scholarly journals KARAKTER MORFOLOGI DAN MOLEKULER ISOLAT Phytophthora palmivora ASAL KELAPA DAN KAKAO

2020 ◽  
Vol 13 (3) ◽  
pp. 111 ◽  
Author(s):  
HIASINTA FJ MOTULO ◽  
MEITY S-SINAGA ◽  
GEDE SUASTIKA ◽  
HAJRIAL ASWIDINNOOR ◽  
ALEX HARTANA
Keyword(s):  
Puerto Rico ◽  
Sequence Analysis ◽  
Theobroma Cacao ◽  
Dna Analysis ◽  
Cocos Nucifera ◽  
Phytophthora Palmivora ◽  
Molecular Characteristics ◽  
Width Ratio ◽  
Length Width ◽  
Black Pod

ABSTRAK<br />Phytophthora palmivora merupakan patogen penyebab penyakit<br />gugur buah pada tanaman kelapa dan busuk buah pada tanaman kakao.<br />Penelitian ini bertujuan untuk membedakan isolat P. palmivora asal kelapa<br />dan asal kakao berdasarkan karakter morfologi dan molekuler.<br />Pengambilan sampel penyakit gugur buah kelapa dan busuk buah kakao<br />dilakukan di Kabupaten Banyuwangi dan Jember, Jawa Timur, Kabupaten<br />Minahasa dan Bolaang Mongondow, Sulawesi Utara, dan Kabupaten<br />Gorontalo, Gorontalo. Analisis morfologi, ekstraksi DNA dan amplifikasi<br />DNA dengan PCR dilakukan di Laboratorium Mikologi dan Laboratorium<br />Virologi, Departemen Proteksi Tanaman, Faperta IPB. Analisis perunutan<br />DNA dilakukan di Laboratorium Biologi Molekuler, Balai Besar<br />Bioteknologi dan Sumberdaya Genetik dan Laboratorium Bioteknologi,<br />LIPI Serpong. Penelitian dilaksanakan pada bulan April 2005 sampai<br />Februari 2007. Berdasarkan karakter morfologi seperti diameter koloni,<br />panjang dan lebar sporangium, tipe koloni, bentuk sporangium, per-<br />bandingan panjang dan lebar sporangium serta runutan DNA ruas ITS<br />menunjukkan bahwa keduapuluh-dua koleksi isolat yang menunjukkan<br />gejala penyakit gugur buah kelapa dan busuk buah kakao adalah P.<br />palmivora. Isolat P. palmivora asal kelapa berbeda dengan isolat P.<br />palmivora asal kakao berdasarkan diameter koloni, panjang dan lebar<br />sporangium serta runutan DNA ruas ITS. Duapuluh-dua isolat P.<br />palmivora asal kelapa dan asal kakao mempunyai sporangium yang mudah<br />lepas dari sporangiospora (caducous), pedikel yang pendek dan papila<br />serta bervariasi dalam bentuk dan ukuran sporangium. Bentuk sporangium<br />terdiri dari 4 tipe yaitu ovoid, limoniform, obturbinate, dan obpyriform.<br />Ukuran sporangium berkisar antara 40 – 62 µm panjang dan 28 – 43 µm<br />lebar. Isolat P. palmivora memiliki tipe koloni rosaceous, stelate dan<br />cottony. Rata-rata diameter koloni isolat asal kelapa 54.8 cm lebih tinggi<br />dari isolat asal kakao 43,4 cm. Hasil perunutan DNA hasil PCR<br />menunjukkan adanya keragaman genetik antar isolat asal kelapa dan kakao<br />di Indonesia. Isolat asal kakao berbeda dengan isolat asal kelapa<br />berdasarkan perunutan DNA ruas ITS. Isolat P. palmivora asal kelapa dan<br />kakao dari Indonesia tidak berada dalam satu kelompok dengan isolat<br />yang berasal dari Thailand, Taiwan, Korea, Puerto Rico, Ghana, dan<br />Cameron.<br />Kata kunci : Kelapa, Cocos nucifera, kakao, Theobroma cacao, penyakit,<br />P. palmivora, morfologi, molekuler, keragaman, runutan<br />DNA-ITS, Jawa Timur, Sulawesi Utara, Gorontalo, Jawa<br />Barat<br />ABSTRACT<br />Morphology  and  molecular  characteristics  of  P.<br />palmivora isolates from coconut and cacao<br />Phytophthora palmivora, is the pathogen of coconut nutfall and<br />cacao black pod diseases. This study was conducted to differentiate the<br />isolates of P. palmivora from coconut and those from cacao fruit based on<br />morphology and molecular characteristics. Samples of nutfall of coconut<br />and black pod of cacao were collected from Banyuwangi and Jember<br />Districts, East Java, Minahasa and Bolaang Mongondow Districts, North<br />Sulawesi, and Gorontalo District, Gorontalo. Morphological analysis,<br />DNA extraction and amplification of PCR-DNA were conducted in<br />Micology Laboratorium and Virology Laboratorium, Plant Protection<br />Division, Faperta IPB. Sequencing DNA analysis was conducted in<br />Molecular Biology Laboratory, Balai Besar Bioteknologi dan Sumberdaya<br />Genetik and Biotechnology Laboratory LIPI Serpong. This research was<br />conducted from April 2005 to February 2007. Comparative morphological<br />evaluated i.e. diameter of colony, length and width of sporangium, l/w<br />ratio, type of colony and sequence Internal Transcribed Sequence (ITS)-<br />DNA showed that all isolates of Phytophthora isolated from coconut and<br />cacao  in  Indonesia  were  Phytophthora  palmivora.  Morphology<br />characteristics of pathogen isolates from cacao were smaller and<br />significantly different in length, width, length/width ratio of<br />sporangium and diameter of colony compared to coconut’s isolates.<br />Sporangia of 22 isolates were caducous with short pedicel, but were<br />variable in shape and size. The culture produced ovoid, limoniform,<br />obturbinate, dan obpyriform sporangia, average 40-62 µm in length and<br />28-43 µm in width. The colony types were stelate, cottony and rossaceous<br />with average diameter of coconut isolates 54.8 cm and cacao isolates 43.4<br />cm. Specific fragment of 900 bp was successfully amplify from coconut<br />and cacao infected by P. palmivora. The DNA sequence analysis of the<br />nuclear ribosomal internal transcribed spacer (ITS) region showed that the<br />coconut isolates were not in the same cluster with the cacao isolates. Based<br />on sequence analysis, the P. palmivora isolates from Indonesia showed<br />different cluster from those of Taiwan, Ghana, Puerto Rico and Costa Rica<br />isolates.<br />Key words :  Coconut, Cocos nucifera, cacao, Theobroma cacao,<br />diseases, P. palmivora, diversity, morphology, molecular,<br />sequencing  ITS-DNA,  East  Java,  North  Sulawesi,<br />Gorontalo, West Jav

scholarly journals Morphological, molecular and pathogenic characterization of Phytophthora palmivora isolates causing black pod rot of cacao in Colombia

2020 ◽  
Vol 18 (2) ◽  
pp. e1003
Author(s):  
Eleonora Rodríguez-Polanco ◽  
Juan G. Morales ◽  
Melissa Muñoz-Agudelo ◽  
José D. Segura ◽  
Martha L. Carrero
Keyword(s):  
Nuclear Dna ◽  
Phenotypic Variability ◽  
Morphological Characteristics ◽  
Phytophthora Palmivora ◽  
Width Ratio ◽  
Rdna Its ◽  
Length Width ◽  
Black Pod ◽  
Molecular Sequencing

Aim of study: To characterize isolates of Phytophthora sp. causing black pod rot (BPR) of cacao (Theobroma cacao L.).Area of study: Eight cocoa-growing regions in Colombia.Material and methods: Sixty isolates of Phytophthora sp. were obtained from tissues of cacao pods showing symptoms of BPR. Isolates were characterized using the morphology of sporangia and chlamydospores, molecular sequencing of regions of nuclear DNA (rDNA-ITS) and mitochondrial (COX) and virulence in different genotypes of cocoa pods.Main results: A high phenotypic variability between the isolates was determined, being the pedicel length and the length/width ratio (L/W) the most stable characters for species identification. Short pedicels with an average of 3.13 μm ± 0.28 and a length/width ratio of sporangia (L/W) with an average of 1.55 μm ± 0.11 were established as the most consistent morphological characteristics within palmivora species.Research highlights: Phytophthora pamivora was the only species associated to BPR, identified using morphology together with sequence analyses.

Genetic diversity of native cultivated cacao accessions (Theobroma cacao L.) in Nicaragua

2012 ◽  
Vol 10 (3) ◽  
pp. 254-257
Author(s):  
Erwin Aragon ◽  
Claudia Rivera ◽  
Helena Korpelainen ◽  
Aldo Rojas ◽  
Paula Elomaa ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Theobroma Cacao ◽  
Phytophthora Palmivora ◽  
Genetic Composition ◽  
Breeding Programmes ◽  
Black Pod ◽  
Black Pod Disease ◽  
Parental Lines ◽  
Resistant Genotypes ◽  
Theobroma Cacao L

A total of 60 farmers' cacao accessions (Theobroma cacao L.) from Nicaragua were investigated using microsatellite markers to reveal their genetic composition and to identify potentially resistant genotypes against the black pod disease caused by Phytophthora palmivora. These accessions were compared with 21 breeders' accessions maintained locally, two Criollo accessions from Costa Rica and two accessions from Ecuador. The analyses showed a low level of differentiation among groups of farmers' accessions (FST = 0.06) and that six Nicaraguan accessions were genetically closely related to the two Criollo accessions used as a reference. In addition, seven distinct genotypes were found to have allelic composition that may indicate linkage to resistance alleles, thus being potential parental lines in future breeding programmes.

scholarly journals The expression of pathogenicity-related genes in Phytophthora palmivora causing black pod rot disease on cacao (Theobroma cacao L.) in Indonesia

2021 ◽  
Vol 16 (1) ◽  
pp. 284-295
Author(s):  
Masanto Masanto ◽  
Arif Wibowo ◽  
Nur Fathurahman Ridwan ◽  
Widhi Dyah Sawitri ◽  
Koji Kageyama ◽  
...  
Keyword(s):  
Theobroma Cacao ◽  
Phytophthora Palmivora ◽  
Black Pod ◽  
Rot Disease ◽  
Theobroma Cacao L

scholarly journals Symbiotic and endophytic fungi as biocontrols against cocoa (Theobroma cacao L.) phytopathogens

2017 ◽  
Vol 43 (2) ◽  
pp. 87-93 ◽  
Author(s):  
Raquel Amanda Villamizar-Gallardo ◽  
Oscar Orlando Ortíz-Rodriguez ◽  
Jhon Wilmer Escobar
Keyword(s):  
Theobroma Cacao ◽  
Trichoderma Viride ◽  
Phytophthora Palmivora ◽  
Mycotoxigenic Fungi ◽  
Black Pod ◽  
Antagonistic Potential ◽  
Native Strains ◽  
Biological Methods ◽  
Theobroma Cacao L

ABSTRACT Cocoa (Theobroma cacao L.) is a tropical tree, seriously affected by fungal diseases. To control several pathogens, biological methods are prescribed since they are friendly to the environment and easy to use. The main objective of this study was to assess the biocontrol effect of two native strains, Trichoderma viride and Botryosphaeria quercum, on phytopathogens such as Phytophthora palmivora and Moniliophtora roreri, causal agents of black pod and frosty pod rot diseases, respectively. In addition, biocontrolers were faced on potential mycotoxigenic fungi such as Aspergills flavus and Fusarium solani, which are very common on cocoa. The Bio-Control Index (BCI) was calculated to determine the in vitro biocontrol effect against the four phytopathogens. Results indicated that the best biocontrol agent of phytopathogens was B. quercum, showing BCI of 82.3%, 80.7%, 63.3% and 59.7% for each tested phytopathogen, respectively. Competition for substrate was the dominant biocontrol strategy. As to the origin of strains, those coming from the Department Norte de Santander and Santander showed the highest average inhibition percentage. This study provides an initial screening to the endophytic and antagonistic potential of fungi, specifically those capable of colonizing cocoa pods and soils. Thus, these strains can be used as an efficient biological control alternative against several known phytopathogens of cocoa in the field.

scholarly journals Observations on the Potential of an Endophytic Fungus Associated with Cacao Leaves against Phytophthora palmivora

2021 ◽  
Vol 12 (3) ◽  
pp. 528-538
Author(s):  
Joan P. Sadoral ◽  
Christian Joseph R. Cumagun
Keyword(s):  
Endophytic Fungus ◽  
Data Distribution ◽  
Phytophthora Palmivora ◽  
Molecular Characteristics ◽  
Homogeneity Of Variances ◽  
Colletotrichum Siamense ◽  
Black Pod ◽  
Asymptomatic Infections ◽  
Rot Disease ◽  
Morphological And Molecular Characterization

A study was conducted to test the pathogenicity of an endophytic fungus associated with asymptomatic cacao leaves, to determine its identity through cultural, morphological and molecular characterization, and to evaluate itsantagonistic ability vs. Phytophthora palmivora causing cacao black pod rot disease. Experiments were carried out under laboratory and shade house conditions. Homogeneity of variances and normal data distribution were determined using Bartlett’s and Shapiro–Wilk tests, respectively. Inoculation of the endophyte in healthy cacao seedlings and pods at 5 × 105 to 1 × 106 conidia per mL by spraying resulted in asymptomatic infections. The endophyte was recovered from artificially inoculated tissues 14 and 26 days after inoculation (DAI) (UF18 seedlings), and at 10 (K9 seedlings) and 14 DAI from cacao pods. The endophyte was identified as Colletotrichum siamense based on its cultural, morphological and molecular characteristics. In vitroanti-pathogen assays showed that C. siamense had the potential to limit pathogen growth by antibiosis. At 3, 5 and 7 days after incubation period (DAIP), growth of the pathogen in co-cultivation with the endophyte measured 60.0, 70.0 and 71.0 mm, respectively, which wasconsiderably lower than the growth of the pathogen alone.

scholarly journals First Report of Phytophthora palmivora, Causal Agent of Black Pod, on Cacao in Puerto Rico

Plant Disease ◽  
2007 ◽  
Vol 91 (8) ◽  
pp. 1051-1051 ◽  
Author(s):  
B. M. Irish ◽  
R. Goenaga ◽  
S. Park ◽  
S. Kang
Keyword(s):  
Puerto Rico ◽  
Germplasm Collection ◽  
Its Region ◽  
Phytophthora Palmivora ◽  
Research Station ◽  
Pathogenicity Test ◽  
Small Surface ◽  
Potato Dextrose Broth ◽  
First Report ◽  
Black Pod

Black pod or Phytophthora pod rot is the most economically important and widespread disease of cacao, Theobroma cacao L. Total losses due to Phytophthora exceed $400 million worldwide (1), and several species are known to attack cacao with P. palmivora (E.J. Butler) E. J. Butler as the most common. All plant parts are infected, but pod infections are particularly damaging. Symptoms resembling those of black pod disease were observed at the National Plant Germplasm Collection System of cacao at the USDA-ARS Tropical Agriculture Research Station (TARS) in Mayaguez, Puerto Rico for a number of years. During May of 2005, to determine the etiology of the disease, small, surface disinfested sections of pod lesions were placed on water agar and incubated for 4 days. The formation of papillate, deciduous, ellipsoidal to ovoid sporangia produced on sympodial sporangiophores on fruits, fit the description of P. palmivora and the identification was confirmed on cultures on water agar (2). Chlamydospores were readily observed in diseased pods and observed in pure cultures on V8 agar (2). Eight, single hyphal tips were transferred to potato dextrose agar (PDA) (Sigma-Aldrich, St. Louis, MO) and maintained as stock cultures. For pathogenicity tests, healthy mature pods were surface disinfested and placed in a humidity chamber lined with moist paper towels. Eight isolates were tested on four fruits per isolate and the pathogenicity test was repeated once. Inoculum was prepared by growing each isolate on PDA for 5 days with irradiation at 24°C, adding approximately 3.0 ml of water to each plate, dislodging the sporangia with a glass rod, mixing the suspension, estimating spore numbers with a hemacytometer, and adjusting to 104 sporangia per ml. A small, sterile scalpel was used to make an approximately 20.0 mm cut on the fruit epidermis, and approximately 0.2 ml of inoculum was placed on the wound. Pods were evaluated daily for 2 weeks. For molecular analysis, each of the eight cultures were grown in 50% potato dextrose broth to produce mycelia for DNA extraction using the FastDNA kit (Q-Biogen1, Irvine, CA). The internal transcribed spacer (ITS) region of the ribosomal RNA gene cluster was amplified, purified, and sequenced for all eight isolates. The ITS sequences of GenBank Accession Nos. DQ987915 to DQ987922 were identical and exhibited strong similarity (>99% identity) to that of three previously described isolates of P. palmivora from cacao (GenBank Accession Nos. AF 228097, AF467093, and AF467089). P. palmivora has been reported on citrus, coconut, black pepper, and Arracacia xanthorrhiza in Puerto Rico (2,3) and inoculum may have originated from these host or imported on cacao planted into the cacao collection before 2000. USDA-ARS-TARS is the official site for the cacao germplasm collection, thus, a detailed integrated pest management plan that includes the evaluation for resistance, sanitation measures, and use of fungicides to reduce disease levels has been implemented. Decreasing incidence and severity of this disease is a top priority. To our knowledge, this is the first report of P. palmivora on cacao in Puerto Rico. References: (1) M. C. T. Braga et al. Agrotropica 1:108, 1989. (2) D. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society. St. Paul, MN, 1996. (3) E. Rosa-Marquez. J. Agric. Univ. P. R. 84:53, 2000.

scholarly journals Phytophthora palmivora Causing Disease on Theobroma cacao in Hawaii

Agriculture ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 396
Author(s):  
Alina Sandra Puig ◽  
Wilber Quintanilla ◽  
Tracie Matsumoto ◽  
Lisa Keith ◽  
Osman Ariel Gutierrez ◽  
...  
Keyword(s):  
Theobroma Cacao ◽  
Temperature Response ◽  
Molecular Data ◽  
Phytophthora Species ◽  
Phytophthora Palmivora ◽  
Growth Responses ◽  
Temperature Growth ◽  
Phytophthora Spp ◽  
Hawaii Island ◽  
Black Pod

Commercial production of cacao in Hawaii has doubled in the past 10 years, and farmers are receiving premium prices for their beans from the expanding local confectionery industry. Black pod, caused by Phytophthora spp., is the only major cacao disease that has been reported in Hawaii but distribution and molecular identification are lacking. To determine the species of Phytophthora affecting Theobroma cacao, a sampling trip was conducted on Hawaii Island and Oahu. Ten isolates of Phytophthora palmivora were obtained from diseased cacao on Hawaii Island, but none from Oahu, despite the presence of symptomatic pods. No other Phytophthora species were found. Laboratory studies showed that all isolates produced lesions on unwounded cacao pods, but they differed in terms of their temperature–growth responses. Fungicide sensitives for a subset of isolates (n = 4) were determined using media amended with a range of fungicide concentrations. The Hawaiian isolates of P. palmivora were more sensitive to mefenoxam, chlorothalonil, and fosetyl-Al, than isolates from Ghana (n = 2) and Mexico (n = 1). This study identifies P. palmivora as a causal agent of black pod in Hawaii based on molecular data and provides valuable preliminary information on fungicide resistance and temperature response that can be used to improve disease management.

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