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All Journal JURNAL PEMULIAAN TANAMAN HUTAN Indonesian Journal of Biotechnology Journal of Tropical Biodiversity and Biotechnology Agrin : Jurnal Penelitian Pertanian Journal of Tropical Crop Science Menara Perkebunan Nusantara Science and Technology Proceedings

Riza Arief Putranto

Unknown Affiliation

Author-ID : 3618383

Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Economics, Econometrics & Finance Engineering Environmental Science Immunology & microbiology Law, Crime, Criminology & Criminal Justice Materials Science & Nanotechnology Medicine & Pharmacology

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Structure-based virtual screening of bioherbicide candidates for weeds in sugarcane plantation using in silico approaches Galuh Wening PERMATASARI; Riza Arief PUTRANTO; Happy WIDIASTUTI
E-Journal Menara Perkebunan Vol 88, No 2 (2020): Oktober,2020
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v88i2.379

Weeds in sugarcane have negatively affected the sugar yield rate. Several approaches have been carried out to overcome the weeds, including the usage of diuron as synthetic herbicide. However, the long-term usage of diuron is known to have a negative effect leads to the production of 3,4- Dichloroaniline responsible for soil leach and bioaccumulation. Therefore, this study aimed to find a potential natural herbicide. By mimicking the diuron's mode of action which inhibits the process of photosynthesis through blocking the Photosystem II protein D1 (psbA) of the weeds, fourteen compounds as potential candidate bioherbicides were virtually docked by PyRx v0.9.5 software to the specific site. Three important species of the weeds were chosen including Eleusine indica, Praxelis clematidea, and Momordica charantia. The binding affinity score was further calculated and ranked to screen the top six compounds as bioherbicide candidates. Interaction of each complex and the biological activity prediction were then performed by Discovery Studio software and PASS server, respectively. Aurachin P, Aurachin A, and Cyanobacterin were placed in the top ranked compounds with high binding affinity score around -6 to -9 kcal mol-1 toward the psbA. The amino acid interaction involved in the complex shows 50-90% similar to the control, psbA and diuron complex. Besides, the biological activity prediction of Aurachin P, Aurachin A, and Cyanobacterin exhibits the terms related to the inhibition of photosynthesis process via enzymatic pathway. Thus, the active compounds might have inhibition action in the photosynthesis process and control the weeds in sugarcane.

Identifikasi famili gen putatif penyandi protease inhibitor dengan pendekatan in silico komparatif pada genom Hevea brasiliensis Muell. Arg (Identification of putative gene family encoding protease inhibitors by in silico comparative analysis in Hevea brasiliensis Muell. Arg genome) Irfan MARTIANSYAH; Riza Arief PUTRANTO; Nurul KHUMAIDA
E-Journal Menara Perkebunan Vol 85, No 2 (2017): Oktober 2017
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

AbstractProtease inhibitors (PIs) are small proteins that form complexes with proteases and inhibits their proteolytic activity. Its potential application as an antimicrobial agent has been studied. Most of PIs' molecule size is around 8-22 kDa depending on their protein families.To date, on the basis of sequence homologies of inhibitor domains, PIs have been classified into 48 families in all organisms. In plant, more than 13 families of PIs have been identified but they were not widely identified in the rubber tree (Hevea brasiliensis Muell.Arg). In the present study, 40 putative HbPI genes, designated as HbPI01 to HbPI36, were identified from whole-genome sequence of rubber tree clone Reyan 7-33-97 using 7453 scaffolds available online in NCBI with the accession code: LVXX01000000. Multiple sequence alignment using MUSCLE algorithm discovered seven conserved motifs (Motifs I-VII) among HbPIs. Phylogenetic analysis of 50 and 36 PI amino acid residues of 32 scaffolds containing putative PI genes from Arabidopsis thaliana and H. brasiliensis showed three clusters (families): LTP-I, SERPIN and LTP-II. LTP-I has 23 putative HbPI genes (HbPI05 to HbPI27) and 12 AtPI genes. SERPIN, a family member of serine protease inhibitor group, has 11 putative HbPI genes (HbPI01 to HbPI04 and HbPI28 to HbPI34) and 22 AtPI genes. LTP-II has 2 putative HbPI genes (HbPI35 to HbPI36) and 16 AtPI genes. In conclusion, this work provides valuable information for further functional characterization of HbPI genes in H. brasiliensis.[Key words: protease inhibitor, genome-wide, scaffold, in silico, Hevea brasiliensis]. AbstrakProtease inhibitor (PI) merupakan protein yang membentuk kompleks dengan protease dan menghambat aktivitas proteolitik dari enzim tersebut. Potensi penggunaan protease inhibitor sebagai agensia antimikroba telah diketahui. Kebanyakan PI memiliki ukuran molekul sekitar 8-22 kDa bergantung pada familinya. Saat ini, PI dapat diklasifikasikan menjadi 48 famili di seluruh organisme berdasarkan kemiripan sekuen dari domain inhibitornya. Pada tanaman, lebih dari 13 famili PI telah diketahui tetapi pada tanaman karet (Hevea brasiliensis Muell.Arg) belum diidentifikasi. Pada penelitian ini, sebanyak 40 gen putatif penyandi PI (HbPI01 hingga HbPI36) telah berhasil diidentifikasi dari 7453 scaffold genom utuh tanaman karet klon Reyan 7-33-97 yang tersedia secara daring dengan kode aksesi LVXX01000000. Penjajaran sekuen menggunakan algoritma MUSCLE memper-lihatkan tujuh konservasi motif (Motif I-VIII) pada famili gen putatif HbPIs. Analisis pohon filogenetik dari tanaman Arabidopsis thaliana dan H. brasiliensis sebanyak 50 dan 36 sekuen residu asam amino dari 32 scaffold yang mengandung gen putatif PI menunjukkan adanya tiga klaster besar, yaitu famili LTP-I, SERPIN dan LTP-II. LTP-I terdiri dari 23 gen putatif HbPI (HbPI05 hingga HbPI27) dan 12 gen AtPI. SERPIN yang merupakan anggota kelas protease inhibitor serin terdiri dari 11 gen putatif HbPI (HbPI01hingga HbPI04 dan HbPI28 hingga HbPI34) dan 22 gen AtPI. LTP-II terdiri dari 2 gen putatif HbPI (HbPI35 hingga HbPI36) dan 16 gen AtPIs. Penelitian ini menghasilkan informasi penting untuk melakukan karakterisasi fungsional lebih mendalam pada gen HbPI tanaman karet ke depannya.[Kata kunci: protease inhibitor, genome-wide,scaffold, in silico, Hevea brasiliensis].

Evaluation of eleven reference genes for Reverse Transcriptase Quantitative PCR of rubber tree under water deficit Evaluasi sebelas gen referensi untuk Reverse Transcriptase Quantitative PCR pada tanaman karet tercekam kekeringan Riza Arief PUTRANTO; Julie LECLERCQ; Pascal MONTORO
E-Journal Menara Perkebunan Vol 83, No 2: Desember 2015
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

AbstrakReverse Transcriptase Quantitative PCR (RT-qPCR) merupakan teknik yang sangat ampuh untuk mendeteksi jumlah mRNA yang rendah dalam sel tanaman. Pengukuran akumulasi transkrip tersebut relatif terhadap kontrol ekspresi seperti gen-gen housekeeping. Keandalan teknik RT-qPCR ber-gantung pada pemilihan kontrol internal yang disebut pula gen referensi. Hal tersebut menjadi alasan kenapa validasi gen referensi disarankan untuk setiap set sampel cDNAs yang akan diguna-kan pada eksperimen RT-qPCR baru. Penelitian ini bertujuan untuk menganalisis stabilitas sebelas gen-gen housekeeping terpilih pada tiga organ Hevea brasiliensis (daun, kulit batang dan akar) tercekam kekeringan moderat selama 15 hari. RNA total diisolasi dari 18 sampel yang terdiri dari tanaman kontrol dan tercekam kekeringan pada hari ke-0 (D0), ke-5 (D5) dan ke-15 (D15). Kualitas cDNA yang disintesis divalidasi dengan amplifikasi PCR menggunakan primer HbActin. Kesebelas pasangan primer penyandi gen-gen housekeeping pada Hevea (HbActin, HbelF1Aa, HbUBC4, HbUBC2b, HbYLS8, HbRH2b, HbRH8, HbUBC2a, HbαTub, Hb40S dan HbUBI) divalidasi dengan amplifikasi PCR. Nilai Crossing-point (Cp) yang diukur dengan metode derivatif kedua pasca analisis RT-qPCR mengungkapkan nilai rerata Cp yang lebih tinggi secara signifikan untuk kesebelas gen housekeeping pada titik sampling D5 dibanding D0 dan D15. Studi ini menyarankan bahwa metode perhitungan koefisien keragaman (CV) sederhana dapat digunakan untuk menentu-kan peringkat gen referensi pada tanaman karet berdasarkan ekspresinya yang stabil. Lima gen housekeeping (HbRH2b, HbRH8, HbUBC4, HbαTUB dan HbActin) dapat digunakan sebagai gen referensi untuk analisis RT-qPCR pada Hevea brasiliensis yang tercekam kekeringan moderat. Gen HbRH2b memiliki ekspresi paling stabil dibanding yang lain.AbstractReverse Transcriptase Quantitative PCR (RT-qPCR) is a powerful technique in order to detect low abundance of mRNA in the plant cell. The measurement of transcript abundance is relative to the control of expression such as housekeeping genes. Therefore, the reliability of RT-qPCR depends essentially to the choice of these internal controls also called reference genes. That is the reason why a prior validation of reference genes is suggested for every set of cDNA samples used in a new RT-qPCR experiment. This study aimed to analyze the stability of eleven selected house-keeping genes in three Hevea brasiliensis tissues (leaf, bark and root) under15 days of moderate water deficit. Total RNA was isolated from 18 samples consisting of control and stressed-plants collected at day-0 (D0), day-5 (D5) and day-15 (D15).The quality of cDNA synthesized was examined by PCR using HbActin primer. The eleventh primers encoding Hevea housekeeping genes (HbActin, HbelF1Aa, HbUBC4, HbUBC2b, HbYLS8, HbRH2b, HbRH8, HbUBC2a, HbαTub, Hb40S and HbUBI) were validated using PCR amplification. The Crossing-point (Cp) values were measured using a second derivative method after RT-qPCR analysis revealing a significantly higher Cp mean values for 11 housekeeping genes at D5 compared to D0 and D15 sampling points. This study suggests that a simple coefficient of variation (CV) method can be used to rank Hevea reference genes based on its stable expression. Five housekeeping genes (HbRH2b, HbRH8, HbUBC4, HbαTUB and HbActin) can be used for RT-qPCR analysis in Hevea brasiliensis under moderate water deficit. The HbRH2b gene was the most stable among others.

Molecular markers and their application for DNA fingerprinting and genetic diversity studies in Coffea species Marka molekuler dan penerapannya untuk studi sidik jari DNA dan keragaman genetik pada spesies Coffea . PRIYONO; Riza Arief PUTRANTO
E-Journal Menara Perkebunan Vol 82, No 1: Juni 2014
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

AbstrakStrategi klasik yang meliputi perbandingan anatomi, fisiologi dan sitogenetika telah banyak diterapkan untuk mengidentifikasi karakter tertentu serta untuk menentukan keragaman dan hubungan antar dan intra spesies. Namun, saat ini penanda molekuler telah melengkapi strategi sebelumnya dengan sangat cepat. Berbagai jenis penanda molekuler digunakan untuk menilai tingkat polimorfisme DNA. Penanda molekuler ini diklasifikasikan sebagai penanda berbasis hibridisasi dan berbasis Polymerase Chain Reaction (PCR). Dalam beberapa tahun terakhir, sistem penanda DNA yang berbeda seperti Restriction Fragment Length Polymorphisms (RFLPs), Random Amplied Polymorphic DNAs (RAPDs), Amplified Fragment Length Polymorphisms (AFLPs), Simple Sequence Repeats (SSRs) yang juga disebut Mikrosatelit, Single Nucleotide Polymorphims (SNPs) dan lain-lain telah dikembangkan dan diterapkan pada berbagai spesies tanaman. Penanda molekuler ini dapat digunakan untuk sidik jari DNA dan studi keragaman genetik. Sidik jari berdasarkan DNA telah banyak digunakan dalam ilmu forensik, juga memiliki berbagai aplikasi dalam pemuliaan tanaman. Tulisan ini memberikan overview tentang berbagai penanda molekuler dan aplikasinya untuk sidik jari dan kajian keragaman genetik tanaman berdasarkan DNA pada berbagai spesies tanaman, dan secara khusus pada Coffea sp.AbstractConventional strategies including comparative anatomy, physiology and cytogenetics were applied to identify the certain character as well as to determine inter- and intra-species diversity and relationships. However, more recently molecular markers have very rapidly complemented the previous strategies. Various types of molecular markers are used to assess DNA polymorphism. They are classified as hybridization-based markers and polymerase chain reaction (PCR) based markers. In recent years, different DNA marker systems such as Restriction Fragment Length Polymorphisms (RFLPs), Random Amplied Polymorphic DNAs (RAPDs), Amplified Fragment Length Polymorphisms (AFLPs), Simple Sequence Repeats (SSRs) which also called as microsatellites, Single Nucleotide Polymorphims (SNPs) and others have been developed and applied to a range of plant species. These molecular markers can be used for DNA fingerprinting and genetic diversity study. DNA fingerprinting has been widely used in forensic science, but is has also a variety of application in plant breeding. This paper provides an overview about various molecular markers and their application for DNA plant fingerprinting and genetic diversity, especially in Coffea sp.

Purification, characterization, and bioassay of putative protease inhibitors from Hevea brasiliensis latex Riza Arief PUTRANTO; . SISWANTO; Agustin Sri MULYATNI; Asmini BUDIANI; Radite TISTAMA
E-Journal Menara Perkebunan Vol 84, No 2 (2016): Desember 2016
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Lateks yang menyerupai cairan susu putih diperoleh dari penyadapan kulit batang tanaman karet (Hevea brasiliensis). Lateks merupakan sitoplasma dari jaringan pembuluh bernama latisifer yang didalamnya terkandung berbagai macam komponen, termasuk protein-protein penting. Berbagai jenis enzim yang memiliki fungsi terkait pertahanan tanaman dari serangan patogen dan pelukaan telah berhasil dideteksi di dalam lateks, di antaranya protease inhibitor (PI). Protease inhibitor memiliki aktivitas senyawa antifungi sehingga berpotensi untuk dimanfaatkan sebagai biofungisida. Pada penelitian ini, protease inhibitor putatif yang berasal dari serum B (lutoid) lateks tanaman karet telah berhasil diisolasi menggunakan teknik Ion Exchange Chroma-tography. Dari total 70 fraksi protein yang diekstrak dari kolom, hanya 26 fraksi yang menunjukkan kadar protein yang terukur. Kandungan protease inhibitor putatif yang di-peroleh berkisar antara 0,0067 hingga 0,022 mL/g serum B dari hasil 3 fraksi terpilih. Aktivitas penghambatan terhadap empat enzim protease (subtilisin A, tripsin, α-kimotripsin, dan papain) menunjukkan karakteristik protease inhibitor putatif tersebut sebagai serine dan/atau cysteine inhibitor protease dengan persentase hambatan di atas 15% terhadap protease target. Hasil SDS-PAGE memperlihatkan pemisahan protein dominan yang diperkirakan merupakan protease inhibitor putatif dengan berat molekul sebesar 21,5 kDa. Uji bioassay aktivitas antifungi secara in vitro dari protease inhibitor memperlihatkan penghambatan pertumbuhan miselium dari fungi Ganoderma boninense, Sclerotium sp., dan Rigidosporus lignosus. [Kata kunci : protease inhibitor, Hevea brasiliensis, lateks, serum B, ion exchange chromatography]AbstractLatex, a milky white liquid, is the main product from rubber tree (Hevea brasiliensis). Latex is the cytoplasm of complex cellular networks named laticifers in which it contains many different components, including important proteins. Various types of enzymes carrying functions associated with plant defense against pathogen and wounding have been detected in latex in which one of these enzymes is protease inhibitor (PI). Plant protease inhibitor has tremendous potential as an antifungal agent which can be developed as biofungicide. In this work, protease inhibitors from B-serum (lutoid) of rubber tree latex were isolated and purified using Ion Exchange Chromatography (IEC) technique. Of the total 70 fractions of proteins extracted from the columns, only 26 fractions showed measurable levels of protein. The concentration of obtained putative protease inhibitors (three fractions of IEC) ranged from 0.007 to 0.022 mL/g B-serum. Inhibitory activity against four protease enzymes (subtilisin A, trypsin, α-chymotrypsin, and papain) showed the characteristics of Hevea putative protease inhibitors from B-serum as serine and/or cysteine protease inhibitors with more than 15% inhibitory activity of target protease. Based on SDS-PAGE visualization, the molecular weight of dominant protein considered as Hevea putative protease inhibitors was 21.5 kDa. In vitro bioassay test of antifungal activity for Hevea putative protease inhibitors showed reduced mycelium growth of Ganoderma boninense, Sclerotium sp., and Rigidosporus lignosus.[Keywords: protease inhibitor, Hevea brasiliensis, latex, B-serum, ion exchange chromatography]

Genetic mapping studies in Coffea sp using molecular marker methods Studi peta genetik pada Coffea sp. menggunakan metode penanda molekuler . PRIYONO; Riza Arief PUTRANTO
E-Journal Menara Perkebunan Vol 83, No 2: Desember 2015
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

AbstrakAnalisis genetik telah menjadi alat yang penting dalam pemuliaan tanaman untuk perbaikan sifat penting tanaman. Salah satu potensi terbesar dari analisis tersebut adalah identifikasi penanda molekuler yang berguna untuk pemetaan genetik. Pemetaan genetik merupakan salah satu langkah penting dari analisis genetik. Intisari dari semua pemetaan genetik adalah menempatkan koleksi pe- nanda molekuler pada posisi tertentu dalam genom. Hal tersebut dapat kemudian digunakan untuk meng- identifikasi lokus sifat kuantitatif (QTLs) dengan memanfaatan keragaman genetik alami yang tersedia dan meningkatkan sifat-sifat penting serta berharga. Sampai saat ini, tiga belas peta genetik telah dipublikasi dan tersedia pada Coffea sp. yang menciptakan database besar untuk kerangka genetik. Sebuah peta genetik terbaru dengan akses terbuka dan berfungsi sebagai referensi telah dibangun oleh International Coffee Genomics Network (ICGN). Peta tersebut tediri dari 3230 lokus, dengan panjang peta 1471 cM (1cm ~ 500 Kb) serta kepadatan satu penanda setiap 220 Kb. Peta-peta genetik pada tanaman kopi telah digunakan dari karakterisasi gen hingga analisis komparatif genom dengan spesies tanaman yang berbeda. Saat ini, pesatnya kemajuan teknologi New Genome Sequencing (NGS) untuk sekuensing DNA dan RNA memungkinkan validasi dari peta-peta genetik untuk prediksi QTLs serta gen-gen yang membawa sifat penting Coffea sp.AbstractGenetic analysis has become an important tool in plant breeding for crop improvement. One of their greatest potential appears to be the identification of molecular markers useful for genetic mapping. Genetic mapping is one of important steps in genetic analysis. The essence of all genetic mapping is to place a collection of molecular markers onto their respective positions on the genome. Thus, it leads to identification of new quantitative trait loci (QTLs) by making benefits of natural available genetic diversity.and to improve important and valuable traits. Until present, thirteen genetic maps were published and available in Coffea sp. creating a huge database for genetic framework. One most recent and open reference genetic map for robusta coffee has been generated by the International Coffee Genomics Network (ICGN) comprising 3230 loci, genetic size 1471 cM (1cM ~500 Kb), with an average density close to one marker every 220 Kb. The Coffea genetic maps have been utilized from gene characterization to genomic comparative analysis with different plant species. Nowadays, the feasibility of NGS for DNA and RNA sequencing allow the validation of genetic map related to the prediction of QTLs and adjacent genes related to important traits for Coffea sp.

In Silico Design and Validation of CRISPR-Cas13a System as a Potential Antiviral for SARS-CoV-2 in Indonesia Alfero Putra Iryanto; Christy; Muhammad Farrel Ewaldo; Anggia Prasetyoputri; Ratih Asmana Ningrum; Riza Arief Putranto; Akhirta Atikana
Nusantara Science and Technology Proceedings 2nd Bioinformatics and Biodiversity Conference
Publisher : Future Science

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.11594/nstp.2022.2107

The novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused a worldwide pandemic of coronavirus disease (COVID-19). Indonesia is one of the countries with large numbers of positive cases in Asia with certain dominant variants. Currently, there are no specific therapeutic agents against SARS-CoV-2. Therefore, the development of specific and effective therapeutic tools is urgently needed to overcome the pandemic. This study designed a CRISPR-Cas13a system strategy as a potential anti-SARS-CoV-2. We utilized comprehensive bioinformatics methods to identify a unique segment in the SARS-CoV-2 consensus sequence from Indonesia that is different from the related segment in the SARS-CoV. This unique segment was used as a specific target for SARS-CoV-2 Spike Protein to design a set of crRNA libraries. Off-target analysis and molecular docking simulation were performed to validate the specificity and to analyze interactions among the crRNA candidates, target RNA, and Cas13a. Our study identified a 17 amino acid unique segment on the Receptor Binding Domain (RBD) region. By using that unique segment, a total of 12 crRNA candidates were selected based on their GC content. Finally, based on the off-target and molecular docking validation, four crRNAs were selected as potential candidates for CRISPR-Cas13a-based antivirals. Although further validation with in vitro assays is important, the present study provides a comprehensive demonstration regarding the potential of CRISPR-Cas13a as a strategy for SARS-CoV-2 antiviral development. Considering the specific property of the CRISPR system, the present methodology can also be utilized to develop novel antiviral candidates for other RNA viruses.

PROFILING AKUMULASI TRANSKRIP GEN PADA AKAR BIBIT KELAPA SAWIT (Elaeis guineensis JACQ.) RENTAN DAN TOLERAN TERHADAP Ganoderma boninense Riza Arief Putranto; Rokhana Faizah; Dini Astika Sari; Vivi Restu Raharti; Irfan Martiansyah; Sapto Nugroho Hadi; Sri Wening; pri yono; Asmini Budiani
Agrin Vol 23, No 2 (2019): Agrin
Publisher : Jenderal Soedirman University

Dalam penelitian ini, analisis akumulasi transkrip menggunakan Reverse-Transcriptase Quantitative PCR (RT-qPCR) diikuti dengan analisis interaksi statistik telah dilakukan pada akar dari dua genotipe tanaman kelapa sawit (A09 dan 23) yang rentan dan toleran terhadap Ganoderma boninense. Tiga gen putatif (Eg#001, Eg#004, dan Eg#007) dari kelapa sawit diprediksi dapat digunakan sebagai biomarker dalam seleksi varietas tanaman kelapa sawit terhadap Ganoderma. Optimasi analisis RT-qPCR dilakukan dengan melakukan serial dilusi cDNA (1/5, 1/10, 1/25 dan 1/50) untuk mendapatkan nilai amplification plot dan melting curve yang ideal. Hasil analisis memperlihatkan serial dilusi 1/10 atau setara konsentrasi cDNA 50 ng/µL mendapatkan nilai Ct pada ambang 22-24 serta nilai melting curve sebesar 83,06. Secara umum, akumulasi transkrip gen pada genotipe 23 lebih melimpah dibandingkan dengan genotipe A09. Gen Eg#007 secara statistik memiliki akumulasi transkrip yang berbeda nyata pada genotipe 23 yang lebih tinggi (5,21E+00) dibandingkan pada genotipe A09 (1,93E+00). Di sisi lain, hasil identifikasi lokus ID pada genom kelapa sawit memperlihatkan bahwa gen Eg#007 merupakan penyandi protein yang tergolong dalam famili sulfotransferase (SOT) yang terkait dengan sistem pertahanan tanaman. Gen tersebut berpotensi mengungkap mekanisme aksi-reaksi antara G. boninense dan akar kelapa sawit. Kata kunci: kelapa sawit, Ganoderma, profil akumulasi transkrip, RT-qPCR, biomarker.

Genes expression analysis of EgUnk1, EgZFP2, and EgIPK2b in oil palm using Ct value correction and two relative quantification approaches Rokhana Faizah; Riza Arief Putranto; Sudarsono Sudarsono; Sri Wening; Dewi Sukma; Asmini Budiani
Indonesian Journal of Biotechnology Vol 29, No 3 (2024)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/ijbiotech.71816

The determination of transcript accumulation values significantly affects gene expression in oil palm. Various genes are involved in pathogen infection, including probable 2‐oxoglutarate‐dependent dioxygenase At5g05600 (EgUnk3), zinc finger protein 2‐like (EgZFP2), and inositol polyphosphate multikinase beta‐like (EgIPK2b). Gene expression is typically measured using relative quantitative methods to calculate differences in quantitative values in the expression levels of targeted genes compared to a reference gene. However, the effectiveness of these methods in assessing the expression of EgUnk3, EgZFP2, and EgIPK2b, which are involved in Ganoderma boninense infection in oil palm seedlings, requires evaluation. This study aimed to establish an effective and straightforward method for analyzing the expression of EgUnk1, EgZFP2, and EgIPK2b genes in oil palm seedlings infected with G. boninense, utilizing Ct value correction through regression coefficients on the 2‐ΔΔCt and E‐ΔΔCt approaches. A correlation regression revealed values of 0.28, ‐0.32, and 0.29 for delta Ct of EgUnk1, EgZFP2, and EgIPK2b, respectively. However, a negative correlation in the Ct mean was corrected by linear regression for the targeted genes: ‐0.55, ‐0.81, and ‐0.29 for EgUnk1, EgZFP2, and EgIPK2b, respectively. The amplification factor (E) and efficiency value (R) using the EgActin gene were 1.95 and 94.92%, respectively. Normalization of log10 on the fold change value 2‐ΔΔCt and 1.95‐ΔΔCt approaches using the regression coefficient yielded consistent results for the EgUnk1, EgZFP2, and EgIPK2b genes. Overall, EgUnk3 and EgIPK2b genes exhibited downregulated expression in susceptible oil palm seedlings (‐0.60 for 2(‐ΔΔCt) and ‐0.58 for 1.95(‐ΔΔCt)), whereas EgIPK2b gene showed up‐regulated and the highest value in inoculated resistant seedlings (1.39 for 2(‐ΔΔCt) and 1.34 for 1.95(‐ΔΔCt)). Basal stem rot disease (BSR) in oil palm decreased EgUnk1 and EgIPK2b expression in susceptible seedlings but increased EgZFP2 gene expression in resistant ones. The results of this research provide valuable corrections to Ct values obtained directly from RT‐qPCR machines using simple linear regression. Consequently, the Ct values of target genes and reference genes exhibit smaller bias values, rendering gene expression levels more reliable.

Sequence-Structure Comparative and Network-Based Prediction of Drought Gene Candidate Regulator in Elaeis guineensis Permatasari, Galuh Wening; Putranto, Riza Arief; Mardhika, Larasati Dena; Aksa, Annisa Aulia; Setiawati, Yuli; Minarsih, Hayati; Riyadi, Imron; Ernayunita, Ernayunita
Journal of Tropical Biodiversity and Biotechnology Vol 9, No 3 (2024): September
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jtbb.90808

Drought poses a significant threat to global food security, particularly impacting crops like oil palm. Selecting genes for genome editing to enhance drought tolerance presents formidable challenges. To ensure that the target gene is chosen correctly and results in the desired character, a pilot study is necessary to determine the target gene for knockout. Two genes drought-related, AtBRL3 and AtOST2, were scrutinized in this context. Aligned with the Elaeis guineensis genome, their neighbouring proteins and gene ontology were analysed to identify potential targets for genome editing. AtBRL3, identified as BRL1 (XP_010913986.1) in E. guineensis, exhibited 58.48% identity and 100% coverage. It interacts with 12 nodes, including BIR1, BRI1, and AT2G20050, crucial for signalling pathways and cellular responses. Molecular function analysis revealed kinase activity. AtOST2 showed high similarity to plasma membrane ATPase/HA1 (XP_010913679.1) in E. guineensis, with 87.46% identity and 100% query cover. It correlated with 14 genes associated with ABA stimulus, stomatal movement, and hormone response. EgBRL1 and EgHA1, resembling AtBRL3 and AtOST2, respectively, emerge as promising targets for developing drought-tolerant oil palm cultivars through gene editing. Nonetheless, further validation through in vitro gRNA target selection and in vivo conversion of OST2/BRL3-containing plasmids in oil palm calluses is indispensable to demonstrate their efficacy in conferring novel drought resistance traits.

Co-Authors . KUSWANHADI . NUHAIMI-HARIS . PRIYONO . SISWANTO . SUMARYONO Afandi, Dadang Agustin Sri MULYATNI Akhirta Atikana Aksa, Annisa Aulia Alfero Putra Iryanto Anggia Prasetyoputri Annisa A Aksa Asmini Budiani Asmini Budiani Asmini Budiani Asmini Budiani Asmini BUDIANI Asmini BUDIANI1 Barahima Abbas Christy Dewi Sukma Dini Astika Sari Dini Astika Sari Dini Astika Sari Ernayunita Ernayunita Ernayunita, Ernayunita Galuh Wening Permatasari Galuh Wening PERMATASARI Galuh Wening PERMATASARI Happy WIDIASTUTI Hayati Minarsih Hayati Minarsih Hayati Minarsih Hayati MINARSIH Imam Bagus Nugroho Imron RIYADI Imron Riyadi Imron Riyadi Imron Riyadi Indra Syahputra Inez Palupi Irfan MARTIANSYAH Irfan Martiansyah Iskandar, Hayati Minarsih Jembar PAMBUDI Julie LECLERCQ Lailia Zubaidah Larasati D Mahardhika Lutfi Anggadhania Mardhika, Larasati Dena MARTIANSYAH, Irfan Masna Maya Sinta Masna Maya Sinta Mose, Windi Muhammad Farrel Ewaldo Mukmin, Restu Prasetya Nurul Khumaida Nurul Khumaida Pambudi, Jembar Pascal MONTORO Pascal MONTORO Pascal MONTORO PERMATASARI, Galuh Wening pri yono Radite TISTAMA Ratih Asmana Ningrum Rizka Tamania Saptari Rizka Tamania SAPTARI Rizka Tamania Saptari Rokhana Faizah Rokhana Faizah Sapto Nugroho Hadi Sapto Nugroho Hadi Sari, Dini Astika Sekar WOELAN Sri Wening Sri Wening Sudarsono Sudarsono Suhandono, Sonny Sustiprijatno Sustiprijatno, Sustiprijatno TATI KRISTIANTI Turhadi Turhadi Vivi Restu Raharti Y.M.M Anita Nugraheni Yuli Setiawati Yuli Setiawati Yuli Setiawati, Yuli

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