Production of Doubled Haploid Embryos from Cork Oak Anther Cultures by Antimitotic Agents and Temperature Stress

Abstract This paper describes a method to obtain haploid and doubled-haploid (DH) embryos using anther cultures of holm oak (Quercus ilex L.). The production of haploids and DH through gametic embryogenesis provides an attractive biotechnological tool for developing homozygous lines from heterozygous parents, which is important in breeding programs, as well as in genetic studies. As a consequence, protocols to produce homozygous plants have a significant impact on forest tree improvement. Anthers were subjected to different temperature treatments for embryo induction: a cold pre-treatment (4°C) from 3 to 7 days was carried out at the beginning, followed by a heat shock (33°C) from 2 to 5 days. Most anthers responding to these stress treatments contained vacuolated microspores, indicating that this developmental stage is responsive to embryogenesis induction in holm-oak microspores. In all cases, embryos grew from the interior of the anthers, breaking through the degenerating anther walls. Under these conditions, embryo formation occurred in 31 anthers between 46 and 95 days after culture initiation. Embryo analysis performed with flow-cytometry and DNA-microsatellite markers showed haploid profiles and/or spontaneous doubling of the chromosomes during early regeneration stages. This is, to our knowledge, the first published report on gametic embryogenesis in holm oak.

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Antimitotic agents increase the production of doubled-haploid embryos from cork oak anther culture

Journal of Plant Physiology ◽

10.1016/j.jplph.2006.11.012 ◽

2007 ◽

Vol 164 (12) ◽

pp. 1595-1604 ◽

Cited By ~ 29

Author(s):

Beatriz Pintos ◽

Jose A. Manzanera ◽

Maria A. Bueno

Keyword(s):

Anther Culture ◽

Doubled Haploid ◽

Cork Oak ◽

Haploid Embryos

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Comparison of methods for obtaining doubled haploids of carrot

Acta Societatis Botanicorum Poloniae ◽

10.5586/asbp.3547 ◽

2017 ◽

Vol 86 (2) ◽

Cited By ~ 3

Author(s):

Waldemar Kiszczak ◽

Urszula Kowalska ◽

Agata Kapuścińska ◽

Maria Burian ◽

Krystyna Górecka

Keyword(s):

Plant Regeneration ◽

Doubled Haploid ◽

Aspartate Aminotransferase ◽

Doubled Haploids ◽

The Other ◽

Plant Production ◽

Comparison Of Methods ◽

Anther Cultures ◽

Induction Stage ◽

Main Factor

Doubled haploid lines of carrot can be obtained through androgenesis in anther cultures and in isolated microspore cultures. The two methods were compared using three carrot cultivars (‘Kazan F1’, ‘Feria F1’, and ‘Narbonne F1’) at the androgenesis induction stage, during plant regeneration from embryos, and during acclimatization of androgenetic plants as well as their characterization. It was found that cultivar was the main factor affecting the efficiency at each stage of plant production in both anther and isolated microspore cultures. The efficiency of androgenesis in anther cultures of ‘Feria F1’ was considerably higher in comparison with isolated microspore cultures, and more plants were obtained from the embryos of androgenesis-cultured plants. In ‘Kazan F1’ and ‘Narbonne F1’, more acclimatized androgenetic plants were produced from anther cultures. Ploidy assessment of acclimatized plants of ‘Narbonne F1’ showed that the majority of the plants in the population derived from anther cultures had a doubled chromosome (DH) set. On the other hand, the majority of plants obtained from isolated microspore cultures were haploids. When assessing homozygosity, it was found among plants obtained in anther cultures that the percentage of homozygotes for phosphoglucose isomerase (PGI) and aspartate aminotransferase (AAT) depended on the cultivar. In contrast, the majority of plants derived from isolated microspore cultures were homozygous regardless of cultivar.

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Factors Affecting Doubled Haploid Plant Production Via Maize Technique in Bread Wheat

Acta Biologica Cracoviensia s Botanica ◽

10.2478/abcsb-2014-0022 ◽

2015 ◽

Vol 56 (2) ◽

pp. 67-73

Author(s):

Ioannis Xynias ◽

Antonios Koufalis ◽

Evdokia Gouli-Vavdinoudi ◽

Demetrios Roupakias

Keyword(s):

Bread Wheat ◽

Doubled Haploid ◽

Embryo Rescue ◽

Plant Production ◽

Haploid Plant ◽

In Planta ◽

Maize Pollen ◽

Doubled Haploid Plants ◽

Haploid Embryos ◽

Haploid Plants

Abstract The effect of two in planta factors (growth conditions, genotype) and two in vitro factors (time of embryo rescue, embryo rescue medium) on doubled haploid (DH) plant production in bread wheat via maize technique was investigated in nine F1 hybrids produced after crossing four bread wheat cultivars. During the first year one group of F1 plants was grown in a field and at the proper stage pollinated with maize pollen (sweet corn popu-lation). In parallel, a second group of F1 plants was grown in a growth chamber and pollinated as in the former group. In the second growing season the experiment was repeated but only field-grown plants were used. All the produced haploid embryos were cultured in three different media and the resulting 146 haploid plants were sub-sequently treated with aqueous solution of colchicine. Finally, 86 doubled haploid plants were obtained. We noted that the growing conditions of the parental plants and the intervening time between day of pollination and day of embryo rescue influenced the percentage of haploid embryo production. Culture medium also influenced haploid and doubled haploid plant production. The two media (MS/2, B5) were found equally effective. Most of the haploid embryos originated from the Penios × Acheloos cross, whereas most of the doubled haploid plants were produced from the KVZ × Penios cross. Doubled haploid plants were produced from all crosses.

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Induction of Parthenogenetic Haploid Embryos after Pollination by Irradiated Pollen in Watermelon

HortScience ◽

10.21273/hortsci.29.10.1189 ◽

1994 ◽

Vol 29 (10) ◽

pp. 1189-1190 ◽

Cited By ~ 21

Author(s):

N. Sari ◽

K. Abak ◽

M. Pitrat ◽

J.C. Rode ◽

R. Dumas de Vaulx

Keyword(s):

In Vitro Culture ◽

Doubled Haploid ◽

Chromosome Doubling ◽

Citrullus Lanatus ◽

Doubled Haploid Lines ◽

Irradiated Pollen ◽

Haploid Embryos ◽

Haploid Plants

Parthenogenetic haploid embryos of `Crimson Sweet', `Halep Karasi', `Sugar Baby' and `Panonia F1' watermelon [Citrullus lanatus (Thunb.) Matsum. & Nakai] were obtained after pollination with γ-irradiated (200 or 300 Gy) pollen. Some globular and heart-shaped embryos were observed in fruit harvested 2 to 5 weeks after pollination. The number of embryos per 100 seeds was highest for `Halep Karasi'. After in vitro culture, 17 haploid plants were obtained and doubled haploid lines were generated after chromosome doubling using colchicine.

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Production of doubled haploid plants from anther cultures of borage (Borago officinalis L.) by the application of chemical and physical stress

Plant Cell Tissue and Organ Culture (PCTOC) ◽

10.1007/s11240-017-1233-4 ◽

2017 ◽

Vol 130 (2) ◽

pp. 369-378 ◽

Cited By ~ 6

Author(s):

Zahra Sanaey Hoveida ◽

Mohammad Reza Abdollahi ◽

Asghar Mirzaie-Asl ◽

Sayyed Saeed Moosavi ◽

Jose M. Seguí-Simarro

Keyword(s):

Doubled Haploid ◽

Physical Stress ◽

Borago Officinalis ◽

Doubled Haploid Plants ◽

Anther Cultures ◽

Haploid Plants

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Strategy for improvement of doubled haploid production in maize

Acta Agronomica Hungarica ◽

10.1556/aagr.53.2005.2.6 ◽

2005 ◽

Vol 53 (2) ◽

pp. 177-182

Author(s):

B. Barnabás ◽

T. Spitkó ◽

K. Jäger ◽

J. Pintér ◽

L. C. Marton

Keyword(s):

Doubled Haploid ◽

Doubled Haploid Line ◽

Breeding Strategy ◽

Breeding Value ◽

Line Production ◽

Hybrid Plants ◽

Heterosis Effect ◽

Anther Cultures ◽

Chinese Germplasm

In the present study the applicability of a self-constructed doubled haploid line (DH 105) in the in vitro breeding of maize was evaluated. This line, which contained only 50% exotic (Chinese) germplasm, could be used to transmit in vitro androgenic ability into non-responsive breeding materials. F1 hybrids resulting from single crosses between the moderately responsive line DH 105 and recalcitrant genotypes with high breeding value showed a considerable heterosis effect in their androgenic responses. Most of the hybrids had favourable morphological and agronomic characters on the basis of “per se” evaluation. The data of the experiments showed that these F1 hybrid plants could be successfully used as anther donors, since numerous fertile DH plants were developed from their anther cultures. By the use of this in vitro breeding strategy the genetic variability can be widened and the effectiveness of inbred line production might be improved.

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