scholarly journals Characterization of South American Snails of the GenusBiomphalaria(Basommatophora: Planorbidae) andSchistosoma mansoni(Platyhelminthes: Trematoda) in Molluscs by PCR-RFLP

2016 ◽  
Vol 2016 ◽  
pp. 1-5 ◽  
Author(s):  
Roberta Lima Caldeira ◽  
Tatiana Maria Teodoro ◽  
Liana Konovaloff Jannotti-Passos ◽  
Pollanah M. Lira-Moreira ◽  
Christiane De Oliveira Goveia ◽  
...  

The identification of snails of the genusBiomphalariacan be done using morphological characteristics which depends on the size of the snails and skill and knowledge of researcher. These methods sometimes are not adequate for identification of species. The PCR-RFLP, using the ITS region of the rDNA, has been used to identify Brazilian species of the genusBiomphalaria. Nevertheless, there is a lack of information about snails from other Latin American countries. In addition, some snails may be infected bySchistosoma mansoniand when submitted to PCR-RFLP they show molecular profiles different from those previously standardized for the other mollusc species. In this work the molecular profiles of 15 species and the subspecies were established by PCR-RFLP of ITS-rDNA with the enzymeDdeI. Moreover, the molecular profiles of host species,B. glabrata,B. straminea,B. tenagophila, andB. prona, infected byS. mansoniwere also established. The molluscs were dissected to permit morphological identification. These results contribute to a correct identification of snails of the genusBiomphalariaand detection of these snails infected byS. mansoni.

Nematology ◽  
2008 ◽  
Vol 10 (5) ◽  
pp. 611-632 ◽  
Author(s):  
Manuel Mundo-Ocampo ◽  
Alberto Troccoli ◽  
Sergei A. Subbotin ◽  
Julio Del Cid ◽  
James G. Baldwin ◽  
...  

Abstract Phylogenetic analysis of five gene fragments: ITS-rRNA, D2 and D3 of 28S rRNA, 18S rRNA, Hsp90 and actin, of Heterodera species and two representative Afenestrata species, A. koreana and A. orientalis, form a clade with H. cynodontis, H. bifenestra and an unidentified Heterodera sp. infecting grasses. Based on these results and the consideration that the key diagnostic characters of Afenestrata are convergent and do not define a clade, synonymisation of Afenestrata with Heterodera is proposed. The following new combinations are made: H. africana comb. n., H. axonopi comb. n., H. koreana comb. n., and H. orientalis comb. n. Furthermore, H. (= Afenestrata) sacchari is renamed as H. saccharophila nom. nov. to avoid homonymy. All these species, together with H. bamboosi, are regarded as members of a paraphyletic ‘Afenestrata group’ within Heterodera. Whilst recognised as artificial, the Afenestrata group is nevertheless an aid to discussion about these similar species. Morphological and molecular characterisation of populations of H. koreana comb. n. from Florida and H. orientalis comb. n. from Florida and Guatemala verify the identification of these populations as valid representatives for molecular studies of the species. Light and SEM observations also provide new detail and a broader understanding of the morphological range of both species. These include a longer stylet for females of H. koreana comb. n. and H. orientalis comb. n. than reported in the original descriptions. In addition, previously unreported tuberculate ridges are noted on the surface of vulval lips of H. orientalis comb. n. The lip region of second-stage juveniles of H. koreana comb. n. and H. orientalis comb. n. both include fused adjacent submedian lips that also fuse with the labial disc and the second lip annulus. The ITS-rRNA gene sequences of H. orientalis comb. n. populations from Florida and Guatemala were similar to those from the Russian type locality. Diagnostic PCR-RFLP of ITS-rRNA profiles with six enzymes for H. orientalis comb. n. and H. koreana comb. n. are given. A key for the morphological identification of species of the Afenestrata group is provided.


Plant Disease ◽  
2014 ◽  
Vol 98 (9) ◽  
pp. 1274-1274 ◽  
Author(s):  
K. Chen ◽  
X.-Q. Wu ◽  
M.-X. Huang ◽  
Y.-Y. Han

Early spring shoot bamboo (Phyllostachys praecox) is profitable for the production of bamboo shoots and is widely cultivated in Jiangsu and Zhejiang provinces in China. In recent years, a new disease named brown culm streak has commonly occurred in bamboo plantations. During initial stages, small yellow or sandy beige spots (1 to 2 mm diameter) develop on the culm. Spots expand and develop into fine brown streaks (10 to 15 × 1 to 2 mm), and the quantity of spots increases greatly at the mid-stage of the disease. The streaks continue to expand (10 to 30 × 2 to 3 mm) and may coalesce into brown patches that cover 70% of the culm surface in late stages of disease. Color of the internal bamboo cavity gradually changes from white to brown without an alcoholic odor, and plants eventually die. Diseased tissues (1 × 1.5 cm) were taken from the edges of brown streaks, surface disinfected in 0.1% HgCl2 for 3 min, rinsed in distilled water three times, and incubated on potato dextrose agar (PDA) at 25°C. Based on colony and conidial morphology, six different fungi (ZBY01, ZBY02, ZBY03, ZBY04, ZBY05, and ZBY06) were isolated. A suspension of hyphal fragments (106 pieces/ml) of each of the fungi was used to inoculate plants in the field. Healthy bamboo culms were stabbed with alcohol-sterilized needles after surface disinfection and wounds of each bamboo were inoculated with 3 ml hypha fragment suspensions by paintbrush, then covered with a piece of sterile plastic film. A control treatment was inoculated with sterile water. Each treatment had two replicates. Twenty days after inoculation, only fungus ZBY01 caused similar symptoms with the diseased bamboo in the field and was re-isolated from necrotic tissue of inoculated plants, thus fulfilling Koch's postulates. Mycelium of fungus ZBY01 covered entire PDA plates (90 mm diameter) after incubating for 6 days at 25°C and was white, flat, and floccose with moderate aerial mycelium. Mycelium consisted of smooth, hyaline, branched, septate hyphae, 2 to 3 μm diameter. Conidiophores were erect, septate, pale brown, smooth, and reduced to conidiogenous cells. Conidiogenous cells were pale brown, smooth, ampulliform, 6 to 9 μm long; the apical neck was 1 to 3 μm long, basal part 5 to 6 μm long. Conidia were 1-celled, dark brown to black, smooth, lenticular, 5 to 7 μm in diameter, 2 to 4 μm wide, lemon-shaped in side view, and spherical from top view with an equatorial ring. Based on morphological characteristics, it was identified as Arthrinium arundinis (Corda) Dyko & B. Sutton (1). The internal transcribed spacer (ITS) region from the isolate ZBY01 was amplified with PCR using ITS1-ITS4 primer pairs and the amplicon sequenced. Size of the sequenced region was 540 bp and had 99% identity with A. arundinis, which was in accord with morphological identification. The sequence was deposited in Gen Bank under accession number KF850624. To our knowledge, brown culm streak of P. praecox is a new disease of bamboo that has not been reported in other countries; however, A. arundinis has been reported as a pathogen on barley in Fairfield, Montana. Reference: (1) P. W. Crous and J. Z. Groenewald. A phylogenetic re-evaluation of Arthrinium. IMA Fungus. 4:133, 2013.


Plant Disease ◽  
2015 ◽  
Vol 99 (2) ◽  
pp. 288-288 ◽  
Author(s):  
T. J. Deng ◽  
Q. L. Li ◽  
X. L. Chen ◽  
S. P. Huang ◽  
T. X. Guo ◽  
...  

Cassia fistula, a member of the Fabaceae, known as the golden shower tree, is native to South Asia. It is now distributed worldwide and is popular as an ornamental plant as well as being used in herbal medicine. In October 2013, symptoms of stem canker were observed on C. fistula in a nursery (108°38′ E, 22°87′ N) in Nanning, Guangxi, China. The symptoms began as small brown lesions, which enlarged over several months to long, striped, slightly sunken lesions, 1 to 9 cm in width and 16 to 135 cm in length. The conspicuous cankers had vertical cracks outlining the canker and evenly spaced horizontal cracks, eventually resulting in whole plants dying back. The cankers were found on 90% of six-year-old plants in this nursery and were also observed in other plantings. On potato dextrose agar (PDA), isolates with similar morphological characteristics were consistently recovered from symptomatic plant tissues after surface sterilization in 75% ethanol for 30 sec and then in 0.1% mercuric chloride for 2 min. Over 100 conidia were examined from three isolates and were found to be elliptical and hyaline when immature, becoming dark brown, one-septate, and longitudinally striate when mature and ranging from 20 to 31 × 11 to 16 μm (average 25.5 × 13.6 μm). The rDNA internal transcribed spacer (ITS) region of isolate LC-1 was sequenced (GenBank Accession No. KM387285), and it showed 100% identity to Lasiodiplodia theobromae (Pat.) Griffon & Maubl. (GenBank KC964548), confirming the morphological identification (2) as L. theobromae (also known as Botryosphaeria rhodina (Cooke) Arx). A culture of this isolate has been preserved in the Guangxi Academy of Agricultural Sciences fungal collection. The pathogenicity of the isolate was tested on healthy twigs and branches of C. fistula trees in a field setting at Guangxi Agricultural Vocational-Technical College, Nanning, Guangxi, in June and August 2014. For each treatment, five green twigs and five 2-year-old branches were used. Five adjacent needle punctures were made on each branch with a sterilized needle. A mycelial plug was then placed on the wound of each branch and wrapped with Parafilm. Control twigs were treated with sterile PDA plugs. One week later, typical lesions were observed on the inoculated branches, with symptoms becoming more extensive after two weeks, but no symptoms were seen on the controls. Koch's postulates were fulfilled by re-isolation of L. theobromae from diseased branches. L. theobromae is recognized as an important wood pathogen and has been reported to cause cankers, dieback, and fruit and root rots in over 500 different hosts, including perennial fruit and nut trees, vegetable crops, and ornamental plants (2). The fungus has been reported on C. fistula in India since the 1970s (1); however, to our knowledge, this is the first report of L. theobromae infecting C. fistula in China. References: (1) R. S. Mathur. The Coelomycetes of India. Bishen Singh Mahendra Pal Singh, Delhi, India, 1979. (2) J. R. Úrbez-Torres et al. Plant Dis. 92:519, 2008.


Plant Disease ◽  
2014 ◽  
Vol 98 (2) ◽  
pp. 282-282
Author(s):  
K. Vrandečić ◽  
J. Ćosić ◽  
D. Jurković ◽  
I. Stanković ◽  
A. Vučurović ◽  
...  

Lavandula × intermedia Emeric ex Loiseleur, commonly known as lavandin, is an aromatic and medicinal perennial shrub widely and traditionally grown in Croatia. The lavandin essential oil is primarily used in perfumery and cosmetic industries, but also possesses anti-inflammatory, sedative, and antibacterial properties. In June 2012, severe foliar and stem symptoms were observed on approximately 40% of plants growing in a commercial lavandin crop in the locality of Banovo Brdo, Republic of Croatia. Initial symptoms on lower leaves included numerous, small, oval to irregular, grayish brown lesions with a slightly darker brown margin of necrotic tissue. Further development of the disease resulted in yellowing and necrosis of the infected leaves followed by premature defoliation. Similar necrotic oval-shaped lesions were observed on stems as well. The lesions contained numerous, dark, sub-globose pycnidia that were immersed in the necrotic tissue or partly erumpent. Small pieces of infected internal tissues were superficially disinfected with 50% commercial bleach (4% NaOCl) and placed on potato dextrose agar (PDA). A total of 10 isolates from leaves and five from stems of lavandin formed a slow-growing, dark, circular colonies with raised center that produced pycnidia at 23°C, under 12 h of fluorescent light per day. All 15 recovered isolates formed uniform hyaline, elongate, straight or slightly curved conidia with 3 to 4 septa, with average dimensions of 17.5 to 35 × 1.5 to 2.5 μm. Based on the morphological characteristics, the pathogen was identified as Septoria lavandulae Desm., the causal agent of lavender leaf spot (1,2). Pathogenicity of one selected isolate (428-12) was tested by spraying 10 lavandin seedlings (8 weeks old) with a conidial suspension (106 conidia/ml) harvested from a 4-week-old monoconidial culture on PDA. Five lavandin seedlings, sprayed with sterile distilled water, were used as negative control. After 5 to 7 days, leaf spot symptoms identical to those observed on the source plants developed on all inoculated seedlings and the pathogen was successfully re-isolated. No symptoms were observed on any of the control plants. Morphological identification was confirmed by amplification and sequencing of the internal transcribed spacer (ITS) region of rDNA (3). Total DNA was extracted directly from fungal mycelium with a DNeasy Plant Mini Kit (Qiagen, Hilden, Germany) and PCR amplification performed with primers ITS1F/ITS4. Sequence analysis of ITS region revealed at least 99% identity between the isolate 428-12 (GenBank Accession No. KF373078) and isolates of many Septoria species; however, no information was available for S. lavandulae. To our knowledge, this is the first report of Septoria leaf spot of lavandin caused by S. lavandulae in Croatia. Since the cultivation area of lavandin plants has been increasing in many continental parts of Croatia, especially in Slavonia and Baranja counties, the presence of a new and potentially harmful disease may represent a serious constraint for lavandin production and further monitoring is needed. References: (1) T. V. Andrianova and D. W. Minter. IMI Descriptions of Fungi and Bacteria, 142, Sheet 1416, 1999. (2) R. Bounaurio et al. Petria 6:183, 1996. (3) G. J. M. Verkley et al. Mycologia 96:558, 2004.


Plant Disease ◽  
2014 ◽  
Vol 98 (8) ◽  
pp. 1154-1154 ◽  
Author(s):  
Y. N. Liu ◽  
N. Nan ◽  
B. H. Lu ◽  
W. Y. Xia ◽  
X. Y. Wu ◽  
...  

Rheum rhabarbarum L., rhubarb, is a perennial herb planted mainly in Hebei, Hubei, Shanxi, Heilongjiang, and Jilin provinces as well as Inner Mongolia, China. The plant grows about 1,000 meters above sea level (4), and is used widely in China to treat constipation and gout. From June to September 2012, a leaf spot was observed on R. rhabarbarum in the medicinal garden of Jilin Agricultural University, Changchun, Jilin Province, causing significant effects on the leaves of all infected plants. In the early stage of disease development, small red lesions were visible on infected leaves, which subsequently developed into irregularly shaped or circular necrotic spots, each with a light colored center, pink-red alternating concentric rings, and surrounded by a chlorotic halo. Some lesions became perforated in the center. Lesions ranged from 1 to 15 mm in diameter. Extensive spotting resulted in general browning and yellowing of entire leaves. As lesions enlarged and coalesced, some leaves died from the margin inwards. Lesions on the stem were fusiform and sunken. Small pieces of diseased leaves and stems were surface-disinfested in 75% ethanol for 60 s, rinsed twice in distilled water, dried, and plated on potato dextrose agar (PDA). A Phoma species was isolated that produced a gray or dark gray colony after 5 to 7 days. The isolate was transferred to oatmeal agar (OA) (3). Pycnidia were dark brown to black, globose to subglobose, and 121 to 354 × 100 to 262 μm. Conidia were ellipsoidal or reniform, colorless, unicellular, and 3.8 to 6.5 × 1.7 to 4.1 μm. On the basis of these characteristics, the fungus was identified as Phoma rhei (1). A PCR assay with the ITS4 and ITS5 primers was used to amplify the internal transcribed spacer (ITS) region of ribosomal DNA (rDNA) (2). The amplified product (567 bp) was sequenced and the sequence submitted to GenBank (Accession No. KF531831). The ITS sequence exhibited 99% identity to that of a P. rhei isolate in GenBank (GU237743.1), confirming the morphological identification. Pathogenicity of eight isolates on rhubarb was confirmed by spraying a spore suspension (1 × 106 spores/ml) produced on PDA on the leaves of each 6-year-old R. rhabarbarum (cv. Boyedahuang) plant. Each isolate was inoculated onto five plants, and five plants were sprayed similarly with sterilized water as a control treatment. The plants were then covered with plastic bags for 48 h, and kept in a greenhouse (20 to 30°C with a 12-h photoperiod/day). Initial symptoms on inoculated leaves were observed 3 to 4 days after inoculation, while the control plants remained healthy. Re-isolations from lesions on the inoculated leaves, using the same protocol as the original isolations, produced fungal colonies with the same morphological characteristics as the original isolates of P. rhei, but no fungi were re-isolated from the control plants. This fungus has been found on R. rhaponticum in New Zealand (1), but to our knowledge this is the first report of P. rhei on R. rhabarbarum in China. References: (1) G. H. Boerema et al. Phoma Identification Manual. Diffferentiation of Specific and Infra-Specific Taxa in Culture. CABI Publishing. Wallingford, UK, 2004. (2) D. E. L. Cooke et al. Mycol. Res. 101:667, 1997. (3) Z. D. Fang. Research Method of Phytopathology. China Agricultural Press (In Chinese), 1998. (4) A. J. Li et al. Flora Reipublicae Popularis Sinicae. Tomus 25:171, 1998.


Author(s):  
Nabilah Mohamad Khairi ◽  
Wilson Thau Lym Yong ◽  
Julius Kulip ◽  
Kenneth Francis Rodrigues

Conservation of plant species plays a vital role in preventing the loss of valuable plant resources. The success of conservation depends on the correct identification and characterization of plant species. Bamboo is one of the most important plants with multiple uses that have contributed to the economy and socio-economy of many people in rural areas. It is under the subfamily of Bambusoideae that includes both woody and herbaceous bamboo. Conventionally, like other plants, bamboo has been classified dependently based on morphological characteristics. However, morphological identification leads to difficulties and misclassification of bamboo species due to their infrequent flowering behaviour and peculiar reproductive biology. Since then, molecular markers have been introduced to overcome the problems associated with bamboo taxonomy and phylogeny. This paper provides an overview of the diverse, predominantly molecular techniques used to assess and determine the genetic diversity of bamboo species.


Zootaxa ◽  
2020 ◽  
Vol 4819 (2) ◽  
pp. 316-334 ◽  
Author(s):  
KIAVASH GOLZARIANPOUR ◽  
MASOUMEH MALEK ◽  
MEHDI GOLESTANINASAB ◽  
ALIMORAD SARAFRAZI ◽  
JUDITH KOCHMANN ◽  
...  

Correct identification of elasmobranch species is crucial for taxonomic and parasitological research. Although molecular barcoding may be the fastest choice to determine the identity of a given species, robust and fast species level identification in the field using morphological characters is essential. During this study, 389 specimens representing seven stingray species (Brevitrygon walga, Himantura leoparda, H. uarnak, Maculabatis randalli, M. arabica, M. gerrardi and Pateobatis fai) were examined from the Persian Gulf and the Gulf of Oman. A 1044 bp fragment of the NADH2 gene was generated for 50 specimens with representatives of all species. To verify the initial morphological identification and to compare intra- and interspecific differences a Neighbor-Joining analysis was conducted using uncorrected p-distances, whereas the Bayesian Inference was used to examine the relationships among taxa. Two species (M. arabica and M. gerrardi) are documented from the Persian Gulf for the first time. The molecular results provide the first known evidence of the sympatric distribution of M. randalli and M. arabica in the north and northwestern Indian Ocean. The results of the Bayesian Inference support the recent divergence of both species. Based on morphological comparisons and molecular support we suggest that the descriptions of M. randalli and M. arabica have been carried out on heterogeneous type series which has led to inconsistency between molecular identification and diagnostic morphological characteristics. Detailed morphological examination revealed that there is a relation between the type and number of denticles on the mid-dorsal surface of the disc and the color pattern of the tail. To address this taxonomic conflict all type materials should be re-examined. The Bayesian Inference tree showed that all specimens from the Persian Gulf and the Gulf of Oman morphologically resembling B. walga were found to group well outside those of the Indian species (B. imbricata) with an average p-distance of 0.097. The low nucleotide differences among the urogymnid taxa (P. fai and H. leoparda) from the Persian Gulf and the Gulf of Oman and their conspecific specimens in the Indo-West Pacific region revealed that philopatric behaviors may cause considerable gene flow among populations.


Plant Disease ◽  
2013 ◽  
Vol 97 (12) ◽  
pp. 1659-1659 ◽  
Author(s):  
M. Vasić ◽  
N. Duduk ◽  
I. Vico ◽  
M. S. Ivanović

Botryosphaeria dothidea (Moug.: Fr.) Ces. & De Not has a worldwide distribution infecting species from over 80 genera of plants (1). Apart from being an important pathogen of apple trees in many countries, B. dothidea can cause pre- and postharvest decay on apple fruit (2). It has been known to cause canker and dieback of forest trees in Serbia (3), but has not been recorded either on apple trees or apple fruit. In December 2010, apple fruit cv. Idared (Malus × domestica Borkh.) with symptoms of white rot were collected from one storage in the area of Svilajnac in Serbia. The incidence of the disease was low but the symptoms were severe. Affected fruit were brown, soft, and almost completely decayed, while the internal decayed tissue appeared watery and brown. A fungus was isolated from symptomatic tissue of one fruit after surface sterilization with 70% ethanol (without rinsing) and aseptic removal of the skin. Small fragments of decayed tissue were placed on potato dextrose agar (PDA) and incubated in a chamber at 22°C under alternating light and dark conditions (12/12 h). Fungal colonies were initially whitish, but started turning dark gray to black after 5 to 6 days. Pycnidia were produced after 20 to 25 days of incubation at 22°C and contained one-celled, elliptical, hyaline conidia. Conidia were 17.19 to 23.74 μm (mean 18.93) × 3.72 to 4.93 μm (mean 4.45) (n = 50). These morphological characteristics are in accordance with those described for the fungus B. dothidea (4). Genomic DNA was isolated from the fungus and internal transcribed spacer (ITS) region of rDNA was amplified with the primers ITS1/ITS4 and sequenced. The nucleotide sequence has been assigned to GenBank Accession No. KC994640. BLAST analysis of the 528-bp segment showed a 100% similarity with several sequences of B. dothidea deposited in NCBI GenBank, which confirmed morphological identification. Pathogenicity was tested by wound inoculation of five surface-sterilized, mature apple fruit cv. Idared with mycelium plugs (5 mm in diameter) of the isolate grown on PDA. Five control fruit were inoculated with sterile PDA plugs. After 5 days of incubation in plastic containers, under high humidity (RH 90 to 95%) at 22°C, typical symptoms of white rot developed on inoculated fruit, while wounded, uninoculated, control fruit remained symptomless. The isolate recovered from symptomatic fruit showed the same morphological features as original isolate. To the best of our knowledge, this is the first report of B. dothidea on apple fruit in Serbia. Apple is widely grown in Serbia and it is important to further investigate the presence of this pathogen in apple storage, as well as in orchards since B. dothidea may cause rapid disease outbreaks that result in severe losses. References: (1) G. H. Hapting Agriculture Handbook 386, USDA, Forest Service, 1971. (2) A. L. Jones and H. S. Aldwinckle Compendium of Apple and Pear Diseases. APS Press, St. Paul, MN, 1990. (3) D. Karadžic et al. Glasnik Šumarskog Fakulteta 83:87, 2000. (4) B. Slippers et al. Mycologia 96:83, 2004.


Plant Disease ◽  
2013 ◽  
Vol 97 (11) ◽  
pp. 1506-1506 ◽  
Author(s):  
P. P. Li ◽  
Z. Y. Cao ◽  
J. G. Dong ◽  
L. H. Zhang ◽  
H. Jia ◽  
...  

Corn is the most important cereal crop in China, with over 34.94 million ha being cultivated in the country annually. However, fungal diseases are a major limiting factor in corn production. In August 2012, 20 ha of corn fields in Anhui Province were found to be heavily infected by fungi. The margin of the lesion was achlorotic, and the middle was yellowish white or off-white, which was similar to the corn Curvalaria leaf spot. The oval lesions were approximately 5 to 7 mm. Lesion tissue was removed from the border between symptomatic and healthy tissue. The surface was sterilized in 75% ethanol for 30 s and 0.1% HgCl2 for 1 min, after which the sample was washed three times in sterile distilled water. The isolate was purified and subcultured on potato dextrose agar (PDA) at 25 ± 2°C. The initial color of the colony was light brown, turning dark brown after being cultured for 7 days. The conidia were boat-shaped or inverted pear-shaped and were clearly bent to one side. The cells of both ends were slightly lighter and respectively ranged from 34.5 to 44.0 μm and 12.0 to 21.0 μm away from the base, with the second cell as the widest. The majority conidia had three or four false septates; isolates produced light brown to medium brown conidiophore, scattered or clustered, often branching, and exhibited bending. These morphological characteristics matched with the description of Bipolaris papendorfii reported by Zhang (3). A pathogenicity test was conducted with the two isolates on each of the 36 corns by spraying 2 ml spore suspension (106 conidia/ml). For the control treatment, 36 corns were inoculated with an equal volume of sterilized water. Inoculated plants were placed in a greenhouse from 29 to 33°C and 95% relative humidity. The typical 5 to 7 mm oval lesions were observed 7 days after inoculation, except on the control samples. Three replications of 36 corns were used for each treatment. The isolate was consistently 100% reisolated from the diseased tissue according to Koch's postulate. The isolate was found to be morphologically similar to B. papendorfii. Preliminary morphological identification of the fungus was confirmed by PCR assay using genomic DNA extracted from the mycelium of a 7-day-old culture on PDA at 25 ± 2°C. A 550-bp amplified region of the internal transcribed spacer (ITS) of rDNA was generated using ITS1 (5′-TCCGTAGGTGAACCTGCGG-3′) and ITS4 (5′-TCCTCCGCTTATTGATATGC-3′) universal primers (1). The ITS region (GenBank Accession No. KC592365) was then sequenced by Sangon Biotech (Shanghai, China), and displayed 99% nucleotide similarity with the rDNA-ITS of B. papendorfii (JQ753972.1) separately after BLASTn research in GenBank. Based on the symptoms, fungal morphology, ITS sequence, and pathogenicity testing, this fungus was identified as B. papendorfii. The pathogen could reportedly infect tobacco and cotton (2). To our knowledge, this is the first study to report that B. papendorfii can infect corn in China. This report will establish a foundation for the further study of B. papendorfii to address the disease effectively. Further studies will be conducted to determine the incidence of the disease and the severity of damage caused by B. papendorfii as well as determine a possible mode for controlling the spread of the disease. References: (1) Y. J. Cao et al. Chin. J. Trop. Crops 31:1098, 2010. (2) H. Deng et al. Mycosystema 21:327, 2002. (3) T. Y. Zhang. Chin. Fungi Chi. 30:21, 2010.


Plant Disease ◽  
2014 ◽  
Vol 98 (6) ◽  
pp. 854-854
Author(s):  
W. Yang ◽  
L. Zheng ◽  
C. Wang ◽  
C.-P. Xie

Serenoa repens [(Bartr) J. K. Small] is an important medicinal plant with their extracts is one of the three most effective drugs to cure benign prostatic hyperplasia (BPH). Also it can be used as an ornamental plant for garden. In November 2010, a new leaf spot disease was found on S. repens in Danzhou, Hainan Province, China. Disease occurred very seriously, with the incidence close or up to 100%, even leading to plant drying and death. Initially, the leaves had circular water-soaked dots, and had an obvious yellow halo on the edge, then expanded into oval, circular, or irregular shaped spots. Eventually the spot was beige and gray in the center and dark brown and slightly concave on the edge. The pathogen was isolated following the method reported by Fang (3) and prepared for further characterization. On potato dextrose agar (PDA) medium, the pathogen formed round and red-brown colonies with neat edges of a sandy beige color. A white powdery substance was formed on the surface of the colony, and it produced reddish-brown pigment on the back. On carnation leaf agar (CLA), only large macroconidium was observed. Macroconidiophores containing a stipe bearing penicillate suites of fertile branches, terminating in a clavate vesicle (5.9-) 6.4 (-6.9) × (33.8-) 39.6 (-46.7) μm. Conidiogenous apparatus had primary branches aseptate or rarely 1-septate and were (21.8-) 28.7 (-38.6) μm long, secondary branches were aseptate and (18.8-) 29.9 (-39.9) μm long, and tertiary branches were aseptate and (14.2-) 17.4 (-19.9) μm long. Macroconidium and microconidium were observed on water agar (WA) at 30 days. Macroconidium was colorless, cylindrical, rounded at both ends, 1 to 3 hyaline septate, but mainly one, and (4.5-) 5.2 (-6.2) × (71.3-) 84.1 (-98.0) μm; microconidium was colorless, cylindrical, both ends obtuse, curved or straight, 1-septate, and (24.8-) 33.2 (-45.2) × (2.5-) 3.5 (-5.0) μm. It could produce microsclerotia on PDA, CLA, and WA media. Morphological characteristics of the specimen examined were similar to Calonectria pteridis. In the genus of Calonectria, only C. pteridis could produce bending microconidium on WA medium (2). To confirm the morphological identification, primer pair ITS1/ITS4 were used for amplification of the ITS region of rDNA. Its sequence (GenBank Accession No. KF994926) showed 99% identity with C. pteridis Crous, M.J. Wingf. & Alfenas. (GQ280617.1). In addition, the translation elongation factor 1-alpha gene sequence was amplified (KF994927) and it showed 100% identify with C. pteridis (FJ918564.1) (1). Thus, the pathogen was identified as C. pteridis. To confirm pathogenicity, conidial suspensions (105 conidia ml−1) of the pathogen were inoculated with healthy leaves of 10 plants by pinprick inoculation method. Control plants were inoculated with water. Plants were maintained at 28°C in a greenhouse with constant humidity (RH 90%) and a 12-h photoperiod of fluorescent light. Symptoms similar to the original ones appeared after 7 days, while the control plants remained healthy. The tests were repeated three times and the pathogen was re-isolated from the leaves of inoculated plants and confirmed to be C. pteridis by both morphology and molecular characterization. To our knowledge, this is the first report of leaf spot caused by C. pteridis on S. repens in China. References: (1) I. Carbone and L. M. Kohn. Mycologia 91:553, 1999. (2) P. W. Crous and M. J. Wingfield. Mycotaxon 51:341, 1994. (3) Z. D. Fang. Plant Disease Research Methods, 3rd edition. China Agriculture Press, Beijing, 1998.


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