Anthracnose of three-leaf akebia (<Emphasis Type="Italic">Akebia trifoliata</Emphasis> Koidzumi) caused by <Emphasis Type="Italic">Colletotrichum acutatum</Emphasis>

In October 2001, anthracnose caused by Colletotrichum acutatum Simmonds ex Simmonds was found on three-leaf akebia (Akebia trifoliata) in Saitama, Japan. This is the first report of anthracnose on three-leaf akebia caused by C. acutatum.     

Phylogenetic relationships and pathogenicity of Colletotrichum acutatum isolates from grape in subtropical Australia

The identity of Colletotrichum acutatum as the causal pathogen of grape ripe-rot, which causes yield loss and a bitter taint that lowers wine quality in Australian subtropical wine-grape regions, was confirmed using species-specific primers. Cultural, morphological and molecular methods (RAPD-PCR and sequencing of parts of the 5·8S-ITS regions and the β-tubulin-2 gene) were used to determine the phylogenetic relationships of Australian C. acutatum isolates from wine grapes and other horticultural crops. A combination of RAPD-PCR and β-tubulin-2 gene data showed that all wine-grape ripe-rot isolates from northern regions of New South Wales (NSW) and Queensland belong to a proposed new C. acutatum group (A9), together with isolates from Australian strawberry, mango, blueberry and olive. The 5·8S-ITS sequences for these grape pathogens were identical to published sequences for an isolate from Cyclamen (the Netherlands) and differed by 1 bp from isolates from Capsicum (Taiwan) and orange (Costa Rica). The grape ripe-rot isolates from the Shoalhaven Valley (southern NSW) were clustered within two other C. acutatum groups: A2 and A5. In vitro infection studies showed that Australian C. acutatum isolates from almond, blueberry, chilli, grape, mango, olive, strawberry and tomato were able to infect grape and could also infect blueberry and strawberry, indicating a lack of host specificity. This lack of host specificity, the genetic similarity with non-grape isolates, and the fact that many of the non-grape hosts were isolated from wine-growing regions, suggest the potential for cross-infection between grape and other horticultural crops.     

Occurrence and development of Colletotrichum acutatum on symptomless blueberry bushes

The anthracnose fungus Colletotrichum acutatum was detected in symptomless blueberry bushes ( Vaccinium spp.) in a Japanese blueberry field. Naturally diseased bushes and their apparently healthy neighbours were selected, and C. acutatum was isolated from the symptomless tissues of each bush from February 2000 to January 2002. Analysis of the diseased bushes during the dormant period revealed that the fungus was able to survive on symptomless tissues, such as shoot bark and bud scales. Furthermore, C. acutatum was consistently isolated from symptomless leaves and shoots of several surrounding symptomless bushes. Arbitrarily primed PCR (ap-PCR) analyses of the fungal isolates obtained from the diseased and symptomless bushes revealed that most C. acutatum isolates were genotypically identical, regardless of their origins. Inoculation tests using leaves of various blueberry cultivars suggested that the presence or absence of symptoms on each bush can not always be explained by differences in cultivar susceptibility, and other factors may be associated with the appearance of symptoms.     

Sclerotinia rot of blueberry caused by <Emphasis Type="Italic">Sclerotinia sclerotiorum</Emphasis>

In 2002, rotted flower clusters and blighted shoot tips and leaves were observed on highbush blueberry (Vaccinium corymbosum L.) and rabbiteye blueberry (V. ashei Reade) plants in Chiba, Japan. The causal fungus isolated from the diseased plants was morphologically identified as Sclerotinia sclerotiorum (Libert) de Bary. The fungus reproduced natural symptoms after inoculation, then reisolated from the symptomatic parts. This is the first report of blueberry sclerotinia rot caused by S. sclerotiorum. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under accession numbers AB269903#(020501) and AB233346 (020505).     

First report of blueberry red ringspot disease caused by <Emphasis Type="Italic">Blueberry</Emphasis><Emphasis Type="Italic">red</Emphasis><Emphasis Type="Italic">ringspot</Emphasis><Emphasis Type="Italic">virus</Emphasis> in Japan

Virus-like symptoms—red ringspots on stems and leaves, circular blotches or pale spots on fruit—were found on commercial highbush blueberry (Vaccinium corymbosum) cultivars Blueray, Weymouth, Duke and Sierra in Japan. In PCR testing, single DNA fragments were amplified from total nucleic acid samples of the diseased blueberry bushes using primers specific to Blueberry red ringspot virus (BRRV). Sequencing analysis of the amplified products revealed 95.7–97.7% nucleotide sequence identity with the BRRV genome. This paper is the first report of blueberry red ringspot disease caused by BRRV in Japan. The nucleotide sequence data reported in this paper are available in the GenBank/EMBL/DDBJ database as accessions AB469884 to AB469893 for BRRV isolates from Japan.     

Inoculum prevalence, host infection and biological control of Colletotrichum acutatum: causal agent of blueberry anthracnose in British Columbia

To identify the causal organism of anthracnose (ripe-rot), which reduces yield and postharvest quality of blueberries grown in British Columbia, Canada, 80 isolates were recovered from diseased fruits collected from commercial blueberry fields during 2002–04 and identified as Colletotrichum acutatum using colony morphology, growth rate and species-specific PCR primers. In vitro incubation of replicated sets of inoculated detached berries at various temperatures produced infection at temperatures of 7–30°C, with an optimum at 20°C. Colletotrichum acutatum could not survive on the soil surface as mummified berries but the pathogen was detected mostly within flower buds and less so in blueberry twigs and fruit trusses. Infection of developing flower buds in May–June of the preceding growing season gave the highest inoculum recovery in the following year. Two commercial fungal biocontrol agents, Prestop ( Gliocladium catenulatum ) and PlantShield ( Trichoderma harzianum ), each reduced anthracnose development in 2003 and 2004 by up to 45% when sprayed three times onto plants between flowering and fruit ripening.     

Responses of avocado to laurel wilt,caused by Raffaelea lauricola

Laurel wilt, caused by Raffaelea lauricola, threatens native and non‐native species in the Lauraceae in the south‐eastern USA. Avocado, Persea americana, is the most important agricultural suscept of laurel wilt. Grafted plants (clonal scions on seedling rootstocks) of 24 cultivars were screened against the disease in the field from 2008 to 2010. Disease was induced with either mycelial plugs or conidial suspensions of R. lauricola. There were significant differences in the severity of disease that developed on different cultivars, and West Indian cultivars were most susceptible (P < 0·05). Simmonds, a West Indian cultivar that comprises 35% of the commercial production in Florida, was consistently susceptible and was used as a standard genotype in different studies. Disease severity increased significantly on cv. Simmonds as plant size (stem diameter) increased (P < 0·0042). In greenhouse studies, internal (sapwood) and external disease severities on cv. Simmonds were correlated (P < 0·0001), and a threshold was evident, in that external symptoms developed only after moderately severe symptoms had developed internally. Latent infection was uncommon; R. lauricola was usually isolated on a semiselective medium or detected via qPCR only from discoloured xylem of inoculated cv. Simmonds. As basipetal movement of the pathogen was common, its movement among trees via root grafts is probable. Greater understanding is needed of the movement of R. lauricola in naturally and artificially infected trees, and whether sufficient tolerance exists in avocado to assist in the management of this important new disease.     

Determination of Festuca filiformis seedbank characteristics,seedling emergence and herbicide susceptibility to aid management in lowbush blueberry (Vaccinium angustifolium)

Festuca filiformis is a common perennial grass in lowbush blueberry fields, but little is known about the general biology, seedbank characteristics, seedling recruitment or susceptibility of seedlings to currently registered herbicides. The objectives of this research were to determine (i) the presence of F. filiformis seedbanks in lowbush blueberry fields, (ii) whether F. filiformis seedbanks accumulate near the soil surface in lowbush blueberry fields, (iii) the dormancy status of fresh F. filiformis seeds, (iv) the temporal patterns of seedling recruitment in established F. filiformis populations, (v) whether F. filiformis has a vernalisation requirement for flowering in lowbush blueberry and (vi) susceptibility of F. filiformis seedlings to various herbicides currently registered in lowbush blueberry. Festuca filiformis formed a seedbank in lowbush blueberry fields, with an average of 1660 ± 272–5680 ± 1409 seedlings m^?2 emerging from soil cores collected from two infested fields. Most seeds were located at the soil surface, providing opportunities for seedbank management through predation or burning. Fresh seeds lacked dormancy and readily germinated, although germination was reduced by dark conditions. New seedlings emerged in spring and autumn and required vernalisation to flower. Seedlings were susceptible to several currently registered herbicides in lowbush blueberry, although mortality rates were highest in plants treated with glufosinate, flumioxazin, glufosinate + flumioxazin and terbacil. Growers should avoid movement of seeds on machinery, and additional research should be conducted to determine the effects of registered herbicides on F. filiformis seedling recruitment under field conditions in lowbush blueberry.     

Novel infection strategies of Colletotrichum acutatum on ripe blueberry fruit

The infection and colonization process of Colletotrichum acutatum on ripe blueberry fruit from two cultivars with different susceptibility to anthracnose were examined using light and confocal laser scanning microscopy. Ripe fruit from susceptible cv. Jersey and resistant cv. Elliott were drop-inoculated with a conidial suspension of C. acutatum, and epidermal peels were evaluated at selected times after inoculation and incubation. Results from pre-penetration studies demonstrated that there were significant differences in the rate of formation of melanized appressoria between the two cultivars, with the rate of formation being faster in the susceptible one. In both cultivars, penetration by the pathogen occurred via appressoria 48 h post-inoculation (hpi). However, in the susceptible cv. Jersey, C. acutatum then adopted an intracellular hemibiotrophic-like infection strategy, whereas in the resistant cv. Elliott subcuticular intramural-like infection occurred. In cv. Jersey by 108 hpi, intracellular growth of the pathogen led to the formation of numerous acervuli, with orange conidial masses. By 120 hpi, the conidial masses had coalesced covering the entire inoculated area. In cv. Elliott, acervuli were not seen until 144 hpi and contained few conidia. These results demonstrate for the first time the ability of C. acutatum to adopt a different infection and colonization strategy depending on the susceptibility of the host tissue being colonized.     

Quantitative detection of <Emphasis Type="Italic">Colletotrichum godetiae</Emphasis> and <Emphasis Type="Italic">C. acutatum sensu stricto</Emphasis> in the phyllosphere and carposphere of olive during four phenological phases

A duplex qPCR detection method was developed to detect and quantify Colletotrichum godetiae and C. acutatum sensu stricto (s.s.) in olive tissues. The method proved highly specific and sensitive with a detection limit of 10 pg for each pathogen. The analysis of green and senescent leaves, fertilized fruitlets with floral residues, green fruit and symptomatic and asymptomatic fruit collected in May, June, October and December revealed a high incidence of both C. godetiae and C. acutatum s.s. in Calabria, southern Italy. In comparison with previous reports, these results highlighted an ongoing population shift from C. godetiae to C. acutatum s.s. Interestingly, C. godetiae was slightly more abundant in terms of number of infected samples, yet the quantity of C. acutatum in infected samples was always higher, suggesting greater aggressiveness and/or sporulation ability of the latter pathogen. The populations of both C. godetiae and C. acutatum s.s. increased sharply in December even though both pathogens were detected widely in asymptomatic samples in May, June and October, confirming an important role of latent infections in the disease cycle. A large quantity of both C. godetiae (1.7 × 10⁸ cells/mg of tissue) and C. acutatum s.s. (7.5 × 10⁸ cells/mg of tissue) was estimated in symptomatic fruit, presenting an enormous inoculum potential for secondary infections. Two other important observations were a high incidence and quantity of both pathogens in senescent leaves and in fertilized fruitlets with floral residues as compared to green leaves.     

Molluscicidal and antifungal activity of Erigeron speciosus steam distillate

The steam-distilled fraction of the aerial parts of Erigeron speciosus (Lindl) DC was tested for activity against strawberry plant pathogenic fungi Botrytis cinerea Pers ex Fr, Colletotrichum acutatum Simmonds, C fragariae Brooks, C gloeosporioides (Penz) Penz & Sacc, and the intermediate host snail Planobdella trivolvis that harbors the trematode, Bolbophorus confusus, that infests and causes severe infections in pond-raised catfish in the Mississippi Delta region of the USA. Bioautography on silica TLC plates demonstrated antifungal activity in the steam distillate. Preliminary bioassays of the steam distillate indicated the presence of phytochemicals toxic to P trivolvis. The bioactive compounds methyl 2Z, 8Z-deca-2,8-diene-4,6-diynoate and its 2E, 8E isomer were isolated by bioassay-guided fractionation and chromatographic techniques and identified by 1H NMR spectroscopy.     

Dehydro-alpha-lapachone isolated from Catalpa ovata stems: activity against plant pathogenic fungi

The methanol extract of stems of Catalpa ovata G Don exhibits potent in vivo antifungal activity against Magnaporthe grisea (Hebert) Barr (rice blast) on rice plants, Botrytis cinerea Pers ex Fr (tomato grey mould) and Phytophthora infestans (Mont) de Bary (tomato late blight) on tomato plants, Puccinia recondita Rob ex Desm (wheat leaf rust) on wheat plants and Blumeria graminis (DC) Speer f. sp. hordei Marchal (barley powdery mildew) on barley plants. An antifungal substance was isolated and identified as dehydro-alpha-lapachone from mass and nuclear magnetic resonance spectral data. It completely inhibited the mycelial growth of B. cinerea, Colletotrichum acutatum Simmonds, Colletotrichum gloeosporioides Simmonds, M. grisea and Pythium ultimum Trow over a range of 0.4-33.3 mg litre(-1). It also controlled the development of rice blast, tomato late blight, wheat leaf rust, barley powdery mildew and red pepper anthracnose (Colletotrichum coccodes (Wallr) S Hughes). The chemical was particularly effective in suppressing red pepper anthracnose by 95% at a concentration of 125 mg litre(-1).     

Glomerella cingulata on olive in India: morphological and pathological notes1

Two strains of a species of Colletotrichum were obtained from infected olives in India. Their morphological characteristics and pathogenicity to various fruits were compared with those of isolates of C. acutatum, C. gloeosporioides, and C. musae. Despite their morphological variability, the two isolates were referred to C. gloeosporioides, the anamorph of Glomerella cingulata. For one isolate, the presence of ascocarps was recorded in culture.     

Demography of Rumex acetosella in lowbush blueberry (Vaccinium angustifolium)

The demography of Rumex acetosella ramets and seedlings was monitored within and between blueberry clones over the two‐year lowbush blueberry production cycle in Nova Scotia, Canada. Overwintering ramets constituted the majority (>70%) of the flowering ramet population. A small proportion of ramets emerging in May and June flowered, but no ramets emerging between July and November flowered in the year of emergence. Emergence of new ramets was season‐long and ramet populations were regulated by a cycle of birth and death in each production year. Ramet populations in blueberry patches had higher growth rates and lower mortality than ramet populations in bare soil patches in the non‐bearing year, resulting in large net gains to ramet populations in blueberry patches during this production year. Ramet population growth rates and mortality were similar in blueberry and bare soil patches in the bearing year and net ramet populations declined during this production year. Survival rates of overwintering and new ramets varied, but ramets from both the overwintering and monthly cohorts contributed to a distinct ramet age structure at the end of each season. Seedling survival ranged from 6 ± 6 to 51 ± 12% across sites, and no seedlings flowered in the year of emergence. Transition probabilities of ramets and seedlings were used to develop a life‐cycle model of R. acetosella for the 2‐year lowbush blueberry production cycle. This model has utility in developing new management strategies for R. acetosella in lowbush blueberry.     

蓝莓枝枯病拮抗细菌HMQAU140045的鉴定和抑真菌活性

以蓝莓毛色二胞枝枯病菌假可可毛色二胞Lasiodiplodia pseudotheobromae为靶标菌,通过稀释平板法从山东青岛的蓝莓种植园根际土壤中分离出20株细菌,采用对峙平板法和菌丝生长速率法筛选得到一株对靶标菌具有明显抑制效果的拮抗菌株HMQAU140045,采用凹玻片法和菌丝生长速率法测定该菌株发酵滤液对靶标菌孢子萌发和菌丝生长的影响。结果表明,稀释50倍的发酵滤液的抑制率可达100%。离体枝条试验结果表明,发酵液不同组分对蓝莓毛色二胞枝枯病的防治效果均可达90%以上。菌丝生长速率法测定结果表明,该菌株抑真菌谱广,对包括4种蓝莓枝枯病菌在内的15种植物病原真菌具有良好的拮抗作用。通过形态观察、生理生化特性以及gyrB的序列分析,确定该菌株为解淀粉芽孢杆菌Bacillus amyloliquefaciens。试验结果表明,解淀粉芽孢杆菌菌株HMQAU140045具有较好的抑真菌活性。     

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。     

Environmental and host requirements for field infection of blueberry fruits by Colletotrichum acutatum in British Columbia

Higher recovery of Colletotrichum acutatum , the causal agent of anthracnose (ripe-rot), from blueberry tissues during the growing seasons of 2002 and 2003 was found at bloom and ripe berry than at other stages of plant development. The effects of leaf-wetness duration and ambient temperature on fruit infection frequency were determined during the growing seasons of 2001–03. Potted 2-year-old blueberry plants were exposed for 1-week periods to prevailing environmental conditions and natural inoculum in a commercial field, and grown to harvest, when fruit infection was assessed. Three peaks of infection were observed: early during bloom, mid-season during the mature green berry stage, and later in the season when berries had ripened. Weather data collected simultaneously indicated that a minimum of 10 h of leaf wetness at 11°C was sufficient for fruit infection. These conditions preceded each peak of infection. To determine whether peaks of infection in the field were also caused by changes in host susceptibility or available inoculum, groups of potted blueberry plants were artificially inoculated at weekly intervals during the growing season of 2004, exposed to prevailing environmental conditions, and fruit infection assessed at harvest. Flowers and developing fruits were found to be susceptible throughout the season, indicating that specific peaks of infection were associated with environmental conditions and availability of inoculum.     

The effect of environmental factors on infection of blueberry fruit by Colletotrichum acutatum

Anthracnose fruit rot of blueberries caused by Colletotrichum acutatum is a serious problem in humid blueberry‐growing regions of North America. In order to develop a disease prediction model, environmental factors that affect mycelial growth, conidial germination, appressorium formation and fruit infection by C. acutatum were investigated. Variables included temperature, wetness duration, wetness interruption and relative humidity. The optimal temperature for mycelial growth was 26°C, and little or no growth was observed at 5 and 35°C. The development of melanized appressoria was studied on Parafilm‐covered glass slides and infection was evaluated in immature and mature blueberry fruits. In all three assays, the optimal temperature for infection was identified as 25°C, and infections increased up to a wetness duration of 48 h. Three‐dimensional Gaussian equations were used to assess the effect of temperature and wetness duration on the development of melanized appressoria (R² = 0·89) on Parafilm‐covered glass slides and on infection incidence in immature (R² = 0·86) and mature (R² = 0·90) blueberry fruits. Interrupted wetness periods of different durations were investigated and models were fitted to the response of melanized appressoria (R² = 0·95) and infection incidence in immature (R² = 0·90) and mature (R² = 0·78) blueberry fruits. Additionally, the development of melanized appressoria and fruit infection incidence were modelled in relation to relative humidity (R² = 0·99 and 0·97, respectively). Three comprehensive equations were then developed that incorporate the aforementioned variables. The results lay the groundwork for a disease prediction model for anthracnose fruit rot in blueberries.     

Characterization and epidemiology of Colletotrichum acutatum sensu lato (C. chrysanthemi) causing Carthamus tinctorius anthracnose

In 2012, Colletotrichum isolates were collected from field‐grown safflower (Carthamus tinctorius) crops in central Italy from plants exhibiting typical anthracnose symptoms. Colletotrichum isolates were also collected from seed surfaces and from within seeds. The genetic variability of these isolates was assessed by a multilocus sequencing approach and compared with those from Colletotrichum chrysanthemi and Colletotrichum carthami isolates from different geographic areas and other Colletotrichum acutatum sensu lato‐related isolates. Phylogenetic analysis revealed that all of the strains isolated from C. tinctorius belonged to the species described as C. chrysanthemi, whereas all of the strains belonging to C. carthami had been isolated from Calendula officinalis. Phenotypic characterization of isolates was performed by assessing growth rates at different temperatures, morphology of colonies on potato dextrose agar (PDA) and the size of conidia. All C. chrysanthemi isolates from safflower had similar growth rates at different temperatures, comparable colony morphologies when grown on PDA and conidial sizes consistent with previously described C. chrysanthemi isolates. Pathogenicity tests were performed by artificially inoculating both seeds and plants and confirmed the seedborne nature of this pathogen. When inoculated on plants, C. chrysanthemi caused the typical symptoms of anthracnose on leaves. This is the first record of this pathogen on C. tinctorius in Italy, and it presents an updated characterization of Colletotrichum isolates pathogenic to safflowers in Europe.     

Significant in vitro antagonism of the laurel wilt pathogen by endophytic fungi from the xylem of avocado does not predict their ability to control the disease

The ascomycete Raffaelea lauricola causes laurel wilt, a lethal vascular disease of avocado, Persea americana, and other members of the Lauraceae plant family. Few effective control measures for laurel wilt exist and new measures are needed. In this study, biological control of the disease with endophytic fungi from avocado was examined. Thirty‐two endophytes (24 operational taxonomic units or OTUs) isolated from the xylem of healthy trees (the infection court of R. lauricola) were evaluated against R. lauricola with in vitro dual‐culture assays. Nine OTUs that showed strong in vitro antagonism of the pathogen were tested in planta against laurel wilt. In three greenhouse experiments, grafted avocado plants of Simmonds or Russell cultivars, which are both susceptible to laurel wilt, were inoculated with endophytes and, after 10–16 days, inoculated with the same isolate of R. lauricola that was used in the in vitro assays. Within 14 days of inoculation with R. lauricola, laurel wilt developed in plants that were not treated with endophytes (positive controls) but also developed in endophyte‐treated plants to the extent observed in the positive controls (P = 0.05). The pathogen colonized plants rapidly and systemically, but endophytes generally did not colonize xylem more than 2 cm above the point at which plants were inoculated. Although the tested endophytes strongly antagonized the pathogen in vitro, this did not translate to an ability to reduce development of laurel wilt. The management of laurel wilt and other plant diseases with fungal endophytes is discussed.