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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Main Authors: López, Luis Antonio, Ochieng, John, Pacheco, Mario, Martinez, Leocadio, Hamilton Amoshe Omar, Gowda, Manje, Prasanna, Boddupalli M., Dhugga, Kanwarpal S., Chaikam, Vijay
Format: Journal Article
Language:Inglés
Published: MDPI 2023
Subjects:
Online Access:https://hdl.handle.net/10568/131234
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author López, Luis Antonio
Ochieng, John
Pacheco, Mario
Martinez, Leocadio
Hamilton Amoshe Omar
Gowda, Manje
Prasanna, Boddupalli M.
Dhugga, Kanwarpal S.
Chaikam, Vijay
author_browse Chaikam, Vijay
Dhugga, Kanwarpal S.
Gowda, Manje
Hamilton Amoshe Omar
López, Luis Antonio
Martinez, Leocadio
Ochieng, John
Pacheco, Mario
Prasanna, Boddupalli M.
author_facet López, Luis Antonio
Ochieng, John
Pacheco, Mario
Martinez, Leocadio
Hamilton Amoshe Omar
Gowda, Manje
Prasanna, Boddupalli M.
Dhugga, Kanwarpal S.
Chaikam, Vijay
author_sort López, Luis Antonio
collection Repository of Agricultural Research Outputs (CGSpace)
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 CGSpace1312342025-12-08T10:29:22Z Effectiveness of R1-nj anthocyanin marker in the identification of in vivo induced maize haploid embryos López, Luis Antonio Ochieng, John Pacheco, Mario Martinez, Leocadio Hamilton Amoshe Omar Gowda, Manje Prasanna, Boddupalli M. Dhugga, Kanwarpal S. Chaikam, Vijay doubled haploids maize breeding programmes inbred lines crops 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. 2023 2023-07-20T20:39:36Z 2023-07-20T20:39:36Z Journal Article https://hdl.handle.net/10568/131234 en Open Access application/pdf MDPI Lopez, L. A., Ochieng, J., Pacheco, M., Martinez, L., Omar, H. A., Gowda, M., Prasanna, B. M., Dhugga, K. S., & Chaikam, V. (2023). Effectiveness of R1-nj Anthocyanin Marker in the Identification of In Vivo Induced Maize Haploid Embryos. Plants, 12(12), 2314. https://doi.org/10.3390/plants12122314
spellingShingle doubled haploids
maize
breeding programmes
inbred lines
crops
López, Luis Antonio
Ochieng, John
Pacheco, Mario
Martinez, Leocadio
Hamilton Amoshe Omar
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 doubled haploids
maize
breeding programmes
inbred lines
crops
url https://hdl.handle.net/10568/131234
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