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

Abstract

Summary 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. We have utilized transgenic switchgrass ( Panicum virgatum L. cv Alamo) as a pollen donor in conjunction with antibiotic or herbicide selection for recovery of intra‐and interspecific F ₁ crosses by using developing ovules from the female parent and selecting for embryogenic cultures derived from the in situ immature embryo. 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 F ₁ 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 F ₁ hybrids. Using genotyping‐by‐sequencing 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 significantlymore » lower between parents and their predicted offspring than between parents and offspring unrelated to them. This 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:

Kausch, Albert P. ^[1]; Tilelli, Michael ^[1]; Hague, Joel ^[1]; Heffelfinger, Christopher ^[2]; Cunha, David ^[1]; Moreno, Maria ^[2]; Dellaporta, Stephen L. ^[2]; Nelson, Kimberly ^[1]

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+ Show Author Affiliations

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:

Wed Jun 01 00:00:00 EDT 2016

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:  

DE‐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