不同寄主来源的灰葡萄孢对番茄的致病力分化研究

Eighteen isolates of Botrytis cinerea were obtained from the diseased plant tissue collected in Hefei, Bengbu, Changfeng and Hexian in Anhui province, by means of tissue isolating method. The pathogenicity of the isolates of B. cinerea from different hosts to the fruits and leaves of tomato were investigated by applying wound inoculation with mycelial blocks. The results showed that all of the tested isolates caused grey mould on tomato fruits, but there was significant difference in the average diameters of the lesions caused by different isolates, suggesting that there was significant differentiation in pathogenicity of B. cinerea strains to tomato fruits among isolates. According to the average diameters of the lesions on tomato fruits, the pathogenicity of the all isolates was classified into three categories: strong, intermediate and weak. In general, the isolates from tomato were more strongly pathogenic to tomato fruits than the isolates from strawberry, grape and capsicum. However, there was difference in pathogenicity among the different isolates from the same host, and the pathogenicity difference was not obviously related to the localities of isolates. After inoculating of tomato leaves, all of the tested isolates except CF3 caused grey mould on tomato leaves, but there was significant difference in the average diameters of the lesions caused by different isolates; and the difference in pathogenicity to tomato leaves was not obviously related to the host and locality of isolates.     

不同寄主来源的灰葡萄孢对番茄的致病力分化研究

从安徽合肥、蚌埠、长丰、和县等市、县的番茄、辣椒、草莓、葡萄等发病寄主上分离鉴定获得18个灰葡萄孢Botrytis cinerea菌株,采用菌丝块创伤接种法,分别测定了上述不同寄主来源的灰葡萄孢菌对番茄果实和叶片的致病力.结果表明,所有供试菌株接种番茄果实后均引起发病,但不同菌株所致病斑的平均直径有显著差异,显示灰葡萄孢菌株间对番茄果实的致病力存在明显分化.按照在番茄果实上所致病斑的平均直径大小可将供试菌株致病力划分为较强、中等和较弱3种类型.总体来说,来自番茄的菌株对番茄果实的致病力较强,来自草莓、葡萄和辣椒的菌株对番茄果实的致病力较弱,但来自相同寄主的菌株间致病力也存在差异,菌株致病力差异与菌株地域来源无明显相关.供试灰葡萄孢菌株接种番茄叶片后,除CF1外,均可引起番茄叶片发病,但不同菌株所致番茄叶片病斑的平均直径也有显著差异;供试菌株对番茄叶片的致病力差异与菌株的寄主和地域来源无显著相关.     

源自不同寄主的灰葡萄孢生物学特性的比较研究

本研究以分离自番茄、辣椒、草莓、葡萄的灰葡萄孢Botrytis cinerea为供试菌株,从生长温度、pH适应性、碳源、氮源营养利用等方面对不同寄主来源的灰葡萄孢菌株的生物学性状进行了比较研究。结果表明,5个不同寄主来源的灰葡萄孢菌株的菌丝生长温度范围相同,均为0-35℃;但它们的最适生长温度和分生孢子致死温度存在差异,来自和县番茄菌株HX12最适生长温度为20℃,分生孢子致死温度为47℃ 10min,其余最适生长温度均为25℃,分生孢子致死温度均为48℃ 10min;不同菌株在相同温度下的生长速率有显著差异。pH对不同寄主来源灰葡萄孢菌株菌丝生长的影响存在差异,来自长丰辣椒的菌株LJ菌丝在pH2-9的范围内均能生长,以在pH3-6.5时生长较快,pH6时最快;其余4个菌株在pH2-12的范围内均能生长,以在pH3-9时生长较快,pH6左右最快。不同碳源、氮源营养对灰葡萄孢菌株菌丝生长和分生孢子产生均有显著影响,不同寄主来源的菌株间在碳源、氮源营养利用差异均极显著。在相同碳源、氮源营养条件下,不同寄主来源的菌株的线性生长、菌丝干重和分生孢子产量均有显著差异。     

Genetic variation, pathogenicity and mycelial growth rate differentiation between Gaeumannomyces graminis var. tritici isolates derived from winter and spring wheat

The variability of Gaeumannomyces graminis var. tritici ( Ggt ) isolates was evaluated at an intravarietal level using non-molecular and molecular methods. Pathogenicity and linear growth rate of the pathogen were estimated. Very high pathogenicity was found in 44% of the isolates, medium in 20% and low only in 8%. Significant differences in mycelial growth rate were observed. The quickest linear growth rate of Ggt mycelium was observed at 25°C. Isolates derived from winter wheat grew faster than those obtained from spring wheat. The correlation between growth rates and pathogenicity was not significant. DNA polymorphism determined by random amplified polymorphic DNA (RAPD)–PCR was used to assess genetic variation among isolates. Thirty-two RAPD markers revealed DNA polymorphism suitable for assessing variability among isolates examined. Cluster analysis of RAPD data identified a few groups of isolates. RAPD markers associated with pathogenicity as well as mycelium growth rate were found.     

枯草芽孢杆菌BS2对葡萄灰霉病菌抑菌机制的初步探索

对葡萄灰霉病菌的生防枯草芽孢杆菌BS2菌液成分及胞外蛋白的抑菌机制进行了初步研究,BS2对葡萄灰霉病菌具有较好的拮抗作用,其菌液成分和胞外蛋白经20°C-120°C处理后,抑菌效果存在差异。BS2的菌液成分及胞外蛋白对灰霉病菌的产孢、萌发和菌丝的生长等方面均具有较好的抑制作用,且对灰霉病菌菌丝的原生质有囊泡和颗粒化现象。由此分析,BS2抑菌活性物质是多种成分共同作用的结果,抑菌物质中含有对温度敏感的高分子蛋白质,且抑菌机制也是从多方面共同起作用。     

Effects of paclobutrazol on Botrytis cinerea isolates obtained from potted plants

The growth of different isolates of Botrytis cinerea, collected from potted plants affected by Botrytis blight in southern Spain during recent years, was studied. These isolates, which show wide phenotypic differences when grown in vitro, are differentially affected by growth temperature, gibberellic acid, and paclobutrazol--an efficient plant growth retardant used widely in nursery potted plants to reduce plant size, favouring compactness, a more intense green foliage and increased stress tolerance to maintain quality prior to sale. In addition, paclobutrazol may have a fungicidal effect since it belongs to the triazole chemical group. However, paclobutrazol is only used as a plant growth retardant in Spain. In this work, we evaluate the effect of paclobutrazol dose (0, 0.05, 0.25, 1.25, and 6.25 mg/plate) on the growth of a collection of different B. cinerea isolates obtained from the following potted plants: Cyclamen persicum, Hydrangea macrophylla, Lantana camara and Lonicera japonica. Mycelial growth curves and growth rates assessed from difference in colony areas during the linear phase, conidiation (measured as time of appearance), conidial length (microm), and sclerotia production (number/plate) were evaluated in the isolates, which were grown at 26 degrees C on Petri dishes containing potato dextrose agar for up to 36 days. Mycelial growth curves fitted a typical kinetic equation of fungus grown on solid media. The B. cinerea isolates showed a high degree of variability in their growth kinetics, depending on the isolate and paclobutrazol dose. This triazole delayed mycelial growth during the linear phase in an isolate-dependent manner, and isolates from C. persicum and L. japonica were more affected by paclobutrazol than H. macrophylla. On the other hand, 0.25 mg of paclobutrazol was the critical dose to significantly reduce the growth rate in all isolates. 6.25 mg paclobutrazol inhibited conidiation in isolates from C. persicum, and reduced the conidial length in isolates from H. macrophylla and L. camara. The sclerotia production process was blocked at paclobutrazol doses higher than 1.25 mg, while no sclerotia were produced in isolates from C. persicum and L. japonica with 0.25 mg. H. macrophylla was the isolate in which sclerotia production was most influenced by paclobutrazol. It was concluded that the exact effect of paclobutrazol on B. cinerea growth depends on the isolate, and new strategies should be considered for evaluating its use as retardant and fungicide.     

Effects of indole-3-acetic acid on Botrytis cinerea isolates obtained from potted plants

We study the growth of different isolates of Botrytis cinerea collected from potted plants which were affected by Botrytis blight in southern Spain during recent years. These isolates, which show widely phenotypic differences when grown in vitro, are differentially affected by growth temperature, gibberellic acid applications and paclobutrazol, an efficient plant growth retardant and fungicide at the same time. In this work, we have evaluated the effect of the auxin indole-3-acetic acid (IAA) dose (0, 1, 10, and 100 mg/plate) on the growth of the collection of B. cinerea isolates obtained from the following potted plants: Cyclamen persicum, Hydrangea macrophylla, Lantona camara, and Lonicera japonica. B. cinerea produces indolacetic acid, but so far the precise biosynthetic pathway and some effects on this fungal species are still unclear, although recent studies have revealed an antifungal activity of IAA on several fungi, including B. cinerea isolated from harvested fruits. Mycelial growth curves and growth rates assessed from difference in colony areas during the both linear and deceleration phase, conidiation (measured as time of appearance), conidia length (microm), and sclerotia production (number/plate) were evaluated in the isolates, which were grown at 26 degrees C on Petri dishes containing potato dextrose agar for up to 35 days. Mycelial growth curves fitted a typical kinetic equation of fungi grown on solid media. B. cinerea isolates showed a high degree of variability in their growth kinetics, depending on the isolate and auxin dose. This plant growth substance delayed mycelial growth during the linear phase in an isolate-dependent manner, thus isolates from C. persicum, H. macrophylla and L. camara were more affected by IAA than L. japonica. On the other hand, 100 mg of IAA was the critical dose to significantly reduce the growth rate in all isolates and to promote brown-striped hyphae development, especially in isolate from C. persicum. 10 and 100 mg IAA delayed conidiation in isolates from H. macrophylla but scarcely effects were found in the conidia length. The sclerotia production process was blocked at IAA doses of 100 mg in isolates from L. camara and L. japonica, and was reduced in isolate from H. macrophylla. However, dose of 100 mg IAA had no effect on sclerotia production in isolate from C. persicum. It was concluded that the effect of IAA on B. cinerea growth depends on the isolate, thus isolates from H. macrophylla and L. camara were the most affected by IAA. B. cinerea reduced its development under IAA applications, depending on the isolate and dose. These results confirm those recently published on the inhibitory effect of IAA on Botrytris species growth.     

灰葡萄孢犬尿氨酸途径关键酶基因的鉴定

【背景】灰葡萄孢是一种重要的植物病原真菌,实验室前期明确了灰葡萄孢犬尿氨酸单加氧酶(kynurenine3-monooxygenase,KMO)基因BcKMO参与调控病菌的生长发育和致病力。犬尿氨酸单加氧酶(KMO)是犬尿氨酸途径的关键酶,但灰葡萄孢是否存在犬尿氨酸途径及其在病菌生长、发育和致病过程中的功能尚未见相关报道。【目的】鉴定灰葡萄孢犬尿氨酸途径中的关键酶基因,确定灰葡萄孢犬尿氨酸途径的存在,为阐明灰葡萄孢生长发育和致病力的分子机理奠定基础。【方法】利用生物信息学方法,对灰葡萄孢犬尿氨酸途径中犬尿氨酸酶(kynureninase,KYN)、吲哚-2,3-双加氧酶(indoleamine-2,3-dioxygenase,IDO)、犬尿氨酸氨基转移酶(kynurenine amino transferase,KAT)的编码基因进行分析;利用Real-time PCR技术,检测灰葡萄孢野生型BC22、BcKMO基因T-DNA插入突变体BCG183、恢复菌株BCG183/BcKMO中犬尿氨酸途径关键酶基因的表达水平;利用真菌犬尿氨酸酶KYN检测试剂盒,测定BcKMO突变体中犬尿氨酸酶(KYN)的含量。【结果】灰葡萄孢中含有2个犬尿氨酸氨基转移酶(KAT)的编码基因、3个吲哚-2,3-双加氧酶(IDO)的编码基因、10个犬尿氨酸氨基转移酶(KAT)的编码基因。灰葡萄孢KYN编码基因、IDO编码基因、KAT编码基因在突变体BCG183中的表达水平显著高于或低于在野生型和恢复菌株。突变体BCG183中犬尿氨酸酶(KYN)的含量显著低于野生型BC22和恢复菌株。【结论】灰葡萄孢中存在犬尿氨酸途径,灰葡萄孢BcKMO基因突变影响KYN、IDO和KAT编码基因的表达以及犬尿氨酸酶(KYN)的含量。     

小花假泽兰精油化学成分及其抑菌活性初步研究

利用GC-MS技术分析了小花假泽兰全株精油的化学成分及其含量,并初步测定了精油的抑菌效果.从小花假泽兰全株精油中共鉴定出49种成分,占色谱峰总面积的57.28%,主要成分为22,23-二羟基豆甾烷醇(7.34%)、氧化丁子香烯(3.34%)、罗汉柏烯(2.92%)、cubenol(2.81%)等.生长速率法测定表明,小花假泽兰精油对番茄灰霉病菌、小麦纹枯病菌和小麦赤霉病菌菌丝生长抑制作用较强,在1 000 mg/L剂量下抑制率均在90%以上;果实针刺法测试表明,该精油对番茄灰霉病菌具明显保护作用和治疗作用,但治疗效果低于保护效果,在2 000mg/L剂量下,后者达70.79%,前者仅51.79%.     

Fumigant activity of essential oils and components of Illicium verum and Schizonepeta tenuifolia against Botrytis cinerea and Colletotrichum gloeosporioides

To develop a natural fungicide against Botrytis cinerea and Colletotrichum gloeosporioides, a total of 25 essential oils were tested for their fumigant activity against post-harvest pathogens. The vaporous phases of oils were treated to each fungus on potato dextrose agar medium in half-plate separated Petri plates at 10 microg per plate. The essential oil of Illicium verum strongly inhibited the mycelial growth of both B. cinerea and C. gloeosporioides by over 90%. On the other hand, the essential oil of Schizonepeta tenuifolia showed inhibitory activity against mycelial growth of only B. cinerea by over 90%. Gas chromatography-mass spectrometry and bioassay indicated trans-anethole in I. verum and menthone in S. tenuifolia as a major antifungal constituent. The essential oils of I. verum and S. tenuifolia and their major constituents could be used to manage post-harvest diseases caused by B. cinerea and C. gloeosporioides.     

灰葡萄孢丝裂原活化蛋白激酶编码基因bmp1和bmp3的功能

【背景】植物病原真菌丝裂原活化蛋白激酶(Mitogen-activated protein kinase,MAPK)信号途径参与病菌有性生殖、细胞壁完整、菌丝侵染、致病力、胁迫响应等过程,灰葡萄孢MAPK信号途径参与病菌生长发育、致病力以及胁迫响应,但MAPK信号途径基因在灰葡萄孢中的功能尚未完全阐明,该信号途径对灰葡萄孢的生长发育和致病力的调控机制尚不明确。【目的】明确灰葡萄孢MAPK编码基因bmp1、bmp3在病菌生长发育、致病力以及氧化胁迫响应过程的功能,为进一步阐明MAPK信号途径调控灰葡萄孢生长发育和致病力的分子机制奠定基础。【方法】利用RNAi技术构建灰葡萄孢MAPK编码基因bmp1和bmp3的RNAi突变体,并以野生型BC22菌株为对照,对bmp1和bmp3基因的RNAi突变体的表型、致病力以及对氧化胁迫的敏感性进行分析。【结果】灰葡萄孢bmp1和bmp3基因的RNAi突变体其菌落形态、菌丝形态均与野生型BC22菌株没有明显差别;bmp1基因的RNAi突变体生长速率明显减慢,分生孢子产量明显降低;bmp3基因的RNAi突变体的生长速率与野生型BC22菌株没有明显差别,不能产生分生孢子。bmp1和bmp3基因的RNAi突变体在番茄果实的表面均不能产生明显的致病症状,而且不能穿透玻璃纸。bmp1基因的RNAi突变体在含有H_2O_2的培养基上受抑制的程度显著低于野生型,而在含甲萘醌的培养基上受抑制的程度显著高于野生型;bmp3基因的RNAi突变体在含有H_2O_2和甲萘醌的培养基受抑制的程度均显著高于野生型。【结论】灰葡萄孢bmp1基因正调控病菌生长、分生孢子形成、致病力和穿透能力,参与调控病菌对氧化胁迫的响应;灰葡萄孢bmp3基因正调控病菌分生孢子形成、致病力、穿透能力以及对氧化胁迫的响应。     

Development of Pathogenicity and AFLP to Characterize Fusarium oxysporum f.sp. momordicae Isolates from Bitter Gourd in China

Bitter gourd (Momordica charantia L.) cultivated in China is regarded as an important vegetable crop and is of considerable economic importance. However, it is susceptible to fusarium wilt, which causes heavy economic losses. Forty‐eight isolates were isolated from diseased bitter gourd plants that displayed typical fusarium wilt symptoms. Based on the morphological features, the rDNA internal transcribed space (ITS) sequences, pathogenicity and host biotypes, all of the isolates tested were pathogenic to the susceptible bitter gourd plants species (cv. ‘Guinongke No. 2’) and were identified as Fusarium oxysporum f. sp. momordicae (FOM). Our results classified different isolates as slightly, moderately or highly virulent. Among the isolates tested, 43 isolates slightly infected bottle gourd (Lagenaria siceraria var. clavata), whereas they did not infect other species from the family Cucurbitaceae. Genetic diversity among 48 isolates was characterized using amplified fragment length polymorphism (AFLP) analysis. The number of bands amplified by each primer pairs ranged from 41 to 66, with sizes ranging from 200 to 500 bp. A total of 366 bands were observed, out of which 363 were polymorphic (99.14%). The Nei's genetic identity of the six geographical populations varied from 0.7362 to 0.9707. The mean Nei's gene diversity index (H = 0.2644) and the mean Shannon's information index (I = 0.4071) at species level were higher than ones at populations level, indicated that the variation within populations was greater than that among populations. The Nei's GST (0.5103) and gene flow (Nm = 0.4923) revealed that genetic differentiation was mainly among populations and few gene exchanges. The dendrogram obtained from AFLP marker showed that there was a good correlation between isolates from different geographical locations and their pathogenicity. The AFLP marker effectively distinguished the high virulent isolates from the less virulent isolates. The highly virulent isolates were distinctly separated in different clusters, which indicated a significantly high correlation with the geographical origin in the AFLP dendrogram. The pathogenicity and molecular marker analysis confirmed the presence of variation in virulence as well as genetic diversity among the FOM isolates studied.     

Intraspecific diversity in the fungal speciesChaunopycnis alba: Implications for microbial screening programs

Intraspecific variation among 84 isolates of the anamorphic fungusChaunopycnis alba from 26 different geographical locations was analyzed by investigating optimal growth temperatures, differences in the production of secondary metabolites and presence or absence of the cyclosporin synthetase gene. The genetic diversity was assessed using random amplified polymorphic DNA (RAPD). Analysis of these data showed high genetic, metabolic and physiological diversity within this species. Isolates from the Antarctic represented the most homogeneous group withinC. alba and together with isolates from the Arctic these polar strains differed from alpine, temperate and tropical strains by low optimal growth temperatures and by low production of secondary metabolites. Isolates from tropical climes were characterized by high optimal growth temperatures and by the production of comparatively diverse metabolite spectra. Most of the isolates that were similar in the combination of their physiological and metabolic characters were also genetically related. Isolates from different geographical origins did not show many similarities, with the exception of the cyclosporin A-producing isolates, and large diversity could be observed even within a single habitat. This leads us to the suggestion that for pharmaceutical screening programs samples should be collected from a diversity of different geographical and climatic locations. For the selection of strains for screening the RAPD assay seems to be the most powerful tool. It reflected the highest intraspecific diversity and the results corresponded well with the other characteristics.     

Elevated genetic variation within virulence-associated Botrytis cinerea polygalacturonase loci

Botrytis cinerea, or gray mold, is a necrotrophic fungal pathogen of hundreds of plant species. The genetic diversity of B. cinerea may contribute to its broad host range; however, the level and structure of genetic variation at pathogenesis-associated loci has not been described. B. cinerea possesses six distinct cell-wall-degrading polygalacturonases (PGs), enzymes of demonstrated importance to pathogenesis and interaction with host plant defenses. Sequencing a collection of 34 B. cinerea isolates at three PG-encoding loci, BcPG1, BcPG2, and BcPG3, revealed limited evidence of host-mediated genetic subdivision within loci, yet suggested differences in the action of evolutionary forces among loci. BcPG1 and BcPG2 are highly polymorphic, particularly when compared with previously published data from nonpathogenicity loci, whereas BcPG3 is relatively conserved. Sequence variation at BcPG1 and BcPG2 did not appear to be associated with virulence on Arabidopsis leaves; however, BcPG2 variation showed a statistically significant association with growth rate on pectin. Rather than providing evidence for host-mediated genetic subdivision at individual PG loci, our data support specialization among PGs and the potential diversification of PGs interacting directly with host defenses.     

双孢蘑菇96．4菌株和单孢后代间的遗传变异

本文研究了野生双孢蘑菇菌株96．4（Agaricus bisporus)和它的10个单孢菌株的生长发育和遗传变异。研究表明,在菌株96．4和它的单孢菌株之间以及在10个单孢菌株之间在菌丝生速度和随机扩增多态DNA（RAPD）揭示的遗传变异等方面都存在着明显的变异。RAPD指纹揭示了96．4菌株通过有性生殖产生的丰富的生物学变异。     

Antifungal Activities of Chelidonium majus Extract on Botrytis cinerea in vitro and Ultrastructural Changes in its Conidia

The extract from the aboveground parts of Chelidonium majus reduced the mycelial growth of Botrytis cinerea on Czapek agar medium. In minimum fungicidal concentration (60  μ l/ml), the extract induced irreversible ultrastructural changes in B. cinerea conidia.     

Competitive ability of the antagonists Ulocladium atrum and Gliocladium roseum at temperatures favourable for Botrytis spp. development

Ulocladium atrum and Gliocladium roseum are fungal antagonists capable of suppressing sporulation of Botrytis spp. on dead plant parts. The effect of temperature (3 to 36 °C) on antagonist conidial germination and mycelial growth was assessed on agar. In addition conidial germination of U. atrum was measured on dead lily leaves. The optimum temperature of both antagonists for both conidial germination and mycelial growth was between 27 and 30 °C. U. atrum was less affected by lower temperatures than G. roseum. At optimum temperature, 50% of conidia of U. atrum and G. roseum germinated within 2.6 and 10.0 hrs, respectively. At low sub-optimal temperatures (6 °C), 50% of conidia germinated within 18 and 96 hours, respectively.In bioassays on dead onion leaves, U. atrum suppressed sporulation of B. cinerea and B. aclada at all temperatures tested (6 to 24 °C) by more than 85%. On dead cyclamen leaves, G. roseum was more efficient than U. atrum at 21 and 24 °C but, in contrast to U. atrum, showed no antagonistic activity at temperatures below 21 °C. On dead hydrangea leaves, U. atrum significantly reduced sporulation of B. cinerea at temperatures as low as 3 and 1 °C. Under Dutch growing conditions, the mean air temperature during leaf wetness periods in onion and lily fields was 15 °C with temperatures only occasionally above 20 °C. In greenhouse crops of cyclamen, the mean temperature during high humidity periods was 17 °C. It is therefore concluded that U. atrum is better adapted than G. roseum to temperatures which occur in the field, in greenhouse crops such as cyclamen, or during cold storage of plant stocks.     

Comparisons of Isolates of Fusarium avenaceum from White Lupin and Other Crops by Pathogenicity Tests,DNA Analyses and Vegetative Compatibility Tests

Isolates of Fusarium avenaceum, mostly from crops of white lupin or wheat, were tested for pathogenicity on white lupin and wheat plants and compared by DNA tests and, in a limited study, vegetative compatibility. Most of the 80 isolates were pathogenic on both plant species after inoculation on shoot bases. Disease severity was greater at higher incubation temperatures that ranged from 15/10°C to 25/20°C (day/night temperatures). Isolates from lupin crops tended to be more pathogenic, on average, on lupins than on cereals. Polymerase chain reaction (PCR)‐restriction fragment length polymorphism (RFLP) analysis of the internal transcribed spacer region of the rDNA distinguished two groups of isolates that occurred in different proportions among isolates from lupins and cereal crops. Random amplified polymorphic DNA (RAPD)‐PCR analyses indicated considerable genetic variation among isolates, but there was some similarity among groups of isolates from populations in the same field. Genetic diversity was confirmed by a high degree of vegetative incompatibility among 20 isolates using nitrate nonutilizing mutants. There were no relationships among pathogenicity, RFLP group, RAPD group and vegetative compatibility group.     

The BMP1 gene is essential for pathogenicity in the gray mold fungus Botrytis cinerea

In Magnaporthe grisea, a well-conserved mitogen-activated protein (MAP) kinase gene, PMK1, is essential for fungal pathogenesis. In this study, we tested whether the same MAP kinase is essential for plant infection in the gray mold fungus Botrytis cinerea, a necrotrophic pathogen that employs infection mechanisms different from those of M. grisea. We used a polymerase chain reaction-based approach to isolate MAP kinase homologues from B. cinerea. The Botrytis MAP kinase required for pathogenesis (BMP) MAP kinase gene is highly homologous to the M. grisea PMK1. BMP1 is a single-copy gene. bmp1 gene replacement mutants produced normal conidia and mycelia but were reduced in growth rate on nutrient-rich medium. bmp1 mutants were nonpathogenic on carnation flowers and tomato leaves. Re-introduction of the wild-type BMP1 allele into the bmp1 mutant restored both normal growth rate and pathogenicity. Further studies indicated that conidia from bmp1 mutants germinated on plant surfaces but failed to penetrate and macerate plant tissues. bmp1 mutants also appeared to be defective in infecting through wounds. These results indicated that BMP1 is essential for plant infection in B. cinerea, and this MAP kinase pathway may be widely conserved in pathogenic fungi for regulating infection processes.