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题名: A protocol for efficient callus induction and stable transformation of Spirodela polyrhiza (L.) Schleiden using Agrobacterium tumefaciens
作者: Yang, Jingjing1, 2, 3; Lia, Gaojie1, 2, 3; Hua, Shiqi1, 2, 3; Bishopp, Antony4; Heenatigala, P. P. M.1, 2, 3; Kumar, Sunjeet1, 2, 3; Duan, Pengfei5; Yao, Lunguang5; Hou, Hongwei1, 2, 3
关键词: Spirodela polyrhiza ; Callus induction ; Transformation ; Agrobacterium tumefaciens ; Cytokinin ; Auxin
刊名: AQUATIC BOTANY
发表日期: 2018-12-01
DOI: 10.1016/j.aquabot.2018.08.004
卷: 151, 页:80-86
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology ; Life Sciences & Biomedicine
类目[WOS]: Plant Sciences ; Marine & Freshwater Biology
研究领域[WOS]: Plant Sciences ; Marine & Freshwater Biology
英文摘要: Spirodela polyrhiza represents the largest specie with the smallest genome of all the members of the Lemnoideae. Its genome features have been delineated, revealing its fewest predicted genes of any known plant genome. It is also ideal system for basic biological researches and various practical applications including toxicity testing, bioreactor, biomonitoring and biofuel. In this study, we reported the successful induction of S. polyrhiza callus coupled with the efficient stable transformation using the Agrobacterium tumefaciens strain LBA4404 by optimizing each step of the process. We found that the highest callus induction efficiency was achieved with 22.62 uM 2,4-D and 8.88 mu M 6-BA, with above 90% of fronds forming calli. We also determined that 100 M acetosyringone in the co-cultivation medium and the maintenance of pH value at 5.2 were crucial for high transformation efficiency (up to 13 1.5%). As proof of concept, we transformed S. polyrhiza with the DR5 and TCS synthetic reporters, which have previously been used to report cytokinin and auxin signaling output in the model plant Arabidopsis thaliana. The cytokinin showed highest accumulation at the initial stage of bud formation and the frond apex of S. polyrhiza whilst the expression of auxin was observed highest at frond with middle size. These transformed lines provide an effective way to investigate the development of S. polyrhiza and may shed light on the interesting way in which this specie reproduces. This is the first report of highly efficient callus induction and Agrobacterium tumefaciens-mediated transformation in S. polyrhiza.
关键词[WOS]: LEMNA-MINOR ; ARABIDOPSIS-THALIANA ; PLANT-REGENERATION ; DUCKWEED ; GIBBA ; GENE ; PHARMACEUTICALS ; RHIZOSPHERE ; EXPRESSION ; COMPONENTS
语种: 英语
项目资助者: State Key Laboratory of Freshwater Ecology and Biotechnology(2016FB04) ; Project of Major Science and Technology Program for Water Pollution Control and Treatment(2015ZX07503-005) ; Major Project of Technology Innovation Program of Hubei, China(2017ABA135)
WOS记录号: WOS:000448091000012
ISSN号: 0304-3770
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内容类型: 期刊论文
URI标识: http://ir.ihb.ac.cn/handle/342005/56036
Appears in Collections:藻类生物学及应用研究中心_期刊论文

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作者单位: 1.Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Hubei, Peoples R China
2.Chinese Acad Sci, Inst Hydrobiol, Key Lab Aquat Biodivers & Conservat, Wuhan 430072, Hubei, Peoples R China
3.Univ Chinese Acad Sci, Wuhan 430072, Hubei, Peoples R China
4.Univ Nottingham, Ctr Plant Integrat Biol, Nottingham, England
5.Nanyang Normal Univ, Coll Agr Engn, Collaborat Innovat Ctr Water Secur Water Source R, Nanyang, Henan, Peoples R China
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