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1.
Long‐term survival of Acidovorax citrulli in citron melon (Citrullus lanatus var. citroides) seeds was investigated. Citron melon seed lots infected with A. citrulli were generated in the field by inoculating either the pistils (stigma) or pericarps (ovary wall) of the female blossoms. Seventeen A. citrulli isolates from 14 different haplotypes belonging to two different groups (group I and II) were used for inoculation. After confirming that 100% of seed lots were infected, they were stored at 4°C and 50% RH for 7 years. After storage, the viability of A. citrulli cells from individual lots was determined by plating macerated seeds on semiselective medium as well as growing seeds for 14 days and scoring for bacterial fruit blotch symptoms. The type of A. citrulli isolate (group I or group II) used did not significantly influence bacterial survival. However, A. citrulli survival was significantly greater in seed lots generated via pistil inoculation (52·9 and 29·4%) than via pericarp inoculation (23·5 and 17·6%). Repetitive extragenic palindrome (rep)‐PCR on A. citrulli isolated from citron melon seed lots after storage displayed similar fingerprinting patterns to those of the reference strains originally used for blossom inoculation, indicating that cross‐contamination did not occur. The results indicate that A. citrulli may survive/overwinter in citron melon seeds for at least 7 years and bacterial survival in seed was influenced more by method of blossom inoculation than by the type of bacterial isolate.  相似文献   

2.
Acidovorax citrulli is a seed-transmitted gram-negative bacterium that can cause substantial economic yield loss in watermelon and melon production worldwide. Four small-molecule libraries containing 4,952 compounds were selected for high-throughput screening against A. citrulli wild-type strain Xu3-14 by evaluation of growth inhibition. One hundred and twenty-seven molecules (2.5% hit rate) were identified as bactericidal or bacteriostatic against A. citrulli at 100 μM. Secondary screens indicated that 27 candidate compounds were more effective against A. citrulli Group II strains than Group I strains (classified using repetitive element PCR). Several compounds were inhibitory to other pathogenic bacteria, including Clavibacter michiganensis subsp. michiganensis and Xanthomonas campestris pv. campestris, but did not affect the growth of plant beneficial bacteria Pseudomonas fluorescens and Bacillus subtilis. More than half of the compounds did not inhibit germination of Arabidopsis or watermelon seeds. The effect of small molecules on A. citrulli seed-to-seedling transmission was evaluated by applying each compound to inoculated watermelon seeds and assessing seedling infection. Nine compounds were chosen for further investigation based on their reduction of percentage seedling infection and compiling scores on their specificity, sensitivity, and phytotoxicity obtained in the secondary screens. The five best compounds were selected (thiamphenicol, nadifloxacin, pipemidic acid, ciclopirox, and zinc pyrithione) for greenhouse tests and were found to effectively reduce the seed-to-seedling transmission of A. citrulli in both artificially and naturally infested seeds. These top five compounds provide a basis for future development of an A. citrulli-specific bactericide.  相似文献   

3.
An Acidovorax citrulli–cucumber pathosystem was established through which A. citrulli mutants with altered pathogenicity, generated by transposon mutagenesis, were identified on cucumber cotyledons. The A. citrulli group I strain FC440 was shown to grow faster in cucumber leaf tissues than a group II strain and was used for Tn5 transposon mutagenesis. A total of 2100 Tn5 insertional mutants were generated, and analysis of the mutant library showed that the transposon insertions were single, independent and stable. A conserved non‐flagellar type III secretion system (NF‐T3SS) ATPase gene hrcN was identified and confirmed to be essential for pathogenicity and functionality of NF‐T3SS in A. citrulli. Comparative sequence analysis of the HrcN protein and its homologues in other representative bacterial plant pathogens revealed that the NF‐T3SS of A. citrulli is close to that of Ralstonia solanacearum and Xanthomonas campestris, but distant from that of Pseudomonas syringae and Erwinia amylovora. The generated Tn5 insertional mutant collection is valuable for identification of genes required for A. citrulli pathogenesis, and the established A. citrulli–cucumber pathosystem will facilitate an improved understanding of A. citrulli biology and pathology.  相似文献   

4.
Acidovorax citrulli is the causal agent of bacterial fruit blotch disease of cucurbits. Strains of this pathogen are distributed into two major groups: Group I strains have been mainly isolated from melon and other non-watermelon cucurbits, while Group II strains have been mainly recovered from watermelon. Here we report the characterization of strains T1 and EP isolated from diseased tomato and eggplant plants, respectively, and further confirmed to belong to A. citrulli species. Based on PCR, PFGE, and rep-PCR, these strains showed high similarity to the Group II strain 7a1. Sequencing and comparative analyses revealed that the genomes of T1 and EP aligned with that of the Group II model strain AAC00-1, over 97.88% and 99.22%, respectively. The virulence of T1, EP, and 7a1 determined on tomato, eggplant, and watermelon was similar and significantly higher than that of Group I strain M6. In contrast, M6 was more virulent on melon. Expression levels of seven virulence genes measured 24 hr after inoculation of tomato, eggplant, watermelon, and melon showed that the expression pattern was generally similar in strains 7a1, T1, and EP, whereas for M6 the expression was high only on melon. Overall, our results indicate that the solanaceous strains belong to Group II. To the best of our knowledge, this is the first study that reports characterization of A. citrulli strains isolated from solanaceous species. The fact that A. citrulli is able to naturally colonize and cause disease in non-cucurbit crops poses additional challenges for management of this important pathogen.  相似文献   

5.
Self‐compatible (SC) plants often deposit pollen on their own stigmas before flowers open. Thus, if we find natural hybrids between a SC species and a closely related self‐incompatible (SI) species, the pollen donor is usually expected to be the SC species. However, in previous studies of naturally occurring mixed populations of invasive Cakile maritima (SI) and Cakile edentula (SC) in Australia, hybridisation was found to have been bidirectional. Here, we explored the floral development of these species in order to test the hypothesis that there might be a short opportunity for pollinators to visit open SC flowers before the self‐pollen is released. Results indicated that both species showed reverse herkogamy during early development, though by the time pollen was ripe the four long anthers always overtopped the stigma; in C. maritima, most of the short anthers remained below the stigma. In both species, ripe, compatible pollen was able to germinate on the stigmatic surface at all stages of floral development. Anther dehiscence commenced earlier in C. edentula and well before flowers opened, in contrast to C. maritima, but the timing of pollen release varied between individual flowers, with a small proportion of uncontaminated stigmas after flowering; therefore, C. edentula can be either pollen recipient or pollen donor when interspecific outcrossing occurs. In summary, our results suggest that the variation between individual flowers of C. edentula contributed to the bidirectional hybridisation despite being a predominant selfer, providing new insights into plant colonisation.  相似文献   

6.
Sexually reproduced, airborne ascospores of Stagonosporopsis citrulli may play a role in its dispersal. S. citrulli causes gummy stem blight (GSB), one of the most important foliar diseases of cucurbits. Four studies were conducted with S. citrulli to investigate for how long ascospores are released and how far they can be dispersed from a source field. In the first study, colonized watermelon debris was sampled during three seasons and samples were assayed for ascospore release. Ascospores were detected 292, 313, and 306 days after inoculation of the source. In the second study, the active release of ascospores from pseudothecia in a Petri dish was monitored for 7 days. The release of ascospores decreased by ≤90% from 1 day after the start of the assay until 7 days after. In the third study, trap plant assays were conducted to measure the dispersal gradient of ascospores up to 366 m from the source. Generally, frequency of pathogen recovery from trap plants decreased with increasing distance from the source. The ascospore dispersal data fitted the exponential model better than the power law model. In the final study, dispersal experiments were conducted under controlled conditions. The incidence of GSB decreased with increasing distance, up to 55 m, from the source. It was concluded that ascospores of S. citrulli can serve as primary inoculum for epidemics and could easily spread among fields. Debris from cucurbit crops can be the source of ascospores for up to 10 months and should be cleared expeditiously.  相似文献   

7.
This study evaluated the direct antifungal activity of tiadinil [N-(3-chloro-4-methylphenyl)-4-methyl-1,2,3-thiadiazole-5-carboxamide], a systemic acquired resistance (SAR) inducer and two formulations of thymol (thymol I and thymol II) against Stagonosporopsis citrulli, the causal agent of gummy stem blight (GSB) disease of watermelon. Tiadinil, thymol I and thymol II completely inhibited the mycelial growth of S. citrulli in vitro at ≥?100 ppm. Conidial germination and germ tube elongation were completely inhibited by tiadinil at ≥?2000 ppm and by thymol-based formulations at ≥?100 ppm. A single foliar application of tiadinil at ≥?10 ppm or a single application of thymol I and II at ≥?1 ppm, 48 h before or after pathogen inoculation, significantly reduced disease severity of watermelon seedlings inoculated with 105/ml conidial suspension of S. citrulli, compared to respective nontreated controls. Plants treated with foliar application of tiadinil at ≥?1000 ppm before pathogen inoculation had significantly lower disease severity than plants that received an equivalent drench application. The efficacy of foliar application of tiadinil was affected by concentration and frequency of application. The study suggests direct antifungal activity of tiadinil, indicating a new mode of action of tiadinil against GSB disease of watermelon. The study also demonstrated direct antifungal action of thymol, a formulated active compound of essential oils, against S. citrulli and GSB disease of watermelon.  相似文献   

8.
为筛选高抗瓜类细菌性果斑病的西瓜资源,以24份西瓜品种资源为试材,采用苗期喷雾接种法分别接种分离自甜瓜上的瓜类嗜酸菌Acidovorax citrulli菌株pslb96和ZZ-1,鉴定各品种资源对瓜类细菌性果斑病的苗期抗性。结果表明,24份西瓜品种资源中未发现有对2株菌株表现免疫的材料,有7份资源对菌株pslb96表现高抗,12份资源对菌株ZZ-1表现高抗;9份资源对菌株pslb96表现中感或感病,7份资源对菌株ZZ-1表现中感和感病;对2株菌株均表现高抗的品种资源有野生型种质资源A9及商品种华欣、申蜜968、申选958和申抗988,占总品种资源的20.83%。部分品种资源A4、A13和申蜜7号对菌株pslb96和ZZ-1的抗性表现出明显的差异,表明相同寄主来源的2株不同菌株致病力存在差异。  相似文献   

9.
Postbloom fruit drop (PFD), an important disease caused by Colletotrichum spp., affects citrus yields in Brazil. PFD is characterised by the presence of necrotic lesions on the petals and stigmas of citrus flowers and by the subsequent abscission of young fruit. PFD epidemics have high disease progress rates, which is unusual for a pathogen that produces acervuli and is dispersed by rain. It is possible that other dispersal agents, such as insects and pollen, are involved in the spread of this disease. The objective of this work was to test whether citrus pollen grains can be colonised by Colletotrichum acutatum. Studies using light and electron microscopy showed that the pollen of Citrus sinensis can be infected by C. acutatum. This pathogen can penetrate and colonise citrus pollen grains 24 h after inoculation with the pathogen. The germ tube of conidia either penetrates the pollen sporodermis directly or passes through pollen germ pores. A single hypha can colonise more than one pollen grain. On the surface of the stigma, conidium formation can be observed. This study shows that Citrus sinensis pollen may, in fact, play a role in the spread of C. acutatum in citrus orchards.  相似文献   

10.
Endozoochory has been recognised as the most important dispersal mechanism in invasive Prosopis species, because their sugary, tasty pods attract animals and because some of their seeds remain intact after passing through some animals' digestive systems. In this study, we evaluated the role of the camel (Camelus dromedarius) as a potential disperser of the seeds of invasive tree Prosopis juliflora. Four camels of similar weight (ca. 400 kg) and age (ca. 2 years) were each fed with 70 fruits (1000 seeds approximately) of P. juliflora, which were retrieved from the camels' dung at 24‐h intervals for 4 days. The seeds retrieved were tested for germination and viability, along with seeds not eaten by the camels, with and without pericarp. Less than 3% of the seeds eaten were retrieved from the camels' dung. Most of the seeds (74%) were retrieved between 24 and 72 h after ingestion. The passage through the camel gut significantly accelerated and increased seed germination of P. juliflora in comparison with uneaten seeds covered with pericarp (48–75% and 15% respectively). While gut passage liberated P. juliflora seeds from the pericarp, increasing and accelerating their germination, the viability of seeds which had not germinated after germination trials were decreased (ca. ~20%) relative to uneaten seeds that had also not germinated. Our results show that, despite the low recorded seed recovery, camels can potentially disperse seeds of P. juliflora, which is in line with field observation showing P. juliflora expansion along the camels' routes in Gebel Elba National Park, south‐east Egypt.  相似文献   

11.
McManus PS  Best VM  Voland RP 《Phytopathology》1999,89(12):1127-1130
ABSTRACT Infection of cranberry flowers by conidia of Monilinia oxycocci, the cottonball pathogen, was investigated using a squash-mount histological method. Conidia germinated on anthers, nectaries, petals, and stigmata, but not styles. The stigma was the only flower part penetrated by the fungus, but no specialized infection structures were noted. Both fungal and pollen germ tubes grew through the stylar canal and made contact with ovules and nucellar tissue by 72 h after inoculation and pollination. Cottonball incidence was greatest when stigmata were inoculated; the low level of cottonball that resulted from inoculation of other flower parts and in noninoculated flowers was attributed to contamination of stigmata. In greenhouse tests, cottonball incidence was 25, 28, 31, and 38% for cvs. Searles, Pilgrim, Ben Lear, and Stevens, respectively, and was greater for M. oxycocci isolate 593 than isolate 591. We conclude that the stigma is the sole floral infection court for conidia of M. oxycocci and that the most popular cranberry cultivars in Wisconsin do not differ in inherent resistance to cottonball. The relevance of these findings to the long-term management of cottonball is discussed.  相似文献   

12.
Bacterial fruit blotch of cucurbits is a destructive disease caused by Acidovorax avenae subsp. citrulli, which is a typical seedborne pathogen. In seed health testing for this disease, we have detected many strains of Acidovorax with some differences from A. avenae subsp. citrulli. Their 16S rRNA sequences were divided into six types. The most common sequence was completely consistent with that of A. avenae subsp. avenae originally isolated from rice. The other sequences were over 99% similar but not identical to those of A. avenae subsp. avenae and A. avenae subsp. citrulli. Some commercialized antibodies against A. avenae subsp. citrulli reacted with several of these strains. Some of these strains incited yellow spots or brownish water-soaked lesions mainly on young true leaves of cucumber and squash after spray inoculation. Histological observations showed that these strains entered the leaf tissues of cucurbit plants through stomata and multiplied in the intercellular spaces of parenchymatous tissues as well as in the vascular tissues. The amount of bacterial multiplication and spread in the tissues differed among the strains, presumably reflecting their ability to induce symptoms. These isolated strains are therefore different from A. avenae subsp. citrulli, and their potential threat to the cultivation of cucurbits is lower than that of A. avenae subsp. citrulli.  相似文献   

13.
Genetic diversity analysis of Acidovorax citrulli in China   总被引：1，自引：0，他引：1  
Acidovorax citrulli has become quite common in China. A collection of 118 strains of A. citrulli was made from throughout China and other countries to determine their genetic relatedness. Strains were identified as A. citrulli by pathogenicity, phenotypic characterization, and PCR. Genetic diversity was determined using pulsed field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). PFGE electrophoresis resulted in nine genotypes, which could be typed into two groups based on host; group 1 included strains mainly from melon and group 2 included strains mainly from watermelon. MLST analysis resulted in 73 sequence types (ST) among the 118 A. citrulli strains. All A. citrulli strains were typed into three groups: group 1 with 82 strains (including type strain Fc247), group 2 with 35 strains and group 3 a singleton (Fc380). Similar to PFGE results, group 1 included strains mainly from melon and group 2 included strains mainly from watermelon. The difference was the 10 watermelon strains (pslbtw1-3, 5–11) from Beijing grouped with melon strains of group 1 based on MLST, suggesting that these 10 watermelon strains had a close relationship with melon. Our study indicated that there was genetic differentiation among A. citrulli strains between watermelon and melon. Also, our study was the first attempt to compare PFGE and MLST on analyzing genetic diversity of A. citrulli strains and proved MLST could better distinguish A. citrulli strains.  相似文献   

14.
Frost occurs in all major areas of cultivation, presenting a threat for the production of kiwifruit crops worldwide. A series of experiments were performed on 1‐year‐old, potted plants or excised twigs of Actinidia chinensis and A. deliciosa to verify whether strict relationships exist between bacterial canker outbreaks from Pseudomonas syringae pv. actinidiae (Psa) attacks and the occurrence of autumn and winter frost events. The association between the occurrence of autumn frost and the sudden outbreak of bacterial canker in A. chinensis in central Italy has been confirmed. Both autumn and winter frosts promote Psa multiplication in the inoculated twigs of both species. The day after the frost, reddish exudates oozing from the inoculation sites were consistently observed in both species, and Psa was re‐isolated in some cases. During the thawing of both A. deliciosa and A. chinensis twigs, the 2‐cm upward and downward migration of Psa from the inoculation site was observed within 3 min, and the leaves were consistently colonized with the pathogen. A consistent brown discoloration, accompanied with a sour‐sap odour, was observed throughout the length of the excised twigs of both Actinidia species after Psa inoculation and winter frost. Psa inoculation induced a remarkably higher necrosis in excised twigs that were not frozen compared with P. s. pv. syringae inoculation. Antifreeze protection using irrigation sprinklers did not influence the short‐term period of Psa and P. s. pv. syringae multiplication in both A. deliciosa and A. chinensis twigs. Thus, the damage from frost, freeze thawing and the accumulation of Psa in Actinidia twigs promotes the migration of the pathogen within and between the orchards. Taken together, the results obtained in this study confirmed that A. deliciosa is more frost tolerant than A. chinensis, autumn frosts are more dangerous to these crops than winter frosts, and in the absence of Psa, young kiwifruit plants remain sensitive to frost.  相似文献   

15.
Plant beneficial bacteria (PBB) have shown potential for disease control and are particularly important in the management of bacterial diseases, which are poorly controlled by conventional methods. In melon, bacterial fruit blotch caused by Acidovorax citrulli is a seedborne disease that is particularly destructive under certain conditions. PBB strains were screened for their ability to protect seeds and leaves from bacterial fruit blotch, and their antibiosis activity and plant colonization were studied. When Bacillus sp. RAB9 was applied to infected seeds, it reduced the area under the disease progress curve (AUDPC) by 47% and increased the incubation period (the time between inoculation and the first visible symptoms) by 35%. Three of the selected strains (JM339, MEN2 and PEP91) displayed antibiosis against A. citrulli. The RAB9Rif-Nal mutant colonized seeds epiphytically and roots and stems endophytically. Paenibacillus lentimorbus MEN2 sprayed on melon seedlings protected leaves, and when challenged with A. citrulli, it reduced the AUDPC (by 88%), disease index (by 81%) and incidence (by 77%). Given that the production of both melon seedlings and commercially grown greenhouse melons is increasing, biocontrol strategies may well be integrated into bacterial blotch management programs.  相似文献   

16.
In this study peroxynitrite (ONOO?) is proposed as an important player in defence responses during the interaction of potato (Solanum tuberosum) and the oomycete pathogen Phytophthora infestans. The potato–avr P. infestans model system exhibited a transient programme of boosted ONOO? formation correlated in time with the burst of nitric oxide (NO) and superoxide during the first 6 h post‐inoculation (hpi). The early ONOO? over‐accumulation was not accompanied by TPx gene expression. In contrast, the compatible interaction revealed a 24 h delay of ONOO? formation; however, an enhanced level of NO and superoxide correlated with TPx up‐regulation was recorded within the earlier stages of pathogen infection. Peroxynitrite over‐accumulation in the susceptible potato coincided with an enhanced level of protein tyrosine nitration starting from 24 hpi. Surprisingly, the nitroproteome profile of the resistant potato did not show any visible difference after inoculation, apart from one band containing subtilisin‐like protease‐like proteins, which appeared 48 h after pathogen attack. An additional pharmacological approach showed that treatment of the susceptible genotype with ONOO? followed by inoculation with P. infestans contributed to slowing down of the colonization of host tissues by the pathogen via a faster and stronger up‐regulation of the key defence markers, including the PR‐1 gene. Taken together, the results obtained indicate that a precise control of emitted NO and superoxide in cooperation with thioredoxin‐dependent redox sensors in sites of pathogen ingress could generate a sufficient threshold of ONOO?, triggering defence responses.  相似文献   

17.
The location of Pepper mild mottle virus (PMMoV) within seeds as they developed on inoculated seedlings of pepper (Capsicum annuum) was followed over time by detecting the viral coat protein using immunofluorescence microscopy. Seedlings were inoculated with PMMoV when the flower buds on the first and second branching nodes were in bloom. Fluorescence indicating the presence of PMMoV was first observed around immature seeds and placentas in the ovaries on the fourth branching node at 20 days post-anthesis (20 DPA), which corresponded to 39 days post-inoculation (39 DPI). The area with fluorescence gradually expanded from the placenta into the integument and the parenchyma, and finally reached the tip of the immature seeds by 34 DPA (53 DPI). The embryo or endosperm beyond the endothelium never fluoresced during the experiment [i.e., ending at 81 DPA (102 DPI)]. For visualizing viral routes of invasion from seeds into new seedlings, PMMoV-infected C. annuum seeds that were heterozygous for the L 3 tobamovirus-resistance gene were sown in soil at 30°C. After ~2 weeks, the cotyledon developed virally induced necrosis. These findings shed light on the infection cycle of PMMoV through vertical transmission in C. annuum.  相似文献   

18.
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19.
Three varieties of the introduced Bidens pilosa (Hairy beggar‐ticks; Asteraceae) co‐exist in Taiwan. Bidens pilosa var. minor and var. pilosa are naturalised, while only var. radiata is invasive. Differences in seed set in bagged versus open‐pollinated capitula, pollen–pistil interactions and the pollen/ovule (P/O) ratio were investigated among these three varieties. In contrast with the resulting 45–55% seed set in each bagged capitulum of var. minor and var. pilosa, almost no mature seeds (1–2 seeds) were found in bagged capitula of var. radiata, showing that var. radiata has a low capability of autonomous seed production. In bagged capitula, only 1.5% of pistils of var. radiata, but 76% and 52% of those of var. minor and pilosa, respectively, had pollen tubes penetrating into their styles, suggesting that var. radiata has strong self‐incompatibility mechanisms preventing autonomous autogamy. The P/O ratio, with mean values of 8189, 2053 and 1613 for var. radiata, var. minor and var. pilosa, respectively, were counted. These results support the hypothesis that the three varieties bear different breeding systems; the invasive var. radiata is highly self‐incompatible, whereas the other two naturalised varieties are self‐compatible.  相似文献   

20.
The effects of root‐knot nematodes on black root rot of watermelon and bottle gourd were studied using field surveys and co‐inoculation tests with Meloidogyne incognita (southern root‐knot nematode) and Diaporthe sclerotioides. The results of the field survey suggested that root‐knot nematodes had little effect on either the severity of black root rot or infection with D. sclerotioides. None of the three fields showed a significant positive correlation between disease severity and nematode gall index, with low correlation coefficients. Co‐inoculation experiments under controlled conditions found no significant effect of root‐knot nematodes on black root rot of watermelon and bottle gourd based on area under disease progress curves (AUDPC). These results were supported by the quantities of DNA of the two agents in root tissues because no significant difference was found between dual‐ and single‐inoculation treatments with M. incognita and/or D. sclerotioides. These findings suggest that root‐knot nematodes probably do not affect the infection of watermelon or bottle gourd roots by D. sclerotioides or the incidence of black root rot in these crops caused by this fungus.  相似文献   

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