Improving <i>in vivo</i> maize doubled haploid production efficiency through early detection of false positives

Choe, Eunsoo; Carbonero, Christine Hayot; Mulvaney, Kelly; Rayburn, A. Lane; Mumm, Rita H.

doi:10.1111/j.1439-0523.2012.01962.x

Cited by 32 publications

(60 citation statements)

References 11 publications

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“…As demonstrated with ten populations in this study, use of seedling traits along with Navajo marker can reduce the FDR by several-fold compared to use of Navajo marker alone. Other methods proposed for identification of false positives include purple stem color marker (Röber et al 2005), differences in guard cell measurements (Choe et al 2012), flow cytometry (Dang et al 2012;de Couto et al 2013), and red root marker (Chaikam et al 2016). Purple stem marker develops only after sufficient vegetative growth (Chaikam et al 2016) and for conducting guard cell measurements and flow cytometry need sufficient growth to collect tissue.…”

Section: Discussionmentioning

confidence: 99%

Identification of in vivo induced maternal haploids in maize using seedling traits

et al. 2017

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In vivo haploid induction in high frequency followed by efficient identification of haploids are important components of deriving completely homozygous doubled haploid (DH) lines in maize. Several genetic marker systems were proposed and/or used for identification of in vivo maternal haploids in maize, such as R1-nj (Navajo), high oil, red root and transgenic markers. In this study, we propose a new method of haploid/diploid identification based on natural differences in seedling traits of haploids and diploids, which can be used in any induction cross independently of the genetic marker systems. Using confirmed haploids and diploids from five different populations, the study established that haploid and diploid seedlings exhibit significant differences for seedling traits, particularly radicle length (RL), coleoptile length (CL), and number of lateral seminal roots (NLSR). In six populations that exhibited complete inhibition of the commonly used R1-nj (Navajo) marker, we could effectively differentiate haploids from diploids by visual inspection of the seedling traits. In the haploid seed fraction identified based on R1-nj marker in ten populations, false positives were reduced several-fold by early identification of haploids at seedling stage using the seedling traits. We propose that seedling traits may be integrated at the haploid identification stage, especially in populations that are not amenable to use of genetic markers, and for improving the efficiency of DH line production by reducing the false positives.

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Section: Discussionmentioning

confidence: 99%

Identification of in vivo induced maternal haploids in maize using seedling traits

et al. 2017

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“…Dang et al (2012) observed rates of duplicated plants ranging from 28% to 54%. Battistelli et al (2013), in turn, obtained a higher percentage of duplicated plants, ranging from 59.1% to 80%, whereas the percentages of plants duplicated by Choe et al (2012) varied from 5% to 57.1% in the evaluated hybrids.…”

Section: Chromosome Duplication Of Haploid Linesmentioning

confidence: 82%

In vivo HAPLOID INDUCTION AND EFFICIENCY OF TWO CHROMOSOME DUPLICATION PROTOCOLS IN TROPICAL MAIZE

Couto

Pinho

et al. 2015

Ciênc. agrotec.

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Artificial chromosome duplication is one of the most important process in the attainment of doubled haploids in maize. This study aimed to evaluate the induction ability of the inducer line KEMS in a tropical climate and test the efficiency of the R1-Navajo marker by flow cytometry to evaluate two chromosome duplication protocols and analyze the development of the doubled haploids in the field. To accomplish this goal, four genotypes (F1 and F2 generations) were crossed with the haploid inducer line KEMS. The seeds obtained were selected using the R1-Navajo marker and subject to two chromosome duplication protocols. Duplication was confirmed using flow cytometry. The percentages of self-fertilized plants after duplication as well as the quantities of doubled haploid seeds obtained after the self-fertilization processes were analyzed. It was observed that the germplasm influences haploid induction but not the duplication rates of the tested protocols. Protocol 2 was more efficient for the duplication of haploids, in the percentage of self-fertilized plants after duplication, and in the attainment of doubled haploid lines. Moreover, the haploid inducer line KEMS can produce haploids in a tropical climate. Other markers, in addition to the R1-Navajo system, should be used in the selection of haploid seeds.Index terms: R-Navajo; colchicine; induction of maternal haploids; Zea mays. RESUMOA duplicação cromossômica artificial está dentre as etapas mais importantes na obtenção de duplo-haploides em milho. Este estudo objetivou avaliar a capacidade de indução da linhagem indutora KEMS em clima tropical e testar a eficiência do marcador R1-navajo por meio de citometria de fluxo; avaliar dois protocolos de duplicação cromossômica e, analisar o desenvolvimento dos haploides duplicados no campo. Para isso, quatro genótipos (gerações F1 e F2) foram cruzados com a linhagem KEMS. As sementes obtidas foram selecionadas pelo marcador R1-navajo e submetidas a dois protocolos de duplicação cromossômica. A duplicação foi confirmada por meio de citometria de fluxo. As porcentagens de plantas autofecundadas após duplicação foram analisadas, bem como as quantidades de sementes duplo-haploides obtidas após as autofecundações. Foi observado que o germoplasma influencia a indução de haploides, mas não na taxa de duplicação dos protocolos testados. O protocolo 2 foi mais eficiente na duplicação de haploides, na porcentagem de plantas autofecundadas após duplicação, e na obtenção de linhagens duplo-haploides. Além disso, a linhagem indutora KEMS pode induzir haploides em clima tropical. Outros marcadores além do sistema do R1-navajo devem ser utilizados na seleção de sementes haploides.Termos para indexação: R-navajo; colchicina; indução de haploides maternos; Zea mays.

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“…This trait could be used to differentiate haploids and DHs from outcrossing plants (Borrino and Powell, 1988;Sood et al, 2003). A recent study in maize revealed that mean stomata length of haploid and DH plants at early stage (through leaf 1 to leaf 8 stage) is significantly smaller than that of the diploid controls (outcrossing seeds), thus indicating that stomata length together with anthocyanin color could be used to detect true haploids and DHs from control plants derived from outcrossing seeds (Choe et al, 2012).…”

Section: Plant Morphology and Stomatal Chloroplast Countmentioning

confidence: 98%

Haploids: Constraints and opportunities in plant breeding

Dwivedi

Britt

Tripathi

et al. 2015

Biotechnology Advances

235

134

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The discovery of haploids in higher plants led to the use of doubled haploid (DH) technology in plant breeding. This article provides the state of the art on DH technology including the induction and identification of haploids, what factors influence haploid induction, molecular basis of microspore embryogenesis, the genetics underpinnings of haploid induction and its use in plant breeding, particularly to fix traits and unlock genetic variation. Both in vitro and in vivo methods have been used to induce haploids that are thereafter chromosome doubled to produce DH. Various heritable factors contribute to the successful induction of haploids, whose genetics is that of a quantitative trait. Genomic regions associated with in vitro and in vivo DH production were noted in various crops with the aid of DNA markers. It seems that F2 plants are the most suitable for the induction of DH lines than F1 plants. Identifying putative haploids is a key issue in haploid breeding. DH technology in Brassicas and cereals, such as barley, maize, rice, rye and wheat, has been improved and used routinely in cultivar development, while in other food staples such as pulses and root crops the technology has not reached to the stage leading to its application in plant breeding. The centromere-mediated haploid induction system has been used in Arabidopsis, but not yet in crops. Most food staples are derived from genomic resources-rich crops, including those with sequenced reference genomes. The integration of genomic resources with DH technology provides new opportunities for the improving selection methods, maximizing selection gains and accelerate cultivar development. Marker-aided breeding and DH technology have been used to improve host plant resistance in barley, rice, and wheat. Multinational seed companies are using DH technology in large-scale production of inbred lines for further development of hybrid cultivars, particularly in maize. The public sector provides support to national programs or small-medium private seed for the exploitation of DH technology in plant breeding.

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Improving in vivo maize doubled haploid production efficiency through early detection of false positives

Cited by 32 publications

References 11 publications

Identification of in vivo induced maternal haploids in maize using seedling traits

Identification of in vivo induced maternal haploids in maize using seedling traits

In vivo HAPLOID INDUCTION AND EFFICIENCY OF TWO CHROMOSOME DUPLICATION PROTOCOLS IN TROPICAL MAIZE

Haploids: Constraints and opportunities in plant breeding

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