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The Hevea brasiliensis AP2/ERF superfamily: from ethylene signalling to latex harvesting and physiological disease response Riza Arief PUTRANTO; Pascal MONTORO
E-Journal Menara Perkebunan Vol 84, No 1: Oktober 2016
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (777.018 KB) | DOI: 10.22302/iribb.jur.mp.v84i1.201

Abstract

Ethylene is a hormone known for its involvement in the process of latex harvesting in Hevea brasiliensis. It facilitates latex flow by activation of endogenous metabolism in the anastomosed latex cells called laticifers. In regard to its ambivalent role, ethylene is both favourable to the latex production and unfavourable, to a certain level, to the apparition of a physiological disease termed as tapping panel dryness (TPD). Comprehensive researches have been carried out to reveal the molecular actors in ethylene biosynthesis and signalling pathways in Hevea brasiliensis. One of the most important superfamily implicated as the last transcription factor known in plant ethylene signalling is the APETALA2/ETHYLENE RESPONSE FACTOR (AP2/ERF). Currently, 114 unique sequences related to the Hevea AP2/ERF gene superfamily have been identified and characterized. Specific characterizations under the condition of harvesting stress and the occurrence of TPD have identified 36 gene expression markers (GEMs). Eighteen of these GEMs were predicted as ortholog with 19 Arabidopsis AP2/ERF genes. The characterization was mainly focused on transcriptional regulation, whilst potential post-transcriptional and post-translational regulations of HbAP2/ERF genes were formerly predicted. Three HbERF groups (HbERF-VII, HbERF-VIII and HbERF-IX) were hypothesized to have an important role in Hevea tolerance during latex production as they highly accumulated in laticifers and in response to multiple abiotic stresses. Further functional analysis of several key genes is suggested in order to fully understand the regulation of HbAP2/ERFs. Finally, the molecular markers for future Hevea breeding could be possibly developed from this superfamily.
The in silico study of the COBRA gene family in sugarcane related to potential biomass content Riza Arief PUTRANTO; Galuh Wening PERMATASARI; Rizka Tamania SAPTARI
E-Journal Menara Perkebunan Vol 90, No 1 (2022): April, 2022
Publisher : INDONESIAN RESEARCH INSTITUTE FOR BIOTECHNOLOGY AND BIOINDUSTRY

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22302/iribb.jur.mp.v90i1.486

Abstract

AbstractSugarcane (Saccharum sp.) is potential as a biofuel and biomaterial source for its high cellulose content. Cellulose is the main constituent of the plant cell wall, as a linear chain arranged in a polysaccharide bundle, called cellulose microfibril. A gene named COBRA has been revealed to play role in the orientation of microfibril and cellulose crystallization. The COBRA gene in the Saccharum spp is under-explored. Therefore, the in silico study was conducted to explore the COBRA gene in Saccharum sp. By comparative genomics methods, the COBRA genes from Arabidopsis sp. (AtCOBLs) were compared to the Saccharum sp. (SoCOBLs). The conserved domain was then identified and the cluster system was constructed under a phylogenetic tree. Furthermore, each SoCOBLs protein was modelled to analyze its structure. According to the analysis, eleven of Saccharum sp. genomic scaffolds were successfully identified. Moreover, conserved domain identification resulted in nine SoCOBLs proteins. The phylogenetic tree showed two main clusters: I and II, differentiating those COBLs families based on the protein sequence, domain motif and amino acid properties. It leads to the variation of SoCOBLs protein structure as the results of the amino acid properties. Overall, the COBRA gene has been identified genomically in Saccharum sp. Yet, the function and tissue-specific expression are still unclear. It was predicted to act as the regulator of microfibril orientation and the cellulose synthesis process. Hence, further analyses by in vitro and in vivo are indispensable.[Keywords: cellulose, comparative genomic, Saccharum sp.]AbstrakTanaman tebu (Saccharum sp.) berpotensi sebagai sumber bahan bakar nabati dan biomaterial karena kandungan selulosanya yang tinggi. Selulosa merupakan komponen utama penyusun dinding sel tanaman, sebagai rantai lurus yang tersusun dalam gugusan polisakarida, yang disebut mikrofibril selulosa. Sebuah gen bernama COBRA telah diketahui berperan dalam menentukan arah mikrofibril dan kristalisasi selulosa. Gen COBRA pada spesies Saccharum spp. belum banyak dipelajari. Oleh karena itu, kajian in silico dilakukan untuk mempelajari gen COBRA pada Saccharum sp. Melalui metode perbandingan genomika, gen COBRA dari Arabidopsis sp. (AtCOBLs) dibandingkan dengan gen COBRA dari Saccharum sp. (SoCOBLs). Domain conserve pada gen kemudian diidentifikasi dan sistem klaster disusun dalam sebuah pohon filogeni. Setelah itu, dibuat model untuk menganalisis struktur dari protein SoCOBL. Dari hasil analisis, sebelas perancah genom Saccharum sp. berhasil diidentifikasi. Kemudian, identifikasi daerah lestari menghasilkan sembilan protein SoCOBL. Pohon filogeni menggambarkan dua klaster utama: I dan II, yang membedakan famili SoCOBLs tersebut berdasarkan sekuens protein, motif domain, dan karakteristik asam amino. Karakteristik asam amino menyebabkan variasi pada struktur protein-protein SoCOBL. Secara umum, gen COBRA telah teridentifikasi pada Saccharum sp., meskipun fungsi dan ekspresi spesifiknya pada jaringan masih belum diketahui. Diperkirakan gen tersebut berperan sebagai pengatur arah mikrofibril dan proses sintesis selulosa. Oleh karena itu, perlu adanya analisis lebih lanjut pada level in vitro dan in vivo.[Kata kunci: selulosa, genomika komparatif, Saccharum 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

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1452.514 KB) | DOI: 10.22302/iribb.jur.mp.v84i2.220

Abstract

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]
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

Abstract

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.