共查询到19条相似文献，搜索用时 140 毫秒
1.
美澳型核果褐腐病菌对甲基硫菌灵和啶酰菌胺的敏感性 总被引：4，自引：1，他引：3
采用传统检测方法测定了我国北方地区美澳型核果褐腐病菌对常用杀菌剂甲基硫菌灵和新药剂啶酰菌胺的敏感性,并采用12条ISSR引物对不同抗性水平的菌株进行了分析.结果显示:74株菌株中检测出5株对甲基硫菌灵表现高抗、2株表现低抗的菌株;我国抗性菌株的突变位点与美国加州抗性菌株的突变位点相同;啶酰菌胺对来自我国和美国、新西兰、法国的45株美澳型核果褐腐病菌的EC50分布频率呈单峰曲线,EC50在0.059～0.320 μg/mL之间,平均值为0.174±0.064μg/mL;基于ISSR分析得到的UPGMA聚类组与菌株的来源地及对甲基硫菌灵的抗性无相关性. 相似文献
2.
3.
本文建立了美澳型核果褐腐病菌(Monilinia fructicola)环介导等温扩增技术,并利用此技术开展了M.fructicola的检测和鉴定。采用M.fructicola SCAR分子标记的MO368特异片段为靶标序列,设计并筛选出M.fructicola的特异性LAMP引物,建立了该病菌快速诊断方法。试验结果表明,仅M.fructicola的菌株DNA扩增后呈天蓝色的阳性反应,而在其他真菌的供试菌株中均呈紫色的阴性反应。检测灵敏度可达到100 pg/μL。利用该方法对无锡机场截获的7份疑似李子褐腐病样本进行检测,结果有3份样品呈LAMP阳性反应。本文建立的M.fructicola LAMP检测方法具有灵敏度高、特异性好,简便易行等优点,为美澳型核果褐腐病菌的快速检测提供了一种新技术。 相似文献
4.
美澳型核果褐腐病菌(Monilinia fructicola(G.Winter)Honey)是核果类褐腐病的重要病原,偶尔也危害仁果。该菌一般发生在北美洲、南美洲、大洋洲和亚洲,在欧洲被列为检疫性有害生物,也是我国的检疫性有害生物。欧洲2001年在法国首次发现该菌,继而在德国、匈牙利、意大利、波兰、罗马尼亚、斯洛文尼 相似文献
5.
苹果壳色单隔孢溃疡病菌(Botryosphaeria stevensii)、美澳型核果褐腐病菌(Monilinia fructicola)和丁香疫霉病菌(Phytophthora syringae)是为害多种水果和其他植物的重要检疫性真菌,均能侵染苹果果实导致烂果,并造成严重的经济损失。本研究设计和筛选了3对特异性引物,建立了同步检测3种检疫性真菌的普通PCR和三重PCR方法。设计的引物Bs-368 F/R、M.cola 986F/R和Psy1/Psy2能分别特异性地扩增出B.stevensii、M.fructicola和P.syringae所有供试菌株的DNA,而对相应多个近似种的DNA则无扩增。灵敏度试验结果表明,上述3对引物分别检测B.stevensii、M.fructicola和P.syringae DNA的灵敏度是41.7 pg、47.4 pg和0.375 pg,三重PCR同时检测到3种病菌DNA的灵敏度是4.44 ng。该方法可以满足对进境苹果果实携带的苹果壳色单隔孢溃疡病菌、美澳型核果褐腐病菌和丁香疫霉病菌的快速初筛,加速进境新鲜水果的快检快放。 相似文献
6.
从新西兰进境苹果的褐腐组织中分离得到1株新丛赤壳属菌株3072,对该菌株进行形态学特征观察、PCR检测、序列比对分析和致病性测定。结果表明,在PDA培养基上菌落乳白色至黄白色,气生菌丝茂密,边缘规则;在MEA培养基上菌落为白色、平铺生长。菌株为异宗配合。利用欧洲枝溃疡病菌(Neonectria ditissima)特异检测引物对Bt-fw135/Bt-rw284扩增菌株的基因组DNA得到150 bp的预期目标条带。菌株的β-tubulin序列与GenBank中N.ditissima的序列相似性为99.43%100%。基于β-tubulin序列构建的系统进化树显示菌株3072与N.ditissima聚在同一个分支,亲缘关系最近。菌株接种苹果,接种处组织14 d后出现典型褐腐症状。根据上述检测结果,将菌株3072鉴定为欧洲枝溃疡病菌(N.ditissima)。 相似文献
7.
果树褐腐病的研究现状及其展望 总被引：1，自引：0，他引：1
《植物病理学报》2017,(2)
褐腐病是一种世界范围内广泛发生、严重危害核果类及仁果类水果生产的真菌性病害。经过一百多年的研究,人们对褐腐病的认识有了长足进展。本文从褐腐病的病原菌种群组成、生活史及危害症状、分布及寄主范围、病原菌生物学特性(菌落形态、分生孢子大小及形状、孢子萌发特征及致病力)、分子检测技术、抗药性及防治方法等方面进行综述,同时对未来的研究进行了展望,以便进一步认识果树褐腐病及其病原菌,提升褐腐病的防治水平,保障果业的安全生产。 相似文献
8.
9.
广州口岸进口水果截获病害概述 总被引：2，自引：0，他引：2
近年来,全国进口水果无论种类还是数量都急剧增加,但很少有截获重要病害的报道.作者对实验室受理的进口水果进行了病害检疫鉴定,共鉴定出病原33种,其中包括美澳型核果褐腐病菌、可可毛色二孢等重要疫情. 相似文献
10.
番石榴焦腐病菌的ITS分析及PCR检测 总被引：3，自引：3，他引：0
番石榴焦腐病是台湾入境大陆水果的重要植物病害,由葡萄座腔菌Botryosphaeria rhodina引起.为建立该病原菌快速、灵敏的检测技术,比较分析了葡萄座腔菌和葡萄座腔菌属其它种的ITS序列,在此基础上设计了1对检测番石榴焦腐病菌的特异性引物BF1/BR1,利用此引物从葡萄座腔菌中特异性扩增出287bp条带,而其余参试的菌株未能获得扩增条带.将真菌通用引物ITS1/ITS4和BF1/BR1进行巢式PCR扩增后,检测灵敏度提高1 000倍,可检测到葡萄座腔菌1pg的基因组DNA.结合快速碱裂解法提取发病组织的DNA,采用该PCR检测技术可从自然感染焦腐病果实中检测到葡萄座腔菌. 相似文献
11.
C.E. Fulton G.C.M. van Leeuwen Averil E. Brown 《European journal of plant pathology / European Foundation for Plant Pathology》1999,105(5):495-500
Nucleotide sequence analysis of the internal transcribed spacer (ITS) regions 1 and 2 of the ribosomal DNA (rDNA) divided the three brown rot pathogens Monilinia laxa, M. fructicola and M. fructigena into four distinct groups. Isolates of M. fructigena received from Japan, which varied by 5 base substitutions in the ITS region from the European M. fructigena isolates, formed the fourth group. Four of five Japanese isolates of M. fructicola tested varied from the New World isolates in that they did not possess a group-I intron in the small subunit (SSU) rDNA. RAPD-PCR data indicated that isolates of M. laxa varied but were randomly distributed worldwide; ITS data indicated no apparent distinction between those from Malus spp. and those from Prunus spp. M. fructigena similarly did not cluster according to geographic origin. In contrast, M. fructicola isolates tended to be clustered according to their origin; Japanese isolates of M. fructicola clustered together and showed similarity to some of the New Zealand isolates. Isolates from USA and Australia were more variable. 相似文献
12.
ABSTRACT Visible and nonvisible quiescent infections of immature and mature fruit are an integral component of the disease cycle of brown rot of sweet cherry in California. Detection of these infections is critical for developing efficient and efficacious fungicide management programs. The previously published DNA amplification primers mfs3 and NS5 for the identification of Monilinia fructicola were very specific in amplifying DNA of M. fructicola only and not M. laxa. This primer set, however, only detected DNA from some of the California isolates of M. fructicola. This genetic diversity was supported by random amplified polymorphic DNA (RAPD) analysis. Using eight 10-mer primers, seven M. fructicola isolates from California were all identified as genetically distinct. Using the same primers, only one polymorphism was detected among seven isolates of M. laxa. The multiple genotypes identified within the small population sample of M. fructicola, but not of M. laxa, using RAPD analysis could be indicative of genetic recombination within M. fructicola but not within M. laxa. To detect early brown rot infections in fruit, two primer sets that were developed from DNA sequences of either ribosomal DNA (MF5/ITS4/ITS3) or a RAPD fragment (X-09intF3/X-09R) specifically amplified DNA from isolates of M. fructicola and Monilinia species, respectively. No amplification products were present when using DNA from Botrytis cinerea or from other fungi commonly found on sweet cherry fruit. Primers X-09intF3 and X-09R were more sensitive and reliable for detecting small amounts of target DNA either extracted from conidia or from laboratory-inoculated cherry fruit with early brown rot infections that showed no visual symptoms or with visible quiescent infections. Furthermore, these primers also were effective for detecting visible quiescent infections in cherry fruit that were collected in the field. 相似文献
13.
ABSTRACT A set of molecular diagnostics was developed for Monilinia fructicola, causal agent of brown rot of stone fruits, capable of sensitive detection of the pathogen in planta. Species-specific repetitive sequences were identified from a partial library of 312 recombinant clones hybridized with total DNA, followed by subsequent screening for specificity. One hundred isolates, comprising 12 fungal species common to California stone fruits, were surveyed for specificity. Three clones hybridized to 60 geographically diverse M. fructicola isolates (California, Michigan, Georgia, Oregon, and Australia) to the exclusion of all other fungi surveyed, including the closely related M. laxa (n = 12). Two clones were identical and of extrachromosomal origin (pMF73 and pMF150), whereas the third (pMF210) migrated with uncut DNA. The sensitivity of all three was comparable and capable of detecting 50 pg of fungal DNA in dot blot hybridizations. Six species-specific primer pair sets were designed. They maintained the same specificity patterns observed in the initial hybridization surveys and were sensitive enough to detect 50 fg of fungal DNA template, approximately equivalent to 10 spores. The species-specific clones were capable of detecting the pathogen in planta, specifically from infected plum flowers and nectarine fruit tissue, using both hybridization- and polymerase chain reaction-based methodologies. 相似文献
14.
Monilinia laxa is a pathogen of brown rot of stone fruit and almond in California, causing blossom blights and fruit rots. In this study, low-level resistance to the benzimidazole fungicides benomyl and thiophanate-methyl was detected in field isolates of M laxa collected from stone fruits and almonds in California. Low-resistant (LR) isolates grew in potato dextrose agar (PDA) plates amended with benomyl and thiophanate-methyl at 1 and 5 microg ml(-1), respectively, but not in plates amended with benomyl at 5 microg ml(-1) or thiophanate-methyl at 50 microg ml(-1). The benzimidazole LR isolates were characterized by temperature sensitivity and the DNA sequence of the beta-tubulin gene. The LR isolates showed high-temperature sensitivity, being sensitive to 1 microg ml(-1) of benomyl at 28 degrees C but resistant at 8-24 degrees C. Analysis of the DNA sequence of the beta-tubulin gene showed that the LR isolates had a point mutation at the amino-acid position 240, causing substitution of leucine by phenylalanine. Based on the point mutation, a pair of allele-specific PCR primers was developed for rapid detection of LR isolates of M laxa. In addition, a pair of PCR primers specific to M laxa was developed on the basis of the differences in the DNA sequence of the intron 6 of beta-tubulin gene from M laxa, M fructicola and other fungal species. The primer pair amplified the expected 376-bp DNA fragment from all M laxa isolates tested, but not from 14 other fungal species isolated from stone fruit and almond crops. The restriction endonuclease BsmA I recognized the sequence GTCTCC in the PCR products from sensitive (S) isolates only, but not the GTTTCC sequence in the PCR products from LR isolates. The endonuclease digested the 376-bp PCR products from S isolates to produce two bands (111 and 265 bp) on agarose gels. Thus, both allele-specific PCR and the PCR-restriction fragment length polymorphism (PCR-RFLP) methods could be useful for rapidly detecting benzimidazole-resistant isolates of M laxa from stone fruit and almond crops in California. 相似文献
15.
Effects of wounding, fruit age and wetness duration on the development of cherry brown rot in the UK 总被引：2，自引：0，他引：2
This research investigated the effects of wounding, fruit age and wetness duration on the development of cherry brown rot. Both Monilinia laxa and M. fructigena infected wounded detached cherry fruits, but M. laxa caused more infections than M. fructigena and only M. laxa efficiently infected intact detached fruits. Results from field monitoring and controlled inoculation in a polyethylene tunnel showed that the susceptibility of fruits to infection by M. laxa increased with fruit maturity. Infection of attached intact fruits by M. laxa was not affected by the length (3–24 h) of the wet period tested. 相似文献
16.
I. R. Van Brouwershaven M. L. Bruil G. C. M. Van Leeuwen L. F. F. Kox 《Plant pathology》2010,59(3):548-555
To prevent the entry and spread of the brown rot fungus Monilinia fructicola in Europe, a fast and reliable method for detection of this organism is essential. In this study, an automated DNA extraction method combined with a multiplex real‐time PCR based on TaqMan chemistry was developed for fast, convenient and reliable detection of both the EU quarantine organism Monilinia fructicola and the three other brown rot fungi M. fructigena, M. laxa and Monilia polystroma. Using the internal transcribed spacer (ITS) region of the nuclear ribosomal RNA gene repeat, a Monilinia genus‐specific primer pair and two differently labelled fluorogenic probes specific for M. fructicola and the group M. fructigena/M. laxa/Monilia polystroma were developed. The analytical specificity of the assay was assessed by testing 33 isolates of the four brown rot fungi and 13 isolates of related fungal species or other fungal species that can be present on stone and pome fruit. No cross‐reactions were observed. The assay was found to have a detection limit of 0·6 pg of DNA, corresponding to 27 haploid genomes or four conidia. Comparison of a manual DNA isolation followed by a conventional PCR with an automated DNA isolation combined with the presently developed real‐time PCR showed that the latter method gave improved results when tested with 72 naturally infected stone fruit samples. The detection rate increased from 65 to 97%. 相似文献
17.
18.
Monilinia fructicola was until very recently a regulated pest in the European Union, and EU countries were requested to monitor its presence on their territories. As accredited laboratories should use validated tests, the mycological laboratory of CRA‐PAV carried out a validation process for the multiplex based PCR test (Coté et al., 2004 ), that is one of the most widely used tests for the identification of M. fructicola, although this test is not described in the EPPO diagnostic protocol PM 7/18 (2) because the validation data were lacking. The performance characteristics of this multiplex PCR test were established according to the EPPO Standard PM 7/98 (1) and the test was compared in a collaborative study with the end point PCR test (Ioos & Frey, 2000 ), considered as the ‘standard test’. The validation data were obtained using different isolates of M. fructicola, M. laxa, M. fructigena and Monilia polystroma, as well as different fruit tissues. Four series of the DNA target at different concentration, repeated three times, were analyzed in four Italian laboratories. The results showed that the multiplex PCR detection test (Coté et al., 2004 ) was fit for diagnostic purpose, although the analytical sensitivity was significantly lower compared to the conventional PCR ‘standard test’. 相似文献
19.
BACKGROUND: Thiophanate-methyl, a member of the benzimidazole class of fungicides, is used in California to control brown rot of stone fruit caused by Monilinia fructicola (G. Wint.) Honey. The goal of this study was to develop a real-time polymerase chain reaction (PCR) assay as an efficient method to quantify the E198A allele of beta-tubulin that confers benzimidazole resistance. RESULTS: Using the real-time PCR assay, the frequency of allele E198A (FEA) in a population was determined from the quantities of DNA amplified with the E198A allele-specific primer pair HRF/HRR and the M. fructicola-specific primer pair MfF6/MfR6. The average proportions of highly resistant isolates determined with the conventional fungicide sensitivity method were within the range of average FEA values determined with the real-time PCR assay. We also determined the FEAs of M. fructicola populations sampled from 21 stone fruit orchards in California. Only one orchard showed a high FEA over 0.20, seven orchards had values between 0.01 and 0.1, and 13 orchards had values less than 0.01. CONCLUSION: The real-time PCR assay developed in this study provides a potentially useful tool to efficiently quantify benzimidazole resistance for large M. fructicola populations. 相似文献