Induced Resistance of Acibenzolar-S-methyl (CGA 245704) to Cucumber and Japanese Pear Diseases

Antifungal activity of the novel compound acibenzolar-S-methyl (CGA245704: benzo[1,2,3]thiadiazole-7-carbothioic acid S-methyl ester) was examined in vitro. No remarkable activity was observed on mycelial growth and conidial germination of almost all fungi tested. Only melon isolates of Didymella bryoniae were sensitive to this compound. On potted plants, acibenzolar-S-methyl showed control efficacy on anthracnose and scab of cucumber and rust of Japanese pear but not on Fusarium wilt of cucumber. In field trials, the occurrence of both rust and scab on Japanese pear was suppressed with this compound. Based on these experiments, it was suggested that acibenzolar-S-methyl induced resistance to some but not all diseases on cucumber and Japanese pear. Induction of disease resistance in cucumber was rapidly triggered after treatment with acibenzolar-S-methyl.     

Plant Resistance Genes: Their Structure, Function and Evolution

Plants have developed efficient mechanisms to avoid infection or to mount responses that render them resistant upon attack by a pathogen. One of the best-studied defence mechanisms is based on gene-for-gene resistance through which plants, harbouring specific resistance (R) genes, specifically recognise pathogens carrying matching avirulence (Avr) genes. Here a review of the R genes that have been cloned is given. Although in most cases it is not clear how R gene encoded proteins initiate pathways leading to disease resistance, we will show that there are clear parallels with disease prevention in animal systems. Furthermore, some evolutionary mechanisms acting on R genes to create novel recognitional specificities will be discussed.     

Study of Bacterial Determinants Involved in the Induction of Systemic Resistance in Bean by Pseudomonas putida BTP1

The ability of Pseudomonas putida BTP1 to induce resistance in bean to Botrytis cinerea was demonstrated in soil experiments on plants pre-inoculated at the root level with the bacteria before challenge with the leaf pathogen. As a first step to characterize the molecules from BTP1 responsible for induction of systemic resistance in bean, heat-killed cells and supernatant from culture in an iron-limited medium were tested for their protective effect. Most of the resistance-eliciting activity of the strain was retained in the crude cell-free culture fluid. In vivo assays with samples from successive fractionation steps of the BTP1 supernatant led, (i) to the conclusion that salicylic acid, pyochelin and pyoverdin, previously identified as Pseudomonas determinants for induced systemic resistance (ISR), were not involved in systemic resistance triggered by BTP1, and (ii) to the isolation of fractions containing one main metabolite that retained most of the resistance-inducing activity in bean. Although this molecule remains to be structurally characterized, its isolation is an addition to the range of determinants from plant growth-promoting rhizobacteria (PGPR) known to stimulate plant defences.     

Evaluation of Induction of Systemic Resistance in Pepper Plants (Capsicum Annuum) to Phytophthora capsici Using Trichoderma harzianum and its Relation with Capsidiol Accumulation

The effect of pepper seed and root treatments with Trichoderma harzianum spores on necrosis caused in stems by Phytophthora capsici inoculation and on the course of capsidiol accumulation in the inoculated sites were studied. The results indicate that seed treatments significantly reduced stem necrosis, which fell by nearly a half compared with the values observed in plants grown from non-treated seeds. Necrosis was also reduced in plants whose roots were drenched with various doses of T. harzianum spores, although the extent of necrosis was not correlated with the dose used. Attempted isolation of P. capsici and T. harzianum from the zones immediately contiguous with the necrotic zones revealed the presence of the former but not of the latter, suggesting that there was no direct contact between them in the zones of isolation, which means that there was no competition for space. The percentage of P. capsici isolated 9 days after inoculation was greater in non-treated inoculated plants than in treated inoculated plants. These results suggest that T. harzianum, introduced into the subterranean part of the plant, induces a systemic defense response against P. capsici in the upper part of the plant. Analysis of capsidiol in the stems of treated inoculated plants by the end of the sixth day after inoculation, revealed that its concentration was more than seven-fold greater than in non-treated and inoculated plants, while after 9 days, the concentration of capsidiol decreased in the treated inoculated plants and increased in the non-treated inoculated plants. The high concentration of capsidiol detected in treated and inoculated stems after 6 days might be one of the contributing factors, but not necessarily the main factor, in delaying lesion development in the stems of pepper plants.     

The Relationship Between Pathogen-induced Systemic Resistance (ISR) and Multigenic (horizontal) Resistance in Plants

Plants have developed mechanisms to successfully co-exist in the presence of pathogenic organisms. Some interactions between plants and pathogens are based on recognition of specific elicitor molecules from avirulent pathogen races (avr gene products), which is described in the gene-for-gene resistance theory. Another type of resistance, multigenic (horizontal) resistance, is a less well-studied phenomenon that depends upon multiple genes in the plant host. All plants possess resistance mechamisms which can be induced upon pre-treatment of plants with a variety of organisms or compounds. This general phenomenon is known as induced systemic resistance (ISR). At least in some plant species, ISR depends on the timely accumulation of multiple gene products, such as hydrolytic enzymes, peroxidases or other gene products related to plant defences. The pre-treatment of plants with an inducing organism or compound appears to incite the plant to mount an effective defense response upon subsequent encounters with pathogens, converting what would have been a compatible interaction to an incompatible one. Our studies in three plant–pathogen systems clearly document that multigenic-resistant plants constitutively express specific isozymes of hydrolytic enzymes that release cell wall elicitors, which in turn may activate other defense mechanisms. ISR induces constitutive accumulation of these and other gene products prior to challenge. ISR is known to function against multiple organisms, and there is no specificity observed in the accumulation patterns of defense-related gene products when ISR is induced. It is therefore hypothesized that the constitutive accumulation of specific isozymes of hydrolytic enzymes, or other defense related gene products, is an integral part of both multigenic resistance and the phenomenon of ISR. Further, plants in which ISR has been activated appear to move from a latent resistance state to one in which a multigenic, non-specific form of resistance is active.     

Induced Systemic Resistance (ISR) by Pseudomonas spp. Impairs Pre- and Post-Infection Development of Pythium aphanidermatum on Cucumber Roots

The effect of induced systemic resistance (ISR) by Pseudomonas rhizobacteria on the pre- and post-infection development of Pythium aphanidermatum on cucumber roots was investigated. Cucumber plants (cv. Corona) were grown in vermiculite, roots were split with one side bacterized with Pseudomonas corrugata strain 13 or P. aureofaciens strain 63-28 (bacterized roots) and the other distant side was treated with water (distant, induced roots). For the non-induced control, roots on the bacterized side were treated with buffer instead of the bacterial treatment. Intact, non-split roots were also treated with the bacteria or buffer as a control. Cucumber root tissue from these treatments were harvested and incubated with a zoospore suspension of P. aphanidermatum for three hours. Most of the zoospores in the suspension were stimulated to encyst or germinate. The numbers of germinated zoospores were significantly decreased on distant induced cucumber roots in comparison to non-induced controls. Germination was also reduced on intact bacterized roots, compared to controls. There was less attachment, germ tube production and penetration on roots bacterized or induced by the rhizobacteria compared to non-induced roots. Effects were significantly greater on bacterized roots (roots colonized by bacteria) compared to distant induced roots (roots with the opposite side bacterized). Systemic resistance induced by the two Pseudomonas spp. also reduced pathogen spread on split cucumber roots in planta. Crown infection from induced or bacterized roots was delayed for four to six days in comparison to the non-induced control. Results indicated that Pseudomonas spp. can exert both an indirect influence on P. aphanidermatum zoospore behaviour and infection via induced systemic resistance (ISR) and a local influence via antibiosis or local induced resistance.     

Effects of Plant Defence Activators on Anthracnose Disease of Cashew

The plant defence activators acibenzolar-S-methyl (Benzo[1,2,3]thiadiazole-7-carbothioic acid-S-methyl ester, ASM), 2,6-dichloro-isonicotinic acid (DCINA), salicylic acid (SA), and dibasic potassium phosphate (K₂HPO₄) were tested for their ability to protect cashew (Anacardium occidentale) seeds and leaves from anthracnose disease caused by Colletotrichum gloeosporioides. No inhibition of the early stages of pathogen development was caused by concentrations equal to or lower than 1.1mM a.i. ASM, 1.2mM a.i. DCINA, 5mM SA and 50mM K₂HPO₄. Maximum reduction of the disease in detached leaves, without phytotoxic effects, was obtained with 0.07mM a.i. ASM and DCINA, 5mM SA, and 50mM K₂HPO₄, with a time interval of at least 72h between application of the activator and inoculation with the pathogen. On attached leaves, foliar sprays were slightly more efficient than soil drench treatments, with 5mM SA being the most effective treatment, while 50mM SA as well as 0.3mM a.i. ASM and DCINA caused phytotoxic effects. In field-grown plants, protection was conferred by a soil drench of concentrations as low as 12.6M a.i. ASM and DCINA and 2.6mM SA. These concentrations were not phytotoxic suggesting that plant defence activators have potential for control of anthracnose disease in the field.     

Effect of treating soybean with 2,6-dichloroisonicotinic acid (INA) and benzothiadiazole (BTH) on seed yields and the level of disease caused by Sclerotinia sclerotiorum in field and greenhouse studies

Field or greenhouse grown soybeans were treated with 2,6-dichloroisonicotinic acid or benzothiadiazole and subsequently assessed for severity of white mold disease caused by Sclerotinia sclerotiorum. Three or four applications of 2,6-dichloroisonicotinic acid to field plots in 1993–1995 reduced severity of white mold after natural infection by 20–70% compared with water-treated controls in soybean cultivars Elgin 87 and Williams 82, which are considered to be highly susceptible to the disease. The effect was not as large in the cultivars Corsoy 79 and NKS19-90 which are more resistant to white mold. Two or four applications of benzothiadiazole to field plots in 1995 and 1996 reduced white mold severity by 20–60%, with the greatest reductions again observed in the more susceptible cultivars. Corresponding yields were increased compared with controls, particularly for the susceptible cultivars under conditions of high disease pressure. In greenhouse trials multiple applications of either compound resulted in significantly smaller lesion diameters following subsequent leaf inoculations with the fungus. The compounds did not result in observable phytotoxicity or inhibit growth of Sclerotinia sp. in vitro. We hypothesize that the decrease in disease severity following treatment with INA or BTH is a result of resistance induction.     

Interaction of Moniliophthora perniciosa biotypes with Micro‐Tom tomato: a model system to investigate the witches' broom disease of Theobroma cacao

The miniature tomato (Solanum lycopersicum) cultivar Micro‐Tom (MT) has become an important platform to investigate plant–pathogen interactions. In the case of the witches' broom disease of Theobroma cacao (cacao), the existence of Moniliophthora perniciosa isolates pathogenic to Solanaceae (S‐biotype) may enable the use of MT to circumvent limitations of the cacao host, whereas the availability of a non‐infective cacao C‐biotype allows the evaluation of contrasting responses of MT. Infection of MT by the S‐biotype led to stem swelling and axillary shoot growth to form broom‐like symptoms similar to the biotrophic phase in cacao, but the infected tissues did not progress to necrosis. Conversely, inoculation with the C‐biotype did not cause typical symptoms, but reduced plant height, appearing as a non‐host interaction. Histopathological characterization of the S‐biotype infection of MT by light and electron microscopy indicated limited germ tube penetration, preferentially through wounds at the base of trichomes or actively through the epidermis. No intracellular mycelium was observed, corroborating the lack of the necrotrophic stage of the pathogen. The analysis of gene expression during the interaction between the S‐ or C‐biotype with MT indicated that expression of plant defence‐associated genes differs for kinetics and intensity between a compatible or incompatible M. perniciosa–MT interaction. The pattern of spore germination and low rate of mycelia penetration suggests that the S‐biotype is not a fully adapted tomato pathogen, but possibly a case of broken non‐host resistance, and evidence suggests the occurrence of a non‐host MT response against the C‐biotype.     

Fusarium crown rot under continuous cropping of susceptible and partially resistant wheat in microcosms at elevated CO2

This study examines the CO₂‐mediated influence of plant resistance on crown rot dynamics under continuous cropping of partially resistant wheat line 249 and the susceptible cultivar Tamaroi. Disease incidence, severity, deoxynivalenol and Fusarium biomass were assessed after each cycle in microcosms established at ambient and 700 mg kg^?1 CO₂ using soil and stubble of these wheat lines from a field experiment with free to air CO₂ enrichment. Monoconidial isolates from wheat stubble were collected initially, and after five cropping cycles, to compare the frequency and aggressiveness of Fusarium species in the two populations. Aggressiveness was measured using a high‐throughput seedling bioassay. At elevated CO₂, the higher initial incidence in Tamaroi increased with cropping cycles, but incidence in 249 remained unchanged. Incidence at ambient CO₂ did not change for either line. Elevated CO₂ induced partial resistance in Tamaroi, but not in 249. Increased Fusarium biomass in wheat tissue at elevated CO₂ matched raised deoxynivalenol of the stem base in both lines. After five cycles of continuous wheat cropping, aggressiveness increased in pathogenic F. culmorum and F. pseudograminearum by 110%, but decreased in weakly pathogenic F. equiseti and F. oxysporum by 50%. CO₂ and host resistance interactively influenced species frequency, and the highly aggressive F. pseudograminearum became dominant on Tamaroi irrespective of CO₂ concentration, while its frequency declined on 249. This study shows that induced resistance at elevated CO₂ will not reduce crown rot severity, or impede the selection and enrichment of Fusarium populations with increased aggressiveness.     

Role of Salicylic Acid in Systemic Resistance Induced by Pseudomonas spp. Against Pythium aphanidermatum in Cucumber Roots

Pseudomonas corrugata strain 13 and P. aureofaciens strain 63-28, applied to roots, induced systemic resistance against Pythium aphanidermatum in cucumber roots. Salicylic acid (SA) from bacterial culture or plant tissues was quantified by high performance liquid chromatography. Both strains produced SA in King's B broth and also induced cucumber root to accumulate endogenous SA one day after bacterial inoculation. Using a split root system, more SA accumulated in roots treated with bacteria than in distant roots on the opposite side of the root system in the first two days, but this difference disappeared after 3–4 days. SA levels were significantly higher in plants treated with bacteria compared to the split control, from one to five days after bacterization. SA did not inhibit mycelial growth of Pythium aphanidermatum at 100–200µgml^–1 in vitro, but higher levels inhibited mycelial growth. Zoospore germination increased at concentrations of 10–500µgml^–1, but decreased at 1000µgml^–1 compared to lower concentrations. Exogenously applied SA failed to induce local or systemic resistance against a challenge infection by the pathogen in planta. The results of this study show that exogenous applied SA does not induce systemic resistance to cucumber root rot caused by P. aphanidermatum, but endogenous SA accumulation in cucumber roots may be involved in induced systemic resistance.     

The potential for the Rapid Screening of Potato Cultivars (Solanum tuberosum) for Resistance to Powdery Scab (Spongospora subterranea) using a Laboratory Bioassay

Powdery scab of potato, once established in a field, is difficult to control because of the longevity of the resting spores (cystosori) of the causal organism, Spongospora subterranea f.sp. subterranea. Host resistance is likely to be the most efficient in a long-term control strategy for preventing build-up of field inoculum and spread of the disease. Resistance screening of potato cultivars is mostly done in laborious field trials where disease development is likely to be unpredictable. A bioassay with potato tissue cultured plantlets and cystosori as inoculum is described and was tested for its potential to screen potato cultivars at an early stage for their relative susceptibility to powdery scab by comparing the lab results with field data. With cystosori inoculum of Swiss origin, the laboratory test showed clear differences between the potato cultivars in the severity of zoosporangial root infection which correlated better with ranked tuber infection data, compared to root galling. There are apparent differences in the relative trends in susceptibility between roots and tubers of five selected cultivars when using naturally infested soil instead of prepared cystosori as inoculum in the lab bioassay. Furthermore, differences in the severity of zoosporangial root infection of two selected cultivars were found when cystosori from different countries where used as inoculum. A possible host genotype × pathogen interaction is discussed. The bioassay has the potential to screen and select for resistant material at an early breeding stage thus making field trials not unnecessary but more economical. It will allow the use of a standard set of pathogen collections and facilitate testing for inoculum virulence in infested soils.     

114种植物提取物诱导烟草抗烟草花叶病毒活性鉴定

本研究以烟草和TMV为供试对象,评价了114种植物提取物诱导烟草抗TMV的活性,同时测试了这些植物提取物对TMV的钝化活性和抑制病毒粒子增殖的活性。结果表明,山苍子、平姜、苦参、八角、白屈菜、余甘子和五倍子共7种植物提取物对烟草具有显著的诱抗活性,对TMV的诱抗效果分别为78.90%、67.23%、66.82%、58.87%、53.92%、51.56%和51.28%,具有作为激发子诱导烟草抗病毒活性的应用潜力。本研究结果为开发植物源免疫诱抗剂生物农药提供了依据。     

A new type of plant activator: thieno[2,3-d][1,2,3]thiadiazole-6-carboxylic acid derivatives

A short and efficient synthesis of a series of the title compounds is presented starting with methylenebutanedioic acid and thioacetic acid. Using the Hurd–Mori reaction in the key step, the optimised reaction sequence allows the large-scale preparation of this new type of plant activator in a few steps with a high overall yield. Additional functionalisation of the 5-position via directed ortho-lithiation methodology is also described. © 1998 Society of Chemical Industry     

沼泽红假单胞菌蛋白Rhp-PSP对烟草花叶病毒的系统抗性诱导及互作蛋白筛选

本文研究了蛋白Rhp-PSP诱导烟草抗TMV过程中相关防御酶的活性以及丙二醛（MDA）含量的变化,同时测定了抗病相关基因的表达情况。试验证明,蛋白Rhp-PSP处理烟草后,叶片中苯丙氨酸解氨酶（PAL）、过氧化物酶（POD）和多酚氧化酶（PPO）的活性显著提高,但降低了MDA含量。同时基因PR-1、PR-3、PR-5、PDF1.2表达显著上调,证明了蛋白Rhp-PSP具有诱导烟草产生抗TMV的活性。为阐明诱导抗性机制,利用酵母双杂交系统从烟草cDNA文库进行互作蛋白的筛选并验证,结果表明GPI锚定蛋白、VAN3结合蛋白及叶绿素结合蛋白A与蛋白Rhp-PSP互作。     

Effect of phenology on susceptibility of Norway spruce (Picea abies) to fungal pathogens

Ceratocystis polonica and Heterobasidion parviporum are important fungal pathogens in Norway spruce (Picea abies). Tree susceptibility to these pathogens with respect to phenology was studied using artificial fungal inoculations at six stages of bud development, and assessed by measuring phloem necroses in the stems of 2‐ and 8‐year‐old trees. Tree capacity for resistance was assessed by measuring phloem nonstructural carbohydrates at each stage. Phloem necroses were significantly larger in trees with fungal versus control inoculations and increased significantly over time. Changes in nonstructural carbohydrates occurred in the trees; a significant decline in starch and a slight but significant increase in total sugars occurred over time. These results suggest that susceptibility to fungal pathogens and carbohydrate levels in the stems of the trees were related to fine‐scale changes in bud development. A trade‐off may occur between allocation of starch (the major fraction of the stem carbohydrate pool) to bud development/shoot growth versus defence of the stem. Previous tests of plant defence hypotheses have focused on herbivory on plants growing under different environmental conditions, but the role of phenology and the effect of pathogens are also important to consider in understanding plant resource allocation patterns.