scholarly journals Identification of Fusarium commune, the Causal Agent of Postharvest Zinnia Meltdown Disease in Tennessee

2021 ◽  
pp. 1-8
Author(s):  
Ravi Bika ◽  
Fulya Baysal-Gurel
Keyword(s):  
Foliar Spray ◽  
Morphological Characterization ◽  
Causal Agent ◽  
Pathogenicity Test ◽  
Conidial Suspension ◽  
Cut Flower ◽  
Cut Flowers ◽  
Flower Bud ◽  
Golden Yellow ◽  
Management Recommendations

The cut flower growers of the eastern and southern United States are threatened with postharvest meltdown of zinnia (Zinnia elegans), which reduces yield and income as well as limiting opportunities for production expansion. Disease symptoms such as bending of the stem just below the flower were visually apparent on zinnia cut flowers. The objective of this study was to identify the causal agent related to zinnia meltdown. A total of 20 symptomatic zinnia cut flower stems were collected from Tennessee. Several Fusarium-like colonies with micro and macroconidia were isolated from the base and bend area of stems on potato dextrose agar (PDA) and Fusarium-selective media. Morphological characterization, polymerase chain reaction, and sequencing of three representative isolates, FBG2020_198, FBG2020_199, and FBG2020_201, were conducted to confirm pathogen identification. The sequence identity of the isolates was >99% identical to Fusarium commune, and a combined phylogenetic tree grouped the isolates with the clade of F. commune from different host and geographical locations. To accomplish Koch’s postulates, a pathogenicity test was performed on ‘Benary’s Giant Golden Yellow’, ‘Benary’s Giant Lime’, and ‘Benary’s Giant Pink’ zinnia plants at vegetative (2 weeks after transplantation) or flower bud stage (1 month after transplantation) by drench, stem injection, and foliar spray of conidial suspension (1 × 105 conidia/mL). Similar symptoms of meltdown (floral axis bending just below the flower) were observed on inoculated zinnia cultivars 2 days after harvesting. Fusarium commune was re-isolated from the infected flower stems of all three cultivars but not from the noninoculated zinnia flower stems. Zinnia stem colonization by F. commune was statistically similar in all three tested cultivars regardless of plant growth stage and method of inoculation. This study confirms F. commune as being the causal agent of postharvest zinnia flower meltdown issue in Tennessee. In the future, possible sources of pathogen will be screened, and disease management recommendations will be developed.

scholarly journals First Report of Colletotrichum boninense Causing Anthracnose on Pepper in China

Plant Disease ◽  
2013 ◽  
Vol 97 (1) ◽  
pp. 138-138 ◽  
Author(s):  
Y. Z. Diao ◽  
J. R. Fan ◽  
Z. W. Wang ◽  
X. L. Liu
Keyword(s):  
Internal Transcribed Spacer ◽  
Severe Disease ◽  
Causal Agent ◽  
Its Sequences ◽  
Universal Primers ◽  
Pathogenicity Test ◽  
Conidial Suspension ◽  
Single Spore ◽  
First Report ◽  
Species Specific

Anthracnose, caused by Colletotrichum spp., is a severe disease and results in large losses in pepper (Capsicum frutescens) production in China (4). Colletotrichum boninense is one of the Colletotrichum species in pepper in China. In August 2011, anthracnose symptoms (circular, sunken lesions with orange to black spore masses) were observed on pepper fruits in De-Yang, Sichuan Province, China. Three single-spore isolates (SC-6-1, SC-6-2, SC-6-3) were obtained from the infected fruits. A 5-mm diameter plug was transferred to potato dextrose agar (PDA); the isolates formed colonies with white margins and circular, dull orange centers. The conidia were cylindrical, obtuse at both ends, and 10.5 to 12.6 × 4.1 to 5.0 μm. The colonies grew rapidly at 25 to 28°C, and the average colony diameter was 51 to 52 mm after 5 days on PDA at 25°C. Based upon these characters, the causal agent was identified as C. boninense. To confirm the identity of the isolates, the internal transcribed spacer (ITS) regions were amplified with the ITS1/ITS4 universal primers (1). The internal transcribed spacer (ITS) sequences (Accession No. JQ926743) of the causal fungus shared 99 to 100% homology with ITS sequences of C. boninense in GenBank (Accession Nos. FN566865 and EU822801). The identity of the causal agent as C. boninense was also confirmed by species-specific primers (Col1/ITS4) (2). In a pathogenicity test, five detached ripe pepper fruits were inoculated with 1 μl of a conidial suspension (106 conidia/mL) or five fruits with 1 μl of sterile water were kept as control. After 7 days in a moist chamber at 25°C, typical anthracnose symptoms had developed on the five inoculated fruits but not on control fruits. C. boninense was reisolated from the lesions, and which was confirmed by morphology and molecular methods as before. There have reports of C. boninense infecting many species of plants, including pepper (3). To our knowledge, this is the first report of C. boninense causing anthracnose on pepper in China. References: (1) A. K. Lucia et al. Phytopathology 93:581, 2002. (2) S. A. Pileggi et al. Can. J. Microbiol. 55:1081, 2009. (3) H. J. Tozze et al. Plant Dis. 93:106, 2009. (4) M. L. Zhang. J. Anhui Agri. Sci. 2:21, 2000.

scholarly journals First Report of Fusarium Wilt in Blueberry (Vaccinium corymbosum) Caused by Fusarium oxysporum in China

Plant Disease ◽  
2014 ◽  
Vol 98 (8) ◽  
pp. 1158-1158 ◽  
Author(s):  
Y. H. Liu ◽  
T. Lin ◽  
C. S. Ye ◽  
C. Q. Zhang
Keyword(s):  
Fusarium Oxysporum ◽  
Infected Plant ◽  
Morphological Characteristics ◽  
Vaccinium Corymbosum ◽  
Morphological Characterization ◽  
Internal Transcribed Spacers ◽  
Fluorescent Light ◽  
Pathogenicity Test ◽  
Conidial Suspension ◽  
First Report

Blueberry (Vaccinium corymbosum) production is developing quickly in China with about 20,000 ha presently cultivated. In 2010 in Lin'an, Zhejiang Province, plants developed an apparently new disease of blueberry (cv. Duke) with symptoms consisting of wilting of foliage, stunting of plants, and reduced fruit yields. Internal vascular and cortical tissues of plant crowns showed a brown to orange discoloration. Approximately 3% of the plants in the commercial plantings were affected and eventually died after 50 to 60 days. Infected plant samples (stems and roots) collected from different fields were surface sterilized with 1.5% sodium hypochlorite for 2 min, rinsed in water, plated on 2% potato dextrose agar (PDA), and incubated at 25°C in the dark for 1 week. Single conidium cultures were consistently isolated and cultured on acidified PDA (APDA) for morphological characterization (1,2). Colonies were light with purple mycelia, and beige or orange reverse colony colors developed after 7 days incubation at 25°C. Colonies producing abundant microconidia and macroconidia. Microconidia were hyaline and oval-ellipsoid to cylindrical (3.9 to 9.6 × 1.1 to 3.4 μm). Macroconidia were 3 to 5 septate and fusoid-subulate with a pedicellate base (28.6 to 37.5 × 3.3 to 4.2 μm). Morphology and development of macroconidia and microconida were consistent with a description of Fusarium oxysporum Schltdl (1,2). The ribosomal internal transcribed spacers ITS1 and ITS2 of eight isolates were amplified using primers ITS1/ITS4 on DNA extracted from mycelium and nucleotide sequences showed 100% similarity to that of F. oxysporum. To confirm pathogenicity, 20 blueberry plants (cv. Duke) were inoculated by dipping the roots into a conidial suspension (107 conidia per ml) for 30 min. The inoculated plants were transplanted into pots containing sterilized peat and maintained at 25°C and 100% relative humidity in a growth chamber with a daily 12-h photoperiod of fluorescent light. The pathogenicity test was conducted twice. Within 40 days, all inoculated plants developed wilt symptoms similar to that observed in the field. No symptoms were observed on plants dipped into distilled water. The fungus was successfully re-isolated from crowns and roots cultured on APDA, exhibiting morphological characteristics identical to F. oxysporum (1,2), confirming Koch's postulates. To our knowledge, this is the first report of blueberry wilt caused by Fusarium. References: (1) P. M. Kirk et al. The Dictionary of the Fungi, 10th edition, page 159. CABI Bioscience, Wallingford, UK, 2008. (2) W. C. Snyder and H. N. Hansen. Am. J. Bot. 27:64, 1940.

scholarly journals First Report of Anthracnose Caused by Colletotrichum gloeosporioides on Soybean (Glycine max) in Malaysia

Plant Disease ◽  
2013 ◽  
Vol 97 (6) ◽  
pp. 841-841 ◽  
Author(s):  
F. Mahmodi ◽  
J. B. Kadir ◽  
M. Y. Wong ◽  
A. Nasehi ◽  
A. Puteh ◽  
...  
Keyword(s):  
Glycine Max ◽  
Morphological Characterization ◽  
Negative Control ◽  
Culture Collection ◽  
Conidial Suspension ◽  
First Report ◽  
Representative Isolate ◽  
Detached Leaves ◽  
Soybean Plants ◽  
Management Recommendations

Soybean (Glycine max L.) is one of the most economically important crops in the world, and anthracnose is known to infect soybean in most countries. Colletotrichum truncatum is the common pathogen causing anthracnose of soybean. However, at least five species of Colletotrichum have been reported on soybean worldwide (2). In July 2010, anthracnose symptoms were observed on soybean in the experimental fields of the agriculture station in Ladang Dua, University Putra Malaysia located in Selangor state of Malaysia. Symptoms were initially observed on a few plants randomly within one field, but after 4 weeks, the disease was found in two additional fields scattered across an area of 1 km2. Pinkish-brown lesions were observed on the pods, and the formation of dark lesions on the leaves and stems was sometimes followed by stem girdling, dieback, and distorted growth. At later stages, numerous epidermal acervuli developed in the lesions, and mucilaginous conidial masses appeared during periods of high relative humidity. Conidia produced in acervuli were straight, cylindric, hyaline, and aseptate, with both ends rounded. Conidia measured (mean ± SD) 14.2 ± 0.6 × 3.6 ± 0.7 μm, and the L/W ratio was 3.95 μm. Six isolates of the fungus were obtained and identified as C. gloeosporioides on the basis of morphological characterization (3). The isolates were deposited in the University Putra of Malaysia Culture Collection (UPMCC). PDA cultures were white at first and subsequently became grayish to pink to reddish-brown. Amplification and sequence analysis of coding and none-coding regions of the ITS-rDNA (GenBank JX669450), actin (JX827430), β-tubulin (JX827454), histone (JX827448), chitin synthase (JX827436), and glyceraldehyde-3-phosphate dehydrogenase (JX827442) obtained from the representative isolate, CGM50, aligned with deposited sequences from GenBank and revealed 99 to 100% sequence identity with C. gloeosporioides strains (JX258757, JX009790, GQ849434, HM575301, JQ005413, and JX00948 from GenBank). One representative isolate, CGM50, was used for pathogenicity testing. Four non-infected detached leaves and pods of 24-day-old G. max var. Palmetto were surface-sterilized and inoculated by placing 10 μl of a conidial suspension (106 conidia ml–1) using either the wound/drop or non-wound/drop method (4), with 10 μl distilled water as a negative control. Leaves and pods were incubated at 25°C, 98% RH. The experiment was repeated twice. Five days after inoculation, the development of typical field symptoms, including acervuli formation, occurred on the leaves and pods of inoculated plants, but not on the negative controls. A fungus with the same colony and conidial morphology as CGM50 was recovered from the lesions on the inoculated leaves and pods. Anthracnose caused by C. gloeosporioides on soybean plants has been reported previously in different countries, but not in Malaysia (3). Geographically, the climate of Malaysia is highly conducive to maintain and cause outbreaks of anthracnose all year round; thus, the development of management recommendations will be inevitable for anthracnose control. To our knowledge, this is the first report of C. gloeosporioides causing anthracnose on soybean in Malaysia. References: (1) U. Damm et al. Fungal Diversity 39:45, 2009. (2) S. L. Chen et al. J. Phytopathol. 154:654, 2006. (3) B. C. Sutton. The Genus Glomerella and its Anamorph Colletotrichum. CAB International, Wallingford, UK, 1992. (4) P. P. Than et al. Plant Pathol. 57:562, 2008. ERRATUM: A correction was made to this Disease Note on May 19, 2014. The author N. Soleimani was added.

scholarly journals Effects of Different Preservatives on Cut Flower of Luculia pinceana: A Novel Fragrant Ornamental Species

HortScience ◽  
2021 ◽  
pp. 1-8
Author(s):  
Lingfang Kong ◽  
Fan Li ◽  
Ronghui Du ◽  
Huaiting Geng ◽  
Shifeng Li ◽  
...  
Keyword(s):  
Water Balance ◽  
Flower Senescence ◽  
Distribution Area ◽  
Vase Life ◽  
Cut Flower ◽  
Cut Flowers ◽  
Water Control ◽  
Flower Bud ◽  
Mda Content ◽  
Bud Opening

Luculia pinceana is a potential cut flower because of its long-term blooming inflorescences and charming fragrance. However, its narrow distribution area and unexplored wild status severely restrict its applications, thus leading to the scientific research of cut L. pinceana flowers. To our knowledge, there is no available published information about the postharvest fresh-keeping of L. pinceana. During this study, the cut flowers of L. pinceana were tested using nine preservatives with different concentrations of sucrose and 8-hydroxyquinoline (8-HQ) to evaluate the fresh-keeping effects. Through the investigation and analysis of vase life, bud opening and abortion rate, water balance, malonaldehyde (MDA) content, and peroxidase (POD) activity, we selected and identified the best vase solution for cut L. pinceana flowers. The results suggested that the preservative of 1% sucrose and 100 mg/L 8-HQ could significantly prolong the vase life of cut L. pinceana flower up to 9 days compared with water control. This solution positively affects flower bud blooming, delays flower senescence, improves the water balance, inhibits the MDA accumulation, and increases POD activity. Therefore, this preservative is suitable for the fresh-keeping of cut L. pinceana flowers. Our study is the first to report the effects of preservatives on cut L. pinceana flower. The results showed that the low-sugar-containing (1% sugar) preservatives can effectively improve the ornamental quality of fresh flowers and demonstrated that the postharvest fresh-keeping of L. pinceana requires low sugar and is insensitive to microorganisms.

scholarly journals First Report of Peronospora parasitica on Wild Rocket (Diplotaxis tenuifolia) in Italy

Plant Disease ◽  
2004 ◽  
Vol 88 (12) ◽  
pp. 1381-1381 ◽  
Author(s):  
A. Garibaldi ◽  
A. Minuto ◽  
M. L. Gullino
Keyword(s):  
New York ◽  
Downy Mildew ◽  
Causal Agent ◽  
Pathogenicity Test ◽  
Conidial Suspension ◽  
Peronospora Parasitica ◽  
First Report ◽  
Downy Mildews ◽  
Air Temperatures ◽  
Wild Rocket

Several species of Diplotaxis (D. tenuifolia, D. erucoides, and D. muralis), known as wild rocket, are now widely cultivated in Italy. Wild rocket is used in Mediterranean cuisine as salad, a component of packaged salad products, and as a garnish for food. During the fall of 2002, a foliar disease of D. tenuifolia was observed in the field or greenhouse on several commercial farms in the Liguria Region of northern Italy. Symptoms appeared as small, irregular, dark brown-to-black speckling on the adaxial surfaces of leaves. The speckled areas sometimes expanded into larger spots. These symptoms were followed by leaf yellowing and the appearance of sporangiophores and sporangia on the lower and upper leaf surfaces. Sporangiophores were dichotomously branched with slender curved tips. Sporangia were ovoid, measuring 20 to 28 (average 22) μm long and 15 to 25 (average 19) μm wide. The causal agent of the disease was identified as Peronospora parasitica (3). Pathogenicity was established by inoculating 10 30-day-old plants of D. tenuifolia grown in pots in a peat/pumice/clay/composted bark mix (60:20:10:10), with a conidial suspension (102 conidia per ml). Ten noninoculated plants maintained under the same conditions served as the control. Plants were maintained in a glasshouse at air temperatures ranging between 10 and 26°C (average 16°C) and relative humidity at 85%. The pathogenicity test was done twice. Downy mildew symptoms developed within 12 days, and the same fungus was observed on inoculated plants. Noninoculated plants did not develop symptoms. To our knowledge, this is the first report of P. parasitica on D. tenuifolia in Italy. P. parasitca has been reported as the causal agent of downy mildew on D. muralis in England (1) and on cultivated rocket (Eruca sativa) in California (2). References: (1) J. Fraymouth. Trans. Br. Mycol. Soc. 39:79, 1956. (2) S. T. Koike. Plant Dis. 82:1063, 1998. (3) D. M. Spencer. The Downy Mildews. Academic Press. New York, 1981.

scholarly journals Fusarium Wilt of Gerbera in Soil and Soilless Crops in Italy

Plant Disease ◽  
2004 ◽  
Vol 88 (3) ◽  
pp. 311-311 ◽  
Author(s):  
A. Garibaldi ◽  
A. Minuto ◽  
D. Bertetti ◽  
M. L. Gullino
Keyword(s):  
The Netherlands ◽  
Vascular Tissue ◽  
Vascular System ◽  
Selective Medium ◽  
Northern Italy ◽  
Pathogenicity Test ◽  
Conidial Suspension ◽  
Cut Flowers ◽  
Planting Material ◽  
Red Bull

In 2002, gerbera (Gerbera jamesonii cv. Kaiki) plants that were grown for cut flowers in a soilless cultivation system (rockwool substrate) at Albenga (Savona) in northern Italy were observed exhibiting symptoms of a wilt disease. During the summer of 2002, in a commercial gerbera farm in the province of Imperia (northern Italy), a similar wilt was also observed on cvs. Red Bull, Anedin, and Gud finger that were grown in soil. In both cases, the planting material originated from the Netherlands. During 2003, wilted plants (cvs. Red Bull, Basic, and Cirill) were repeatedly observed in other commercial greenhouses located in the same area. Affected plants were stunted and developed yellowed leaves with initially brown and eventually black streaks in the vascular system. The vascular streaks in the yellow leaves were continuous with a brown discoloration in the vascular system of the crown and upper taproot. In some cases, the leaves of affected plants turned red. From these plants, Fusarium spp. were consistently and readily isolated from symptomatic vascular tissue onto a Fusarium-selective medium (2). Colonies were identified as F. oxysporum after subculturing on potato dextrose agar. Healthy rooted 30-day old plants (cv. Dino) were inoculated by dipping roots into a conidial suspension (5 × 107 conidia per ml) in one of six test isolates of F. oxysporum. Plants were transplanted (1 plant per pot) into pots (3.5 l vol) containing rockwool-based substrate. Noninoculated plants served as control treatments. Plants (21 per treatment) were grown in a glasshouse with an average day temperature of 31°C and night temperature of 25°C (minimum of 20°C and maximum of 42°C). Wilt symptoms and vascular discoloration in the roots, crown, and veins developed within 30 days on each inoculated plant, while noninoculated plants remained healthy. F. oxysporum was consistently reisolated from infected plants. The pathogenicity test was conducted twice. To our knowledge, this is the first report of the presence of F. oxysporum on gerbera in Italy. A wilt of gerbera was described in the Netherlands in 1952 (1) but its presence was not confirmed in further observations (3). Reference: (1) J. Arx and J. A. von Tijdschr. PlZiekt. 58:5, 1952 (2) H. Komada. Rev. Plant Prot. Res. 8:114, 1975. (3) G. Scholten. Neth. J. Plant Pathol. 76:212, 1970.

scholarly journals Fusarium Wilt of Gerbera in Spain in Soilless Crops

Plant Disease ◽  
2007 ◽  
Vol 91 (5) ◽  
pp. 638-638 ◽  
Author(s):  
A. Garibaldi ◽  
A. Minuto ◽  
A. M. Prados-Ligero ◽  
J. M. Melero-Vara ◽  
M. L. Gullino
Keyword(s):  
Vascular Tissue ◽  
Vascular System ◽  
Casein Hydrolysate ◽  
Personal Communication ◽  
Selective Medium ◽  
Fluorescent Light ◽  
Pathogenicity Test ◽  
Conidial Suspension ◽  
Cut Flowers ◽  
Brown Discoloration

In 2004, gerbera (Gerbera jamesonii cv. Excellence) plants, grown for cut flowers, were observed in a soilless cultivation system (coconut fiber substrate) in one farm in the Cadiz area (southwestern Spain) exhibiting symptoms of a wilt disease. Gerbera represents a relevant crop for the industry in the region, after rose and carnation. Affected plants were stunted and developed yellow leaves with initially brown and eventually black streaks in the vascular system. The vascular streaks in the yellow leaves were continuous with a brown discoloration in the vascular system of the crown and upper taproot. In some cases, the leaves of affected plants turned red. Fusarium spp. was consistently and readily isolated from symptomatic vascular tissue of infected plants onto a Fusarium-selective medium (3). Colonies were identified as F. oxysporum after subculturing on potato dextrose agar on the basis of morphological observations. Pathogenicity tests were carried out by using two monoconidial isolates, compared with an Italian one, obtained from wilted gerbera plants. Each isolate of F. oxysporum was grown in shake culture (90 rpm) for 10 days on casein hydrolysate at 25°C with 12 h of fluorescent light per day. Healthy rooted 30-day-old plants (cv. Jaska), were inoculated by dipping roots into a conidial suspension (5 × 107 conidia/ml) in one of the three test isolates of F. oxysporum. Plants were transplanted (1 plant per pot) into pots (3.5 liter vol.) containing rockwool-based substrate. Noninoculated plants served as control treatments. Plants (15 per treatment) were grown in a glasshouse at an average day temperature of 30°C and night temperature of 24°C (minimum of 22°C and maximum of 41°C). Wilt symptoms and vascular discoloration in the roots, crown, and veins developed within 30 days on each inoculated plant, while noninoculated plants remained healthy. F. oxysporum was consistently reisolated from infected plants. The pathogenicity test was conducted twice. A wilt of gerbera was described in the Netherlands in 1952 (1) but its presence was not confirmed in further observations (4). Gerbera wilt was recently reported in Italy (2) and identified as F. oxysporum f. sp. chrysanthemi (A. Garibaldi, personal communication). Currently, the wilt of gerbera in Spain is limited to a few farms and a very limited percent (2 to 3%) of plants. References: (1) J. Arx and J. A. von Tijdschr. PlZiekt. 58:5, 1952. (2) A. Garibaldi et al. Plant Dis. 88:311, 2004. (3) H. Komada. Rev. Plant Prot. Res. 8:114, 1975. (4) G. Scholten. Neth. J. Plant Pathol. 76:212, 1970.

scholarly journals Longevity of Oncidium varicosum (Orchidaceae) inflorescences treated with 1-methylciclopropene

2012 ◽  
Vol 42 (6) ◽  
pp. 987-992 ◽  
Author(s):  
Claudia Fabrino Machado Mattiuz ◽  
Ben-Hur Mattiuz ◽  
Teresinha de Jesus Deléo Rodrigues ◽  
Júlia de Pietro ◽  
Ramilo Nogueira Martins ◽  
...  
Keyword(s):  
Large Scale ◽  
Reducing Sugars ◽  
Soluble Carbohydrates ◽  
Vase Life ◽  
Cut Flower ◽  
Cut Flowers ◽  
Efficient Inhibitor ◽  
Respiration Rates ◽  
Golden Yellow ◽  
Postharvest Treatment

Oncidium varicosum belongs to Orchidaceae family and nowadays it is commercialized on large scale due to its potential as cut flower. The species distinguishes decoratively in function of the high number of flowers golden yellow that compose its great and delicate inflorescence. The hormone ethylene performs an important function in the processes related with the senescence of cut flowers, and especially in relation to orchids anti-ethylene treatments are recommended to extend the vase life. Among chemicals used today for the postharvest treatment of flowers the 1-methylciclopropene (1-MCP) is an efficient inhibitor of autocatalytic production of ethylene. This research aimed to evaluate the effect of different concentrations of 1-methylciclopropene (Control and 1-MCP: 250ppb, 500ppb, 1000ppb) upon physiological aspects of cut inflorescences of Oncidium varicosum. The best treatment was 1-MCP 1000ppb and the flowers presented larger values of water content, soluble carbohydrates, reducing sugars, carotenoids and the respiration rates were lower. These results contributed to higher quality and longer life of inflorescences.

scholarly journals First Report of Colletotrichum aenigma Causing Anthracnose of Grape in Korea

Plant Disease ◽  
2021 ◽  
Author(s):  
Ju Sung Kim ◽  
Oliul Hassan ◽  
Taehyun Chang
Keyword(s):  
South Korea ◽  
Disease Incidence ◽  
Juglans Regia ◽  
Morphological Characteristics ◽  
Causal Agent ◽  
Effective Control ◽  
Pathogenicity Test ◽  
Conidial Suspension ◽  
First Report ◽  
Colletotrichum Species

Grape (cv. Kyoho) is one of the most popular dessert fruits in South Korea. Anthracnose caused by Colletotrichum species is a common and very destructive disease of grape in the country. In 2019, severe outbreaks of anthracnose was observed in different grape orchards in Gimcheon (36º09´N, 128º00´ E), South Korea. The disease incidence on fruit was up to 50% in the orchards with most severe outbreaks and infected fruit displayed typical anthracnose symptoms including sunken necrotic lesions with orange-like conidial mass. For isolation of putative causal agents, nine diseased fruits were collected from three commercial orchards. A total of nineisolates were made from nine of the infected fruit by spreading spore masses (1x106 conidia mL-1) from each fruit on water agar and collecting single germinated spores after incubation at 25 ºC overnigh. The single germinated spores were transferred on to fresh potato dextrose agar (PDA) (Difco, Becton Dickinson) and incubated at 25ºC in the dark. Seven day old colonies were cottony white on the upper side and gray at the center on the reverse side. Conidia were cylindrical with round ends and measured 13.9 – 20.1 × 5.4 – 8.1 μm (mean = 16.5 × 6.6 μm, n = 30). Appressoria were brownish, sub-cylindrical with a few lobes and 10.3 –16.7 × 6.6 – 10.9 μm (mean = 13.1 × 8.1 μm, n = 30). The morphological characteristics of the solates resembled those of Colletotrichum species within the C. gloeosporioides complex (Weir et al. 2012). DNA was amplified using the following primer pairs: ITS1/ITS4, GDF / GDR, ACT-512F / ACT-783R, Bt2a/ Bt2b, and CHS79-F/CHS-354R (Weir et al. 2012). Accession numbers, LC586811 to LC586825 were obtained after depositing all the resulting sequences in GenBank. A 50% majority rules phylogenetic tree (Bayesian phylogenic analysis) was constructed based on concatenated sequences of ITS, GAPDH, ACT, TUB, and CHS using MrBayes 3.2.10. The present isolates formed a single clade with the reference isolates of C. aenigma (isolate ICMP 18608 and ICMP 18686). For a pathogenicity test, healthy grapefruits were collected from an orchards, surface sterilized by dipping in 1% sodium hypochlorite, rinsed with sterilized water and dried by blotting. A conidial suspension (1×106 conidia mL-1) in sterilized water were prepared from one week old colonies of isolates GRAP10 and GRAP12. A small wound was made on sterilized detached fruit by punching with a sterile pin. A drop of the conidial suspension was placed on the wound, while the control fruit received a drop of sterile water. Similarly, unwounded fruit were also inoculated with a single droplet of conidial suspension. For each isolate and method (wounded and unwounded), ten fruit were inoculated, and ten non-inoculated fruit were used as control. All the treated fruit were kept in a plastic box containing moist tissue and incubated at 25º C in the dark. Typical anthracnose lesions appeared on all inoculated wounded fruit while non-inoculated and inoculated unwounded fruits remained asymptotic. Koch postulates were fulfilled by re-isolating and re-identifying the causal agent from inoculated fruit. Colletotrichum aenigma has been reported as the causal agent of anthracnose on Juglans regia, Camellia sinensis and Actinidia arguta in China (Weir et al. 2012; Wang et al. 2016; Wang et al. 2018). Previous studies reported four Colletotrichum species (C. acutatum, C. gloeosporioides, C. fructicola, and C. viniferum) to cause this disease on grapes in South Korea (Oo and Oh 2017; Lim et al. 2020). To the best of our knowledge, this is the first report on grape anthracnose caused by C. aenigma in South Korea. This finding may help to take effective control measures of this disease.

scholarly journals First Report of Colletotrichum cereale Causing Anthracnose on Avena nuda in China

Plant Disease ◽  
2020 ◽  
Author(s):  
Na Zhao ◽  
Junyu Yang ◽  
Xiaoli Fang ◽  
lingrui Li ◽  
Hongfei Yan ◽  
...  
Keyword(s):  
Morphological Characteristics ◽  
Causal Agent ◽  
Post Inoculation ◽  
Pathogenicity Test ◽  
Conidial Suspension ◽  
Single Spore ◽  
First Report ◽  
Fungal Isolates ◽  
Colletotrichum Cereale ◽  
Avena Nuda

Naked oats (Avena nuda L.) is rich in protein, fat, vitamin, mineral elements and so on, and is one of the world's recognized cereal crops with the highest nutritional and healthcare value. In July 2019, leaf spot was detected on A. nuda in Zhangbei experimental station of Hebei Agricultural University. The incidence of disease is 10% to 20%. The symptoms were similar to anthracnose disease, the infected leaves had fusiform or nearly fusiform yellowish-brown spots, yellow halo around the spots. Numerous acervuli with black setae diagnostic of fungi in the genus Colletotrichum were present on necrotic lesions. To identify the pathogen, ten symptomatic leaves were collected, and only one disease spot was isolated from each leaf. Small square leaf pieces (3 to 5 mm) were excised from the junction of diseased and healthy tissues with a sterile scalpel and surface disinfested with 75% alcohol for 30s, 0.1% corrosive sublimate for 1 min, rinsed three times in sterile water. Plant tissues were then transferred on potato dextrose agar (PDA), and incubated at 25°C for 7 days. Two fungal isolates were obtained and purified by single-spore isolation method. All fungi have the same morphology and no other fungi were isolated. The aerial mycelium was gray black. The conidia were colorless and transparent, falcate, slightly curved, tapered toward the tips, and produced in acervuli with brown setae. The length and width of 100 conidia were measured and size ranged from 1.86 to 3.84 × 8.62 to 29.81 μm. These morphological characteristics were consistent with the description of Colletotrichum cereale (Crouch et al. 2006). To further assess the identity of the species, the genomic DNA of two fungal isolates (LYM19-4 and LYM19-10) was extracted by a CTAB protocol. The ribosomal DNA internal transcribed spacer (ITS) region as well as, the glyceraldehyde-3-phosphate dehydrogenase (GAPDH), actin (ACT), and the beta-tubulin 2 (Tub2) partial genes were amplified and sequenced with primers ITS4/5, GDF/GDR, ACT-512F/ACT-783R, and T1/Bt2b, respectively (Carbone et al. 1999; Templeton et al. 1992; O'Donnell et al. 1997; Glass et al. 1995). The sequences of the ITS-rDNA region (MW040121, MW040122), the GAPDH sequences (MW052554, MW052555), the ACT sequences (MW052556, MW052551) and the Tub2 sequences (MW052552, MW052553) of the two single-spore isolates were more than 99% identical to C. cereale isolate CGMCC3.15110 (JX625159, KC843517, KC843534 and JX625186). Maximum likelihood tree based on concatenated sequences of the four genes were constructed using MEGA7. The results showed the strains isolated from A. nuda were closely related to C. cereale, as supported by high bootstrap values. A pathogenicity test of the C. cereale isolates was performed on first unfolding leaves of A. nuda. Koch's postulates were carried out with isolates by spraying a conidial suspension of 106 conidia/mL on leaves of healthy A. nuda. Four replicated pots were inoculated at a time, 10 leaves each pot, while sterile distilled water was used as the control. All treated plants were placed in a moist chamber (25°C, 16-h light and 8-h dark period). Anthracnose symptoms developed on the inoculated plants 7 days post inoculation while all control plants remained healthy. Microscopic examination showed the surface of infected leaves had the same acervuli, setae, and conidia as the original isolate. The pathogenicity test was repeated three times. C. cereale was previously reported as the causal agent of anthracnose on feather reed grass in US (Crouch et al. 2009). To our knowledge, this is the first report of C. cereale as the causal agent of A. nuda anthracnose in China.

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