2020•Biotech StudiesOpen access

Development of doubled haploid maize lines by using in vivo haploid technique

Rahime Cengiz, Kayıhan Zahit Korkut

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Abstract

The doubled haploid technology is now an integral component of modern maize breeding programs.In this study, the maternal haploid induction (gynogenesis) method was used to derive Doubled-Haploid (DH) lines from elite maize germplasm adapted to Turkey.Temperate haploid inducers (RWS, RWK-76, RWS x RWK-76 and WS14) were used as pollinators, and a set of 30 single-crossses (in FAO 650-700 maturity groups) were used as source materials.Putative haploid seeds were selected based on expression of R1-nj anthocyanin color marker.Highest haploid induction rate (20.42%) was recorded by using RWK-76 as inducer line, and the lowest haploid induction rate (17.75%) was obtained through WS14.Putative haploid seeds were germinated and seedlings were treated with 0.06% colchicine + 0.5% dimethylsulfoxide solution.Following transfer of seedlings into the field, 2178 D0 plants were obtained out of a total of 3012 treated haploids.Live plants were from 89% of 2178 seedlings which are planted to the field.Fertile plants were formed 57% of live plants.Inbreeding was succeeded in 31.23% of fertile plants and only 7.8% of inbreeding plants were able to produce seeds.Consequently, 27 doubled haploid lines were developed.

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The doubled haploid technology is now an integral component of modern maize breeding programs.In this study, the maternal haploid induction (gynogenesis) method was used to derive Doubled-Haploid (DH) lines from elite maize germplasm adapted to Turkey.Temperate haploid inducers (RWS, RWK-76, RWS x RWK-76 and WS14) were used as pollinators, and a set of 30 single-crossses (in FAO 650-700 maturity groups) were used as source materials.Putative haploid seeds were selected based on expression of R1-nj anthocyanin color marker.Highest haploid induction rate (20.42%) was recorded by using RWK-76 as inducer line, and the lowest haploid induction rate (17.75%) was obtained through WS14.Putative haploid seeds were germinated and seedlings were treated with 0.06% colchicine + 0.5% dimethylsulfoxide solution.Following transfer of seedlings into the field, 2178 D0 plants were obtained out of a total of 3012 treated haploids.Live plants were from 89% of 2178 seedlings which are planted to the field.Fertile plants were formed 57% of live plants.Inbreeding was succeeded in 31.23% of fertile plants and only 7.8% of inbreeding plants were able to produce seeds.Consequently, 27 doubled haploid lines were developed.

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Available abstract

The doubled haploid technology is now an integral component of modern maize breeding programs.In this study, the maternal haploid induction (gynogenesis) method was used to derive Doubled-Haploid (DH) lines from elite maize germplasm adapted to Turkey.Temperate haploid inducers (RWS, RWK-76, RWS x RWK-76 and WS14) were used as pollinators, and a set of 30 single-crossses (in FAO 650-700 maturity groups) were used as source materials.Putative haploid seeds were selected based on expression of R1-nj anthocyanin color marker.Highest haploid induction rate (20.42%) was recorded by using RWK-76 as inducer line, and the lowest haploid induction rate (17.75%) was obtained through WS14.Putative haploid seeds were germinated and seedlings were treated with 0.06% colchicine + 0.5% dimethylsulfoxide solution.Following transfer of seedlings into the field, 2178 D0 plants were obtained out of a total of 3012 treated haploids.Live plants were from 89% of 2178 seedlings which are planted to the field.Fertile plants were formed 57% of live plants.Inbreeding was succeeded in 31.23% of fertile plants and only 7.8% of inbreeding plants were able to produce seeds.Consequently, 27 doubled haploid lines were developed.

Key concepts: Doubled haploidy, Ploidy, Biology, Agronomy, Biotechnology, Genetics, Gene

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