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Title: In situ embryo rescue for generation of wide intra- and interspecific hybrids of Panicum virgatum L.

Abstract

Wide crosses have been used for decades as a method for transferring novel genetic material and traits in plant breeding. Historically, many products of wide crosses require tedious and inefficient surgical embryo rescue prior to embryo abortion to recover single plantlets. Here, we utilize transgenic switchgrass (Panicum virgatum L. cv Alamo) as a pollen donor in conjunction with antibiotic or herbicide selection for recovery of intra-and interspecific F1 crosses by using developing ovules from the female parent and selecting for embryogenic cultures derived from the in situ immature embryo. Furthermore, by using this approach, several intravarietial crosses were generated between transgenic Alamo and the switchgrass varieties Kanlow, Blackwell and Cave-in-Rock as well as an interspecific cross with Atlantic coastal panicgrass. This procedure selected F1 embryogenic callus produced from the developing embryo contained within isolated immature ovules. Several clonal plants were successfully regenerated from each cross. Southern blot, PCR, phenotypic analyses and genomic analysis confirmed F1 hybrids. Using genotyping-bysequencing shows the hybridization of the recovered plants by determining the ratio of transgressive markers to total compared markers between parents and their potential offspring. The ratio of transgressive markers to total compared markers was significantly lower between parents and their predicted offspringmore » than between parents and offspring unrelated to them. Our approach provides the possibility to move useful transgenes into varieties that are recalcitrant to direct transformation which can be optionally segregated thus useful to create new hybrids, as well as recovery of wide crosses that are either difficult or impossible using traditional techniques.« less

Authors:
 [1];  [1];  [1];  [2];  [1];  [2];  [2];  [1]
  1. Department of Cell and Molecular Biology, University of Rhode Island, Kingston RI USA
  2. Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven CT USA
Publication Date:
Research Org.:
Univ. of Rhode Island, Kingston, RI (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1255171
Alternate Identifier(s):
OSTI ID: 1255172; OSTI ID: 1361514
Grant/Contract Number:  
FG-36-08GO88070; FG36-08GO88070
Resource Type:
Published Article
Journal Name:
Plant Biotechnology Journal
Additional Journal Information:
Journal Name: Plant Biotechnology Journal Journal Volume: 14 Journal Issue: 11; Journal ID: ISSN 1467-7644
Publisher:
Wiley-Blackwell
Country of Publication:
United Kingdom
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; 59 BASIC BIOLOGICAL SCIENCES; wide crosses; switchgrass; tissue culture; transgenic plants; genetic transformation

Citation Formats

Kausch, Albert P., Tilelli, Michael, Hague, Joel, Heffelfinger, Christopher, Cunha, David, Moreno, Maria, Dellaporta, Stephen L., and Nelson, Kimberly. In situ embryo rescue for generation of wide intra- and interspecific hybrids of Panicum virgatum L.. United Kingdom: N. p., 2016. Web. doi:10.1111/pbi.12573.
Kausch, Albert P., Tilelli, Michael, Hague, Joel, Heffelfinger, Christopher, Cunha, David, Moreno, Maria, Dellaporta, Stephen L., & Nelson, Kimberly. In situ embryo rescue for generation of wide intra- and interspecific hybrids of Panicum virgatum L.. United Kingdom. doi:10.1111/pbi.12573.
Kausch, Albert P., Tilelli, Michael, Hague, Joel, Heffelfinger, Christopher, Cunha, David, Moreno, Maria, Dellaporta, Stephen L., and Nelson, Kimberly. Wed . "In situ embryo rescue for generation of wide intra- and interspecific hybrids of Panicum virgatum L.". United Kingdom. doi:10.1111/pbi.12573.
@article{osti_1255171,
title = {In situ embryo rescue for generation of wide intra- and interspecific hybrids of Panicum virgatum L.},
author = {Kausch, Albert P. and Tilelli, Michael and Hague, Joel and Heffelfinger, Christopher and Cunha, David and Moreno, Maria and Dellaporta, Stephen L. and Nelson, Kimberly},
abstractNote = {Wide crosses have been used for decades as a method for transferring novel genetic material and traits in plant breeding. Historically, many products of wide crosses require tedious and inefficient surgical embryo rescue prior to embryo abortion to recover single plantlets. Here, we utilize transgenic switchgrass (Panicum virgatum L. cv Alamo) as a pollen donor in conjunction with antibiotic or herbicide selection for recovery of intra-and interspecific F1 crosses by using developing ovules from the female parent and selecting for embryogenic cultures derived from the in situ immature embryo. Furthermore, by using this approach, several intravarietial crosses were generated between transgenic Alamo and the switchgrass varieties Kanlow, Blackwell and Cave-in-Rock as well as an interspecific cross with Atlantic coastal panicgrass. This procedure selected F1 embryogenic callus produced from the developing embryo contained within isolated immature ovules. Several clonal plants were successfully regenerated from each cross. Southern blot, PCR, phenotypic analyses and genomic analysis confirmed F1 hybrids. Using genotyping-bysequencing shows the hybridization of the recovered plants by determining the ratio of transgressive markers to total compared markers between parents and their potential offspring. The ratio of transgressive markers to total compared markers was significantly lower between parents and their predicted offspring than between parents and offspring unrelated to them. Our approach provides the possibility to move useful transgenes into varieties that are recalcitrant to direct transformation which can be optionally segregated thus useful to create new hybrids, as well as recovery of wide crosses that are either difficult or impossible using traditional techniques.},
doi = {10.1111/pbi.12573},
journal = {Plant Biotechnology Journal},
number = 11,
volume = 14,
place = {United Kingdom},
year = {2016},
month = {6}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record
DOI: 10.1111/pbi.12573

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