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All Journal HAYATI Journal of Biosciences Jurnal Agronomi Indonesia (Indonesian Journal of Agronomy) Jurnal Ilmu Pertanian Indonesia Jurnal Fitopatologi Indonesia Jurnal Pengolahan Hasil Perikanan Indonesia Jurnal Akuatika Jurnal Ilmu Dasar Microbiology Indonesia BIOTROPIA - The Southeast Asian Journal of Tropical Biology BERKALA SAINSTEK Biota: Jurnal Ilmiah Ilmu-Ilmu Hayati Indonesian Journal of Agricultural Science Indonesian Journal of Biotechnology Jurnal Perlindungan Tanaman Indonesia Journal of Tropical Biodiversity and Biotechnology UNEJ e-Proceeding JURNAL HAMA DAN PENYAKIT TUMBUHAN TROPIKA ANNALES BOGORIENSES Marine Research in Indonesia Biosaintifika: Journal of Biology & Biology Education Menara Perkebunan International Journal of Environment, Engineering, and Education International Journal of Environment, Engineering & Education International Journal of Oil Palm
Antonius Suwanto
Department Of Biology, Faculty Of Mathematics And Natural Science, IPB University, Bogor, Indonesia
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Civil Engineering, Building, Construction & Architecture Control & Systems Engineering Decision Sciences, Operations Research & Management Earth & Planetary Sciences Economics, Econometrics & Finance Education Engineering Environmental Science Immunology & microbiology Materials Science & Nanotechnology Mathematics Medicine & Pharmacology Physics Veterinary

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Konstruksi pustaka cDNA dari daun klon karet AVROS 2037 yang diinfeksi patogen Corynespora cassiicola Construction of a cDNA library from leaf of AVROS 2037 rubber clone infected by Corynespora cassiicola pathogen . NURHAIMI-HARIS; Hajrial ASWIDINNOOR; Antonius SUWANTO; Maggy T. SUHARTONO; Nurita TORUAN-MATHIUS; Agus PURWANTARA
Menara Perkebunan Vol. 73 No. 2: 73 (2), 2005
Publisher : INDONESIAN OIL PALM RESEARCH INSTITUTE

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v73i2.156

Abstract

SummaryConstruction of cDNA library derived fromtranscripts made under certain condition is animportant first step to understand diseaseresistant mechanisms. To identify rubber genesor transcripts involved in defense responsetoward Corynespora cassiicola, cDNA librarywas constructed using rubber clone AVROS2037, one of resistant clone to this pathogen.cDNA library was constructed based on thestrategy of leaves infection using conidia, withthe assumption that transcript expression relatedto defense response would be induced bypathogen infection. RNA was isolated from leavesthree days after inoculation with conidia ofC. cassiicola. Steps involved in the cDNA libraryconstruction were RNA isolation, mRNApurification, cDNA synthesis, vector modifcation,cDNA insert ligation, plasmid transformation andclone verifications. Each gram of leaf producedapproximately 300  g RNA, and 0.25% of themwas mRNA. The mRNA was used to synthesizedcDNA. Ligation of cDNA and modified vectorwas facilitated by restriction enzyme SfiI. Theconstructs were transformed into the E. coliDH5 competent cells. A total of 8000 colonieswere produced. Random examination of 270colonies showed that approximately 93% of thesecolonies carried plasmid vector with DNA insertsize of 200 – 2000 bp, with average size of 500 –800 bp. cDNA library construction of rubberleaves from AVROS 2037 clone as well as somenecessary modification steps are presented in thispaper.RingkasanKonstruksi pustaka cDNA yang me-ngandung transkrip yang diekspresikan dalamkondisi tertentu merupakan tahap awal yangsangat penting dalam berbagai studi biologi.Untuk mengidentifikasi gen karet atau transkripyang berperan dalam respons pertahanan tanamankaret terhadap Corynespora cassiicola, pustakacDNA dibuat dengan menggunakan daun klonAVROS 2037 yang merupakan salah satu klonresisten terhadap patogen tersebut. PustakacDNA dibuat berdasarkan strategi menginfeksidaun dengan konidia C. cassiicola denganpertimbangan bahwa ekspresi transkrip yangberperan dalam respons pertahanan akandiinduksi oleh adanya infeksi patogen. Dengandemikian pustaka cDNA yang dibuat diharapkanmengandung gen atau bagian gen yang ber-hubungan dengan respons pertahanan. RNAdiisolasi dari daun setelah daun diinokulasiselama tiga hari dengan konidia C. cassiicola.Beberapa tahapan telah dilakukan, dimulaidengan isolasi RNA, pemurnian mRNA, sintesiscDNA, modifikasi vektor kloning, ligasi fragmencDNA utas ganda dengan vektor kloning sertatransformasi hasil ligasi ke bakteri Escherichiacoli DH5 kompeten. Dari setiap gram jaringandaun berhasil diisolasi RNA sekitar 300 g, dandari jumlah tersebut sekitar 0,25% mRNA dapatdiisolasi. mRNA yang diisolasi digunakan untuksintesis cDNA. cDNA dipotong dengan enzimrestriksi SfiI dan diligasi ke vektor plasmid yangdimodifikasi dengan menyisipkan situs enzimSfiI. cDNA-vektor rekombinan ditransformasi kedalam sel bakteri E. coli DH5 kompeten meng-gunakan metode standar. Transformasi konstrukini menghasilkan 8.000 koloni. Pengujian PCRterhadap 270 koloni yang dipilih secara acakmengindikasikan bahwa sekitar 93% kolonitersebut membawa cDNA sisipan dengan ukuranfragmen cDNA yang menyisip berkisar antara200 sampai 2000 bp. cDNA sisipan terbanyakterdapat pada ukuran antara 500 – 800 bp. Dalamtulisan ini dibahas tahap demi tahap proses yangdilakukan untuk membuat pustaka cDNA asaldaun karet klon AVROS 2037 serta beberapamodifikasi yang diperlukan.
Engineering of LysR-type Regulator DmlR in Burkholderia ubonensis CP01 to Enhance Its Antifungal Production against Ganoderma boninense Budinarta, Widyah; Purnamasari, Maria Indah; Hermosaningtyas, Anastasia Aliesa; Arifudin Rafif , Muhammad Ghildan; Prihatna , Cahya; Suwanto, Antonius
HAYATI Journal of Biosciences Vol. 33 No. 1 (2026): January 2026
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.33.1.28-43

Abstract

The utilization of an antifungal substance, occidiofungin and burkholdine, derived from Burkholderia ubonensis CP01 has displayed promising results in the management of basal stem rot caused by Ganoderma boninense. The study aims to further enhance the antifungal production of B. ubonensis CP01 through genetic modification. Through comparative genetic analysis, we identified the dmlR gene in B. ubonensis CP01, which is homologous to the scmR gene, a LysR-type transcriptional regulator (LTTR), in B. thailandensis. Deleting the dmlR gene in CP01 resulted in a complete loss of antifungal synthesis. In contrast, overexpression of this gene led to a substantial increase in antifungal production, as determined by an agar well diffusion assay. These findings suggest that dmlR acts as a positive regulator of antifungal gene expression in B. ubonensis CP01. RP-HPLC analysis revealed that the mutant strain overexpressing the dmlR gene (mutant WB12) produced a higher peak at the 24-25 minute elution time. Previous high-resolution mass spectrometry analysis by our group identified the compound at this peak as six analog compounds with monoisotopic masses similar to those of cyclic lipopeptides, including occidiofungin and burkholdine. The WB12 mutant exhibited approximately 15% higher concentrations of antifungal compounds than the wild type. Additionally, whole genome sequencing confirmed that the introduced dmlR gene had been integrated into the locus on chromosome 2 of B. ubonensis CP01. LTTRs play a pivotal role in regulating the production of antifungal agents in CP01. Furthermore, it highlights the potential for manipulating LTTRs to enhance the desirable characteristics of the Burkholderia genus in regard to the production of secondary metabolites.
Co-Authors . EFRIWATI . Melani . NURHAIMI-HARIS A A Hermosaningtyas A A Hermosaningtyas, A A ADI YULANDI Agus PURWANTARA Agus Purwito Agustin Wydia Gunawan Agustina, Delia Alfred Michael ALINA AKHDIYA RUSMANA AMARILA MALIK AMARILA MALIK Andi Khaeruni Andi Khaeruni Andreas Adhi Satya Andreas Adhi Satya ANJA MERYANDINI Anja Meryandini ARI SUSILOWATI Arifudin Rafif , Muhammad Ghildan Arild Ranlym Arifin Arina Amalia Putri Aris Tjahjoleksono Aris Tri Wahyudi ARTINI PANGASTUTI Asmini Budiani Basil J NIKOLAU BIBIANA W LAY BUDI TJAHJONO BUDI TJAHJONO Budi Tjahjono Budi Tjahjono Budinarta, Widyah C Hanny Wijaya CAHYA PRIHATNA Cahya Prihatna Cahya Prihatna CECILIA ANNA SEUMAHU CECILIA ANNA SEUMAHU DEBORA HADISUSANTO Delia Agustina Desniar - - DIAH ISKANDRIATI DIANA ELIZABETH WATURANGI Djoko Santoso Dondin Sajuthi DWI SURYANTO Dyah Kusuma Anggraini EDI HUSEN Edi Husen Edi Husen ERNIN HIDAYATI Esti Puspitasari ESTI PUSPITASARI ESTI PUSPITASARI Esti Utarti Eunice Limantara EVELINE AYU Felicia Felicia Felicia Kartawidjajaputra Ferymon Mahulette Ferymon Mahulette Ferymon Mahulette, Ferymon G. A. Wattimena Gayuh Rahayu Griselda Griselda Griselda Herman Natadiputri Griselda, Griselda Hajrial ASWIDINNOO HAJRIAL ASWIDINNOOR Hariyatun Hariyatun Hariyatun, Hariyatun Hermosaningtyas, Anastasia Aliesa Iman Rusmana Inez H.S. Loeddin Suharsono It Jamilah IVAN HANJAYA JOANITA SADELI KATHARINA JESSICA Kusharyoto, Wien Lilis Nuraida Linda Wati LISTYA UTAMI KARMAWAN Ludovika Jessica Virginia MAGGY T HENAWIDJAJA Maggy T Suhartono Maggy T Suhartono MAGGY T. SUHARTONO Maggy T. Suhartono MAGGY T. SUHARTONO MAGGY THENAWIJAYA SUHARTONO Maria Dita Febriani Lumban Gaol Maria Indah Purnamasari Maria Indah Purnamasari Maria Indah Purnamasari Maria Sugiharti Meity S. Sinaga MEITY SURADJI SINAGA Muhamad Azwar Syah Muhammad Agus Muljanto MUHAMMAD ZAIRIN Jr. Ni Nyoman Tri Puspaningsih NISA RACHMANIA MUBARIK NURITA TORUAN-MATHIUS NURUL AINI PRIHASTO SETYANTO Prihasto Setyanto Prihatna , Cahya Puspitasari, Esti QURROTA A’YUN R. Sapto Hendri Boedi Soesatyo Raden Ajie Syahbarie RAHAYU WIDYASTUTI RASTI SARASWATI RASTI SARASWATI Rasti Saraswati Rasti Saraswati RATIH DEWI HASTUTI RATNA SETYANINGSIH RATNA SETYANINGSIH RIDWAN AFFANDI Rika Indri Astuti ROB HARLING ROHANI CINTA BADIA GINTING Rustam, Yepy Hardi Satya, Andreas Adhi Sheila Sutanto Sheila Sutanto, Sheila SUMARDI Suryo Wiyono SUSAN SOKA SUSILOWATI1 SUSILOWATI1 T RESNAWATI P URWADARIA Tan Watumesa Agustina TARUNI SRI PRAWAST MIEN KAOMINI ANY ARYANI DEDY DURYADI SOLIHIN Tati Barus TEMMY DESILIYARNI TRESNAWATI PURWADARIA VICKY MEICY WALTER ERDELEN WIDANARNI WIDANARNI Widanarni Widanarni Wien Kusharyoto Yasinta Ratna Esti Wulandari Yepy Hardi Rustam Yogiara Yogiara YULIN LESTARI Yusminah Hala