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Delivering of Over-Expression Construct OsWRKY76 Candidate Gene in Rice cv. Nipponbare through Agrobacterium tumefaciens Apriana, Aniversari; Sisharmini, Atmitri; Enggarini, Wening; Sudarsono, Sudarsono; Khumaida, Nurul; Trijatmiko, Kurniawan R.
Jurnal AgroBiogen Vol 7, No 1 (2011): Jurnal AgroBiogen
Publisher : Jurnal AgroBiogen

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Plant genetic improvement can be done through classical breeding or genetic engineering. WRKY is a transcription factor involved in regulating plant defense responses. OsWRKY76 gene is located in a narrow segment of chromosome 9 which is identified previously to be related to wide spectrum resistance in rice. A sequence of OsWRKY76 (+1.200 bp) has available in the gene bank and it makes possible to isolate, clone, and construct the gene into over-expression vector. The aim of this research was to assemble an over-expression construct of OsWRKY76 candidate gene and introduce it into rice through Agrobacterium-mediated transformation. A construct of pCAMBIA-1301::35S::OsWRKY76 has been successfully assembled and transformed into embryogenic calli of rice cv. Nipponbare using A. tumefaciens strain Agl-1 and EHA 105. A number of 126 independent lines has been produced, in which Agl-1 showed 3.8 times more efficient than EHA 105. PCR analysis of randomly selected 25 independent lines showed that all of them positively contained hptII gene, a selectable marker used in the over-expression construct of the OsWRKY76 candidate gene. Based on the result, it could be concluded that the over-expression construct of OsWRKY76 candidate gene have been successfully introduced into the tissue of Nipponbare.

Identification Of Aquatic Plant Species From Bantimurung Waterfall And Their Phytochemical Compounds Analysis Nugraha, Media Fitri Isma; Maharani, Fasya Hadaina; Novita, Hessy; Rajamuddin, Muh Alias L; Yunita, Rossa; Reflinur, Reflinur; Enggarini, Wening; Julzarika, Atriyon; Elya, Berna Elya
ANNALES BOGORIENSES Vol 23, No 1 (2019): Annales Bogorienses
Publisher : Research Center for Biotechnology - Indonesian Institute of Sciences (LIPI)

A total of 21 aquatic plant species were collected from Bantimurung waterfall areas, South Sulawesi, Indonesia, in October 2017. These plant materials were subjected to both species and phytochemicals identification. The aims of this study were to determine the species or taxonomic rank of Indonesian aquatic plants collected from Bantimurung waterfall, South Sulawesi, Indonesia and to identify their chemical compounds (phytochemicals) as a candidate for new herbal medicine. Plant genetic materials used in this study were collected from Bantimurung Bulusaraung waterfall and were then identified based on standard botanical techniques for species identification in the Herbarium Bogoriense, Research center for Biology Indonesian Institute of Science (LIPI), Cibinong, West Java. The samples were subjected to the phytochemistry screening such as alkaloids, flavonoids, tannins, saponins, glycosides, terpenoids and anthraquinone followed the procedures of Indonesian Materia Medika and Harborne. Results showed that all collected aquatic plant samples were able to be identified, including their species names. Phytochemical screening of each sample revealed the presence of glycoside in all of the tested species. However, no alkaloids, anthraquinones, and terpenoids were observed in those tested plant samples. Of the total 21 aquatic plants, 14 species contained flavonoids, 8 species contained phenol compound, and 10 species contained saponins. Among these species Donnax canniformis possessed good antioxidant activity, which correlated to its total phenolic and flavonoid contents. Our results would be beneficial for any future effort in the development of new herbal drugs derived from aquatic plants.

Resistance Analysis of CRISPR/Cas9 Genome-Edited Chili M2 Mutant Lines against Pepper Yellow Leaf Curl Viral Disease Prasetya, Wandy Murti; Hadiarto, Toto; Enggarini, Wening; Polosoro, Aqwin; Suharsono, Suharsono
Jurnal AgroBiogen Vol 17, No 1 (2021): JUNE
Publisher : Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21082/jbio.v17n1.2020.p1-10

Pepper yellow leaf curl virus (PepYLCV) infection transmitted by silverleaf whitefly (Bemisia tabaci [Gennadius]) can decrease chili pepper yield up to 100%. At this moment, there is no chili pepper variety resistant to PepYLCV available. Genome editing approach through CRISPR/Cas9 is an effort to develop variety resistance to the viral infection. The purpose of this study was to obtain M2 lines developed by CRISPR/Cas9 system on proliferating cell nuclear antigen (PCNA) gene for resistance to PepYLCV. A total of four M2 lines (C47-7, L84-2, L84-23, and L120-19) consisting of 60 chili plants were tested for their resistance to PepYLCV. PCR analysis was performed to detect the presence (infection) of the virus. The results showed that a total of 35 plants derived from the four lines were resistant to PepYLCV. They consisted of 7 plants from C47-7 line, 11 plants from L84-2 line, 9 plants from L84-23 line, and 8 plants from L120-19 line. PCR analysis confirmed that the resistant plants obtained from this study were negatively infected by the virus. Since not all tested plants were resistant to virus infection, the PCNA gene allele in these resistant lines were most likely heterozigotes. Sequencing of PCNA gene of the resistant lines is needed to confirm that the resistance phenotypes obtained was due to mutation of the gene. Therefore, further selection needs to be performed to obtain stable and PepYLCV-resistant lines.

Identification of A Major Quantitative Trait Locus for Grain Weight In Rice Using Microsatellite Marker Susilowati, Mariana; Aswidinnoor, Hajrial; Enggarini, Wening; Trijatmiko, Kurniawan Rudi
Makara Journal of Science Vol. 21, No. 4
Publisher : UI Scholars Hub

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Rice is one of the major staple foods in the world, especially in Asia. Improving yield potential of superior cultivars is important to meeting the demand for rice production, which is increasing due to human population increase, climate change, and degradation of agricultural resources, such as land and water. In this study, a BC3F2 population developed from an intraspecific cross between Ciherang and a new plant type line (B11143D) was used in a quantitative trait locus (QTL) analysis. Ciherang is a high yielding rice cultivar with good grain quality which has been planted in 37% of the irrigated rice area in Indonesia. The objective of this study was to identify QTL(s) for yield components on chromosome 12, which can be used to improve the elite cultivar Ciherang or other popular cultivars through marker-assisted breeding. A total of two hundred BC3F2 lines were evaluated in the greenhouse during this study. The population was observed for eight agronomic traits including days to heading (dth), plant height (ph), flag leaf length (fll), panicles per plant (ppl), panicle length (pl), grains per panicle (gpp), 1000-grain weight (gw), and yield (yld). Four simple sequence repeats (SSR) markers (RM3472, RM28048, RM28195, and RM1986) were used for targeted mapping on chromosome 12. Linkage analysis identified a QTL for 1000-grain weight located on chromosome 12 at position 53.5 cM–73 cM.

Phytochemical analysis and antibacterial activity of Staurogyne sp. in fish disease bacteria Nugraha, Media Fitri Isma; Novita, Hessy; Rajamuddin, Muh Alias; Yunita, Rossa; Enggarini, Wening; Reflinur, Reflinur; Maharani, Fasya Hadaina; Elya, Berna
Torani Journal of Fisheries and Marine Science VOLUME 2 NOMOR 2, JUNI 2019
Publisher : Hasanuddin University

This study was aimed to identify the potential use of active compounds extracted from Staurogyne sp as an antibacterial agent to control disease-causing bacteria in fish. Staurogyne sp, an aquatic plant collected from Bantimurung, South Sulawesi were subjected to allelochemical compound. Plant extracts were tested to five bacteria including four types of bacterial pathogen, such as Aeromonas hydrophilla, Edwardsiella ictaluri, Streptococcus agalactiae, and Flavobacterium columnare, and one bacterial sensing quoroum, Chromobacterium violaceum. Based on phytochemical analysis, Staurogyne’s extracts derived from both stem and root contained flavonoids, phenols, and antioxidants compounds with the concentration of 0.018 mgQE/g, 0.3471 mgGAE/g, and 1004,391 IC50 µg/mL, respectively. In addition, Staurogyne plant extracts derived from both their stem and root has also produced both saponins and glycosides compounds. Plant extracts derived from Staurogyne’s leaves revealed three major chemical compounds such as flavonoids (0.77 mgQE/g), phenol (0.0629 mgGAE/g), and glycosides (+). However, no antioxidants and saponin compounds were detected. Applying plant extracts as an antibacterial on five disease-causing bacteria causing diseases in aquaculture, such as Aeromonas Hydrophilla, Edwardsiella ictaluri, Streptococcus agalactiae, Flavobacterium columnare, and Chromobacterium violaceum bacterials revealed that at concentration of 0.1 g the Staurogyne sp extracts did not influence all the bacterials growth. However, by application of plant extract at concentration of 0.2 g on the growth media,three out of 5 tested bacterials (Aeromonas hydrophilla, Edwardsiella ictaluri, and Streptococcus agalactiae) showed intermediate inhibition responses on bacterial growth. The growth of remaining two bacterials pathogen, Flavobacterium columnare and Chromobacterium violaceum at 0.2 g plant extract of Staurogyne sp was not affected. This study revealed that the use of active compound derived from Staurogyne sp would be potential to be used in inhibiting disease-causing bacterial in fish in future. However, optimum concentration of the plant extracts, in particular on the inhibition of the growth of disease-causing bacteria in fish is still needed to adjust.Key words: Antibacterial, aquatic plant, Staurogyne sp, fish disease

Study of aquatic plants and ecological- physics Tempe Lake, Sulawesi Selatan Nugraha, Media Fitri Isma; Julzarika, Atriyon; Radjamuddin, Alias; Reflinur, Reflinur; Yunita, Rossa; Enggarini, Wening; Novita, Hessy
Torani Journal of Fisheries and Marine Science VOLUME 2 NOMOR 2, JUNI 2019
Publisher : Hasanuddin University

Aquatic plants are an indicator of the fertility of an aquatic region. The waters of Lake Tempe are the largest waters of the lake area in South Sulawesi. Lake Tempe is located in the western part of Wajo District, precisely in Tempe District, about 7 km from Sengkang City towards the banks of the Walanae River in southern Sulawesi. The area is about 13,000 ha with a maximum depth of 5.5 m and can reach more than 30,000 ha during floods, and during the dry season, the inundation area reaches only 1,000 ha with a maximum depth of 1 m, located above the continental and Australian and Asian plates. This lake is one of the tectonic lakes in Indonesia. Every year silting the lake occurs. The Tempe hydro vegetation and eco-physical research were carried out in October 2017. The purpose of this study was to record aquatic plant species that live in Tempe Lake and observe ecological changes and physical properties of Lake Tempe. Aquatic plants are expected to be able to filter lake water. The results obtained are physical conditions of sharp-smelling water, unpleasant taste, dark brown, and cloudy color. Chemical indicators of NH3-N waters (0.2976-0.0634), PO4-P (0.0172-0.0844) NO2-N (undetectable), NO3-N (1.7131-1.9335), Sulphate (27.761900 - 37.047620), DO (6.88-7.18) and pH (7.88-8.02). There are 14 species of aquatic plants found in these waters. The most dominant species is water hyacinth. In the case of Tempe lake water vegetation results in siltation of the lake area.Keywords: Aquatic plant, Biodiversity, Species, Tempe lake,

Performance Test of Vegetative Characteristics of Crossed Rice (Oryza sativa L.) Lines of Ciherang Variety X B11143D Line in Telagasari, Karawang Regency, Indonesia Damayanti, Utari; Lestari, Ani; Subardja, Vera Oktavia; Hadiarto, Toto; Enggarini, Wening
Agro Bali : Agricultural Journal Vol 7, No 2 (2024)
Publisher : Universitas Panji Sakti

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.37637/ab.v7i2.1721

One way to increase the genetic diversity of Ciherang as the superior variety is to cross Ciherang with the donor B11143D line as a New Plant Type (NPT) rice. This study aimed to obtain Ciherang X B11143D lines with the best vegetative characteristics in the field. The research was conducted in rice fields in Talagasari Village, Telagasari District, Karawang Regency, from May to September 2023. This experiment used a single-factor Randomized Block Design (RBD) with three replications of 22 treatments, consisting of 19 Ciherang X B11143D lines and three comparison varieties. The effect of treatment was studied using analysis of variance. The results showed that the rice lines significantly influenced the vegetative characters of Ciherang X B11143D lines in Telagasari, Karawang Regency. Based on the observed characters, several lines were selected i.e. 124.2.3, 94.3.3, and 20.4.4 lines. Those three lines chosen as backcrossing lines performed similarly compared to Ciherang as recurrent parent and inherited several important traits for rising productivity from the B11143D line as donor parent, namely the length and area of flag leaf, the total number of tillers, and the number of productive tillers, which were significantly higher than Ciherang.

Optimasi Ekstraksi RNA dan Teknik Kloning: Studi Kasus Kloning Gen Heading Date 3a pada Kelapa Sawit Polosoro, Aqwin; Enggarini, Wening; Kusumanegara, Kusumawaty; Hadiarto, Toto; Miftahudin, Miftahudin; Supena, Ence Darmo Jaya
Vegetalika Vol 13, No 2 (2024)
Publisher : Universitas Gadjah Mada

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/veg.92085

Pembungaan memegang peranan penting bagi tumbuhan karena memfasilitasi rekombinasi genetik, sehingga mendukung perkembangan keragaman genetik yang penting. Keluarga protein phosphatidylethanolamine binding proteins (PEBP) memainkan peran penting dalam mengatur waktu pembungaan dan dormansi benih di beragam spesies tanaman. Penelitian ini bertujuan untuk merancang vektor biner dengan membangun pCAMBIA1300 yang menggabungkan rangkaian gen EgHd3a dari kelapa sawit. Proses konstruksi gen meliputi ekstraksi RNA, sintesis cDNA, amplifikasi gen EgHd3a, kloning gen menjadi vektor kloning, subkloning ke dalam vektor biner pCAMBIA1300, dan diakhiri dengan validasi gen melalui analisis sekuens. Pada ekstraksi RNA, metode PCL-Chisam telah terbukti efektif melalui ekstraksi berulang, meningkatkan kualitas dan kuantitas total RNA. Dalam proses kloaning, metode konvensional menghadapi tantangan dalam memilih lokasi pembelahan yang tepat. Untuk mengatasi kendala ini, penggunaan enzim dengan overhang yang kompatibel diusulkan sebagai solusi potensial. Secara khusus, penggantian BamHI dari BglII telah secara efektif mengatasi tantangan ini. Konfirmasi integrasi fragmen gen ke dalam plasmid pCAMBIA1300 dicapai melalui pengurutan. Meskipun perbedaan diidentifikasi dalam rangkaian EgHd3a-2, perubahan ini tidak berdampak pada asam amino yang dikodekan, sehingga menjaga integritas rangkaian protein

Perbandingan Metode Ekstraksi DNA pada Tanaman Ulin (Eusideroxylon zwageri) Ridzqya, Salsabila Al Alya; Polosoro, Aqwin; Enggarini, Wening; Kusumanegara, Kusumawaty; Helmanto, Hendra; Magandhi, Mahat; Satyawan, Dani; Hadiarto, Toto; Yuniaty, Alice
Buitenzorg: Journal of Tropical Science Vol 1 No 1 (2024): Buitenzorg: Journal of Tropical Science
Publisher : Innovation Centre for Tropical Sciences

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.70158/buitenzorg.v1i1.7

DNA isolation is a routine procedure in molecular analysis. The method of plant genome DNA extraction has been widely available throughout global laboratories. Several labs made some modifications to obtain optimal results. This research was aimed to analyse two types of DNA extraction methods for ironwood plant (Eusideroxylon zwageri), one tropical rain forest woody plant known for its high strength and durability. The results indicate that isolation DNA kit produced high DNA purity with lower concentrations while CTAB methods generated lower DNA purity with higher concentrations. This may be used as consideration of DNA isolation for downstream molecular analyses. Keywords: DNA extraction, ironwood, CTAB, genomic DNA

Identification of Aquatic Plant Species from Bantimurung Waterfall and Their Phytochemical Compounds Analysis Nugraha, Media Fitri Isma; Maharani, Fasya Hadaina; Novita, Hessy; Rajamuddin, Muh Alias L; Yunita, Rossa; Reflinur, Reflinur; Enggarini, Wening; Julzarika, Atriyon; Elya, Berna
Annales Bogorienses Vol. 23 No. 1 (2019): Annales Bogorienses
Publisher : BRIN

Show Abstract | Download Original | Original Source | Check in Google Scholar

A total of 21 aquatic plant species were collected from Bantimurung waterfall areas, South Sulawesi, Indonesia, in October 2017. These plant materials were subjected to both species and phytochemicals identification. The aims of this study were to determine the species or taxonomic rank of Indonesian aquatic plants collected from Bantimurung waterfall, South Sulawesi, Indonesia and to identify their chemical compounds (phytochemicals) as a candidate for new herbal medicine. Plant genetic materials used in this study were collected from Bantimurung Bulusaraung waterfall and were then identified based on standard botanical techniques for species identification in the Herbarium Bogoriense, Research center for Biology Indonesian Institute of Science (LIPI), Cibinong, West Java. The samples were subjected to the phytochemistry screening such as alkaloids, flavonoids, tannins, saponins, glycosides, terpenoids and anthraquinone followed the procedures of Indonesian Materia Medika and Harborne. Results showed that all collected aquatic plant samples were able to be identified, including their species names. Phytochemical screening of each sample revealed the presence of glycoside in all of the tested species. However, no alkaloids, anthraquinones, and terpenoids were observed in those tested plant samples. Of the total 21 aquatic plants, 14 species contained flavonoids, 8 species contained phenol compound, and 10 species contained saponins. Among these species Donnax canniformis possessed good antioxidant activity, which correlated to its total phenolic and flavonoid contents. Our results would be beneficial for any future effort in the development of new herbal drugs derived from aquatic plants.

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