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1.
进境哈萨克斯坦向日葵种子中向日葵黑茎病菌的鉴定 总被引：1，自引：1，他引：0
自进境哈萨克斯坦向日葵种子中,采用常规平板分离法获得1株疑似向日葵黑茎病菌Plenodomus lindquistii菌株LL4,通过生物学性状、ITS序列测定和比对分析及致病性测定等方法对其进行了鉴定。结果表明,该病菌在PDA平板上培养初期菌丝无色,分隔分枝多,后期菌丝褐色或深褐色,常膨大;分生孢子器暗褐色,分散或聚集,球形、近球形或不规则形,大小为71.2~361.8μm;分生孢子形态变化较大,无色,单孢,肾形至椭圆形,大小为3.6~8.2μm×2.2~3.7μm,常含有2个油球;培养期间未见有性阶段。基于ITS序列分析比对发现,菌株LL4与Gen Bank中多个向日葵黑茎病菌分离物序列同源性达99%以上,系统进化树显示其与其它向日葵黑茎病菌分离物聚在同一个分支。菌株LL4可侵染向日葵根茎部,造成典型黑茎病症状。表明分离获得的菌株LL4为向日葵黑茎病菌P.lindquistii。 相似文献
2.
正大豆是一种重要的粮食作物和食用油原料,国内的市场需求量日益增长,目前我国已成为大豆净进口国。进口大豆虽能满足国内市场需求,但加大了外来有害生物传入的风险。向日葵黑茎病是一种毁灭性的向日葵病害,其病原菌向日葵黑茎病菌Plenodomus lindquistii属于我国进境检疫性植物病原真菌(段维军等,2015;2017)。2017年12月,宁波口岸自进境美国大豆夹杂的向日葵种子上分离获得1株菌株72298-10,为明确其分类地位,本研究通过形态学观察、ITS序列分析和致病性测定对其进行鉴定,以期为口岸疫情检测、防控和后续处理等工作提供参考。 相似文献
3.
《植物检疫》2015,(1)
分别对来自比利时和荷兰的水曲柳原木进行了病原菌分离,结果在原木的树皮中各分离到1株疑似苹果壳色单隔孢溃疡病菌的菌株Bs-1268和Bs-8212。对其进行了病原菌形态学观察、致病性测定并结合分子生物学方法检测,实验结果表明,菌株Bs-1268和Bs-8212在PDA和OA培养基上均能产生一定量的分生孢子器和分生孢子,分生孢子器通常为球形;经真菌通用引物(ITS4/ITS5)扩增和测序,菌株Bs-1268和Bs-8212与Gen Bank中登录号KF766158.1的菌株序列同源性达到100%;病菌分别接种苹果和梨6 d后发病。根据上述实验结果,将分离获得的菌株Bs-1268和Bs-8212鉴定为苹果壳色单隔孢溃疡病菌(Botryosphaeria stevensii Shoemaker)。 相似文献
4.
为准确快速筛查进境油葵种子中携带的向日葵黑茎病菌Plenodomus lindquistii,采用常规PCR方法比较了3对向日葵黑茎病菌PCR检测引物320FOR/320RVE、LEPB/LEPF和LLF/LLR的灵敏度,对哈萨克斯坦进境120批油葵种子样品进行PCR检测,并对检测为阳性的样品进行产物序列测定及病原菌分离。结果显示,引物320FOR/320RVE的检测灵敏度最高,可达10-5ng/μL,引物LEPB/LEPF和LLF/LLR分别为10-4ng/μL和10-3ng/μL。利用引物320FOR/320RVE、LEPB/LEPF和LLF/LLR对120批进境油葵种子样品进行检测,阳性检出率分别为65%、50%和45%,阳性检出率高低与引物的检测灵敏度相一致。阳性样品扩增的产物序列与Gen Bank中向日葵黑茎病菌的序列同源性最高;分离的病原菌菌落为乳白色至灰白色,分生孢子器黑褐色、球形并产生透明状分生孢子液,分生孢子单胞、无色、肾型、两端有油球,与已报道的向日葵黑茎病菌的病原特征描述一致,初步鉴定哈萨克斯坦进境油葵种子中存在向日葵黑茎病菌。 相似文献
5.
采用常规平板分离法, 从进境澳大利亚大麦中夹杂的油菜籽上获得1株疑似油菜茎基溃疡病菌的菌株01829?通过致病性测定?形态学观察?特异性引物扩增?ITS序列比对分析, 对01829进行了种类鉴定?结果表明:菌株01829在PDA培养基上生长较慢, 菌落边缘不整齐, 产生大量分生孢子器和分生孢子; 采用特异性引物对LMR1-D和Lmb分别进行PCR检测, 结果均有预期扩增片段产生; 基于ITS序列构建的系统发育树中, 菌株01829和GenBank中其他油菜茎基溃疡病菌相关序列聚在同一分支; 菌株01829接种油菜子叶和茎基部, 在子叶和茎基部接种部位分别引起叶斑和凹陷溃疡斑?根据上述试验结果, 将菌株01829鉴定为油菜茎基溃疡病菌, 这是我国口岸首次从进境澳大利亚大麦中截获油菜茎基溃疡病菌? 相似文献
6.
2013年12月,甘肃省白银市水川镇日光温室中的西葫芦发生了严重的根腐和茎基腐,部分棚室病株率达50%,从病根和病茎上分离得到拟漆斑菌属真菌3株,病株分出率27.3%。采用胚根和茎基部接种法测定了菌株FG-62对西葫芦的致病性:茎基部接种后27 d,植株开始出现凋萎;接种后40 d,两种接种法的西葫芦苗均呈现严重根腐和茎基腐症状,茎基部接种的西葫芦凋萎株率达30%;从病根和病茎上均可再分离出原接种菌。菌株FG-62在PDA平板上25℃培养14 d,产生大量墨绿色至黑色分生孢子座,分生孢子无色至淡榄黑色,单胞,杆状或腰鼓状,两端钝圆,大小为(7.04~9.15)μm ×(1.97~2.46)μm,聚集的分生孢子呈黑色。BLASTn分析结果显示,菌株FG-62(GenBank 登录号 MK252098)的rDNA-ITS序列与露湿拟漆斑菌Paramyrothecium roridum分离物E-469 (GenBank 登录号KY582183.1)和CBS 357.89(源自模式材料,GenBank 登录号NR_145077.1)的序列相似性分别达99.65%和96.83%。依据病原菌形态学和rDNA-ITS序列,将其鉴定为露湿拟漆斑菌P. roridum (Basionym:Myrothecium roridum)。这是露湿拟漆斑菌引起西葫芦根腐和茎基腐的首次报道。 相似文献
7.
2020年,在由斯里兰卡入境的甘蔗种茎中发现多条发病蔗茎。通过分离培养获得菌株,对菌株进行形态学鉴定、序列比对、系统进化分析和致病性测定。结果表明,该菌株形态与暗色座腔孢菌Phaeocytostroma sacchari相符。用引物ITS1和ITS4进行核酸序列扩增,获得约536 bp的目的片段,所获菌株km-1序列与P. sacchari一致性为100%;经系统进化分析发现,菌株km-1与暗色座腔孢菌聚在同一个进化分支。致病性接种结果表明,接种后的甘蔗具茎腐病典型症状,内部组织变为橙红色,并再次分离出暗色座腔孢菌。根据形态学、分子鉴定和致病性测定结果,确证分离到的甘蔗茎腐病的病原菌是给南非和印度甘蔗种植国家带来严重损失的暗色座腔孢菌。 相似文献
8.
火龙果赤斑病病原菌的分离与鉴定 总被引：1，自引：0，他引：1
通过对广东省湛江南亚热带植物园的火龙果病害调查,发现1种新的病害。其症状表现为病斑红色或褐色,圆形或近圆形,少数不规则形,稍凹陷,外缘有黄色晕圈,严重时可呈现褐色坏死斑。从田间感病火龙果茎的病健交界处用PDA培养基分离,获得了菌株BJ28,并对其进行了致病性测定、形态学观察和分子序列分析。结果表明,经柯赫氏法则验证菌株BJ28为致病病原菌。采用引物ITS1和ITS4对致病菌的ITS1-5.8S-ITS2基因片段进行扩增结果显示,该致病菌的ITS1-5.8S-ITS2序列与HQ608063(Nigrospora sphaerica)同源性最高,达到100%。通过培养形状、形态学观察和ITS1-5.8S-ITS2序列分析,将火龙果赤斑病病原菌鉴定为球黑孢菌[Nigrospora sphaerica(Sacc.)E.W.Mason]。 相似文献
9.
采用常规组织分离法对广西南宁、河池两地的葡萄黑痘病菌进行分离、纯化,分别得到37和31株分离菌。经菌落形态观察及r DNA ITS序列分析,南宁37株分离菌为同一菌株,河池31株分离菌为同一菌株,以NN和HC分别代表两地菌株,对它们进行形态学、致病性鉴定及r DNA ITS区域序列分析。结果显示,两地菌株形态学与致病性存在较大差异,但都符合黑痘病菌生长形态。NN菌落为红棕色、近圆形,边缘光滑。菌落中心位置丘状凸起,表面有白色菌丝和透明粘稠的小液滴,周围边缘有较规则的褶皱。HC菌落呈浅橙色、近圆形,边缘光滑。菌落中心位置丘状凸起,表面有少量白色菌丝,无液滴,周围边缘有不规则褶皱隆起。人工接种葡萄后均能引起典型的黑痘病症状,NN致病性强于HC。使用r DNA ITS区域通用引物ITS1F/ITS4进行PCR扩增后,NN和HC分别得到1 124 bp和818 bp的片段。比对结果显示1 124 bp与Elsinoe ampelina(AY826763.1)序列覆盖率达92%,序列一致性达99%;818 bp与Elsinoe ampelina(AY826762.1)序列覆盖率达75%,序列一致性达99%。因此,NN和HC均是引起广西葡萄黑痘病的病原菌。 相似文献
10.
广东梅州蜜柚柑橘黑斑病菌的检验鉴定 总被引：1，自引：0，他引：1
近年,柑橘黑斑病成为阻碍我国柑橘鲜果出口贸易的瓶颈,广东梅州名优鲜果蜜柚生产基地也遇到类似情况。该病发生后,病果果皮表面产生许多圆形病斑,初呈红褐色,中间凹陷,直径多为2~3mm,后期病斑逐渐转为褐色至黑褐色,病斑中部明显可见细小的黑色粒点状分生孢子器。病果分离物的培养性状与形态特征与柑橘黑斑病菌(柑橘球座菌,Guignarida citricarpa)一致。采用真菌ITS通用引物和柑橘球座菌特异性引物,从病斑组织基因组DNA及分离物基因组DNA中分别扩增得到541bp和396bp的片段。BLAST分析表明:扩增到的片段均与GenBank中已知柑橘球座菌菌株的rDNAITS序列相似性最高,分别为97%和98%。根据分离菌株的培养性状和形态特征及ITS区序列分析结果,将梅州蜜柚上的柑橘黑斑病菌鉴定为柑橘球座菌(Guignarida citricarpa Kiely)。 相似文献
11.
Quantitative trait loci associated with isolate specific and isolate nonspecific partial resistance to Phoma macdonaldii in sunflower 总被引：1，自引：0，他引：1
R. Darvishzadeh S. Poormohammad Kiani G. Dechamp-Guillaume L. Gentzbittel A. Sarrafi 《Plant pathology》2007,56(5):855-861
Black stem, caused by Phoma macdonaldii , is one of the most important diseases of sunflower in the world. Quantitative trait loci (QTLs) implicated in partial resistance to two single pycnidiospore isolates of P. macdonaldii (MP8 and MP10) were investigated using 99 recombinant inbred lines (RILs) from the cross between sunflower parental lines PAC2 and RHA266. The experimental design was a randomized complete block with three replications. High genetic variability and transgressive segregation were observed among RILs for partial resistance to P. macdonaldii isolates. QTL-mapping was performed using a recently developed high-density SSR/AFLP sunflower linkage map. A total of 10 QTLs were detected for black stem resistance. The phenotypic variance explained by each QTL (R2 ) was moderate, ranging from 6 to 20%. Four QTLs were common between two isolates on linkage group 5 and 15 whereas the others were specific for each isolate. Regarding isolate-specific and isolate-nonspecific QTLs detected for partial resistance, it is evident that both genetic effects control partial resistance to the disease isolates. This confirms the need to consider different isolates in the black stem resistance breeding programmes. The four SSR markers HA3700, SSU25, ORS1097 and ORS523_1 encompassing the QTLs for partial resistance to black stem isolates could be good candidates for marker assisted selection. 相似文献
12.
Two experiments were undertaken to determine the partial resistance of sunflower genotypes to seven isolates of Phoma macdonaldii . In the first experiment, 28 genotypes, including recombinant inbred lines and their parents, M6 mutant lines developed by gamma irradiation, and some genotypes from different geographical origins, were used. The experiment consisted of a split-plot design with three replications, each with 12 seedlings per genotype per isolate, in controlled conditions. Seven days after inoculation, plantlets were scored on a 1–9 scale for percentage necrotic area. Highly significant differences were observed among genotypes, isolates and their interactions. The presence of a differential interaction between genotypes and P. macdonaldii isolates was confirmed in a second experiment using 12 genotypes representing large variability for partial resistance to P. macdonaldii isolates, as identified in the first experiment. Inbred lines B454/03, ENSAT-B5 and LC1064C were the most susceptible sunflower genotypes, whereas two American lines SDR19 and SDR18 presented high partial resistance to all P. macdonaldii isolates studied. The least and most aggressive isolates were MA6 and MP3, respectively. Isolates interacted differentially with sunflower genotypes. This study identified two genotypes (AS613 and PAC2) presenting specific resistance to isolate MP8. The results also showed a wide range of isolate-nonspecific partial resistances among the lines tested. The information presented here could assist sunflower breeders to choose parents of crosses for breeding of durable resistance to phoma black stem disease. 相似文献
13.
Growth of 17 isolates of Phoma macdonaldii , the causal agent of sunflower black stem, was investigated for response to pH and temperature, and for morphology and asexual morphogenesis (pycnidiogenesis and pycnidium size). For all isolates, the optimum pH for growth was between 4 and 5, and the optimum temperature varied between 20 and 30°C and radial growth was slowest at 5 and 35°C. Significant differences in the number and size of pycnidia were observed between isolates. Pycniospore germination was investigated under various conditions in five isolates chosen for their geographical origins, pigmentation, optimum growth temperature and pycnidiogenesis. Increasing the concentration from 106 to 107 pycniospores per mL decreased the germination rate. The optimum temperature for pycniospore germination varied between 15 and 30°C, depending on the isolate, and the optimum and maximum pH values were 5 and 7, respectively. The optimum and minimum relative humidities allowing pycniospore germination were 100 and 95%, respectively. Pycniospore germination was photo-independent. An artificial inoculation method was developed and the aggressiveness of the pathogen was assessed on a susceptible sunflower cultivar, using a 1–9 scale that integrated the percentage of necrotic area on the cotyledon petiole at the stage when the first pair of leaves was fully developed. Significant differences in aggressiveness were observed among the 17 isolates. The parameters investigated clearly suggest the occurrence of a wide phenotypic variability in Phoma macdonaldii . 相似文献
14.
ABSTRACT Eighty-six isolates of Botryosphaeria dothidea, the causal agent of Botryosphaeria panicle and shoot blight of pistachio, were collected from pistachio and other plant hosts in California. The isolates were characterized by microsatellite-primed polymerase chain reaction (MP-PCR), sequences of the nuclear ribosomal DNA internal transcribed spacer region (ITS1, 5.8S gene, and ITS2), morphological and cultural characters, osmotic and fungicide sensitivity, and pathogenicity on pistachio. Three groups of these isolates were identified based upon analysis of MP-PCR data and ITS sequences. Group I contained 43 pycnidiospore-derived isolates collected from pistachio and other hosts. Group II consisted of 20 ascosporic isolates obtained from a single sequoia plant. Group III consisted of 20 ascosporic isolates from three shoots on a single blackberry plant, two pycnidiospore-derived isolates from incense cedar, and one from pistachio. Group I predominated over the other two groups in California pistachio orchards. B. dothidea isolates of group III grew faster on acidified potato dextrose agar (APDA) than the isolates of groups I and II. Isolates of group III produced pycnidia on both APDA and autoclaved pistachio shoots, but the isolates of the other two groups produced pycnidia on only autoclaved pistachio shoots. Additionally, significant differences in osmotic and fungicide sensitivities were observed among these three groups. Results from lathhouse inoculations demonstrated that the representative isolates for each of the three groups were all capable of infecting pistachio and producing characteristic disease symptoms of Botryosphaeria blight. The virulence of group II isolates on pistachio was, however, significantly lower than that of group I isolates. 相似文献
15.
为明确芝麻叶斑病病原菌及其致病力,本研究以河南不同地区芝麻上分离所得叶斑病分离物为研究对象,通过形态观察和分子鉴定探讨这些分离物的分类属性,并且进行离体叶片和活体茎杆接菌,进一步探究了不同菌株的致病力差异。结果显示分离所得的6株芝麻叶斑病分离物的菌落形态、生长速率和产孢量存在差异,分生孢子多数呈倒棍棒状,具隔膜,6个菌株的ITS序列与GenBank中Corynespora cassiicola的一致性达99%以上,结合形态特征与ITS序列将其鉴定为山扁豆生棒孢。室内人工接菌芝麻发现,这6个菌株的致病力存在显著的差异;在芝麻叶片上,20180909-03属强致病力菌株,20180821-01、20180909-05和20180824-01属中等致病力菌株,20180904-02-02和SF1-1属弱致病力菌株;在芝麻茎杆上,20180824-01致病力最强,其次是20180821-01、20180909-03和20180909-05,SF1-1和20180904-02-02致病力最弱。研究结果证明山扁豆生棒孢不同菌株在芝麻上的致病力存在明显的分化。 相似文献
16.
为明确芝麻叶斑病病原菌及其致病力,本研究以河南不同地区芝麻上分离所得叶斑病分离物为研究对象,通过形态观察和分子鉴定探讨这些分离物的分类属性,并且进行离体叶片和活体茎杆接菌,进一步探究了不同菌株的致病力差异。结果显示分离所得的6株芝麻叶斑病分离物的菌落形态、生长速率和产孢量存在差异,分生孢子多数呈倒棍棒状,具隔膜,6个菌株的ITS序列与GenBank中Corynespora cassiicola的一致性达99%以上,结合形态特征与ITS序列将其鉴定为山扁豆生棒孢。室内人工接菌芝麻发现,这6个菌株的致病力存在显著的差异;在芝麻叶片上,20180909-03属强致病力菌株,20180821-01、20180909-05和20180824-01属中等致病力菌株,20180904-02-02和SF1-1属弱致病力菌株;在芝麻茎杆上,20180824-01致病力最强,其次是20180821-01、20180909-03和20180909-05,SF1-1和20180904-02-02致病力最弱。研究结果证明山扁豆生棒孢不同菌株在芝麻上的致病力存在明显的分化。 相似文献
17.
从进境的美国大豆豆秆样品中分离到2株疑似大豆茎褐腐病菌Phialophora gregata的分离物247-8和8300-5,2株分离物在PGM培养基上菌落圆形,边缘规则,白色至暗褐色,表面隆起,粗糙,轮纹状。分生孢子卵形至椭圆形,无色,平滑,单胞,平均大小4.3 μm×1.9 μm。分生孢子梗具有瓶梗状结构,无色,无隔膜或有隔膜,大小(5~16)μm×(2~3) μm,呈桶型或长颈瓶型。特异性引物BSRIGS1/2、BSR1/2和Pgl/4分别扩增分离物247-8的DNA得到预期1 022 bp、483 bp和499 bp的产物;分离物8300-5的DNA经PCR扩增分别得到834 bp、483 bp和499 bp的预期条带。2株分离物的ITS区序列完全一致,与GenBank中大豆茎褐腐病菌(登录号AB190396、DQ459387、DQ459386和AF132804)的序列相似性为100%。人工接种大豆幼嫩植株茎基部均产生大豆茎褐腐病菌的典型症状。根据分离物形态特征、PCR检测、序列分析以及致病性测试结果,将进境美国大豆样品中的分离物247-8和8300-5鉴定为大豆茎褐腐病菌P. gregata。 相似文献
18.
蓝莓枝枯病病原菌鉴定 总被引：6，自引：0，他引：6
蓝莓 (Vaccinium spp.),是一类属于杜鹃花科 (Ericaceae) 越橘属 (Vaccinium) 的多年生小浆果类果树,广泛地应用于医药、保健、化妆品和环境保护等各方面,联合国粮农组织将其果实列为人类世界五大健康食品之一,也是我国南方酸性土丘陵地区值得发展的经济作物。全国已超过 10 个省份开始大面积的蓝莓商业化栽培,预计未来 10 年将超过 10 万 hm2 [1]。但是,随着生产面积的不断扩大,蓝莓病害也日趋严重,严重影响和制约蓝莓产业的发展。2010年至今,在云南省昆明市周边及丽江市蓝莓种植区发生一种病害,主要为害嫩枝、枝条和主干,发病初期,感病枝条上产生红褐色圆或椭圆形坏死斑,随后逐渐扩大,造成枝条干枯死亡,并可扩展造成主干枯萎。调查发现,该病发病率通常在 10％～20％,重者可达 25％ 以上,可引起蓝莓整株枯萎死亡。本文通过病原菌分离、致病性测定及传统、分子鉴定,明确了该病的病原,以期为病害防治提供理论依据。 相似文献