Production of double haploids in oat (Avena sativa L.) by pollination with maize (Zea mays L.)

Abstract

Production of double haploids in oat (Avena sativa L.) by pollination with maize (Zea mays L.) Abbreviations 2,4-D -2,4-dichlorophenosyacetic acid; dicamba -3,6-dichloro-2-methoxybenzoic acid; DH -double haploid. IntroductionModern crop breeding has a number of advantages when using DH plants, such as shortening of the production time of new varieties and the possibility of using them in research. Classical methods of production of new varieties, based on a selection in every generation, take six to ten years. Biotechnological methods allow for a shortening of this time, even to one vegetative season. Breeders can make new cultivars with features from DH lines and be confident about completely homozygous plants. Moreover, DH lines can be used in the production of genetic maps, which help in the search for quantitative traits loci (QTL), conjugated with molecular markers and connected with many physiological processes, such as receptivity to the effectiveness of androgenesis or tolerance to biotic and abiotic stresses [1,2]. Haploids can be obtained using a number of methods, namely: from male gametophyte by androgenesis, using anther cultures or isolated microspores cultures [3][4][5][6][7][8], or from female gametophytes by gynogenesis or wide crossing, usually with maize (a method also known as chromosomes elimination) [1,9,10]. In comparison to anther culture, pollination with maize has an advantage because all regenerated haploid embryos are green, whereas in anther culture many of them are albino. The method of chromosome elimination that has been used in these experiments includes the following stages: after emasculation of cereal flowers the stigma is pollinated by maize, the pollen then germinates, a pollen tube is formed [11,12] and grows into the cereal embryo sac, where the cereal egg is fertilised by the maize sperm nuclei. A hybrid zygote is produced. The hybrid zygotes Keywords: Wide crossing • 2,4-D • Dicamba • Oat haploid embryosAbstract: The aim of the study was to optimize the method of oat haploid production by pollination with maize. Seventeen oat genotypes were used in the experiment. Various factors influencing the growth and development of ovaries and embryo production were investigated: genotype, time of pollination, growth regulators and time of their application. Emasculated before anthesis, oat florets were pollinated with maize pollen after 0, 1 or 2 days. Next, one of two auxins analogues (2,4-D or dicamba) were applied to oat pistils. These auxins had no significant influence on the number of enlarged ovaries and embryos. The time of application of these growth regulators had a significant influence on embryo production. Haploid embryos were obtained from all used genotypes, although the frequency of enlarged ovaries and obtained embryos did not differ markedly between the genotypes. On average, 85% of ovaries were enlarged and 11.7% of them produced haploid embryos. Depending on the regeneration medium, 24-41% of embryos were germinated, of which 12% had developed into green plant...

Application of chosen factors in the wide crossing method for the production of oat doubled haploids

“…They have also tested the medium without amino acids and medium supplemented with filtered coconut milk. are consistent with the study of Marcińska et al [10], where haploid plants were also obtained from embryos cultivated on medium with maltose. A unique part of our experiment was the isolation of haploid embryos at different ages.…”

“…Five oat genotypes (F 1 progeny) derived from Strzelce Plant Breeding Ltd., selected from the previous experiment as the most responsive to haploidization [10] …”

Application of photochemical parameters and several indices based on phenotypical traits to assess intraspecific variation of oat (Avena sativa L.) tolerance to drought

“…Plant materials and growth conditions DH lines were obtained by pollination of oat with maize at the Institute of Plant Physiology in Cracow, according to the method of Marcińska et al (2013a). F 1 oat (Avena sativa L.) generation (Table 1) and sweet corn Waza (Zea mays) derived from the Strzelce Plant Breeding Ltd. served as the source of plant material.…”

“…Oat doubled haploids (DH) are one of the technologies currently being used in the programs aimed at developing new varieties with a combination of desirable traits in a shorter time frame. In our previous study, we have obtained a number of DH lines of oat (Marcińska et al 2013a), while in the current study we have tested their susceptibility to drought stress in relation to the abovedescribed valuable characteristics of those types of lines.…”

The effect of light intensity on the production of oat (Avena sativaL.) doubled haploids through oat × maize crosses

“…The most commonly applied DH systems in oat (Avena sativa L.) are wide hybridization through interspecific crosses (Rines 2003;Sidhu et al 2006;Marcińska et al 2013;Ishii et al 2013;Nowakowska et al 2015;Warchoł et al 2016) and the use of androgenesis in anther culture (Kiviharju 2009) or isolated microspore culture (Sidhu and Davies 2009). Wide crossing with maize (Zea mays L.) is most extensively applied in the commercial production of oat DH lines.…”