A seed resource for screening functionally redundant genes and isolation of new mutants impaired in CO2 and ABA responses

The identification of homologous genes with functional overlap in forward genetic screens is severely limited. Here, we report the generation of over 14000 artificial microRNA (amiRNA)-expressing plants that enable screens of the functionally redundant gene space in Arabidopsis. A protocol was devel...

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Veröffentlicht in:Journal of experimental botany. - 1985. - 70(2019), 2 vom: 07. Jan., Seite 641-651
1. Verfasser: Hauser, Felix (VerfasserIn)
Weitere Verfasser: Ceciliato, Paulo H O, Lin, Yi-Chen, Guo, DanDan, Gregerson, J D, Abbasi, Nazia, Youhanna, Diana, Park, Jiyoung, Dubeaux, Guillaume, Shani, Eilon, Poomchongkho, Nusra, Schroeder, Julian I
Format: Online-Aufsatz
Sprache:English
Veröffentlicht: 2019
Zugriff auf das übergeordnete Werk:Journal of experimental botany
Schlagworte:Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. MicroRNAs Carbon Dioxide 142M471B3J Abscisic Acid 72S9A8J5GW
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520 |a The identification of homologous genes with functional overlap in forward genetic screens is severely limited. Here, we report the generation of over 14000 artificial microRNA (amiRNA)-expressing plants that enable screens of the functionally redundant gene space in Arabidopsis. A protocol was developed for isolating robust and reproducible amiRNA mutants. Examples of validation approaches and essential controls are presented for two new amiRNA mutants that exhibit genetically redundant phenotypes and circumvent double mutant lethality. In a forward genetic screen for abscisic acid (ABA)-mediated inhibition of seed germination, amiRNAs that target combinations of known redundant ABA receptor and SnRK2 kinase genes were rapidly isolated, providing a strong proof of principle for this approach. A new ABA-insensitive amiRNA line that targets three avirulence-induced gene 2(-like) genes was isolated . A thermal imaging screen for plants with impaired stomatal opening in response to low CO2 exposure led to the isolation of a new amiRNA targeting two essential proteasomal subunits, PAB1 and PAB2. The seed library of 11000 T2 amiRNA lines (with 3000 lines in progress) generated here provides a new platform for forward genetic screens and has been made available to the Arabidopsis Biological Resource Center (ABRC). Optimized procedures for amiRNA screening and controls are described 
650 4 |a Journal Article 
650 4 |a Research Support, N.I.H., Extramural 
650 4 |a Research Support, Non-U.S. Gov't 
650 4 |a Research Support, U.S. Gov't, Non-P.H.S. 
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700 1 |a Ceciliato, Paulo H O  |e verfasserin  |4 aut 
700 1 |a Lin, Yi-Chen  |e verfasserin  |4 aut 
700 1 |a Guo, DanDan  |e verfasserin  |4 aut 
700 1 |a Gregerson, J D  |e verfasserin  |4 aut 
700 1 |a Abbasi, Nazia  |e verfasserin  |4 aut 
700 1 |a Youhanna, Diana  |e verfasserin  |4 aut 
700 1 |a Park, Jiyoung  |e verfasserin  |4 aut 
700 1 |a Dubeaux, Guillaume  |e verfasserin  |4 aut 
700 1 |a Shani, Eilon  |e verfasserin  |4 aut 
700 1 |a Poomchongkho, Nusra  |e verfasserin  |4 aut 
700 1 |a Schroeder, Julian I  |e verfasserin  |4 aut 
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