Inducer line generated double haploid seeds for combined waxy and opaque 2 grain quality in subtropical maize (Zea mays. L.)

Dang, Ngoc-Chi; Munsch, Magali; Aulinger, I. E.; RenLai, Wen; Stamp, P.

doi:10.1007/s10681-011-0423-0

Cited by 29 publications

(32 citation statements)

References 14 publications

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“…Therefore, cytometric analysis was performed on all plants that survived the colchicine treatment and remained alive until sample collection (N = 918), which was approximately 62% of all treated plants. Dang et al (2012) observed similar seedling survival results (65.7%) in colchicine-treated waxy and opaque maize haploids. The authors noted that the surviving plants had twisted leaves with stunted growth during the first stage of development.…”

Section: Discussionsupporting

confidence: 70%

Production and identification of doubled haploids in tropical maize

Battistelli

Pinho²,

Justus³

et al. 2013

Genet. Mol. Res.

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ABSTRACT. The production of maize doubled haploid (DH) lines is a technique commonly used by private companies, but not by Brazilian public institutions. Research on this technique is essential to develop and improve the production of DH lines grown under tropical conditions. We assessed the ability of a gynogenetic haploid inducer system to induce haploids in a tropical environment, assessed the induction rate of haploids identified using the R-navajo morphological marker, checked for interference from the generation of hybrid donors on haploid induction, measured the ability of flow cytometry, and simple sequence repeat marker techniques to identify doubled haploids. Seeds from the inducer Krasnodar Embryo Marker Synthetic (KEMS) line were sown in Ponta Grossa, PR, and Cravinhos, SP, and the plants were crossed to produce six hybrids and their F 2 generations. The seeds were separated according to the R-navajo morphological marker indicator of haploidy (purple endosperm and white embryo) and germinated in a controlled environment. Chromosomal duplication was performed in seedlings selected as putative haploids. We performed subsequent confirmation of ploidy and the success of duplication using flow cytometry and SSR 4231©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 12 (4): 4230-4242 (2013) Production of doubled haploid lines in tropical maize marker techniques. We concluded that DH lines can be obtained from hybrids crossed with the inducer KEMS line. The generation of inbred hybrids did not affect the induction rate or chromosomal duplication in haploids. The use of flow cytometry and SSR markers was effective in verifying chromosomal duplication in haploids.

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Section: Discussionsupporting

confidence: 70%

Production and identification of doubled haploids in tropical maize

Battistelli

Pinho²,

Justus³

et al. 2013

Genet. Mol. Res.

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“…Flow cytometry is advantageous in experiments involving chromosome duplication (Choe et al, 2012;Dang et al, 2012;Battistelli et al, 2013;Couto et al, 2013), as the sample is not degraded because only a piece of the leaf is sufficient for the analyses and little time is required for cytometry measurements (Loureiro and Santos, 2004). In this study, we analyzed all doubled plants in less than one week.…”

Section: Discussionmentioning

confidence: 99%

“…Because flow cytometry can be used to directly evaluate DNA content and indirectly determine the ploidy level, this technique allows for the analyses of haploid induction as well as the chromosome duplication (Dang et al, 2012;Battistelli et al, 2013;Couto et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

Verification and characterization of chromosome duplication in haploid maize

Couto¹,

Pinho²,

Pinho³

et al. 2015

Genet. Mol. Res.

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ABSTRACT. Doubled haploid technology has been used by various private companies. However, information regarding chromosome duplication methodologies, particularly those concerning techniques used to identify duplication in cells, is limited. Thus, we analyzed and characterized artificially doubled haploids using microsatellites molecular markers, pollen viability, and flow cytometry techniques. Evaluated material was obtained using two different chromosome duplication protocols in maize seeds considered haploids, resulting from the cross between the haploid inducer line KEMS and 4 hybrids (GNS 3225, GNS 3032, GNS 3264, and DKB 393). Fourteen days after duplication, plant samples were collected and assessed by flow cytometry. Further, the plants were transplanted to a field, and samples were collected for DNA analyses using microsatellite markers. The tassels were collected during anthesis for pollen viability analyses. Haploid, diploid, and mixoploid individuals were detected using 7000 E.G. de Oliveira Couto et al.©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 14 (2): 6999-7007 (2015) flow cytometry, demonstrating that this technique was efficient for identifying doubled haploids. The microsatellites markers were also efficient for confirming the ploidies preselected by flow cytometry and for identifying homozygous individuals. Pollen viability showed a significant difference between the evaluated ploidies when the Alexander and propionic-carmin stains were used. The viability rates between the plodies analyzed show potential for fertilization.

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“…The hybrids did not differ significantly between each other regarding the duplicated plant rate. 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: 98%

“…Generally, these haploid inducer lines have a temperate origin, which hampers the handling and development of these lines in tropical conditions. According to Rotarenco et al (2010) and Dang et al (2012), some haploid inducer lines have been developed and have relatively high haploid induction rates, such as: MHI with 7.2% induction, the modern European line RWS with 8% induction, and the 436 Ciênc. Agrotec., Lavras, v. 39, n. 5, p. 435-442, set./out., 2015 PHI line with 12.8% induction.…”

Section: Introductionmentioning

confidence: 99%

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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Inducer line generated double haploid seeds for combined waxy and opaque 2 grain quality in subtropical maize (Zea mays. L.)

Cited by 29 publications

References 14 publications

Production and identification of doubled haploids in tropical maize

Production and identification of doubled haploids in tropical maize

Verification and characterization of chromosome duplication in haploid maize

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

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