. Giyanto
Department Of Plant Protection, Faculty Of Agriculture, IPB University Jln. Kamper, Kampus IPB Dramaga, Bogor, West Java 16680

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Isolation, identification, and detection of ACC deaminase gene-encoding rhizobacteria from rhizosphere of stressed pineapple Dori Kusuma Jaya; Giyanto Giyanto; Novik Nurhidayat; Sarjiya Antonius
Indonesian Journal of Biotechnology Vol 24, No 1 (2019)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1615.339 KB) | DOI: 10.22146/ijbiotech.39018

Abstract

ACC deaminase is a microbial cytoplasmic enzyme that cleaves ACC, a precursor of ethylene, in the stressed plant. The aims of this study were to isolate, identify, and detect the presence of ACC deaminase gene-encoding rhizobacteria from the rhizospheric soil of pineapple plants that have been exposed to abiotic and biotic stress, specifically herbicide, flooding, and Phytophthora spp. stress. A total of 49 rhizobacterial isolates were obtained, seven of which were observed for their growth on DF medium containing 3 mM L-1 ACC. The four best-growing isolates were selected for genomic DNA extraction. They were molecularly identified as Stenotrophomonas maltophilia (3), Burkholderia territorii (2A), Pseudomonas oryzihabitans (5B), and Bacillus tropicus (1E). A set of primers, 105F-acdS 5’-TGCCAAGCGTGAAGACTGC-3’ and 244R-acdS 5’-GGGTCTGGTTCGACTGGAT-3’, were constructed to amplify the ACC deaminase gene (acdS). Based on melt peak curve analysis, four products appeared to show a specific single peak at 86, 89, 87, and 89.5°C, indicating a single product was produced. In addition, a Blast search showed that these four products met the ACC deaminase feature and their acdS sequences were clustered into an ancestral group compared with the bacterial strains deposited in GenBank. These results suggest that ACC deaminase gene-encoding rhizobacteria from a pineapple plantation of tropical origin may affect the acdS sequences and may contribute to the host plant’s stress tolerance.
Morphological and Molecular Characteristics of Pratylenchus coffeae from the Origin of Robusta Coffee Plantation in Malang, East Java Aris Budiman; Supramana Supramana; Giyanto Giyanto
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.42481

Abstract

Pratylenchus coffeae is the most important plant-parasitic nematode in Robusta coffee plantations. Information regarding morphology, morphometric and molecular characters of P. coffeae has not been reported in Indonesia. This study is aimed to describe those characters of P. coffeae that attack Robusta coffee. Root samples were taken from Robusta coffee plantation in Malang, East Java. Nematode extractions was conducted using a mist chamber method. Morphology and morphometric characters were observed from the permanent nematode slides. Single nematode DNA extract was amplified at the D2D3 segment of 28S rRNA and ITS1-5.8S-ITS2 rRNA with universal primers. Amplicon was sequenced and analysed for phylogenetic tree relationships. Female morphological key character of P. coffeae observed are: lip with two annulations, four lateral lines, esophageal overlap with intestine ventrally, monodelphic, and truncated tail shape. Male spicules curved ventrally. Female morphometrics are: n=26, L = 556.4 μm, DGO = 2.4 μm, anterior gonad = 174.8 μm, a = 28.5, b = 6.1, b’ = 4.1, c = 20.1, c’ = 2.3, V = 81.7. A Male is smaller than a female with n=24, L = 505.9 μm,  a = 32.3, b = 5.5, b '= 3.9, c = 15.3, c' = 2.8 and T = 40.6. The molecular characters of P. coffeae were investigated for two isolates, namely SA1 and SA2.  Based on the D2D3 and ITS1-5.8S-ITS2 regions, isolate SA1 has similarity level of 99% and 97% to the P. coffeae  from NCBI. Similar result was shown by Isolate SA2 with similarity of 100% and 100% respectively. Phylogenetic tree analysis using Maximum Likelihood at the D2D3 segment of 28S rRNA and ITS1-5.8S-ITS2 regions showed that P. coffeae in this study was included in one clade with P. coffeae from several countries.
Phytonematodes Community and Polyphasic Character of Aphelenchoides varicaudatus on Garlic Plants in Tegal Regency, Central Java Mokhammad Danang Kusuma; Supramana Supramana; Giyanto Giyanto
Jurnal Perlindungan Tanaman Indonesia Vol 24, No 2 (2020)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jpti.49779

Abstract

The interception of Ditylenchus dipsaci, D. destructor, and Aphelenchoides fragariae, which are classified as quarantine pests, has been reported to occur on imported garlic bulbs used for consumption at several traditional markets in Bogor. This finding should increase awareness of the chance of garlic contamination in Indonesia’s garlic fields. This study aimed to evaluate the nematode community and determine polyphasic characters of Aphelenchoides species from garlic plantations. The study was conducted by sampling soil and plants from garlic plantations in Tuwel and Rembul Villages, Bojong District, Tegal Regency, Central Java in December 2018. Nematode extraction was done using a mist chamber for plant materials and sugar flotation – centrifugation methods for soil samples. Parameters measured included nematode species identity, absolute population and prominence indexes. Nematode identification was conducted based on their morphological characters. Further identification based on morphometric and molecular characters (polyphasic) conducted for nematodes species suspected as quarantine pests. Six generas of nematodes identified were Helicotylenchus, Aphelenchoides, Rotylenchulus, Aphelenchus, Criconemoides, and Tylenchus. Aphelenchoides sp. had the highest prominence value from plant tissue with a value of 6.32 and is categorized as a quarantine pest genus (A. fragariae) in garlic and was further identified to the species level. Further identification based on polyphasic characters showed the Aphelenchoides sp. found was A. varicaudatus.
Utilization of Liquid Smoke to Suppress Blood Diseases on Bananas and Its Effects on the Plant Growth Imas Aisyah; Meity Suradji Sinaga; Abdjad Asih Nawangsih; Giyanto Giyanto; Gustan Pari
AGRIVITA, Journal of Agricultural Science Vol 40, No 3 (2018): OCTOBER
Publisher : Faculty of Agriculture University of Brawijaya in collaboration with PERAGI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.17503/agrivita.v40i3.1390

Abstract

Liquid smoke is reported to be effective to inhibit some bacteria. The study aimed to evaluate the effects of liquid smoke (LS) from coconut shell (CS-LS), pinecone (P-LS), and oil palm branch (OPB-LS) on the incidences of blood disease, induced resistance to Ralstonia syzygii subsp. celebesensis, and plant growth. Two days after banana seedlings were treated with LS, ethylene, auxin, lignin, activities of the enzymes of peroxidase (POD) and phenylalanine ammonia lyase (PAL) on the root of the banana seedlings were measured. Then, the seedlings were inoculated by R. syzygii subsp. celebesensis. The plant growth and incidence of blood diseases were observed daily and the 31st day after the application of LS and this bacteria inoculation. The results showed that the CS-LS, P-LS, and OPB-LS at all tested concentration could suppress the incidence of blood diseases up to 100 %, induce resistance of banana seedlings to the R. syzygii subsp. celebesensis with increased levels of ethylene, auxin, lignin, activities of POD and PAL, and plant growth, significantly. Based on the effectiveness of this bacterial control and the ability to promote the growth of banana seedlings test, it is recommended that the most effective treatment is P-LS 0.5 %.
Identification of Quorum Quenching Bacteria and Its Biocontrol Potential Against Soft Rot Disease Bacteria, Dickeya dadantii Syaiful Khoiri; Tri Asmira Damayanti; Giyanto Giyanto
AGRIVITA, Journal of Agricultural Science Vol 39, No 1 (2017): FEBRUARY
Publisher : Faculty of Agriculture University of Brawijaya in collaboration with PERAGI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.17503/agrivita.v39i1.633

Abstract

Dickeya dadantii is one of newly found bacteria causing soft rot on orchids in Indonesia. Infected plants showed severe rot rapidly only in few days. An effort to control the bacteria was conducted by utilizing selected quorum quenching (QQ) inducer bacteria which produce AHL-lactonase by aiiA gene. The aims of this research were to screen and identify of quorum quenching bacteria, and also assayed their biocontrol potential ability against D. dadantii in laboratory. The screening of QQ bacteria was achieved using the anti-QS test, anti-microbial activity, and detection of aiiA gene using specific primer. The determination of the ability against D. dadantii was done using the soft rot assay on potato and orchid. Among thirty one bacteria isolates screened, four isolates (in succession namely B37, BT2, GG3, and GG6) were selected to control D. dadantii. All of these bacteria showed QQ ability to suppress the virulence of D. dadantii infection on orchids, significantly. Based on nucleotide sequences of 16S ribosomal RNA, those of bacteria isolates had the highest identity with Brevibacillus brevis, Bacillus cereus ATCC14579, Bacillus cereus ATCC14579 and Bacillus thuringiensis ATCC 10792. Brevibacillus brevis was reported for the first time as QQ bacteria in this study.
Detection of Wolbachia Endosymbiont in Several Agriculturally Important Insect Parasitoids in Bogor, Indonesia Mahardika Gama Pradana; Giyanto Giyanto; Seiichi Furukawa; Satoshi Nakamura; Damayanti Buchori
AGRIVITA, Journal of Agricultural Science Vol 41, No 2 (2019)
Publisher : Faculty of Agriculture University of Brawijaya in collaboration with PERAGI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.17503/agrivita.v41i2.1855

Abstract

Wolbachia that have been found to be maternally inherited in arthropods, including insects behave primarily as a reproductive parasite by inducing feminization and/or death among genetic males; causing cytoplasmic incompatibility, and activating parthenogenesis, in host insects. This research aims to detect the presence of Wolbachia in several parasitic insects and to determine the diversity of Wolbachia at the supergroup level. Insect genetic samples were collected, amplified using wsp (Wolbachia surface protein) gene and with the sequences then analyzed using species homologues drawn from the Genbank database. These activities were dedicated to detect intracellular presence of Wolbachia and assembled the phylogenetic tree of the respected bacteria. Phylogenetic analysis confirmed that Wolbachia existed in the samples belonged to the supergroup A (found in Cherapron sp.) and supergroup B (found in E. japonica). Wolbachia were detected in seven out of nine wasps i.e. Anagyrus lopezi, Ceraphron sp., Microplitis manilae, Brachymeria lasus, Scelionidae sp01, Trichogramma sp, and Exorista japonica. Single infection by Wolbachia supergroup A was detected in A. lopezi, Ceraphron sp., M. manilae, and Scelionidae sp01. Meanwhile single infection by Wolbachia supergroup B was detected only in E. japonica. Double infection by both supergroups occurred in B. lasus and Trichogramma sp. samples.
The Endophytic Bacteria, Salicylic Acid, and their Combination as Inducers of Rice Resistance Against Xanthomonas oryzae pv. oryzae Christoffol Leiwakabessy; Meity Suradji Sinaga; Kikin H. Mutaqien; Trikoesoemaningtyas Trikoesoemaningtyas; Giyanto Giyanto
AGRIVITA, Journal of Agricultural Science Vol 40, No 1 (2018): FEBRUARY
Publisher : Faculty of Agriculture University of Brawijaya in collaboration with PERAGI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.17503/agrivita.v40i1.1029

Abstract

Bacterial leaf damage or blight brought by bacteria Xanthomonas oryzae pv. oryzae (X. oryzae pv. oryzae) is considered as an extremely serious disease of rice worldwide, including Indonesia. Induced resistance using chemical and biological agents was considered as a method to control the disease. The objectives of this research were to analyze of endophytic bacteria (Lysinibacillus sphaericus/L.sphaericus) and salicylic acid as the inducers of rice resistance against X. oryzae pv. oryzae. This study used three-factorial incompletely randomized designs to study the effect of endophytic bacteria and salicylic acid on three varieties of rice. The results of the study showed that endophytic bacteria and salicylic acid induced rice resistance to X. oryzae pv. oryzae. This endophytic bacterium was also able to increase the plant height (AUPHGC), the number of tillers (AUNTGC), and the number of productive tillers. The application of salicylic acid at the concentrations of 10 mM was able to suppress the BLB disease and increased the number of tillers and 1000-grain weight. The result of study showed that the endophytic bacteria and salicylic acid could induce resistance of rice varities against BLB disease and increased the number of productive tillers.