.4.5.9.11 and a set of previously described races 2, 4, 5, 2.4., 2.4.5 and 2.4.5.9. Upon injection of the apoplastic fluids into leaves of the near-isogenic lines Cf0, Cf2, Cf4, Cf5 and Cf9 and the genotypes carrying Cf6 and Cf11 chlorosis or necrosis was observed in all plants that were resistant to the race that initially had been used to raise the apoplastic fluid, except in the genotype Cf11 when the races 4, 2.4, 2.4.5 and 2.4.5.9 from our own collection were involved. The latter races may contain virulence gene all as our collection had not yet been checked for presence of this virulence gene. Apoplastic fluids of all races induced chlorosis on the genotype carrying resistance gene Cf6, a gene which has not yet been overcome by any of the presently described races ofC. fulvum.Low pH non-denaturing polyacrylamide gel electrophoresis showed that apoplastic fluids of compatible interactions with the new races 2.5.9, 2.4.9.11 and 2.4.5.9.11 carrying the virulence gene a9, like the previously described race 2.4.5.9, did not contain the necrosis-inducing peptide, while the peptide was present in all races carrying avirulence gene A9. This indicates that this race-specific elicitor is strongly associated with avirulence gene A9 and is possibly its product as has been suggested previously.     

Identification of race 2.5 of leaf mold (Passalora fulva,syn. Cladosporium fulvum) on tomato

Journal of Plant Diseases and Protection - Leaf mold caused by the fungus Passalora fulva has been a serious disease in greenhouse tomatoes in Turkey. This study was conducted to determine races of...     

Further identification of races of Cladosporium fulvum (Fulvia fulva) on tomato originating from the Netherlands France and Poland

Races ofCladosporium fulvum, which can overcome the resistance of the genes Cf₂, Cf₄, Cf₅, Cf₈, Cf₉ and Cf₁₁ have appeared in the Netherlands, France and Poland. Known isolates from the Netherlands and France and three new isolates from Poland have been investigated for the presence of virulence genes using a set of genotypes carrying resistance genes Cf₂ to Cf₁₁.Several Dutch isolates of races, earlier designated as 2.4, 2.4.5 and 2.4.5.9, were found to break down the resistance gene Cf₁₁. These races must therefore be designated as 2.4.11, 2.4.5.11 and 2.4.5.9.11 respectively. In the new Polish isolates virulence genes, overcoming the resistance genes Cf₂, Cf₄, Cf₈, Cf₉ and Cf₁₁ were found. Since all races able to grow on genotypes with Cf₄, could also grow on genotypes carrying Cf₈, it was impossible to discriminate between the genes Cf₄ and Cf₈. These Polish isolates were designated as races 4.11, 2.4.11 and 2.4.9.11. The consequences of the occurrence of these races for tomato breeding are discussed.Samenvatting Fysio's vanCladosporium fulvum, die de resistentie-genen Cf₂, Cf₄, Cf₅, Cf₈, Cf₉ en Cf₁₁ kunnen doorbreken, zijn in Nederland, Frankrijk en Polen opgetreden. Met behulp van een groep genotypen, die de resistentie genen Cf₂ tot en met Cf₁₁ dragen, zijn Nederlandse, Franse en enkele nieuwe Poolse isolaten onderzocht op de aanwezigheid van virulentiegenen. Enkele Nederlandse isolaten, eerder aangeduid met 2.4, 2.4.5. en 2.4.5.9, bleken het resistentie-gen Cf₁₁ te kunnen doorbreken. Deze moeten daarom aangeduid worden als respectievelijk 2.4.11, 2.4.5.11 en 2.4.5.9.11. In de nieuwe Poolse isolaten werd virulentie gevonden voor Cf₂, Cf₄, Cf₈, Cf₉ en Cf₁₁. Alle fysio's die op genotypen met Cf₄ konden groeien, groeiden ook op genotypen met Cf₈. Daarom kon geen ondersheid gemaakt worden tussen Cf₄ en Cf₈. De Poolse isolaten behoren tot de fysio's 4.11, 2.4.11 en 2.4.9.11. De gevolgen van het voorkomen van deze fysio's voor de tomateveredeling worden besproken.     

A biochemical mechanism of natural resistance of apple to Venturia inaequalis

Samenvatting In cultuurfiltraten vanVenturia inaequalis, de verwekker van appelschurft, werden stoffen gevonden die toxisch zijn voor blad van appelrassen welke tegen dit physio resistent zijn. Blad van vatbare rassen is ongevoelig. De resistentie komt tot stand doordat deze toxische werking in het blad de vorming veroorzaakt van fungicide oxydatie-producten van phloridzine. Op grond van deze waarnemingen kan een mogelijke interpretatie van de gen-om-gen relatie bij deze plant-parasietverhouding worden opgesteld.     

The first report on the occurrence race 9 of the tomato leaf mold pathogen Cladosporium fulvum (syn. Passalora fulva) in Cuba

Leaf mold caused by the fungus Cladosporium fulvum (syn. Passalora fulva) has been a serious disease in greenhouse tomatoes in Cuba, but less damage is seen in outdoor grown plants. This study was conducted to identify races of the causal agent of tomato leaf mold. A total of 36 single conidial isolates obtained from different provinces in Cuba and the reference strain race 0 were inoculated on a differential set of tomato cultivars carrying different Cf resistance genes. Further, a polymerase chain reaction (PCR) was developed using Avr9 gene-specific primers. As result, all isolates could overcome the Cf-9 resistance gene only, but not resistance genes Cf-2, Cf-4 and Cf-5 and must therefore be coined race 9. Race identity was also confirmed by PCR analysis, which showed that the Avr9 gene was not amplified in any of 36 single conidial isolates studied proving that they all lack this gene. This is the first report of the occurrence of race 9 of C. fulvum in Cuba. Pyramiding other Cf genes in hybrid of tomato may provide a more durable level of resistance to C. fulvum.

     

The relationship of host-mediated induced resistance to polymorphism in gene-for-gene relationships

Gene-for-gene relationships are a common feature of plant-parasite interactions. Polymorphism at host resistance and parasite avirulence loci is maintained if there is negative, direct frequency-dependent selection on alleles of either gene. More specifically, selection of this kind is generated when the disease is polycyclic with frequent auto-infection. When an incompatible interaction occurs between a resistant host and an avirulent parasite, systemic defenses are triggered, rendering the plant more resistant to a later attack by another parasite. However, induced resistance (IR) incurs a fitness cost to the plant. Here, the effect of IR on polymorphism in gene-for-gene interactions is investigated. First, in an infinite population model in which parasites have two generations per host generation, increasing the fitness cost of IR increases selection for susceptible plants at low disease severity, while increasing the effectiveness of IR against further parasite attacks enhances selection for resistant plants at high disease severity. This reduces the possibility of polymorphism being maintained in host and parasite populations. In finite population models, the number of plants varies over time as a function of the disease burden of the population. Polymorphism in gene-for-gene relationships is then more stable at high disease prevalence and severity if IR reactions are more costly when there is competition for resources between plants.     

Carbohydrate composition of apoplastic fluids isolated from tomato leaves inoculated with virulent or avirulent races of Cladosporium fulvum (syn. Fulvia fulva)

Inoculation of tomato (Lycopersicon esculentum) with virulent races ofCladosporium fulvum (compatible interactions), resulted in substantial changes of the carbohydrate composition of apoplastic fluids isolated from the leaves, during the course of the infection process. In addition to a decrease in the concentration of the translocation sugar sucrose, a transient accumulation of the hexoses glucose and fructose and an accumulation of the polyol mannitol were observed. The latter coincided with a rising level of mannitol dehydrogenase, an enzyme that reduces fructose to mannitol. Only minor changes were detected in the carbohydrate composition of apoplastic fluids isolated from leaves of uninoculated control plants or plants inoculated with avirulent races ofC. fulvum (incompatible interactions). The fungal metabolite mannitol was not detected in apoplastic fluids isolated from the latter plants.These results suggest that, upon colonization of the intercellular spaces by virulent races ofC. fulvum, apoplastic sucrose is hydrolyzed by a host and/or fungal invertase and the resulting hexoses, glucose and fructose, are converted into mannitol by the fungus. In incompatible tomato-C. fulvum interactions a functional nutritional relationship between plant and fungus is prevented by plant defense responses, which might explain why in these interactions the carbohydrate composition of apoplastic fluids is similar to that of uninoculated control plants.Samenvatting Inoculatie van tomaat (Lycopersicon esculentum) met virulente fysio's vanC. fulvum (compatibele interacties), leidde tot aanzienlijke veranderingen in de koolhydraatsamenstelling van apoplastische vloeistoffen die uit de bladeren werden geïsoleerd in de loop van het infectieproces. Naast een sterke daling van de concentratie van de transportsuiker saccharose, vond er ophoping van de hexoses glucose en fructose en de polyol mannitol plaats. De accumulatie van mannitol ging gepaard met een toename in de activiteit van mannitol dehydrogenase, een enzym dat fructose reduceert tot mannitol. In de koolhydraatsamenstelling van apoplastische vloeistoffen geïsoleerd uit bladeren van niet geïnoculeerde controleplanten, of planten geïnoculeerd met avirulente fysio's vanC. fulvum (incompatibele interacties), werden slechts kleine veranderingen waargenomen. De schimmelmetaboliet mannitol kon niet worden aangetoond in de apoplastische vloeistoffen die uit deze planten werden geïsoleerd.Deze resultaten suggereren dat bij de kolonisatie van de intercellulaire ruimtes door virulente fysio's vanC. fulvum, saccharose uit de apoplast wordt gehydrolyseerd door invertase afkomstig van de plant of de schimmel waarna de ontstane hexoses, glucose en fructose, door de schimmel worden omgezet in mannitol. Bij incompatibele tomaatC. fulvum interacties wordt een functionele voedingsrelatie tussen plant en schimmel voorkomen door het optreden van afweerreacties van de plant, hetgeen kan verklaren waarom in deze interacties de koolhydraatsamenstelling van apoplastische vloeistoffen vergelijkbaar is met die van niet geïnoculeerde controleplanten.     

A biochemical mechanism of resistance to pirimicarb in two glasshouse clones of Aphis gossypii

The susceptibility of three clones of Aphis gossypii Glover to 15 insecticides was established by bioassay. A high level of resistance towards pirimicarb was confirmed for a clone from Holland (Dutch R) and a clone from Japan (Jap R), while the susceptible clone (S) was killed by very low doses of the insecticide. However, only limited cross-resistance was shown towards other carbamates and organophosphates, and no marked resistance to the pyrethroids tested. The acetylcholinesterase (AChE) of both resistant clones hydrolysed acetyl-choline faster than that of susceptible aphids, with greatest enzyme activity shown by the Dutch R clone. Inter-clone differences in these rates were consistent with differences in catalytic centre activities. Inhibition (I₅₀) of AChE by pirimicarb was approximately 900-fold higher for the resistant clones than for the S clone. First-order kinetics revealed that resistance to pirimicarb in Dutch R and Jap R involved a modified AChE which had a reduced (approximately 350-fold) affinity (K_d) for pirimicarb. The marked change in AChE affinity for pirimicarb was not repeated with the other carbamates tested, ethiofencarb and aldicarb. It was considered that the resistant aphids would not require mechanisms in addition to insensitive AChE in order to show the high level of resistance to pirimicarb shown in the bioassay.     

褐飞虱对氟啶虫胺腈的抗性监测与生化抗性机制

为明确我国褐飞虱田间种群对氟啶虫胺腈的抗性现状及生化抗性机制,2017年-2019年采用稻茎浸渍法测定了采集自7省共13个褐飞虱田间种群对氟啶虫胺腈的抗性,并研究了氟啶虫胺腈抗性种群与其他杀虫剂的交互抗性以及增效剂对氟啶虫胺腈的增效效果。结果表明：近3年来褐飞虱对氟啶虫胺腈产生了中等水平抗性(RR=10.3～30.9)。氟啶虫胺腈抗性品系对呋虫胺、噻虫嗪和烯啶虫胺分别产生了9.1倍、7.9倍和4.1倍的低水平交互抗性,与噻嗪酮、毒死蜱、吡蚜酮、三氟苯嘧啶和吡虫啉不存在交互抗性。增效剂PBO对氟啶虫胺腈抗性品系和浙江龙游19(Longyou-19)田间种群分别具有4.2倍和3.8倍的明显增效作用。综上,褐飞虱田间种群已对氟啶虫胺腈产生中等水平抗性。多功能氧化酶参与了褐飞虱对氟啶虫胺腈的代谢抗性。     

Use of detached tomato leaves to test for resistance to tomato mosaic virus

Samenvatting Een methode werd ontwikkeld om de resistentie van in de kas geteelde tomateplanten tegen tomatemozaïek virus (ToMV) te bepalen. Bladeren van een vatbare en een resistente cultivar werden afgesneden en geïnoculeerd met ToMV. Na 6, 10 en 17 dagen werden de geïnoculeerde bladeren getoetst op de aanwezigheid van virus met ELISA en door inoculatie van bladeren vanNicotiana glutinosa. Met beide toetsmethoden kon de virustoename in de vatbare cultivar al vroeg na inoculatie duidelijk worden aangetoond. In de bladeren van de resistente cultivar was een zeer kleine hoeveelheid virus pas laat na de inoculatie aantoonbaar. Met deze methode is het mogelijk om de resistentie tegen ToMV te bepalen, tevens zaad te winnen en landbouwkundige eigenschappen te evalueren, zonder de plant te infecteren.     

Diastereomer-selective resistance in Cladosporium cucumerinum to triazole-type fungicides

Diastereomers of triazole-type fungicides behaved differently with respect to their fungitoxicity towards wild-type strains of Cladosporium cucumerinum and strains resistant to ergosterol biosynthesis inhibitors. With increasing resistance to the fungicide ‘as a whole’, the difference in resistance to the two diastereomers of triadimenol became progressively smaller, whereas for the other triazole derivatives, this difference became progressively larger. The possible consequences of such phenomena for the practical use of diastereomeric fungicides are discussed.