Commercial scale genetic transformation of mature seed embryo explants in maize
A novel, efficient maize genetic transformation system was developed using -mediated transformation of embryo explants from mature seeds. Seeds from field grown plants were sterilized and crushed to isolate embryo explants consisting of the coleoptile, leaf primordia, and shoot apical meristem which...
Saved in:
Published in | Frontiers in plant science Vol. 13; p. 1056190 |
---|---|
Main Authors | , , , , , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
Switzerland
Frontiers Media S.A
29.11.2022
|
Subjects | |
Online Access | Get full text |
Cover
Loading…
Summary: | A novel, efficient maize genetic transformation system was developed using
-mediated transformation of embryo explants from mature seeds. Seeds from field grown plants were sterilized and crushed to isolate embryo explants consisting of the coleoptile, leaf primordia, and shoot apical meristem which were then purified from the ground seed bulk preparation. The infection of relevant tissues of seed embryo explants (SEEs) by
was improved by the centrifugation of the explants. Transgenic plants were obtained by multiple bud induction on high cytokinin media, followed by plant regeneration on hormone-free medium. Three different selectable markers (
, and
) were successfully used for producing transgenic plants. Stable integration of transgenes in the maize genome was demonstrated by molecular analyses and germline transmission of the inserted transgenes to the next generation was confirmed by pollen segregation and progeny analysis. Phenotypic evidence for chimeric transgenic tissue was frequently observed in initial experiments but was significantly reduced by including a second bud induction step with optimized cytokinin concentration. Additional improvements, including culturing explants at an elevated temperature during bud induction led to the development of a revolutionary system for efficient transgenic plant production and genome editing. To our knowledge, this is the first report of successful transgenic plant regeneration through
-mediated transformation of maize mature SEEs. This system starts with mature seed that can be produced in large volumes and the SEEs explants are storable. It has significant advantages in terms of scalability and flexibility over methods that rely on immature explants. |
---|---|
Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Edited by: Phanikanth Jogam, Kakatiya University, India Reviewed by: Vijay Sheri, East Carolina University, United States; Sahil Mehta, University of Delhi, India; Dhirendra Fartyal, Agricutural Research Organization, Israel These authors have contributed equally to this work This article was submitted to Technical Advances in Plant Science, a section of the journal Frontiers in Plant Science |
ISSN: | 1664-462X 1664-462X |
DOI: | 10.3389/fpls.2022.1056190 |