Detoxification of α-tomatine by tomato pathogens Alternaria alternata tomato pathotype and Corynespora cassiicola and its role in infection

In tomato plants, α-tomatine, a steroidal glycoalkaloid saponin, inhibits fungal growth. Tomato pathogens that produce host-specific toxins, Alternaria alternata tomato pathotype causing Alternaria stem canker and Corynespora cassiicola causing Corynespora target spot, were investigated for sensitivity to α-tomatine. Although spore germination of A. alternata pathogenic and nonpathogenic to tomato and of C. cassiicola pathogenic to tomato was not affected by 0.1 mM α-tomatine, spore germination of C. cassiicola nonpathogenic to tomato was significantly inhibited. This result showed that A. alternata, regardless of its pathogenicity, and only the C. cassiicola pathogenic to tomato are resistant to α-tomatine. Germinating spores of A. alternata and C. cassiicola resistant to α-tomatine detoxified α-tomatine by degrading it to a less polar product. After inoculation of tomato leaves, spores of A. alternata and C. cassiicola nonpathogenic to tomato germinated and formed appressoria, but did not form infection hyphae in host tissues. When a host-specific toxin (CCT-toxin) produced by C. cassiicola pathogenic to tomato was added to nonpathogenic spores, colonization within leaves was observed in A. alternata, but not in C. cassiicola. On the other hand, when spores of C. cassiicola nonpathogenic to tomato were suspended in spore germination fluid of nonpathogenic A. alternata with α-tomatine detoxification activity, the fungus could be induced to colonize leaves in the presence of CCT-toxin. These results indicate that A. alternata tomato pathotype and C. cassiicola pathogenic to tomato detoxify α-tomatine during infection and that this detoxification is essential for host colonization by pathogens that produce host-specific toxins.     

重庆市烟草赤星病菌致病力测定

利用离体叶片悬滴接种法,将2005年在重庆市各烟区采集分离的烟草赤星病24个代表菌株进行了致病力测定。结果显示,所有测试菌株均具有致病力,但存在明显的致病力差异,其中强致病力菌株5个,较强菌株8个,中等致病力菌株10个,弱致病力菌株1个,分别占供试菌株的20.8%、33.3%、41.7%和4.2%。这表明试验分离获得的大多数菌株致病力居中或偏强。对来源性不同的菌株的致病力进行分析比较,发现重庆各烟区菌株的致病力有所不同,而且同一烟区的菌株致病力也存在一定的差异。离体叶片试验表明,赤星病菌不同病原菌株对烟草叶片的穿透生长能力与致病力呈明显的正相关。     

苹果叶片应答斑点落叶病菌胁迫的蛋白质组学分析

In order to elucidate key proteins related to disease-resistance in apple leaves,the total proteins were extracted from control and infected apple leaves, and separated by two-dimensional electrophoresis. The differential expressed proteins were then identified by MALDI-TOF-TOF/MS.In total, 25 differential expressed proteins were detected by Image Master Software. After tryptic digestion, MALDI-TOF-TOF/MS analysis and database searching, 20 protein spots were finally identified, including 11 functional proteins related to photosynthesis, energy metabolism, stress and defense responses. The pathogenesis-related proteins involved in defense responses such as APX, GPX and Mal d1 were differentially expressed in apple leaves. It indicates that these proteins may play a key role in the resistance to A. alternata apple pathotype.     

UV-B辐射增强对链格孢菌(Alternaria alternata)生长、生理及致病力的影响

研究UV-B辐射增强对灯盏花(Erigeron breviscapus)叶斑病致病菌—链格孢菌(Alternaria alternata)的生长、生理及其致病力的影响,并探讨链格孢菌对UV-B辐射的响应机制。结果表明,UV-B辐射增强可延缓链格孢菌的生长,导致菌丝生长率、产孢量、纤维素酶活性及可溶性蛋白含量均显著降低(p0.05),链格孢菌的致病力显著下降(p0.05)。同一UV-B辐射强度下,20～60min的辐射处理对链格孢菌的生长、生理及致病力的影响没有显著性差异(p0.05)。链格孢菌表现出形态学和生理生化方面的UV-B适应机制,来减轻UV-B辐射的损伤。经UV-B辐射处理后的病菌积累较多黑色素,菌落颜色加深,菌丝分布更加致密,菌丝干重显著增加(p0.05);并且,链格孢菌的过氧化氢酶(CAT)活性显著升高(p0.05)。     

温度对烟草赤星病菌生物学特性及代谢表型的影响

为了解温度对烟草赤星病菌致病力及代谢表型的影响, 本研究采用菌丝生长速率法和离体叶片法, 分别测定不同温度下烟草赤星病菌的生长速率和致病力, 同时采用Biolog代谢表型技术测定了其在22?25?30℃和35℃下不同碳源?渗透压和pH下的代谢表型?结果表明, 烟草赤星病菌在15~35℃下均可生长, 30℃时菌丝生长最快?致病力和产孢能力最强, 35℃时孢子萌发率最高?在22?25?30℃和35℃时, 赤星病菌均可代谢Biolog FF代谢板上的95种碳源, 且随着温度升高对碳源的代谢能力逐渐增强?22℃和25℃时烟草赤星病菌对渗透压的适应范围最广, 其次为30℃, 35℃时适应范围最窄?在22?25?30℃和35℃时, 赤星病菌在pH 3.5~10范围内均可正常代谢, 在22?25℃和30℃下, 赤星病菌表现出强脱羧酶活性和弱脱氨酶活性, 在35℃下, 其脱羧酶和脱氨酶活性均相对较弱?研究结果揭示了烟草赤星病菌在不同温度下的适应能力, 为赤星病菌与环境互作研究提供了参考?     

Evolution of pathogenicity controlled by small,dispensable chromosomes in Alternaria alternata pathogens

Alternaria alternata includes seven pathogenic variants, called pathotypes, which produce host-selective toxins (HSTs) as determinant factors for pathogenicity. The gene clusters for HST biosynthesis were identified from six pathotypes (Japanese pear, strawberry, tangerine, apple, tomato and rough lemon) and were found to reside on small chromosomes of <2.0 Mb in most strains tested. We isolated mutants lacking the small chromosomes from the strawberry, apple and tomato pathotypes and showed that the small chromosomes are dispensable for growth. In this review, we summarize our current understanding of the evolution of pathogenicity controlled by small, dispensable chromosomes in Alternaria alternata pathogens.     

DNA transposon fossils present on the conditionally dispensable chromosome controlling AF-toxin biosynthesis and pathogenicity of Alternaria alternata

The strawberry pathotype of Alternaria alternata produces host-specific AF-toxin and causes Alternaria black spot of strawberry. Previously, we isolated cosmid clones pcAFT-1 and pcAFT-2 from strain NAF8 of the strawberry pathotype that contain AF-toxin biosynthetic genes, named AFT genes. In a molecular characterization here of pcAFT-1 and pcAFT-2, 11 AFT genes and five transposon-like sequences, named TLS-S1 to TLS-S5, were detected. The nucleotide sequences of TLS-S1 and TLS-S4 share high homology, and their putative products have similarity to transposases of the hAT family transposons. Thus, TLS-S1 and TLS-S4 were renamed TLS-S1-1 and TLS-S1-2, respectively. Amino acid sequences deduced from TLS-S2, TLS-S3, and TLS-S5 have similarity to transposases of the Fot1/Pogo family transposons, but they are significantly different. All five sequences have incomplete open reading frames (ORFs) for transposases owing to deletions, termination codons, and/or frameshifts, indicating that they are inactivated elements. Analysis of genomic distribution of these sequences revealed that they are specifically distributed on a 1.05-Mb chromosome of NAF8, which has been identified as a conditionally dispensable (CD) chromosome encoding AFT genes. The presence of three, four, and three copies of TLS-S1, TLS-S2, and TLS-S3, respectively, and a single copy of TLS-S5 on the CD chromosome were estimated by DNA gel blot analysis. The remaining copy of TLS-S1 and the three copies of TLS-S2 were isolated and identified to also encode incomplete ORFs. Thus, it appears that all copies of the transposon-like sequences identified are inactivated elements (fossils) unique to the CD chromosome in the genome of the strawberry pathotype. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the accession numbers AB236733 (TLS-S1-1), AB236734 (TLS-S1-2), AB236735 (TLS-S1-3), AB236736 (TLS-S2-1), AB236737 (TLS-S2-2), AB236738 (TLS-S3), and AB236739 (TLS-S5)     

Identification and pathogenicity of Alternaria alternata causing leaf spot disease on sweet cherry in Province of Turkey

Journal of Plant Diseases and Protection - Sweet cherry leaf spot, caused by Alternaria spp., causes significant yield losses to sweet cherry production globally. This study was aimed at...     

Effect of Zataria multiflora essential oil on Alternaria alternata in vitro and in an assay on tomato fruits

Journal of Plant Diseases and Protection - The present study aimed to evaluate the effects of Zataria multiflora essential oil at different concentrations (0, 100, 200, 300 and 500 ppm) on the...     

Microscopic detection of reactive oxygen species generation in the compatible and incompatible interactions of Alternaria alternata Japanese pear pathotype and host plants

 Reactive oxygen species (ROS) generation was examined in the interaction of Alternaria alternata Japanese pear pathotype and host plants using three methods: nitro blue tetrazolium (NBT) method for microscopic detection of O₂ ⁻, diaminobenzidine (DAB) methods for microscopic detection of H₂O₂, and cerium chloride methods for ultrastructural detection of H₂O₂. ROS generation was detected by NBT and DAB methods at appressoria on leaves of susceptible cultivars and heat-shocked leaves of resistant cultivars but not in leaves of resistant cultivars. Ultrastructural detection by the cerium chloride method identified ROS generation at cell walls of appressoria and penetration pegs in susceptible, resistant leaves and heat-shocked leaves. These differences in the ultrastructural and microscopic data in resistant areas were due to the restriction of ROS generation in limited areas, the side facing the plant surface, of appressoria and penetration pegs. Therefore, ROS generation was apparently induced regardless of the resistance or susceptibility of the cultivar with the difference being in the volumes generated. After evaluating the pathological role of ROS generation in fungal structures, such generation was found to be associated with early penetration of cell walls in pear plants. Additionally, ROS generation in plants was also found in degrading pectin layers near infected hyphae and in plasma membrane modification sites in susceptible leaves but not in resistant leaves. ROS generation in susceptible leaves might be accompanied with plasma membrane damage, although the role of ROS generation in the pectin layers is not clear. ROS generation in both fungal and plant cells during their interaction was likely associated with the expression of susceptibility. Received: June 3, 2002 / Accepted: July 31, 2002     

Alternaria Leaf Spot on Three Species of Pelargonium Caused by Alternaria alternata in Japan

A new disease of pelargonium (Pelargonium domesticum Bailey), ivy geranium (P. peltatum (L.) L'Hér. ex Ait.) and scented geranium (P. graveolens L'Hér.), primarily causing brown spots on leaves, was found in Kawasaki-shi in Kanagawa Prefecture and Tachikawa-shi in Tokyo. An Alternaria sp. was consistently isolated from these diseased leaves, and the isolates were pathogenic to their host leaves. Based on morphological characteristics, the causal fungus in all three cases was identified as Alternaria alternata (Fr.) Keissler. Because Alternaria leaf spot of geranium by A. alternata has already been reported, the pathogenicity of isolates from four groups of genus Pelargonium was investigated. The isolates from scented geranium were pathogenic only to their original host, but the isolates from pelargonium, ivy geranium and geranium were pathogenic to all groups of pelargonium. This is the first report of this disease on pelargonium, ivy geranium and scented geranium caused by A. alternata in Japan. We propose the names for these diseases as Alternaria leaf spot of pelargonium (kappan-byo), Alternaria leaf spot of ivy geranium (kappan-byo) and Alternaria leaf spot of scented geranium (kappan-byo). Received 11 December 2000/ Accepted in revised form 19 July 2001