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Effectiveness of R1-nj Anthocyanin Marker in the Identification of In Vivo Induced Maize Haploid Embryos

Doubled haploid (DH) technology has become integral to maize breeding programs to expedite inbred line development and increase the efficiency of breeding operations. Unlike many other plant species that use in vitro methods, DH production in maize uses a relatively simple and efficient in vivo hapl...

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Autores principales: Lopez, Luis Antonio, Ochieng, John, Pacheco, Mario, Martinez, Leocadio, Omar, Hamilton Amoshe, Gowda, Manje, Prasanna, Boddupalli M., Dhugga, Kanwarpal S., Chaikam, Vijay
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10302338/
https://www.ncbi.nlm.nih.gov/pubmed/37375939
http://dx.doi.org/10.3390/plants12122314
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author Lopez, Luis Antonio
Ochieng, John
Pacheco, Mario
Martinez, Leocadio
Omar, Hamilton Amoshe
Gowda, Manje
Prasanna, Boddupalli M.
Dhugga, Kanwarpal S.
Chaikam, Vijay
author_facet Lopez, Luis Antonio
Ochieng, John
Pacheco, Mario
Martinez, Leocadio
Omar, Hamilton Amoshe
Gowda, Manje
Prasanna, Boddupalli M.
Dhugga, Kanwarpal S.
Chaikam, Vijay
author_sort Lopez, Luis Antonio
collection PubMed
description Doubled haploid (DH) technology has become integral to maize breeding programs to expedite inbred line development and increase the efficiency of breeding operations. Unlike many other plant species that use in vitro methods, DH production in maize uses a relatively simple and efficient in vivo haploid induction method. However, it takes two complete crop cycles for DH line generation, one for haploid induction and the other one for chromosome doubling and seed production. Rescuing in vivo induced haploid embryos has the potential to reduce the time for DH line development and improve the efficiency of DH line production. However, the identification of a few haploid embryos (~10%) resulting from an induction cross from the rest of the diploid embryos is a challenge. In this study, we demonstrated that an anthocyanin marker, namely R1-nj, which is integrated into most haploid inducers, can aid in distinguishing haploid and diploid embryos. Further, we tested conditions that enhance R1-nj anthocyanin marker expression in embryos and found that light and sucrose enhance anthocyanin expression, while phosphorous deprivation in the media had no affect. Validating the use of the R1-nj marker for haploid and diploid embryo identification using a gold standard classification based on visual differences among haploids and diploids for characteristics such as seedling vigor, erectness of leaves, tassel fertility, etc., indicated that the R1-nj marker could lead to significantly high false positives, necessitating the use of additional markers for increased accuracy and reliability of haploid embryo identification.
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spelling pubmed-103023382023-06-29 Effectiveness of R1-nj Anthocyanin Marker in the Identification of In Vivo Induced Maize Haploid Embryos Lopez, Luis Antonio Ochieng, John Pacheco, Mario Martinez, Leocadio Omar, Hamilton Amoshe Gowda, Manje Prasanna, Boddupalli M. Dhugga, Kanwarpal S. Chaikam, Vijay Plants (Basel) Article Doubled haploid (DH) technology has become integral to maize breeding programs to expedite inbred line development and increase the efficiency of breeding operations. Unlike many other plant species that use in vitro methods, DH production in maize uses a relatively simple and efficient in vivo haploid induction method. However, it takes two complete crop cycles for DH line generation, one for haploid induction and the other one for chromosome doubling and seed production. Rescuing in vivo induced haploid embryos has the potential to reduce the time for DH line development and improve the efficiency of DH line production. However, the identification of a few haploid embryos (~10%) resulting from an induction cross from the rest of the diploid embryos is a challenge. In this study, we demonstrated that an anthocyanin marker, namely R1-nj, which is integrated into most haploid inducers, can aid in distinguishing haploid and diploid embryos. Further, we tested conditions that enhance R1-nj anthocyanin marker expression in embryos and found that light and sucrose enhance anthocyanin expression, while phosphorous deprivation in the media had no affect. Validating the use of the R1-nj marker for haploid and diploid embryo identification using a gold standard classification based on visual differences among haploids and diploids for characteristics such as seedling vigor, erectness of leaves, tassel fertility, etc., indicated that the R1-nj marker could lead to significantly high false positives, necessitating the use of additional markers for increased accuracy and reliability of haploid embryo identification. MDPI 2023-06-14 /pmc/articles/PMC10302338/ /pubmed/37375939 http://dx.doi.org/10.3390/plants12122314 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Lopez, Luis Antonio
Ochieng, John
Pacheco, Mario
Martinez, Leocadio
Omar, Hamilton Amoshe
Gowda, Manje
Prasanna, Boddupalli M.
Dhugga, Kanwarpal S.
Chaikam, Vijay
Effectiveness of R1-nj Anthocyanin Marker in the Identification of In Vivo Induced Maize Haploid Embryos
title Effectiveness of R1-nj Anthocyanin Marker in the Identification of In Vivo Induced Maize Haploid Embryos
title_full Effectiveness of R1-nj Anthocyanin Marker in the Identification of In Vivo Induced Maize Haploid Embryos
title_fullStr Effectiveness of R1-nj Anthocyanin Marker in the Identification of In Vivo Induced Maize Haploid Embryos
title_full_unstemmed Effectiveness of R1-nj Anthocyanin Marker in the Identification of In Vivo Induced Maize Haploid Embryos
title_short Effectiveness of R1-nj Anthocyanin Marker in the Identification of In Vivo Induced Maize Haploid Embryos
title_sort effectiveness of r1-nj anthocyanin marker in the identification of in vivo induced maize haploid embryos
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10302338/
https://www.ncbi.nlm.nih.gov/pubmed/37375939
http://dx.doi.org/10.3390/plants12122314
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