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1.
Phytophthora ramorum is a plant pathogen with a wide host range including many ornamental hosts and tree species. In Ireland and the UK P. ramorum is known to cause sudden larch death. There are four distinct genetic lineages of P. ramorum, with the fourth lineage (EU2) described in 2012 and only present in Northern Ireland and Scotland. In this work, experiments that compare all four lineages of P. ramorum using several phenotypic characters are described. A total of 166 isolates (EU1: 116, EU2: 40, NA1: 8, NA2: 2) from several EU countries and the United States and several hosts were amassed, and a selection of isolates were compared according to standard phenotypic tests. The EU1 and EU2 isolates tested were all A1 mating type. Regarding linear growth rate, we found the isolates ranked as follows EU2 > NA2 > EU1 > NA1, with all lineages growing fastest at 20 °C. The lineages ranked as NA2 > EU1 > EU2 > NA1 based on their in-vitro aggressiveness on detached wounded Rhododendron leaves, all lineages most aggressive at 20 °C. At 20 °C, we found that there was no significant difference between the EU1 and EU2 lineage based on their linear growth rate or in-vitro aggressiveness. Temperature, host ramet and P. ramorum lineage all had statistically significant effects on the observed aggressiveness of the isolates. From an experimental point of view, our results are broadly in agreement with other phenotypic studies of P. ramorum, finding variation between the lineages, but also variation within the lineages. From an applied perspective, our work on Rhododendron indicates that the EU1 and EU2 lineages pose similar levels of threat to plant health in Ireland and the UK, however, how these results transfer to other hosts (e.g. Larix kaempferi) needs more study.  相似文献   

2.
Invasive oomycete pathogens have been causing significant damage to native ecosystems worldwide for over a century. A recent well‐known example is Phytophthora ramorum, the causal agent of sudden oak death, which emerged in the 1990s in Europe and North America. In Europe, this pathogen is mainly restricted to woody ornamentals in nurseries and public greens, while severe outbreaks in the wild have only been reported in the UK. This study presents the results of the P. ramorum survey conducted in Swiss nurseries between 2003 and 2011. In all 120 nurseries subjected to the plant passport system, the main P. ramorum hosts were visually checked for above ground infections. Phytophthora species were isolated from tissue showing symptoms and identified on the basis of the morphological features of the cultures and sequencing of the ribosomal ITS region. Phytophthora was detected on 125 plants (66 Viburnum, 58 Rhododendron and one Pieris). Phytophthora ramorum was the most frequent species (59·2% of the plants), followed by P. plurivora, P. cactorum, P. citrophthora, P. cinnamomi, P. cactorum/P. hedraiandra, P. multivora and P. taxon PgChlamydo. The highest incidence of P. ramorum was observed on Viburnum × bodnantense. Microsatellite genotyping showed that the Swiss P. ramorum population is highly clonal and consists of seven genotypes (five previously reported in Europe, two new), all belonging to the European EU1 clonal lineage. It can therefore be assumed that P. ramorum entered Switzerland through nursery trade. Despite sanitation measures, repeated P. ramorum infections have been recorded in seven nurseries, suggesting either reintroduction or unsuccessful eradication efforts.  相似文献   

3.
This study assesses the survival of Phytophthora ramorum in the root ball of Rhododendron container plants as well as in different rootless forest substrates and a horticultural potting medium. Following inoculation of the root balls, the aboveground plant parts stayed symptomless, whilst the pathogen could be recovered with a novel non‐destructive baiting assay from the root balls until at least 8 months post‐inoculation. Plating of surface‐sterilized roots and direct microscopic analysis confirmed the presence of P. ramorum in the roots. Phytophthora ramorum could also be baited from the root balls of symptomless Rhododendron plants from commercial nurseries, even 2 years after acquisition. Survival of P. ramorum in rootless media was assessed after burying disks of infected leaf material below the soil surface in columns filled with four different undisturbed forest substrates or a potting medium, and incubated at an outdoor quarantine facility. Phytophthora ramorum could be recovered at least 33 months after burial from all substrates, with a significant increase in recovery after the winter period. These data suggest the possibility for long‐term symptomless presence of P. ramorum in root balls of commercial Rhododendron plants as well as survival in potting medium and different forest substrates under western European climate conditions. Symptomless presence in root balls can contribute to latent spread of this pathogen between nurseries. The novel baiting test, being non‐destructive, simple and applicable to a relatively large number of plants, can offer a valuable tool to test plants for the presence of Phytophthora species in root balls.  相似文献   

4.
Two genetically distinct evolutionary lineages of the oomycete pathogen Phytophthora ramorum are responsible for the major epidemic on larch (Larix spp.) in the UK: EU1 (historically widespread) and the recently identified EU2 (reported only from Northern Ireland and a small area in southwest Scotland). Methods for lineage discrimination have required pure cultures of P. ramorum but, as the pathogen is challenging to isolate from infected larch tissue, only limited data have been available on the distribution of EU2. In this study a protocol was developed to determine the lineage of P. ramorum in infected larch tissue without the need for isolation. The protocol was applied to 134 UK samples collected during 2013–14. In addition, lineage testing was applied to over 300 P. ramorum isolates cultured from a wide range of hosts between 2002 and 2012. Combined data confirmed that EU2 is restricted to Northern Ireland and a small area of southwest Scotland where it is the dominant lineage. There was no evidence of EU2 spread into England and Wales where only EU1 was found. However, EU2 was more widely distributed in southern and eastern parts of Scotland than previously reported. Furthermore, EU1 and EU2 were detected <10 km apart in larch plantations. This study provides the first reports of natural EU2 infection on European larch (Larix decidua), hybrid larch (Larix × eurolepis), beech (Fagus sylvatica), noble fir (Abies procera) and western hemlock (Tsuga heterophylla).  相似文献   

5.
The objective of this study was to determine whether genetically differentiated groups of Puccinia triticina are present in Europe. In total, 133 isolates of P. triticina collected from western Europe, central Europe and Turkey were tested for virulence on 20 lines of wheat with single leaf rust resistance genes, and for molecular genotypes with 23 simple sequence repeat (SSR) markers. After removal of isolates with identical virulence and SSR genotype within countries, 121 isolates were retained for further analysis. Isolates were grouped based on SSR genotypes using a Bayesian approach and a genetic distance method. Both methods optimally placed the isolates into eight European (EU) groups of P. triticina SSR genotypes. Seven of the groups had virulence characteristics of isolates collected from common hexaploid wheat, and one of the groups had virulence characteristics of isolates from tetraploid durum wheat. There was a significant correlation between the SSR genotypes and virulence phenotypes of the isolates. All EU groups had observed values of heterozygosity greater than expected and significant fixation values, which indicated the clonal reproduction of urediniospores in the overall population. Linkage disequilibria for SSR genotypes were high across the entire population and within countries. The overall values of RST and FST were lower when isolates were grouped by country, which indicated the migration of isolates within Europe. The European population of P. triticina had higher levels of genetic differentiation compared to other continental populations.  相似文献   

6.
Phytophthora ramorum, the cause of sudden oak death, is an invasive pathogen present in parts of coastal California and south-western Oregon forests. The majority of these forest infestations have been caused by the NA1 clonal lineage. In 2015, the EU1 lineage of P. ramorum was isolated from a tanoak (Notholithocarpus densiflorus) tree located in a mixed-conifer forest of Curry County, Oregon. In order to evaluate the threat to Oregon forests of the EU1 lineage relative to the established NA1 lineage, a series of experiments was conducted comparing aggressiveness and sporulation of NA1 and EU1 isolates on logs and seedlings in the growth chamber and forest. There was no significant difference in lesion size on logs inoculated with NA1 and EU1 isolates for any of the tree species tested. Across all seedling experiments differences among isolates within lineage, in terms of both aggressiveness and sporulation, were more commonly observed than differences among lineages. Site to site variation in tanoak sporulation, as measured by rain bucket baiting, appears to be correlated with the number of P. ramorum-positive seedlings detected at each site.  相似文献   

7.
Temperature and exposure time effects on Phytophthora kernoviae and Phytophthora ramorum viability were examined in flasks of compost and in a large‐scale composting system containing plant waste. Cellophane, rhododendron leaf and peat‐based inoculum of P. kernoviae and P. ramorum isolates were used in flasks; naturally infected leaves were inserted into a large‐scale system. Exposures of 5 and 10 days respectively at a mean temperature of 35°C in flask and large‐scale composts reduced P. kernoviae and P. ramorum inocula to below detection limits using semi‐selective culturing. Although P. ramorum was undetectable after a 1‐day exposure of inoculum to compost at 40°C in flasks, it survived on leaves exposed to a mean temperature of 40·9°C for 5 days in a large‐scale composting system. No survival of P. ramorum was detected after exposure of infected leaves for 5 days to a mean temperature of ≥41·9°C (32·8°C for P. kernoviae) or for 10 days at ≥31·8°C (25·9°C for Phytophthora pseudosyringae on infected bilberry stems) in large‐scale systems. Fitted survival probabilities of P. ramorum on infected leaves exposed in a large‐scale system for 5 days at 45°C or for 10 days at 35°C were <3%, for an average initial infection level of leaves of 59·2%. RNA quantification to measure viability was shown to be unreliable in environments that favour RNA preservation: high levels of ITS1 RNA were recovered from P. kernoviae‐ and P. ramorum‐infected leaves exposed to composting plant wastes at >53°C, when all culture results were negative.  相似文献   

8.
Phytophthora ramorum came to the Netherlands in 1993. Despite initially not seeming to pose a high risk, findings in California showed its potential destructive impact on ecosystems. A programme began in the Netherlands to eliminate P. ramorum from nurseries and surveys in the natural environment were held to obtain information to determine a strategy for dealing with the disease. About 1100 nurseries are inspected annually by NAKtuinbouw under the auspices of the Plant Protection Service and measures are taken according to EC directives. The percentage of infected nurseries decreased steadily during recent years, from 4% in 2002/2003 to 0.5% in 2004/2005. Surveys in the natural environment show that P. ramorum occurs on 2% of the sites with Rhododendron and therefore it was concluded that an elimination scenario is not realistic. A programme based on containment measures supported by an extension programme was put into place with its effects being monitored by the Plant Protection Service. 12 years of observing P. ramorum show that the risk for indigenous trees and shrubs in the Netherlands is very limited. Spread from infected rhododendrons to other potential hosts, even at heavily infected Rhododendron sites, hardly takes place although some infected Quercus rubra trees have been found. Recently several new Phytophthora species were found in natural environments in Europe and California, mainly as a result of intensive P. ramorum surveys. As well as P. ramorum, the Phytophthora spp. P. kernoviae, P. numerosa and P. pseudosyringae pose risks, indicating the need for a more general approach against Phytophthora diseases. As a result, a new protocol for detection and identification of Phytophthora spp. both as a group and individually is being developed and workers are asking whether these Phytophthora species could be managed together.  相似文献   

9.
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10.
Botrytis cinerea isolates from pear blossoms (Pyrus communis) in South Africa were collected from four orchards in two production areas in the Western Cape. The cryptic species status based on vegetative‐incompatibility alleles of the Bc‐hch gene indicated that all the isolates belonged to B. cinerea. A microsatellite analysis of B. cinerea populations was performed to assess the genetic population structure. Total gene diversity (H) was high, with a mean of 0.69 across all populations. Some genotype flow was evident between orchards as indicated by the spread of microsatellite multilocus genotypes, in agreement with the moderate, but significant population differentiation among orchards (mean φPT = 0.118, P = 0.001). Index of association analyses (IA and r?d) suggest that the populations reproduce mostly asexually, even though mating type distribution did not differ significantly from a 1:1 ratio, suggesting frequency‐dependent selection. Isolates resistant to benomyl were evident in one orchard only. This orchard was also significantly differentiated from all other populations, suggesting infrequent localized selection for benomyl resistance. Overall, the findings of this study highlight the dangers of a mixed reproduction system, and stress the importance of regularly monitoring fungicide resistance levels towards developing more efficient management practices.  相似文献   

11.
Phytophthora ramorum causes sudden oak death (SOD) in western coastal forests of the USA. In Europe, the pathogen is mainly present in the nursery industry, particularly on Rhododendron. Because of the primary role of Rhododendron as a host and potentially as a vector, the effect of Rhododendron host factors on P. ramorum susceptibility and sporulation was investigated. Inoculation methods using either wounded or non‐wounded detached leaves were applied to 59 Rhododendron cultivars and 22 botanical species, replicated in three separate years. All Rhododendron species and cultivars were susceptible when using wounded leaves, but not when using non‐wounded leaves, suggesting a resistance mechanism operating at the level of leaf penetration. Using a regression tree analysis, the cultivars and species were split into four susceptibility classes. Young leaves were more susceptible than mature leaves when wounded, but less susceptible when non‐wounded. This effect was not correlated with leaf hydrophobicity or the number of leaf hairs. The presence or the type of rootstock did not affect the cultivar susceptibility level. Sporangia and chlamydospore production in the leaf lesions varied widely among Rhododendron cultivars and was not correlated with the susceptibility level. The susceptibility to P. ramorum correlated well with the susceptibility to P. citricola and P. hedraiandra × cactorum, suggesting that the resistance mechanisms against these species are non‐specific. Susceptibility to P. kernoviae was low for most cultivars. These findings have implications for detection, spread and disease control, and are therefore important in pest risk assessment.  相似文献   

12.
Y. Tian  J. Sun  H. Li  G. Wang  Y. Ma  D. Liu  J. Quan  W. Shan 《Plant pathology》2015,64(1):200-206
Late blight caused by Phytophthora infestans is the most serious disease of potato worldwide. To understand the P. infestans population structure in northern Shaanxi, an emerging potato production region in China, 125 single‐lesion isolates were randomly collected from farmers' fields in 2009 and characterized phenotypically and genotypically. A mating type assay showed that 94 isolates were A1 mating type. Virulence determination of selected isolates on a set of differential potato lines containing R1 to R11, respectively, showed the presence of two pathotypes, of which the pathotype lacking avirulence genes Avr3, Avr4 and Avr10 was dominant. Isolates lacking all avirulence factors Avr1 to Avr11 were detected but at lower frequency (13·6%). Analysis for mtDNA haplotype showed all 61 examined isolates were IIa. A total of seven multilocus genotypes were distinguished among 125 isolates, as determined with seven polymorphic microsatellite markers. The genotype SG‐1 was dominant in the population with a frequency of 75·2% and was present throughout the region. Analysis of the phenotypic and genotypic structures of P. infestans populations indicated strict clonal reproduction of the pathogen and suggested that sexual reproduction probably does not occur. Potential implications for disease management are discussed.  相似文献   

13.
Phytophthora ramorum, a recently described North American and European pathogen, has three clonal lineages. The NA1 and NA2 lineages are found in North American forests and nurseries, while the EU1 lineage appears mainly in European nurseries. P. ramorum is heterothallic, having two mating types A1 and A2. All NA1 and NA2 isolates are of A2 mating type. When first collected, all EU1 isolates were of A1 mating type, with the exception of one A2 isolate collected in Belgium in 2002. Screening 410 other Belgian isolates for mating type revealed two additional EU1-A2 isolates collected in 2002 and 2003. PCR-RFLP, AFLP and SSR markers were used to determine the nature of the mating type change. The three isolates show no indications of sexual recombination or mitotic crossing over, indicating that mutation or mitotic gene conversion is the most likely explanation for the mating type change. We compared the pathogenicity and sporulation characteristics of the EU1-A2 isolates to those of EU1-A1 and NA1-A2 isolates on four host plants. Despite small differences in pathogenicity on some hosts, the EU1-A2 isolates were similarly aggressive to each other and to the EU1-A1 isolates and more aggressive than the NA1-A2 isolates. Sporulation characteristics were also comparable among EU1-A2 isolates and between EU1-A1 and EU1-A2 isolates, except for EU1-A2 isolate BBA 26/02. The limited genotypic and phenotypic differences between EU1-A2 isolates probably evolved after the mating type change, which may have occurred several years before the isolates were detected. There are strong indications that the EU1-A2 population has been eradicated from Belgium.  相似文献   

14.
The antimicrobial activity of heat-treated woodchips of three woody host species against the invasive oomycete plant pathogen Phytophthora ramorum was investigated to assess the potential of heated woodchips to suppress disease. Results demonstrated that heat-treated woodchips of pine (Pinus sylvestris), Japanese larch (Larix kaempferi) and rhododendron (Rhododendron ponticum) inhibited recovery of P. ramorum spores and mycelium compared with similar material that had only been air-dried. Effects were most evident with pine and larch; inhibition was maintained even when larch woodchips were diluted with soil. In vitro assays using methanol crude extracts from woodchips of the three species showed they all had an inhibitory effect on P. ramorum zoospores and reduced chlamydospore germination compared with air-dried wood extracts. Chemical analysis of the extracts revealed several induced compounds were present but in different concentrations for each species. Coniferaldehyde was the most active inhibitor against spores and mycelium, while the dominant resin acids, dehydroabietic and abietic acid, decreased the minimum inhibitory concentration of phenolic compounds tested against P. ramorum but were ineffective when used alone. An array of compounds, including dehydroabietic acid, methyl abietate, α-pinene and 3-carene, occurred at elevated levels in the living tissue of Japanese larch bark attacked by P. ramorum. These compounds may be part of the induced resistance response of larch to P. ramorum. Results of a field trial using heat-treated and air-dried woodchips were consistent with the crude extract bioassay results, suggesting that heat-treated woody materials have potential to reduce the survival of P. ramorum under natural conditions.  相似文献   

15.
Limited knowledge is available on Phytophthora infestans populations in Sub‐Saharan Africa (SSA). Therefore, and in response to recent severe late blight epidemics, P. infestans isolates from potato, tomato and Petunia × hybrida from eight SSA countries were characterized. Isolates were characterized with ‘old’ markers, including mating type (176 isolates), mitochondrial DNA haplotype (mtDNA) (281 isolates), glucose‐6‐phosphate isomerase (Gpi) (70 isolates), restriction fragment length polymorphism analysis with probe RG‐57 (49 isolates), and by metalaxyl sensitivity (64 isolates). Most isolates belonged to the US‐1 genotype or its variants (US‐1.10 and US‐1.11). The exceptions were genotype KE‐1 isolates (A1 mating type, mtDNA haplotype Ia, Gpi 90/100 and unique RG‐57 genotype), identified in two fields in Kenya, which are related to genotypes previously identified in Rwanda (RW‐1 and RW‐2), Ecuador and Europe. Metalaxyl‐resistant P. infestans isolates from potato were present in all the countries except Malawi, whereas all the isolates from tomato were sensitive. Genotyping of 176 isolates with seven simple sequence repeat (SSR) markers, including locus D13 that was difficult to score, revealed 79 multilocus genotypes (MLGs) in SSA. When this locus was excluded, 35 MLGs were identified. Genetic differentiation estimates between regional populations from SAA were significant when locus D13 was either excluded (P = 0·05) or included (P = 0·007), but population differentiation was only low to moderate (FST = 0·044 and 0·053, respectively).  相似文献   

16.
Early detection provides the best way to prevent introduction and establishment of alien plant pathogens. Amplification of DNA by PCR has revolutionized the detection and monitoring of plant pathogens. Most of those assays rely on the amplification of a fraction of the genome of the targeted species. With the availability of whole genomes for a growing number of fungi and oomycetes it is becoming possible to compare genomes and discover regions that are unique to a target organism. This study has applied this pipeline to develop a set of hierarchical TaqMan real-time PCR detection assays targeting DNA of all four Phytophthora ramorum lineages, and a closely related species, P. lateralis. Nine assays were generated: three targeting DNA of all P. ramorum lineages, one for each lineage of P. ramorum, one for P. lateralis and one targeting DNA of P. ramorum and P. lateralis. These assays were very accurate and sensitive, ranging from 98.7% to 100% detection accuracy of 2–10 gene copies of the targeted taxa from pure cultures or inoculated tissues. This level of sensitivity is within the lowest theoretical limit of detection of DNA. It is expected that these assays will be useful because of their high level of specificity and the ease with which they can be multiplexed because of the inherent flexibility in primer and probe design afforded by their lack of conservation in non-target species.  相似文献   

17.
Exserohilum turcicum is the causal agent of northern leaf blight, a devastating foliar disease of maize and sorghum. Specificity of E. turcicum to either maize or sorghum has been observed previously, but molecular evidence supporting host specialization is lacking. The aim of this study was to compare the genetic structure of E. turcicum isolates collected from adjacent maize and sorghum fields in Delmas and Greytown in South Africa. In addition, the mode of reproduction of this pathogen was investigated. Isolates from maize (N = 62) and sorghum (N = 64) were screened with 12 microsatellite markers as well as a multiplex mating type PCR assay. No shared haplotypes were observed between isolates from different hosts, although shared haplotypes were detected between isolates from maize from Delmas and Greytown. Population structure and principal coordinate analyses revealed genetic differentiation between E. turcicum isolates from maize and sorghum. Analysis of molecular variance indicated higher among‐population variation when comparing populations from different hosts, than comparing populations from different locations. Lack of shared haplotypes, high proportion of private alleles, greater among‐population variance between hosts than locations and significant pairwise population differentiation indicates genetic separation between isolates from maize and sorghum. The high haplotypic diversity in combination with unequal mating type ratios and significant linkage equilibrium indicates that both sexual and asexual reproduction contributes to the population genetic structure of E. turcicum in South Africa.  相似文献   

18.
In Europe, Phytophthora ramorum basically infects Rhododendron and Viburnum. To prevent the spread of the new quarantine organism in nurseries more knowledge about the transmission biology of this pathogen is necessary. For this reason the pathways of spread for P. ramorum on the two main host plants have been studied for the first time. Under practical field conditions inoculated plants were placed as sources of infections in a larger stock. Over 3?months the development of infestation was recorded. The pathogen showed a poor potential of spread. At the end of the trial only 1.0% of Rhododendron and 0.3% of Viburnum were infested with P. ramorum. Typical symptoms could be observed. On Rhododendron the pathogen caused a branch dieback. Stems showed a brown discoloration, which starts usually at the tip of the twig and moved towards the base. Infected Viburnum showed a stem base rot with wilting symptoms. Additionally rhododendrons were natural infested with Phytophthora citricola. This pathogen caused the same symptoms like P. ramorum and spread much faster. Investigations of leaf litter showed that both Phytophthora species had colonized the ground. This observation and the pattern of spread indicate that inoculum on the ground has been transmitted to arial plant parts via rainsplash. There is little evidence that P. ramorum has been transmitted directly from plant to plant via splashwater or air.  相似文献   

19.
The genetic structure of Mycosphaerella musicola has never been investigated in Brazil to address epidemiologically related questions associated with yellow sigatoka. A total of 223 single‐conidium isolates from four regions (North, South, Zona da Mata and Triângulo Mineiro) of Minas Gerais State, Brazil, were used to assess the population genetic structure of M. musicola. Isolates were characterized regarding the frequency of the MAT1‐1‐1 or MAT1‐2‐1 idiomorphs and polymorphism at nine microsatellite loci. The mating‐type ratio in three of the four subpopulations was c. 1:1. A total of 87 alleles and 216 multilocus genotypes were identified. The overall population was in linkage equilibrium. Most (93.9%) genetic variation was detected within the subpopulations and there was weak differentiation between them. In total, eight genetic groups were detected and isolates of seven groups were present in all regions. The population of M. musicola in Minas Gerais seems to have high evolutionary potential: it is panmictic and both sexual reproduction and gene flow affect genetic variability. Strategies to avoid fungicide resistance should be enforced and breeding programmes need to consider quantitative resistance in the banana cultivars.  相似文献   

20.
In order to improve understanding of its diversity, 338 isolates of Cryphonectria parasitica, the causal agent of chestnut blight, were sampled from 10 chestnut populations throughout chestnut‐growing coastal and continental areas of Croatia. Eighteen vegetative compatibility (VC) types were identified. The VC type EU‐1 was the most widespread, comprising 42·9% of the isolates, followed by EU‐2 (21%) and EU‐12 (14·2%). In respect to the occurrence of the main VC types, the C. parasitica populations in Croatia combined features of both northwestern and southeastern European populations. Perithecia and mating‐type ratios of approximately 1 : 1 were found in all populations, suggesting that sexual reproduction of the fungus is common in Croatia. Natural hypovirulence was also evident in all populations, with incidence of hypovirus‐infected isolates ranging from 12·7% in Istria‐Buje to 66·6% in the continental part of the country. A total of 36 hypovirus‐infected isolates sampled throughout Croatia were analysed in ORF‐A and ORF‐B by RT‐PCR/RFLP analysis. All viral isolates belonged to the Italian subtype of Cryphonectria hypovirus 1 (CHV‐1) and were closely related to the isolates found in other European countries. The RFLP patterns found were also identical or similar to the patterns of three isolates collected in Croatia 22 years ago, suggesting a slow evolution of the hypovirus.  相似文献   

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