Accelerating haploid induction rate and haploid validation through marker-assisted selection for qhir1 and qhir8 in maize

Kanogporn Khammona; Abil Dermail; Khundej Suriharn; Thomas Lübberstedt; Samart Wanchana; Burin Thunnom; Wasin Poncheewin; Theerayut Toojinda; Vinitchan Ruanjaichon; Siwaret Arikit

doi:10.3389/fpls.2024.1337463

Accelerating haploid induction rate and haploid validation through marker-assisted selection for qhir1 and qhir8 in maize

Front Plant Sci. 2024 Mar 5:15:1337463. doi: 10.3389/fpls.2024.1337463. eCollection 2024.

Authors

Kanogporn Khammona^#¹, Abil Dermail^#², Khundej Suriharn^{2

3}, Thomas Lübberstedt⁴, Samart Wanchana⁵, Burin Thunnom⁵, Wasin Poncheewin⁵, Theerayut Toojinda⁵, Vinitchan Ruanjaichon⁵, Siwaret Arikit^{1

6}

Affiliations

¹ Department of Agronomy, Faculty of Agriculture at Kamphaeng Saen, Kasetsart University, Nakhon Pathom, Thailand.
² Department of Agronomy, Faculty of Agriculture, Khon Kaen University, Khon Kaen, Thailand.
³ Plant Breeding Research Center for Sustainable Agriculture, Faculty of Agriculture, Khon Kaen University, Khon Kaen, Thailand.
⁴ Department of Agronomy, Iowa State University, Ames, IA, United States.
⁵ National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Pathum Thani, Thailand.
⁶ Rice Science Center, Kasetsart University, Nakhon Pathom, Thailand.

^# Contributed equally.

Abstract

Doubled haploid (DH) technology becomes more routinely applied in maize hybrid breeding. However, some issues in haploid induction and identification persist, requiring resolution to optimize DH production. Our objective was to implement simultaneous marker-assisted selection (MAS) for qhir1 (MTL/ZmPLA1/NLD) and qhir8 (ZmDMP) using TaqMan assay in F₂ generation of four BHI306-derived tropical × temperate inducer families. We also aimed to assess their haploid induction rate (HIR) in the F₃ generation as a phenotypic response to MAS. We highlighted remarkable increases in HIR of each inducer family. Genotypes carrying qhir1 and qhir8 exhibited 1 - 3-fold higher haploid frequency than those carrying only qhir1. Additionally, the qhir1 marker was employed for verifying putative haploid seedlings at 7 days after planting. Flow cytometric analysis served as the gold standard test to assess the accuracy of the R1-nj and the qhir1 marker. The qhir1 marker showed high accuracy and may be integrated in multiple haploid identifications at early seedling stage succeeding pre-haploid sorting via R1-nj marker.

Keywords: doubled haploid; haploid identification; haploid induction; hybrid breeding; molecular assay.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. The National Science and Technology Development Agency (NSTDA) (Grant No. P-20-52286, P-21-50610 and P-23-51489). Also, the National Science, Research and Innovation Fund, Thailand Science Research and innovation (TSRI).