Management and cross-infectivity potential of Colletotrichum acutatum causing anthracnose on bell pepper in Florida

Early anthracnose caused by Colletotrichum acutatum has become an increasingly serious disease on green, unripe bell pepper fruit in Florida. This contrasts with earlier reports of anthracnose occurring on bell pepper primarily as a ripe-rot disease of mature, colored pepper fruit caused by Colletotrichum gloeosporioides. Management of anthracnose on green bell pepper fruit using fungicides and a commercial inducer of systemic acquired resistance, acibenzolar-S-methyl (ASM), was evaluated during three seasons. In two of the three trials, all the fungicides tested including azoxystrobin, fludioxonil + cyprodinil, mancozeb, famoxadone + cymoxanil, copper hydroxide, and ASM significantly increased the number of marketable fruit compared with control plants. These trials identified fungicides that could contribute to a successful pest management program on pepper for controlling anthracnose caused by C. acutatum. The cross-infectivity potential of C. acutatum was investigated on tomato and strawberry by in vitro and field inoculation. Anthracnose lesions formed readily on wound-inoculated detached fruits of all hosts in in vitro assays. Under field conditions, after inoculation, anthracnose lesions occurred on pepper fruit but no lesions of anthracnose were found on either ripe or unripe tomato or strawberry fruit in adjacent plots.     

云南葡萄产区葡萄炭疽病病原鉴定及致病力分析

为了明确引起云南葡萄产区炭疽病的病原种类,利用形态鉴定和特异性引物分子检测相结合的方法对从云南省主要葡萄产区采集的60株炭疽病菌菌株进行了鉴定。葡萄炭疽病菌菌株的菌落形态和生长速率与对照菌株尖孢炭疽菌Colletotrichum acutatum差异不明显,但其分生孢子大小显著小于尖孢炭疽菌,附着胞深褐色,球形或不规则形。胶孢炭疽菌Colletotrichum gloeosporioides特异性引物CgInt/ITS4从供试葡萄炭疽病菌菌株基因组DNA中扩增出1条约500 bp的特异性条带,而尖孢炭疽菌特异性引物CaInt2/ITS4对葡萄炭疽病菌无扩增条带。研究表明,引起云南葡萄主产区炭疽病的病原为胶孢炭疽菌;供试胶孢炭疽菌对红提葡萄均有致病力,但菌株致病力差异较大,对番茄和草莓存在交叉侵染的能力,且对多菌灵的敏感性较尖孢炭疽菌高。     

Anthracnose of sweet pepper caused by Colletotrichum scovillei in Japan

Severe fruit rot of sweet pepper was found in Shimane, Hyogo, Chiba, Toyama, and Nagano prefectures, Japan from 2005 to 2011. Dark, sunken spots with concentric rings of orange conidial masses appeared on fruits. Pathogenic isolates from diseased fruits in the prefectures were identified as Colletotrichum scovillei. This species was added to the pathogens of sweet pepper anthracnose in Japan. The representative isolate was pathogenic to sweet pepper, tomato and chili pepper fruits, kidney bean pod, azuki bean, pea and strawberry leaves, but a caused no symptoms on cucumber or carrot in inoculation tests.     

Identification and characterization of <Emphasis Type="Italic">Colletotrichum</Emphasis> spp. associated with chili anthracnose in peninsular Malaysia

Anthracnose fruit rot caused by Colletotrichum spp. is a serious post-harvest disease of chili fruits (Capsicum spp.). One hundred-thirty isolates of Colletotrichum spp. were isolated from anthracnose of green and red cayenne pepper (Capsicum annuum) and bird’s eye chili (Capsicum frutescens). The isolates were morphologically identified as Colletotrichum acutatum sensu lato (62 isolates), Colletotrichum truncatum (54 isolates), and Colletotrichum gloeosporioides sensu lato (14 isolates). Molecular identification and phylogenetic analyses were based on internal transcribed spacer regions, β-tubulin, actin, and glyceraldehyde-3-phosphate dehydrogenase genes, and the isolates were re-identified as C. scovillei (58 isolates), C. truncatum (54 isolates), C. siamense (11 isolates), C. fioriniae (four isolates), and C. fructicola (3 isolates). Maximum likelihood trees using combined sequences showed that isolates of the same species grouped in the same main clade with the isolates used for comparison. Pathogenicity testing showed that the tested isolates from each species were pathogenic towards green and red Capsicum annuum and Capsicum frutescens upon treatment of wounded fruit, using both the mycelial plug and conidial suspension methods. Only five isolates of C. truncatum and seven isolates of C. scovillei were found to be pathogenic upon treatment of unwounded fruit. The occurrence of five Colletotrichum spp. (C. siamense, C. fructicola, C. scovillei, C. fioriniae, and C. truncatum) associated with chili anthracnose in Peninsular Malaysia indicates that correct species identification is important to formulate not only effective disease management, but also effective quarantine policy.     

The potential of fludioxonil for anthracnose control on China chili fruit

Anthracnose, caused by Colletotrichum scovillei, is one of the most destructive disease which causes massive yield losses in chili. Our preliminary study has demonstrated that fludioxonil had high activity against mycelial growth and spore germination of C. scovillei but it has not been labeled on chili. The aim of this study was to investigate the impact of fludioxonil on C. scovillei infection. In greenhouse and field trials, fludioxonil was applied to chili plants to determine the duration of protection of fruit provided by this compound against chili anthracnose. Fludioxonil may have impact on the early stage of chili anthracnose infection by inhibiting the germ tube elongation and appressorium formation. Moreover, it inhibited the secondary infection of C. scovillei by inhibiting the sporulation, germination of spores and formation of appressoria. Fludioxonil provided high level of protective activity for up to 96 h, as well as excellent curative activity of within 24 h in vivo tests. In field trials, fludioxonil can reduce the incidence and severity of chili anthracnose while giving a higher chili yield. These results suggest that fludioxonil could be a promising fungicide for anthracnose control in chili production.     

<Emphasis Type="Italic">Colletotrichum acutatum</Emphasis> occurs asymptomatically on sweet cherry leaves

Leaves of sweet cherry, exposed to either paraquat or freezing to quickly senesce the leaf tissue, were incubated in about 100% RH at 25°C for 6 d. Sporulating colonies of Colletotrichum acutatum, the cause of anthracnose, developed on up to 100% of the paraquat-treated and frozen leaves, and on none of the untreated controls. Number of leaves and leaf area containing C. acutatum on naturally infected leaves increased over time from May to September. Mean incidence of C. acutatum on leaf blades on fruit spurs and vegetative shoots from eight orchard/year samplings were 41 and 33%, respectively. Secondary conidiation (formation of short hyphae and new conidia) from conidia applied to detached leaves took place 6 h after inoculation, but only up to 3% of the conidia formed new conidia. It may be concluded that asymptomatic sweet cherry leaves frequently host C. acutatum and may be a potential inoculum source for cherry fruit.     

1种辣椒新炭疽病的初步鉴定及室内药剂筛选

炭疽病是辣椒上的重要病害,影响了辣椒的产量与品质。生产中,在“辛香15号”辣椒上发现了1种新炭疽病,仅在辣椒果实上发生,典型病斑为椭圆形,周围水渍状,中间呈黑色同心轮纹,无小黑点出现,对常用杀菌剂不敏感;保湿后镜检发现,该病菌产生分生孢子盘,无刚毛,分生孢子近长椭圆形,无色,单孢,大小为12．5μm×3．75μm,有油球,一端稍尖;进一步分离培养病原菌并通过柯赫氏法则验证后,初步将其鉴定为尖孢炭疽菌（Collectotrichum acutatum Simmonds）。为了获得对该病效果好的化学药剂,采用牛津杯法测定了8种化学药剂对病菌的抑制效果,结果表明：250g/L丙环唑乳油3000倍液抑制效果最好且药效持续时间长,可作为该病的候选药剂。本试验结果可为防治尖孢炭疽菌引起的辣椒炭疽病提供理论依据和用药参考。     

Anthracnose on almond in Australia:disease progress and inoculum sources of Colletotrichum acutatum

Anthracnose, caused by Colletotrichum acutatum, is an important disease of almond and has caused significant economic losses in California, Israel and Australia. Anthracnose development was monitored for three growing seasons in an almond orchard in South Australia on two almond cultivars, Price and Nonpareil, with up to 80 % of fruit affected in 2004. Lesions, typical of anthracnose, formed on young developing fruit and symptoms continued to appear until the fruit were ca 20 mm long, after which no further lesions developed. Symptoms were observed on leaves, woody tissue showed signs of dieback, but blossom blight was not observed. Maximum disease incidenceperfor, man and Relative Area Under the Disease Progress Curve (RAUDPC) were significantly larger for Price than Nonpareil for each season, but differences in the apparent rates of infection for both cultivars were insignificant for the three growing seasons. The apparent rates of infection were correlated with rainfall and daily temperature for the three years combined but there was no correlation between maximum disease incidence or RAUDPC and these environmental parameters. Considerably more mummified fruit remained on the trees of cv. Price than Nonpareil each year; however, there was no correlation between the number of mummified fruit in one season and maximum disease incidence, RAUDPC or apparent rate of infection, in the following season. C. acutatum was recovered from mummified fruit, peduncles and bark, from both Price and Nonpareil, every month throughout a year-long sampling period. C. acutatum was also recovered from asymptomatic leaves, fruit, bark, buds and blossom, however, less frequently and at lower rates than from mummified fruit and peduncles. Recovery was consistently greater from Price than from Nonpareil for all tissues.     

Serratia marcescens associated with bell pepper (Capsicum annuum L.) soft-rot disease under greenhouse conditions

Soft-rotting bacteria affecting bell peppers crops represent an economically destructive disease of growing importance worldwide. In Venezuela since 2006, soft-rot symptoms have been occasionally observed in bell pepper fruits grown under greenhouse conditions. Affected fruits presented water-soaked lesions that progressed to complete fruit maceration. Bacteria were isolated from water-soaked lesions in order to identify the causal agent of this disease. Of 13 bacterial isolates recovered from affected fruits, only isolate AGPim1A was able to produce a hypersensitive reaction in tobacco plants and to reproduce soft-rot symptoms in bell peppers fruits. Several methods, including classical bacteriological tests and carbon utilization profiling, alongside with sequence analysis of 16S rRNA and housekeeping genes gyrB and groES-groEL, allowed identification of the soft-rotting bacterium as Serratia marcescens. To our knowledge, this is the first report showing a S. marcescens strain associated with soft-rot disease in bell pepper fruits.     

The effect of climatic factors on Colletotrichum gloeosporioides, causal agent of mango anthracnose, in the Philippines

The epidemiology of the anthracnose pathogen of mango, Colletotrichum gloeosporioides, was studied over two growing seasons in the Philippines. This pathogen spreads within tree canopies as water-borne conidia during rainfall, and causes symptoms on young flush leaves, flowers and fruit. Infection studies with leaves and fruit incubated at different temperatures and humidities led to the derivation of a logistic regression model of the percentage of conidia forming appressoria. This model was compared with a similarly derived model from work in Australia; it appeared that the Philippine isolates of C. gloeosporioides were adapted to the higher mean temperatures of the Philippines. Conidia were observed to germinate and form appressoria at relative humidities (RH) between 95 and 100%, even though free surface moisture was only visible at 100% RH. This model was used retrospectively to estimate infection levels in two field trials which had been established to compare pre- and post-harvest practices in the control of the disease. Using this information to plan applications of a curative fungicide might have resulted in four fewer sprays in the first trial and one less in the second, compared with the standard protective spray programme employed.     

Colletotrichum species associated with chili anthracnose in Australia

Phylogenetic relationships were determined for 45 Colletotrichum isolates causing anthracnose disease of chili in Queensland, Australia. Initial screening based on morphology, ITS and TUB2 genes resulted in a subset of 21 isolates being chosen for further taxonomic study. Isolates in the C. acutatum complex were analysed using partial sequences of six gene regions (ITS, GAPDH, ACT, CHS‐1, TUB2 and HIS3), and in the C. gloeosporioides complex were analysed using four gene regions (ITS, TUB2, ApMat and GS). Phylogenetic analysis delineated four Colletotrichum species including C. siamense, C. simmondsii, C. queenslandicum, C. truncatum and a new Colletotrichum species, described here as C. cairnsense sp. nov. This is the first reported association of C. queenslandicum, C. simmondsii and C. siamense with chili anthracnose in Australia; these species were previously associated with anthracnose on papaya and avocado. Furthermore, the dominant species causing anthracnose of chili in Southeast Asia, C. scovillei, was not detected in Australia. Inoculations on chili fruit confirmed the pathogenicity of C. cairnsense and the other four species in the development of chili anthracnose in Australia.     

湖南芷江辣椒上一种新炭疽病的病原鉴定

 2009 年8 月湖南省芷江县线果型辣椒品种红秀2003 未成熟果实上发生了一种新炭疽病。典型病斑长椭圆状,其中央呈成片黄褐色粉末,外围粉红色小点呈同心圆排列,边缘水渍状。8个分离菌株（HNZJ001 HNZJ008）培养时菌落初呈白色,渐变为浅红色,后期加深, 具明显灰黑色同心轮纹。分生孢子盘无刚毛, 分生孢子单胞,大小为15. 8 μm × 4. 1 μm,含2～7 个油球,一端稍尖。分离菌株HNZJ001 针刺接种,在离体的成熟果和未成熟果上均产生病斑,而作为对照的胶孢炭疽菌Collectotrichum gloeosporioides菌株LSQ1 不能侵染未成熟果。核糖体RNA 基因内转录间隔区(ITS)的PCR 产物经〖JP2〗测序后做BLAST 分析,结果表明该菌与尖孢炭疽菌C. acutatum(有性阶段为尖孢小丛壳Glomerella acutata)的ITS 序列100% 相同,系统进化分析显示置信度高达100%,据此确定该病原为尖孢炭疽菌。这是国内辣椒上尖孢炭疽菌的首次报道。     

Morphological and molecular diversity of Colletotrichum spp. causing pepper spot and anthracnose of lychee (Litchi chinensis) in Australia

Since the 1980s a new disease has been affecting Australian lychee. Pepper spot appears as small, black superficial lesions on fruit, leaves, petioles and pedicels and is caused by Colletotrichum gloeosporioides, the same fungus that causes postharvest anthracnose of lychee fruit. The aim of this study was to determine if a new genotype of C. gloeosporioides is responsible for the pepper spot symptom. Morphological assessments, arbitrarily‐primed PCR (ap‐PCR) and DNA sequencing studies did not differentiate isolates of C. gloeosporioides from anthracnose and pepper spot lesions. The ap‐PCR identified 21 different genotypes of C. gloeosporioides, three of which were predominant. A specific genotype identified using ap‐PCR was associated with the production of the teleomorph in culture. Analysis of sequence data of ITS and β‐tubulin regions of representative isolates did not group the lychee isolates into a monophyletic clade; however, given the majority of the isolates were from one of three genotypes found using ap‐PCR, the possibility of a lychee specific group of C. gloeosporioides is discussed.     

影响柿树炭疽菌孢子萌发、附着胞形成及致病性的环境因子

 在凹玻片上测定了不同浓度的葡萄糖溶液对柿树炭疽菌(Colletotrichum gloeosporioides)的分生孢子萌发和附着胞形成率的影响,结果表明:分生孢子萌发率随葡萄糖浓度升高而增加,但附着胞形成率下降,芽管长度增加,附着胞的直径几乎没有变化;随着时间的延长,孢子萌发率和附着胞形成率都有所增加。不同pH对分生孢子萌发和附着胞形成率的影响结果显示,分生孢子在pH 2.0~9.0的溶液中可以萌发,并产生附着胞;最适分生孢子萌发和附着胞形成的pH是在5.0~6.0。不同pH处理的致病试验结果说明。在23℃时病斑在pH 4.0~8.0条件下都可产生;致病试验结果也发现,17℃时,pH6.0处理能发病,但不形成分生孢子团,pH 5.0处理不发病;15℃时,pH 5.0和pH 6.0的处理都不能发病。温度对菌丝生长的影响显示,菌落生长最适温度是25℃左右,高温抑制菌落生长。寄主表面侵染结构扫描电镜观察结果表明,芽管长度变化很大,芽管可以纵向沿着脊或沟延伸,也可横向通过脊沟;附着胞均在沟底或近底部形成。     

Phylogenetic and morphological identification of Colletotrichum boninense: a novel causal agent of anthracnose in avocado

In recent years, anthracnose has become a significant disease affecting avocado fruit in the state of Michoacan, Mexico, where it significantly reduces fruit quality and commercial yield. Anthracnose has been assumed to involve Colletotrichum gloeosporioides and C. acutatum as causal agents. However, because of the increasing incidence of anthracnose, a more precise identification of the Colletotrichum spp. involved in this disease has become desirable. During the years 2004–2007, avocado fruits of different sizes exhibiting brown‐black and reddish spots on the pericarp and soft rot in the mesocarp, were gathered from orchards in nine counties. Fungal isolates were cultured on potato dextrose agar, and among these, 31 were selected for molecular, morphological and pathogenicity analyses. The molecular approaches used sequence typing of the internal transcribed spacer region and the partial nuclear large ribosomal subunit, allowing the unequivocal identification of C. gloeosporioides (71%), C. acutatum (16%) and C. boninense (13%). This last species has not been previously reported as being associated with anthracnose symptoms in avocado fruits anywhere in the world. Various morphological characteristics such as the size and shape of conidia were determined, as well as the conidial mass colour. Pathogenicity tests performed with all three species were conducted by inoculating healthy fruits. In each case, identical symptoms developed within 3 days of inoculation. Knowledge of the Colletotrichum populations in the Michoacan state, including the newly encountered avocado pathogen C. boninense, will facilitate further studies addressing the relationships between these Colletotrichum spp. and their avocado host.     

Effect of humidity and temperature on conidial germination and appressorium development of two Philippine isolates of the mango anthracnose pathogen Colletotrichum gloeosporioides

A comparison of rates of germination and appressorium formation by an isolate of Colletotrichum gloeosporioides on mango leaves, fruit surfaces and cellophane membranes showed that behaviour was broadly similar on all three substrates. Frequency of appressorium formation was slightly higher on cellophane membranes, and both hyaline and melanized appressoria were formed. Only melanized appressoria were formed on mango surfaces. In vitro experiments on membranes showed comparative differences in physiological behaviour with temperature for two Philippine isolates of C. gloeosporioides . The most stimulatory temperature for production of appressoria differed in isolates I-2 and I-4 (25 and 20°C, respectively). At 30°C more appressoria became melanized than at lower temperatures, but the frequency of formation of penetration pegs was highest at 25°C. Conidia of C. gloeosporioides germinated on cellophane membranes at relative humidities as low as 95%, but the percentage of conidia germinating and forming appressoria increased as the RH approached 100%. Approximately 18% of conidia of C. gloeosporioides I-2 held at 62 and 86% RH for 4 weeks retained viability, and some were capable of forming appressoria when placed at 100% RH. These results have implications for epidemiological models for disease control.     

Assessment of the baseline sensitivity and resistance risk of <Emphasis Type="Italic">Colletotrichum acutatum</Emphasis> to fludioxonil

Anthracnose, which is caused by Colletotrichum acutatum, is a destructive disease of pepper. A preliminary study demonstrated that fludioxonil (a phenylpyrrole fungicide) has good activity against C. acutatum and thus has potential to be used as an alternative fungicide for the management of pepper anthracnose. However, there is no information regarding the baseline sensitivity and resistance risk of C. acutatum to fludioxonil. Thus, the sensitivities of 205 isolates of C. acutatum to fludioxonil were determined. The results showed that the frequency distributions of the EC₅₀ values were unimodal, and the mean EC₅₀ values for the inhibition of mycelial growth and spore germination were 0.031 μg/mL and 0.035 μg/mL, respectively. Three stable mutants with high resistance to fludioxonil were obtained in the laboratory. Two parameters, namely in vitro sporulation and the in vitro and in vivo germination of spores, showed significant difference (P < 0.01) when the mutants were compared to the sensitive isolates. Moreover, the mutants were more sensitive to osmotic stress compared to the parents. No significant differences (P ≥ 0.05) were detected in colony diameter, mycelia weight, pathogenicity or sporulation in vivo between the fludioxonil-resistant mutants and their corresponding parents. Cross-resistance occurred between fludioxonil, iprodione and procymidone. Overall, resistance risk of C. acutatum to fludioxonil was low to medium, and thus resistance management should be considered.     

Comparative colonisation by virulent versus avirulent <Emphasis Type="Italic">Pyricularia oryzae</Emphasis> on wild <Emphasis Type="Italic">Oryza australiensis</Emphasis>

Pyricularia oryzae (rice blast) conidial development at pre-penetration stage determines success or otherwise of infection inside the rice host plants. Studies on conidial germination and growth on the leaf surface in commercial rice (Oryza sativa) report differently, dependent upon host type and level of blast resistance. Although wild rice (O. australiensis) is known to be an alternative host of blast, the interaction between P. oryzae conidia and wild O. australiensis on its leaf surface has not been previously studied. We found significant (P?<?0.001) differences in conidial development between two blast isolates with different virulence in terms of conidial germination, germ tube growth and appressoria formation on both wild and cultivated rice. Conidial germination at 6 h post-inoculation (hpi) for the virulent isolate was significantly (P?<?0.001) delayed. Germ tubes of the avirulent isolate conidia grew significantly (P?<?0.001) faster and with significantly (P?<?0.001) longer germ tubes than from virulent conidia. Appressoria development for the virulent isolate was significantly (P?<?0.001) faster at its later growth stages of 12 and 18 hpi when approximately 100% of germ tubes formed appressoria. In contrast, formation rate of appressoria for the avirulent isolate was significantly (P?<?0.001) slower and only reached 76% of germ tubes forming appressoria. Appressoria formation on O. australiensis was significantly (P?<?0.001) greater than the formation on O. sativa for both virulent and avirulent P. oryzae at 12 hpi, a clear indication that host type influences the extent of appressoria formation.     

Stomatal penetration of cowpea (Vigna unguiculata) leaves by a Colletotrichum species causing latent anthracnose

The infection process of a Colletotrichum species causing latent infection and anthracnose in cowpea ( Vigna unguiculata ) was studied in seedlings by light and confocal microscopy. Leaf surfaces were extensively colonized by an anastomosing network of germ-tubes and conidia. This epiphytic mycelium produced abundant secondary conidia on short conidiophores. Although melanized appressoria were developed, the host surface was not penetrated directly. The fungus only gained ingress into leaves through stomatal openings, by means of undifferentiated germ-tubes, and slowly colonized the mesophyll by intercellular hyphae, without initially producing visible symptoms. Anthracnose lesions with multisetate acervuli appeared on senescent leaves after a prolonged symptomless period of host colonization lasting > 2 weeks. Analysis of the nucleotide sequences of the amplified D2 and ITS-2 regions of rDNA revealed close similarities (95–96%) between this cowpea pathogen and isolates of C . gloeosporioides from Aeschynomene virginica, Stylosanthes scabra and Mangifera indica. These results, in addition to other morphological and growth attributes, identify this endophytic anthracnose pathogen of cowpea as a Colletotrichum species distinct from C. capsici and C. destructivum .     

Two subpopulations of Colletotrichum acutatum are responsible for anthracnose in strawberry and leatherleaf fern in Costa Rica

Strawberry, Fragaria × ananassa, and leatherleaf fern, Rumohra adiantiformis, are two important crops in Costa Rica. One of the most severe diseases affecting these crops is anthracnose, caused by members of the fungal genus, Colletotrichum (teleomorph; Glomerella). Eighty single-spore isolates from strawberry and leatherleaf fern were identified as Colletotrichum acutatum by species-specific PCR, and were further characterised by Universally Primed PCR (UP-PCR) fingerprinting analysis, and sequence analysis of the ribosomal internal transcribed spacer (ITS) region. Morphological differences, genotypic variation revealed by UP-PCR fingerprinting analysis, and a single sequence polymorphism within the ITS2 region were found between the isolates from strawberry and leatherleaf fern, respectively. The UPGMA cluster analysis of the fingerprints clearly separated the isolates derived from strawberry and leatherleaf fern into two different clusters. Pathogenicity assays on detached strawberry fruits confirmed the apparent difference between the two groups of isolates. It is therefore suggested that the pathogens responsible for strawberry anthracnose fruit rot and leatherleaf fern anthracnose in Costa Rica, belong to two distinct subpopulations of C. acutatum.