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MOLECULAR IDENTIFICATION OF Pepper yellow leaf curl Indonesia virus ON CHILI PEPPER IN NUSA PENIDA ISLAND Selangga, Dewa Gede Wiryangga; Listihani, Listihani
JURNAL HAMA DAN PENYAKIT TUMBUHAN TROPIKA Vol. 21 No. 2 (2021): SEPTEMBER, JURNAL HAMA DAN PENYAKIT TUMBUHAN TROPIKA
Publisher : Universitas Lampung

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.23960/jhptt.22197-102

Abstract

Molecular identification of Pepper yellow leaf curl Indonesia virus on chili pepper in Nusa Penida Island. Pepper yellow leaf curl Indonesia virus (PYLCV) has been reported as caused yellow leaf curl disease in Bali Island since early 2012. Dominant symptoms of PYLCV infection in chili pepper were yellowing, leaf curl, yellow mosaic, and mottle. Bemisia tabaci, has been known to vector on the case yellow leaf curl disease. Observations on the Nusa Penida Island in 2020 showed symptoms such as yellow leaf curl disease, however, identification of PYLCV in Nusa Penida Island has not been studied. Molecular identification was conducted using polymerase chain reaction and sequence analysis. Data collected in this study was disease symptoms and disease incidence. The results showed that dominant disease symptoms caused by virus from Nusa Penida were yellow mosaic, yellowing, and mottle. Universal DNA fragments of 912 bp were successfully amplified from 50 leaf samples using Begomovirus degenerate primers SPG 1 (5’-CCCCKGTGCGWRAATCCAT-3’) and SPG 2 (5’ATCCVAA YWTYCAGGGAGCT-3’). Sequence analysis showed that the isolate from Nusa Penida was a Pepper yellow leaf curl Indonesia virus with a 98–100% homology with several reference isolates.
NATURAL INFECTION OF Tobacco mosaic virus ON BUTTERNUT SQUASH IN BALI, INDONESIA Listihani, Listihani; Selangga , Dewa Gede Wiryangga; Sutrawati, Mimi
JURNAL HAMA DAN PENYAKIT TUMBUHAN TROPIKA Vol. 21 No. 2 (2021): SEPTEMBER, JURNAL HAMA DAN PENYAKIT TUMBUHAN TROPIKA
Publisher : Universitas Lampung

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.23960/jhptt.221116-122

Abstract

Natural infection of Tobacco mosaic virus on butternut squash in Bali, Indonesia. Tobacco mosaic virus (TMV) was a newly emerging virus infecting cucumbers in Indonesia since 2017. The mosaic disease caused by TMV potentially caused yield loss cucumber in Java. In 2019, mosaic symptoms were observed in butternut squash plants in Bali and molecular detection using universal primer of Tobamovirus indicated the presence of TMV infection. Further research was conducted to determine molecular characteristics of TMV on butternut squash plants in Bali. Specific DNA bands of Tobamovirus were amplified using reverse transcription polymerase chain reaction method, followed by DNA sequencing. The DNA were successfully amplified from CP Tobamovirus using universal primers from several butternut squash samples, i.e. Denpasar, Gianyar, Buleleng, and Karangasem Districts. The homology analysis of nucleotide and amino acid sequences of TMV among isolates from Denpasar, Gianyar, Buleleng, and Karangasem Districts was ranged between 95.6 – 97.7% and 98.1 – 99.4%, respectively. This indicated that low genetic diversity of TMV among Bali isolates. The highest homology of corresponding sequences of TMV isolates from Denpasar, Gianyar, Buleleng, and Karangasem Districts was closely related to TMV Kediri-Indonesia isolate on cucumber plant. Correspondingly, the phylogenetic analysis showed that TMV Bali isolates were categorized into same cluster with Kediri-Indonesia isolates. This was the first report of TMV on butternut squash in Indonesia.
Karakterisasi Molekuler Papaya ringspot virus tipe P pada Tanaman Mentimun di Jawa Listihani Listihani; Tri Asmira Damayanti; Sri Hendrastuti Hidayat; Suryo Wiyono
Jurnal Fitopatologi Indonesia Vol. 14 No. 3 (2018)
Publisher : The Indonesian Phytopathological Society (Perhimpunan Fitopatologi Indonesia)

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (526.327 KB) | DOI: 10.14692/jfi.14.3.75

Abstract

Moleculer Characterization of Papaya ringspot virus type P on Cucumber in JavaInfection of Papaya ringspot virus (PRSV) on cucumber plants showing mosaic symptom was detected using specific antibody.  Further investigation was conducted to determine molecular characters and status of PRSV infecting cucumber in Java.  Infection of PRSV was detected from leaf samples collected from the field using dot immunobinding assay (DIBA).  Disease frequency caused by PRSV infection reached 81.11%, 95.86%, 91.66%, and 92.3% in East Java, Central Java, Yogyakarta, and West Java, respectively.  Characterization of PRSV isolates was conducted by reverse transcription polymerase chain reaction (RT-PCR) using specific primers for PRSV-P and PRSV-W, followed by cloning, and DNA sequencing.  DNA fragment of 470 bp was successfully amplified using specific primers for PRSV-P from several samples from Nganjuk, Brebes, Kulon Progo, and Subang; but no amplification was achieved using specific primers for PRSV-W.  Nucleotide and amino acid analysis showed high homology among PRSV-P isolates from Nganjuk, Brebes, Kulon Progo, and Subang, i.e. 98.6%-99.7% and 99.3%-100%, respectively.  This is an indication of a low genetic variation among PRSV-P from Java. Further phylogenetic analysis indicated that PRSV-P isolate cucumber is in the same cluster with PRSV-P isolate papaya from Bali, Indonesia.  This is the first report of PRSV-P infecting cucumber in Indonesia.
Isolasi cendawan endofit pada tanaman padi serta potensinya sebagai pemacu pertumbuhan tanaman Hermanu Triwidodo; Listihani Listihani; Dewa Gede Wiryangga Selangga
Agrovigor Vol 14, No 2 (2021): September
Publisher : Universitas Trunojoyo Madura

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21107/agrovigor.v14i2.10166

Abstract

Cendawan endofit banyak mendapat perhatian karena potensinya dalam memacu pertumbuhan dan kemampuannya dalam meningkatkan ketahanan tanaman terhadap penyakit dan hama tanaman. Penelitian ini bertujuan mengisolasi cendawan endofit pada tanaman padi di Bali dan menguji potensinya sebagai pemacu pertumbuhan tanaman. Isolasi bakteri endofit dilakukan dengan metode sterilisasi permukaan dengan alkohol dan natrium hipoklorit pada medium PDA. Lebih dari 15 isolat cendawan endofit berhasil diisolasi dan dikoleksi dari beberapa bagain tanaman padi. Isolat tersebut diseleksi untuk melihat kemampuan antibiosisnya terhadap cendawan patogen secara in vitro dan memacu pertumbuhan tanaman. Hasil seleksi menunjukkan cendawan endofit non pathogen yang memacu pertumbuhan tanaman yaitu 3 isolat asal pelepah, 3 isolat asal batang, dan 1 isolat asal daun padi. Hasil identifikasi cendawan endofit menunjukkan bahwa isolate asal pelepah dan batang padi dinamakan Phoma sp, sedangkan asal daun padi dinamakan Acremonium. Hal ini mengindikasikan bahwa beberapa cendawan endofit non patogen asal padi berpotensi sebagai pemacu pertumbuhan tanaman.
Molecular Characterization of Begomovirus on Cucumber in Java Listihani Listihani; Tri Asmira Damayanti; Sri Hendrastuti Hidayat; Suryo Wiyono
Jurnal Perlindungan Tanaman Indonesia Vol 23, No 2 (2019)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jpti.41402

Abstract

A survey on several cucumber cultivation areas in West Java, Central Java, Yogyakarta, and East Java found many plants showing typical Begomovirus symptoms such as yellow mosaic, cupping, and vein banding. This study was aimed to determine disease frequency, detection and molecular characterization of the causal virus of those symptoms on cucumber in Java. Sampling was conducted by purposive sampling by collecting 50 symptomatic plants from each location in West Java (Indramayu, Subang, and Bogor), Central Java (Brebes and Klaten), Yogyakarta (Kulon Progo), and East Java (Nganjuk, Kediri, and Tulungagung). The detection and disease frequency was determined based on DIBA test using a specific antiserum of Tomato leaf curl New Delhi virus (ToLCNDV) and Squash leaf curl virus (SLCV). The identification of nucleic acid was conducted by PCR using specific primer of ToLCNDV and SLCV, DNA cloning, and sequencing. The results of serological detection showed the disease frequency of ToLCNDV and SLCV ranged from 92.77-100% and 78.33-93.3%, respectively. PCR using specific primer of ToLCNDV successfully amplified the coat protein gene at a size of 600 bp from all samples. Homology nucleotide and amino acid sequences among ToLCNDV Java isolate ranging from 95.6-99.2% and 99.7-100%. ToLCNDV isolates Java had highest nucleotide and amino acid sequences similarity with cucumber isolate from Klaten, Indonesia (AB613825) ranging from 96.1-98.1% and 99.7-100%, and was considered as “Indonesia” strain. SLCV not amplified on all samples by PCR using specific primer, indicating it might not present yet on cucumber in Java.
Biology and the Statistic Demographic of Aphis glycines Matsumura (Hemiptera: Aphididae) on the Soybean with Plant Growth Promoting Rhizobacteria (PGPR) Treatment Hermanu Triwidodo; Anggun Agustini; Listihani Listihani
Jurnal Perlindungan Tanaman Indonesia Vol 24, No 1 (2020)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jpti.49846

Abstract

A correction has been published:Erratum to “Biology and the Statistic Demographic of Aphis glycines Matsumura (Hemiptera: Aphididae) on the Soybean with Plant Growth Promoting Rhizobacteria (PGPR) Treatment” [Jurnal Perlindungan Tanaman Indonesia, 24(1), 54 ̶ 60]Plant Growth Promoting Rhizobacteria (PGPR) applied to different plants may suppress pests population developments. This research was to study the capability of a commercial PGPR product contained Bacillus polymyxa and Pseudomonas fluorescens in suppressing population developments of Aphis glycines Matsumura (Hemiptera: Aphididae). The biology and demographic statistics of A. glycines reared on soybean with and without the PGPR applications were compared. The PGPR suspensions of 5 g formulation per liter water were used to soak soybean seed for 15 minutes and to water soybean plant 2 weeks after transplanting. Cohorts of 65 first instar A. glycines of each treatment were observed daily and individual mortality, molting, and fecundity were recorded until the last individual dead. Second instar stadium of A. glycines reared on treated plant lasted longer than those reared on untreated plant, i.e. 1.4 and 1.1 days, respectively. These resulted on a longer life cycle for A. glycines reared on treated plant than on untreated plant, i.e. 4.9 and 4.5 days, respectively. In turn, it caused the A. glycines population to experienceslower growth on treated plants than on untreated plants.  The values of A. glycines GRR, Ro, rm, T and DT on treated plants were 71.834, 57.780, 0.557, 7.287 and 1.245, consecutively; whilst that of untreated plants were 104.861, 63.326, 0.586, 7.084 and 1.184, respectively.
Molecular Identification of Sweet potato virus C on Sweetpotato in Bali, Indonesia Listihani Listihani; Dewa Gede Wiryangga Selangga
Jurnal Perlindungan Tanaman Indonesia Vol 25, No 1 (2021)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jpti.64545

Abstract

A survey was conducted in several sweet potato cultivations in Bali Province. Survey found that many plants exhibited potyvirus symptom, such as chlorosis blotches. This study was to determine disease incidence, detection and identification of the virus causing these symptoms on sweet potato plants in Bali. Samples were collected by purposive sampling of 10 plants from each location in Bali (Denpasar, Gianyar, Badung, Buleleng, Tabanan, Klungkung, Karangasem, Jembrana, Bangli). Disease insidence was observed based on viral symptoms in the field. Identification of nucleic acids was done using Potyvirus universal primer and DNA sequencing. Disease incidence in Bangli, Buleleng, and Denpasar Regencies was > 50%. RT-PCR and CiFor/CiRev Potyvirus universal primers successfully amplified ± 700 bp of CI genes from all samples from Bangli, while samples from 8 other districts were not amplified using the same primers. The SPVC isolate of sweet potato showed nucleotide and amino acid homology similarities with the sweet potato isolate from East Timor (MF572066), 96.8% and 97.4%, respectively and these were referred to the "Asian" strain. This indicates that SPVC has spread in East Java and Bali.
The Existence of Papaya ringspot virus-Papaya Strain on Cucumber in Gianyar, Bali Dewa Gede Wiryangga Selangga; I Ketut Widnyana; Listihani Listihani
Jurnal Perlindungan Tanaman Indonesia Vol 25, No 1 (2021)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jpti.64703

Abstract

Yellow mosaic symptoms were identified from cucumber production systems in Gianyar and were similar to symptoms of PRSV infection. Further research was conducted to determine diseases incidence and molecular characteristic of PRSV. Ninety leaf samples were collected from Gianyar by purposive sampling and disease incidence calculations were based on symptoms in the field. Detection and identification were done using a RT-PCR with specific primers of CP PRSV-P, CP PRSV-W and DNA sequencing. Disease incidences in the fields ranged between 5.81–66.87%. Specific DNA band 470 bp was successfully amplified from several cucumber leaf samples collected from Ubud, Payangan, Tegallalang, Sukawati, Gianyar, and Blahbatuh; but no DNA were amplified from all samples when using CP PRSV-W specific primer. Nucleotide and amino acid analysis showed nucleotides homology to isolates from Ubud, Payangan, Tegallalang, Sukawati, Gianyar, and Blahbatuh, i.e. 98.9–99.5% and 99.1–100%, respectively. Results indicated that genetic variation of PRSV-P from Gianyar was low. Furthermore, the nucleotides homology of PRSV-P isolates from Ubud, Payangan, Tegallalang, Sukawati, Gianyar, and Blahbatuh were with PRSV-P isolates which infected cucumbers from Nganjuk (LC311783) and Brebes (LC311784), while from native papaya collected in Bali Bali (LC223115) were 97.2–98.4% and 98.6–100%, respectively. Phylogenetic analysis confirmed that PRSV-P isolates from Indonesia were in the same cluster with Thailand isolates. The results showed that sources of PRSV-P inoculums spreading into new areas.
Screening of Endophytic Bacteria Isolated from Mimosa pudica in Bali Island Dewa Gede Wiryangga Selangga; Listihani
SEAS (Sustainable Environment Agricultural Science) Vol. 5 No. 1 (2021)
Publisher : Warmadewa University Press

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (512.158 KB) | DOI: 10.22225/seas.5.1.3303.50-57

Abstract

This study aims to determine the screening of endophytic bacteria isolatd from Mimosa pudica in Bali Island. This research was conducted from September 2020 to January 2021 at the Plant Disease Laboratory, Faculty of Agriculture, Udayana University, Denpasar. Isolation of endophytic bacteria was carried out by taking a sample of the root of the shy daughter plant which was then washed and dried on a tissue.. Hypersensitivity test followed Klement and Goodman (1967) by growing bacteria in a petri dish containing 100% TSA and NA.Tests were carried out by growing endophytic bacteria in 100% NA and TSA media. Isolation of the roots of the Mimusa pudica plant resulted in 43 isolates. The resulting isolates then passed the hypersensitive test so that 27 isolates were selected. A total of 27 isolates were then tested for hemolysis on blood agar. The final results obtained were 12 endophytic bacterial isolates which were then used in the next practicum. The results showed that the endophytic bacterial isolates were not able to inhibit the fusarium sp. and Phytophthora sp., when using the inhibition percentage formula. This is because the average diameter of the pathogenic fungi colonies in the control treatment (R1) with the average diameter of the pathogenic fungi colonies in the endophytic treatment (R2) has the same size, so that when calculated by the percentage inhibition formula it results in the number 0 which means no. there is a zone of inhibition.
The Isolation, Selection and Determination of Endophytic Bacteria from Bamboo, Gamal, Tulsi, and Alamanda Hermanu Triwidodo; Listihani listihani
SEAS (Sustainable Environment Agricultural Science) Vol. 5 No. 2 (2021)
Publisher : Warmadewa University Press

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (235.557 KB) | DOI: 10.22225/seas.5.2.4068.151-162

Abstract

Endophytic bacteria have many benefits, including increasing plant growth by producing phytohormones, increasing the production of mineral absorption, nitrogen fixation, reducing damage due to weather changes and increasing plant resistance to disease. Based on the above, it is necessary to select endophytic bacteria from various plants to be used as biocontrol agents. This study aims to obtain endophytic bacterial isolates that have the potential as biocontrol agents and plant growth supporters from bamboo shoots, Gamal, Tulsi, Lotus, and Alamanda. This research method includes sampling, endophytic bacteria isolation, hypersensitive, hemolysis, phosphate solvent, chitinolytic, proteolytic, and antagonist tests. Isolation of endophytic bacteria in 5 plants using 22 plant parts had a diversity of isolates. The isolated plant parts produced 1 to 7 isolates that had different morphology. The total isolates obtained were 59 isolates. In antagonistic observations, there was one isolate of endophytic bacteria that showed a clear zone when tested together with S. rolfsii, namely the isolate with code A24 from allamanda flower. From the data obtained, it is known that the endophytic bacterial isolates had an effect on inhibiting the growth of the pathogenic fungus S. roflsii, the endophytic bacterial isolates Consortium, A21 and the endophytic bacterial isolates A22 had no incidence of disease, while the bacterial isolates T00 (Bx) with an average disease incidence of 40% and 30% disease intensity. Meanwhile isolates A23, A24 and A25 had an average disease incidence ranging from 13.3%-26.6%, while controls had the highest disease incidence, namely 53.3% and disease intensity 66.6%.