Application of CRISPR/Cas9 genome editing system for molecular breeding of orchids
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article-39485 |
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<dc schemaLocation="http://www.openarchives.org/OAI/2.0/oai_dc/ http://www.openarchives.org/OAI/2.0/oai_dc.xsd"><title lang="en-US">Application of CRISPR/Cas9 genome editing system for molecular breeding of orchids</title><creator>Semiarti, Endang</creator><creator>Nopitasari, Sri</creator><creator>Setiawati, Yuli</creator><creator>Lawrie, Muhammad Dylan</creator><creator>Purwantoro, Aziz</creator><creator>Widada, Jaka</creator><creator>Yoshioka, Yasushi</creator><creator>Matsumoto, Shogo</creator><creator>Ninomiya, Kana</creator><creator>Asano, Yuuki</creator><subject lang="en-US">CRISPR/Cas9; Genome editing; Molecular breeding; Orchid; Phytoene desaturase (PDS)</subject><description lang="en-US">Orchid is an important ornamental plant in Indonesia due to their natural beauty of flowers. In the tropical forest, orchids are being acquired for trading and commercial market. Thus, the effort is required to proliferate orchid in large quantities for conservation and improve the floral variation for plant breeding. The purpose of this study is to develop a firmed methodology of molecular breeding of orchids using CRISPR/Cas9 KO system. The plant material used was Phalaenopsis amabilis protocorms growth on NP medium+pepton (2 g/L). Protocorm were submerged in the culture of Agrobacterium tumefaciens that Ti‐plasmid had been filled with a T‐DNA construct of a pRGEB32 vector harboring sgRNA with PDS3 sequence. Detection for transformants was confirmed by PCR using HPT primers (545 bp), Cas9 primers (402 bp), PDS primers (280 bp) and trnL‐F (1200 bp) as an internal control. The results showed that 0.96% PDS transformants were obtained from PDS3T2 lines. Several transformant showed pale leaf color compared to non‐transformant plants. This study suggests that the target gene has successfully edited by CRISPR/Cas9 system and could be applied for that functional gene editing in orchids.</description><publisher lang="en-US">Universitas Gadjah Mada</publisher><contributor lang="en-US">Basic Research Grant from the Indonesian Ministry of Research, Technology and Higher Education (RISTEK DIKTI)</contributor><contributor lang="en-US">Japan Society for Promotion of Science (JSPS)</contributor><date>2020-06-30</date><type>Journal:Article</type><type>Other:info:eu-repo/semantics/publishedVersion</type><type>Journal:Article</type><type>File:application/pdf</type><identifier>https://jurnal.ugm.ac.id/ijbiotech/article/view/39485</identifier><identifier>10.22146/ijbiotech.39485</identifier><source lang="en-US">Indonesian Journal of Biotechnology; Vol 25, No 1 (2020); 61-68</source><source>2089-2241</source><source>0853-8654</source><language>eng</language><relation>https://jurnal.ugm.ac.id/ijbiotech/article/view/39485/28051</relation><relation>https://jurnal.ugm.ac.id/ijbiotech/article/downloadSuppFile/39485/5992</relation><relation>https://jurnal.ugm.ac.id/ijbiotech/article/downloadSuppFile/39485/10947</relation><relation>https://jurnal.ugm.ac.id/ijbiotech/article/downloadSuppFile/39485/10948</relation><relation>https://jurnal.ugm.ac.id/ijbiotech/article/downloadSuppFile/39485/10949</relation><relation>https://jurnal.ugm.ac.id/ijbiotech/article/downloadSuppFile/39485/10950</relation><relation>https://jurnal.ugm.ac.id/ijbiotech/article/downloadSuppFile/39485/10951</relation><relation>https://jurnal.ugm.ac.id/ijbiotech/article/downloadSuppFile/39485/10952</relation><relation>https://jurnal.ugm.ac.id/ijbiotech/article/downloadSuppFile/39485/10953</relation><relation>https://jurnal.ugm.ac.id/ijbiotech/article/downloadSuppFile/39485/10954</relation><rights lang="en-US">Copyright (c) 2020 The Author(s)</rights><rights lang="en-US">http://creativecommons.org/licenses/by-sa/4.0</rights><recordID>article-39485</recordID></dc>
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| language |
eng |
| format |
Journal:Article Journal Other:info:eu-repo/semantics/publishedVersion Other File:application/pdf File Journal:eJournal |
| author |
Semiarti, Endang Nopitasari, Sri Setiawati, Yuli Lawrie, Muhammad Dylan Purwantoro, Aziz Widada, Jaka Yoshioka, Yasushi Matsumoto, Shogo Ninomiya, Kana Asano, Yuuki |
| author2 |
Basic Research Grant from the Indonesian Ministry of Research, Technology and Higher Education (RISTEK DIKTI) Japan Society for Promotion of Science (JSPS) |
| title |
Application of CRISPR/Cas9 genome editing system for molecular breeding of orchids |
| publisher |
Universitas Gadjah Mada |
| publishDate |
2020 |
| topic |
CRISPR/Cas9 Genome editing Molecular breeding Orchid Phytoene desaturase (PDS) |
| url |
https://jurnal.ugm.ac.id/ijbiotech/article/view/39485 https://jurnal.ugm.ac.id/ijbiotech/article/view/39485/28051 https://jurnal.ugm.ac.id/ijbiotech/article/downloadSuppFile/39485/5992 https://jurnal.ugm.ac.id/ijbiotech/article/downloadSuppFile/39485/10947 https://jurnal.ugm.ac.id/ijbiotech/article/downloadSuppFile/39485/10948 https://jurnal.ugm.ac.id/ijbiotech/article/downloadSuppFile/39485/10949 https://jurnal.ugm.ac.id/ijbiotech/article/downloadSuppFile/39485/10950 https://jurnal.ugm.ac.id/ijbiotech/article/downloadSuppFile/39485/10951 https://jurnal.ugm.ac.id/ijbiotech/article/downloadSuppFile/39485/10952 https://jurnal.ugm.ac.id/ijbiotech/article/downloadSuppFile/39485/10953 https://jurnal.ugm.ac.id/ijbiotech/article/downloadSuppFile/39485/10954 |
| contents |
Orchid is an important ornamental plant in Indonesia due to their natural beauty of flowers. In the tropical forest, orchids are being acquired for trading and commercial market. Thus, the effort is required to proliferate orchid in large quantities for conservation and improve the floral variation for plant breeding. The purpose of this study is to develop a firmed methodology of molecular breeding of orchids using CRISPR/Cas9 KO system. The plant material used was Phalaenopsis amabilis protocorms growth on NP medium+pepton (2 g/L). Protocorm were submerged in the culture of Agrobacterium tumefaciens that Ti‐plasmid had been filled with a T‐DNA construct of a pRGEB32 vector harboring sgRNA with PDS3 sequence. Detection for transformants was confirmed by PCR using HPT primers (545 bp), Cas9 primers (402 bp), PDS primers (280 bp) and trnL‐F (1200 bp) as an internal control. The results showed that 0.96% PDS transformants were obtained from PDS3T2 lines. Several transformant showed pale leaf color compared to non‐transformant plants. This study suggests that the target gene has successfully edited by CRISPR/Cas9 system and could be applied for that functional gene editing in orchids. |
| id |
IOS1080.article-39485 |
| institution |
Universitas Gadjah Mada |
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19 |
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library:university library |
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Perpustakaan Pusat Universitas Gadjah Mada |
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488 |
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Indonesian Journal of Biotechnology |
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1080 |
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Bioteknology medicine and health agriculture biology |
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SLEMAN |
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DAERAH ISTIMEWA YOGYAKARTA |
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IOS1080 |
| first_indexed |
2020-07-01T19:59:54Z |
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2020-08-04T21:11:53Z |
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dc |
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1722525147798175744 |
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17.60897 |