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All Journal Vegetalika Torani Journal of Fisheries and Marine Science Jurnal AgroBiogen ANNALES BOGORIENSES Agro Bali: Agricultural Journal Makara Journal of Science Buitenzorg: Journal of Tropical Science Annales Bogorienses
Wening Enggarini
Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian, Jl. Tentara Pelajar 3A, Bogor 16111 Telp. (0251) 8337975
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemical Engineering, Chemistry & Bioengineering Chemistry Earth & Planetary Sciences Economics, Econometrics & Finance Environmental Science Health Professions Immunology & microbiology Medicine & Pharmacology Social Sciences

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Journal : Jurnal AgroBiogen

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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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Abstract

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.
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

Abstract

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.
Co-Authors Alice Yuniaty Ani Lestari Aniversari Apriana Atmitri Sisharmini Atriyon Julzarika Berna Elya Damayanti, Utari Dani Satyawan Hadiarto, Toto HAJRIAL ASWIDINNOOR Helmanto, Hendra Kurniawan R. Trijatmiko Kurniawan Rudi Trijatmiko, Kurniawan Rudi Kusumanegara, Kusumawaty Maharani, Fasya Hadaina Mahat Magandhi Mariana Susilowati, Mariana Miftahudin . Novita, Hessy Nugraha, Media Fitri Isma Nurul Khumaida Polosoro, Aqwin Prasetya, Wandy Murti Radjamuddin, Alias Rajamuddin, Muh Alias Rajamuddin, Muh Alias L REFLINUR REFLINUR Ridzqya, Salsabila Al Alya ROSSA YUNITA Sudarsono Suharsono Suharsono Supena, Ence Darmo Jaya Toto Hadiarto Vera Oktavia Subardja