THE PHYTOTOXIC PROPERTIES OF ALTERNARIC ACID IN RELATION TO THE ETIOLOGY OF PLANT DISEASES CAUSED BY ALTERNARIA SOLANI (ELL. & MART.) JONES & GROUT

Alternaric acid, a metabolic product of some strains of Alternaria solani , is highly phytotoxic. If introduced into the vascular system of cut shoots of tomato or potato, it travels upwards and causes necrotic lesions of the stem, petioles and leaf blades, very similar in appearance to the lesions appearing in some phases of natural attack by A. solani. Alternaric acid also produces similar lesions on plants outside the host-range of A. solani. Associated with the production of visible lesions is a disturbance of the water-balance of shoots. Transpiration is increased and, in spite of a simultaneous but smaller increase in water uptake, this eventually leads to complete desiccation of the shoots. A strain of A. solani known to produce alternaric acid in synthetic culture media produces a substance with similar biological properties when inoculated into tomato fruits. Since other workers have shown that some of the symptoms of A. solani attack are toxigenically induced, it is tempting to suggest that alternaric acid is the toxin concerned. On the other hand, there is no correlation between virulence of strains of A. solani and their ability to produce alternaric acid in synthetic culture media or in tomato fruits. Some highly virulent strains produce little or no alternaric acid under any conditions tested. There is some evidence that other toxins beside alternaric acid are produced by A. solani , not necessarily in the same proportions by all strains.     

菊芋叶片提取物抑菌活性与化学成分的研究

为了开发新型植物源杀菌剂和充分利用菊芋(Helianthus tuberosus L.)资源,本文尝试用石油醚、乙醚、乙酸乙酯和水等4种溶剂对菊芋叶片进行平行提取,采用生长速率法测定菊芋叶片提取物对水稻纹枯菌(Rhizoc-tonia solaniKühn)、小麦赤霉菌[Gibberella zeae(Schw.)Petch]、番茄早疫菌[Alternaria solani(Ellis et Martin)Jones et Grour]和番茄灰霉菌(Botrytis cinerea Pers.)生长量的抑制活性;并通过试管法和滤纸法对菊芋叶片内化学成分进行初步预试。抑菌实验结果表明:(1)菊芋叶片各溶剂提取物处理与对照处理相比差异显著;(2)菊芋叶片水提取物处理与各有机溶剂提取物处理差异也基本显著;(3)菊芋叶片各溶剂提取物同浓度处理对水稻纹枯菌、番茄早疫菌和番茄灰霉菌抑制效果较好;(4)菊芋叶片乙酸乙酯提取物抑菌效果最为显著,浓度为20mg/mL时对番茄早疫菌和番茄灰霉菌已达到完全抑制,对水稻纹枯菌抑制率也达到77.91%。初步化学预试结果说明,菊芋叶片中含有蛋白质、氨基酸、还原糖类、有机酸、酚类和鞣质、黄酮类、内酯类、强心甙以及油脂等化学成分。     

三种多菌灵耐药性链格孢属真菌β-tubulin基因的序列特征

应用两对引物,从对多菌灵具有耐药性的链格孢属3个真菌中扩增了与多菌灵耐药性相关的β-微管蛋白基因,基因长度分别为1,419bp（瓜链格孢）,1,426bp（茄链格孢）,1,419bp（链格孢）,均包含3个内含子,编码398个氨基酸。3个链格孢属真菌与其他对多菌灵敏感的植物病原真菌β-微管蛋白氨基酸具有高度的同源性。但是,3个真菌167位氨基酸均为酪氨酸,而其他对多菌灵敏感的真菌167位均为苯丙氨酸,研究结果表明链格孢属真菌对多菌灵的耐药性可能与167位酪氨酸有关。     

人参(Panax ginseng)根系分泌物成分对人参致病菌的化感效应

采用室内培养结合生物学测定的试验方法,研究了不同浓度人参(Panax ginseng)根系分泌物成分苯甲酸、邻苯二甲酸二异丁酯、十六酸和2,2-二(4-羟苯基)丙烷对人参立枯丝核菌(Rhizoctonia solani)、黑斑菌(Alternaria panax)、疫病菌(Phytophthora cactorum)、菌核菌(Sclerotinia schinseng)、锈腐菌(Cylindrocarpon destructans)和绿色木霉菌(Trichoderma viride)菌落生长及孢子萌发的化感效应.结果显示,不同浓度人参根系分泌物成分对人参致病菌及绿色木霉菌的化感效应存在显著差异.苯甲酸浓度与人参立枯丝核菌、菌核菌和锈腐菌菌落生长以及人参黑斑菌、锈腐菌孢子萌发呈负相关,与人参黑斑菌、绿色木霉菌菌落生长呈正相关;对人参疫病菌菌落生长的化感效应表现为低浓度和高浓度抑制,中浓度促进.邻苯二甲酸二异丁酯浓度与人参立枯丝核菌、黑斑菌、菌核菌和绿色木霉菌菌落生长以及人参黑斑菌孢子萌发呈负相关;对人参锈腐菌菌落生长和孢子萌发表现为低浓度和高浓度抑制,中浓度促进;对人参疫病菌菌落生长表现为低浓度和中浓度抑制,高浓度促进.2,2-二(4-羟苯基)丙烷浓度与人参立枯丝核菌、黑斑菌、疫病菌、绿色木霉菌菌落生长以及人参黑斑菌、锈腐菌孢子萌发呈负相关;对人参菌核菌、锈腐菌菌落生长表现中浓度促进,高浓度抑制.十六酸浓度与人参锈腐菌、疫病菌和绿色木霉菌菌落生长呈正相关,与人参锈腐菌孢子萌发呈负相关,对黑斑菌孢子萌发表现为中浓度抑制.4种根系分泌物的等量混合物浓度与人参致病菌及拮抗木霉菌菌落生长速率呈负相关.     

Isolation and characterization of microsatellite markers from the phytopathogenic fungus Alternaria dauci

Eleven polymorphic microsatellite markers were isolated from the necrotrophic phytopathogenic fungus Alternaria dauci based on enriched genomic libraries. In order to assess allelic variability, the microsatellite loci were analysed in a collection of 43 isolates. The number of detected alleles in 11 loci ranged from two to 24 (mean 10.4). Test of cross-species amplification and sequencing of the resulting amplicons showed that some of these microsatellites could be used in different species such as Alternaria solani, Alternaria bataticola and Alternaria zinniae.     

Isolation of two antifungal compounds fromNeolitsea cuipala and their activity against some plant pathogenic fungi

Zeylanidine and zeylanicine isolated fromNeolitsea cuipala fruit peels are found to be potential antifungal agents againstHelminthosporium oryzae, Fusarium oxysporum, Alternaria solani, Curvularia varuciformis andColletotrichum gleosporioides.     

INVESTIGATIONS ON FUNGICIDES

Twenty (aryloxythio)trichloromethanes were examined for in vitro fungicidal activity against six fungi. All compounds showed a direct fungistatic effect and some exhibited a marked fumigant action. Good protectant action against Alternaria solani on tomato and Botrytis fabae on broad bean was obtained with certain compounds, but none was better than the three standard protectants used for comparison. When supplied to plants through their roots, eight conferred significant systemic fungicidal protection against Alternaria solani in tomato, but there was no significant protection against Botrytis fabae in broad bean. In preliminary tests the 2:4:5-trichlorophenoxythio analogue gave promising results when examined as a fumigant fungicide for eradication of downy mildew in lettuce plants, for reducing lenticel rot in stored apples and for preventing blue-mould in oranges.     

瑞香狼毒根的抑菌活性研究（Ⅰ）

以苹果干腐病菌、小麦赤霉病菌、番茄早疫病菌、南瓜枯萎病菌、玉米大斑病菌、烟草赤星病菌和辣椒疫霉病菌为供试菌,采用生物活性跟踪法、生长速率测定法和系统溶剂提取法对瑞香狼毒根中的杀菌活性物质进行了筛选和分离。结果表明,杀菌活性物质主要集中在乙酸乙酯提取物和甲醇粗提物中。从乙酸乙酯提取物中分离出两种杀菌活性物质AF1-4和AF1-5,同时还发现瑞香狼毒根中还存在增菌物质。     

培养条件对茄链格孢产孢的影响

茄链格孢Alternaria solani在体外培养条件下不产生或产生极少量的分生孢子。研究了培养基、菌丝损伤、紫外线照射和温度变化对7个茄链格孢菌株产孢的影响,发现这些处理方法对供试大多数菌株的产孢都有显著影响,确立了茄链格孢大量产孢的最优条件。利用优化后的条件对160株采自不同地区的茄链格孢进行体外诱导产孢,发现120株可大量产孢,占总数的75%,产孢量为1.26×104–5.03×104个/cm2。     

Comparison of Infection by W-type and R-type Isolates of the Eyespot Pathogen, Pseudocercosporella herpotrichoides, in Relation to Host Species and Cultivar, Temperature and Watering Regime in Controlled Environments

Abstract The spore-germination fluid of Alternaria solani was examined for factors which are important in the early stages of infection and probably explain the selectivity of A. solani to tomato and potato as host plants. The following activities of the spore-germination fluid were found: 1. Host-selective toxicity at low concentrations. 2. Infection promotion and enhancement of A. solani infections on host plants when spore-germination fluid was added to the inoculum. 3. An induction of susceptibility which enabled a non-pathogenic A. alternata isolate to cause necrotic spots on the leaves of the host plants of A. solani when its inoculum contained spore-germination fluid of A. solani. Properties 1 and 2 seemed to, be attributed to alternaric acid. This phytotoxin was detectable in effective concentrations in the spore-germination fluid. A factor, responsible for the susceptibility-induction for A. alternata on the host plants of A. solani was characterized as a water-soluble fraction of the spore-germination fluid which was free of alternaric acid and not toxic.     

ERIC and REP-PCR Banding Patterns and Sequence Analysis of the Internal Transcribed Spacer of rDNA of Stemphylium solani Isolates from Cotton

The genetic diversity of the Stemphylium solani isolates from cotton was assessed by Enterobacterial Repetitive Intergenic Consensus (ERIC) and Repetitive Extragenic Palindromes (REP)-PCR fingerprinting. Twenty eight monosporic isolates of S. solani from cotton were used along with five isolates from tomato and one isolate of Alternaria macrospora from cotton for comparison. The dendrogram obtained revealed clear differences between the cotton and tomato isolates as well as between the tomato isolates from different geographic regions. The genetic relationships among S. solani isolates were also analyzed by sequencing the internal transcribed spacer (ITS) region of four isolates representing the three ERIC and REP groups. The tomato isolate from the State of S?o Paulo showed a distinct ITS sequence from that of the cotton isolates and tomato isolate from the State of Goiás, giving evidence that it belongs to a different genotype of S. solani. This is the first report of the entire sequence of the ITS1-5.8S-ITS2 regions of S. solani.     

链格孢霉醇和链格孢霉醇单甲醚产生菌株的筛选

本文对分离自小麦、马铃薯、番茄和茄子上链格孢霉属(Alternaria)2个种(链格孢和茄链格孢)的96个菌株,用枯草杆菌生长抑制试验筛选链格孢霉醇(AOH)和链格孢霉醇单甲醚(AME)的产生菌株,有48株产生毒性作用(占所测菌株的50％)。18株产强、中毒性菌用高效液相色谱分析,有13株产AOH和AME(占所测菌株的72.2％)。链格孢的产毒素菌株率比茄链格孢低。但产毒素含量却是前者明显高于后者。其中产AOH和AME的最高含量,链格孢菌株XA-8分别为280和5140mg/kg,而茄链格孢菌株SA-10分别为95.9和94.3mg/kg。     

Fungicidal Activity of Some Common Weed Extracts Against Different Plant Pathogenic Fungi

Aqueous extract effects of 64 weed species on growth and development of Alternaria solani Sorauer. Helminthosporium satirum King & Bakke and Rhizodonia solani Kuhn. plant pathogenic fungi were studied in vitro . Extracts varied in the strength and persistence of their antifungal effects against the three fungi species. Some stimulated, others inhibited or had no effect. Among all species tested, extracts of Chenopodium murale, Falearia vulgaris. Ranunculus asiaticus and Sisymbrium irio were the most toxic to A. solani. Anagallis arlensis. Atriptex leucoclada, Crepis aspera. Notobasis syriaca. R. asiatieus, Rumex crispu. S. irio. Sonehu.s oleraceous and Vieia narhonensis to H. satirum and R. asiatieus. S. oleraceous and Mercurialis annua to R. solani. However. R. asiatieus extract was the most effective and completely inhibited growth and sporulation of the three fungi species at all incubation periods.     

PHOTO-INHIBITION OF SPORULATION IN ALTERNARIA SOLANI

Lukens, R. J. (Connecticut Agric. Expt. Sta., New Haven.) Photo-inhibition of sporulation in Alternaria solani. Amer. Jour. Bot. 50(7): 720–724. Illus. 1963.—Day-old conidiophores from starved cultures of Alternaria solani require a 12-hr dark period to produce conidia. If the cultures are illuminated during the dark period, conidial production is inhibited. The action spectrum of light inhibiting conidial formation contains 2 maxima, a sharp one at 450 mμ and a broad one extending from 375 to 425 mμ. The entire action spectrum corresponds approximately to the absorption spectra of riboflavin-5-phosphate mononucleotide (FMN) and of 6-carotene. FMN nullifies the effect of light in inhibiting sporulation, but b-carotene does not. Flavins appear to be essential for conidial formation and are photo-inactivated. It is likely that flavins are photo-receptors through which light inhibits conidial formation in A. solani.     

链格孢属一新种

在陕西洛南烟田发现一个链格孢属新种一烟草链格孢（AlternarianicotianaJ.L.Cheng）,该新种与烟草赤星病[A.alternata（Fries）Keissler]的分生孢子形态、大小明显不同,引起的病害症状相近,但前者病斑大,色泽深,有光泽,发病时间早,在烟叶生长中后期发病。新种的模式标本存放在陕西省烟草研究所植保室。     

Mycoflora and Potential for Mycotoxin Production of Freshly Harvested Black Bean from the Argentinean Main Production Area

A mycological survey was carried out, for the first time, on black bean samples from the northwestern Argentinean province of Salta in the 1999 harvest season. Ten varieties of black beans were evaluated at three locations. Species of the genus Alternaria were the most prevalent component of the black bean mycoflora. Species of Fusarium, Sclerotinia, Rhizoctonia and Acremonium were also recorded. The predominant species of the genera isolated were Alternaria alternata, Sclerotinia sclerotiorum, Rhizoctonia solani, Fusarium semitectum and Acremonium strictum. An analysis of variance was applied to determine possible differences between black bean varieties. Variety FT88/519 was the most susceptible to Sclerotinia sclerotinia infection, while the variety DOR 604 was the least susceptible. As toxigenic species were recovered, Alternaria toxins, zearalenone and trichothecenes may pose a contamination risk for black bean.     

河北省马铃薯早疫病菌群体遗传结构的研究

通过对2009－2010年河北省145株及外省30株马铃薯早疫病菌对代森锰锌的敏感性、产孢量两个表型性状和AFLP基因型的测定,揭示了河北省早疫病菌群体的遗传结构。毒力测定表明所有供试菌株对代森锰锌均表现敏感,其EC50范围介于1.04－4.86μg/mL之间,平均为2.93μg/mL。被测河北省47株早疫病菌产孢量平均为85个孢子/mm2,大大高于30个对照菌株的产孢量,对照菌株的平均产孢量为50个孢子/mm2,菌株间产孢量存在明显差异。AFLP聚类分析揭示出了马铃薯早疫病菌丰富的遗传多样性,每个菌株都具有独特的AFLP基因型。早疫病菌AFLP基因型与地理来源存在着密切的相关性,与菌株产孢量有一定的相关性,但与代森锰锌敏感性无相关性。     

Alternaric acid stimulates phosphorylation of His-tagged RiCDPK2, a calcium-dependent protein kinase in potato plants

Calcium-dependent protein kinases (CDPK) are an essential component of plant defense mechanisms against pathogens. We investigated the effect of alternaric acid, a host-specific toxin produced by the plant fungal pathogen Alternaria solani (Pleosporaceae), on a putative plasma membrane and cytosolic kinase RiCDPK2 of potato (Solanum tuberosum) and on hypersensitive cell death of host potato cells. Alternaric acid, in the presence of Ca(2+) and Mg(2+), stimulated in vitro phosphorylation of His-tagged RiCDPK2, a Ca(2+)-dependent protein kinase found in potato plants. We concluded that Ca(2+) and Mg(2+) play an important role in the interaction between alternaric acid and RiCDPK2. Based on our observations, alternaric acid regulates RiCDPK2 kinase during the infection process in an interaction between host and A. solani, leading to the inhibition of hypersensitive cell death in the host. We suggest that alternaric acid is a primary determinant by which A. solani stimulates CDPK activity in the host, suppressing hypersensitive cell death.     

欧美107杨树提取物体外对植物病原真菌的抑制活性

制备了欧美107杨树枝条和叶的乙醇粗提物及不同极性溶剂的萃取部分,并测定了它们对植物病原真菌的抑制活性。枝条和叶的乙醇粗提物对棉花枯萎病菌、小麦纹枯病菌、番茄早疫病菌、番茄枯萎病菌、黄瓜枯萎病菌、小麦赤霉以及玉米弯孢等7种植物病原真菌均具有一定的抑制作用。而枝条的乙醇粗提物对杨树溃疡病菌的菌丝生长有一定的促进作用。抗菌活性成分主要集中在乙酸乙酯萃取部分。     

地表球囊霉诱发番茄抗早疫病的机理

丛枝菌根可以改善植物营养状况,提高宿主植物的抗病性.本文研究了番茄幼苗预先接种丛枝菌根真菌(AMF)地表球囊霉后对番茄植株保护酶活性和防御反应基因表达,以及对番茄早疫病抗性的影响.结果表明:被AMF侵染的番茄植株在接种早疫病病原菌茄链格孢菌后,其叶片内的超氧化物歧化酶(SOD)和过氧化物酶(POD)活性迅速提高.其中SOD酶活性在接种后18h达到最高,比只接种地表球囊霉(G)、茄链格孢菌(A)以及未接种AMF和病原菌的对照(CK)分别高28.6％、79.2％和82.8％;POD酶活性在接种后65 h达到最高,分别比G、A处理和CK高762％、18.3％和1710％.经荧光定量PCR检测表明,AMF侵染后的番茄植株再接种病原菌,其叶片中PR1(病程相关蛋白基因)、PR2(β-1,3-葡聚糖酶基因)和PR3(几丁质酶基因)基因的最高转录水平达到CK的9.67、8.54和13.4倍.与CK相比,先接种地表球囊霉再接种茄链格孢菌的番茄植株(GA)的早疫病发病率和病情指数分别降低了36.3％和61.4％.预先接种AMF的番茄植株在遇到病原菌袭击时诱导的防御反应强而迅速,诱发(priming)可能是菌根真菌提高宿主植物抗病性的重要机制.