Methods for Performing Crosses in Setaria viridis, a New Model System for the Grasses
Abstract
Setaria viridis is an emerging model system for C4 grasses. It is closely related to the bioenergy feed stock switchgrass and the grain crop foxtail millet. Recently, the 510 Mb genome of foxtail millet, S. italica, has been sequenced 1,2 and a 25x coverage genome sequence of the weedy relative S. viridis is in progress. S. viridis has a number of characteristics that make it a potentially excellent model genetic system including a rapid generation time, small stature, simple growth requirements, prolific seed production 3 and developed systems for both transient and stable transformation 4 . However, the genetics of S. viridis is largely unexplored, in part, due to the lack of detailed methods for performing crosses. To date, no standard protocol has been adopted that will permit rapid production of seeds from controlled crosses. The protocol presented here is optimized for performing genetic crosses in S. viridis, accession A10.1. We have employed a simple heat treatment with warm water for emasculation after pruning the panicle to retain 20-30 florets and labeling of flowers to eliminate seeds resulting from newly developed flowers after emasculation. After testing a series of heat treatments at permissive temperatures and varying the duration of dipping,more »
- Authors:
-
- Donald Danforth Plant Science Center, St. Louis, MO (United States)
- Boyce Thompson Institute for Plant Research, Ithaca, NY (United States)
- Publication Date:
- Research Org.:
- Donald Danforth Plant Science Center, St. Louis, MO (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Biological and Environmental Research (BER). Earth and Environmental Systems Science Division; National Science Foundation (NSF)
- OSTI Identifier:
- 1628634
- Grant/Contract Number:
- SC0008769; IOS-1127017
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Journal of Visualized Experiments
- Additional Journal Information:
- Journal Issue: 80; Journal ID: ISSN 1940-087X
- Publisher:
- MyJoVE Corp.
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 54 ENVIRONMENTAL SCIENCES; science & technology; environmental sciences; hybridization; genetics; plants; Setaria viridis; crosses; emasculation; flowering; seed propagation; seed dormancy
Citation Formats
Jiang, Hui, Barbier, Hugues, and Brutnell, Thomas. Methods for Performing Crosses in Setaria viridis, a New Model System for the Grasses. United States: N. p., 2013.
Web. doi:10.3791/50527.
Jiang, Hui, Barbier, Hugues, & Brutnell, Thomas. Methods for Performing Crosses in Setaria viridis, a New Model System for the Grasses. United States. https://doi.org/10.3791/50527
Jiang, Hui, Barbier, Hugues, and Brutnell, Thomas. Tue .
"Methods for Performing Crosses in Setaria viridis, a New Model System for the Grasses". United States. https://doi.org/10.3791/50527. https://www.osti.gov/servlets/purl/1628634.
@article{osti_1628634,
title = {Methods for Performing Crosses in Setaria viridis, a New Model System for the Grasses},
author = {Jiang, Hui and Barbier, Hugues and Brutnell, Thomas},
abstractNote = {Setaria viridis is an emerging model system for C4 grasses. It is closely related to the bioenergy feed stock switchgrass and the grain crop foxtail millet. Recently, the 510 Mb genome of foxtail millet, S. italica, has been sequenced 1,2 and a 25x coverage genome sequence of the weedy relative S. viridis is in progress. S. viridis has a number of characteristics that make it a potentially excellent model genetic system including a rapid generation time, small stature, simple growth requirements, prolific seed production 3 and developed systems for both transient and stable transformation 4 . However, the genetics of S. viridis is largely unexplored, in part, due to the lack of detailed methods for performing crosses. To date, no standard protocol has been adopted that will permit rapid production of seeds from controlled crosses. The protocol presented here is optimized for performing genetic crosses in S. viridis, accession A10.1. We have employed a simple heat treatment with warm water for emasculation after pruning the panicle to retain 20-30 florets and labeling of flowers to eliminate seeds resulting from newly developed flowers after emasculation. After testing a series of heat treatments at permissive temperatures and varying the duration of dipping, we have established an optimum temperature and time range of 48 °C for 3-6 min. By using this method, a minimum of 15 crosses can be performed by a single worker per day and an average of 3-5 outcross progeny per panicle can be recovered. Therefore, an average of 45-75 outcross progeny can be produced by one person in a single day. Broad implementation of this technique will facilitate the development of recombinant inbred line populations of S. viridis X S. viridis or S. viridis X S. italica, mapping mutations through bulk segregant analysis and creating higher order mutants for genetic analysis.},
doi = {10.3791/50527},
journal = {Journal of Visualized Experiments},
number = 80,
volume = ,
place = {United States},
year = {Tue Oct 01 00:00:00 EDT 2013},
month = {Tue Oct 01 00:00:00 EDT 2013}
}
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