Anatomical tool in the identification of ploids in maize seedlings and potential use in initial stage of double haploides obtainment process

Author:

Pires Raquel M. O.,Souza Genaina A.,Vilela Danielle R.ORCID,Santos Heloisa O.,C.Vasconcellos Renato C.,Von Pinho Édila V. R.

Abstract

AbstractStudies that optimize the haploid technique in the removal of maize lines are necessary. Between the stages that mostly requires attention and it is directly related to the success of the technology is the correctly separation of induced haploids and diploids. Morphological markers are commonly used but have strong influence of the environment, and laboratory methods have been developed and may be more efficient. Thus, the objective was to study the use of the anatomical analysis tool, through the analysis of young maize leaf for use as the indirect markers in the identification of ploidys. The hybrids were crossed with the KEMS haploid inducer. The seeds crossed, were selected according to the R-navajo marker and submitted to two different protocols of chromosome duplication. Plants that survived to the duplication protocols were acclimated in greenhouse and then transferred to the field. After the self-polinization of the DH0 plants, the DH1 seeds were taken to the field, divided into treatments according to the parentals and duplication protocols. At the vegetative stage V4 of the plants, leaf tissue samples were collected to the evaluation of the amount of DNA and identification of ploidys and anatomical analysis. The nuclear DNA review of each sample was performed for the comparison in histograms of the position of G1 peak to the G1 peak of the internal or external reference standard. A high accuracy came to validate an anatomical tool, through the variables studied in this work, as a marker in the differentiation of ploidis in maize plants, and it can be used in selection programs. The anatomy made in some letters is a non-destructible technique and, together with a flow cytometry technique, can be used as an indirect method in haploid cutting programs at the initial stage of the identification of seedlings.

Publisher

Cold Spring Harbor Laboratory

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