Cucumber Gynogenesis : Effects of 8 Different Media on Embryo and Plant Formation (original) (raw)
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Canadian Journal of Plant Science
The effects of the genotypes of donor plants and three steps of culture media (induction, differentiation, and regeneration media) were evaluated in unpollinated ovary culture of cucumber (Cucumis sativus L.) to obtain the best combination of culture media for doubled haploid (DH) production of recalcitrant cultivars. Embryo-like structure (ELS) formation was significantly affected by differentiation and regeneration media. The highest percentages of ELS formation (59.89%) were obtained when the ovaries were cultured on the D2 second step culture medium and transferred to the newly developed MS medium supplemented with several plant growth regulators and additives, MST3+, the third step culture medium. However, there was no significant effect of the genotypes and induction (first step culture) media on ELS and callus formation. The ELSs developed into shoots when transferred to the third step culture media and complete plantlets were obtained after rooting on the MS medium with no supplement (MS0). Among the 10 regenerated plantlets that survived after transplanting to soil, three were identified as haploid (H) plantlets (2n = x = 7), one was a triploid plantlet (2n = 3x = 21), and six were diploid plantlets (2n = 2x = 14). Inter-simple sequence repeat analysis confirmed that these six diploids were DH plantlets. These results suggest that the newly developed media are useful for the future production of cucumber DHs.
Review of research on haploid production in cucumber and other cucurbits
Folia Horticulturae, 2000
This review provides a summary of haploid induction methods and factors affecting the efficacy of specific methodologies as applied to cucumber (Cucumis sativus L.), melon (Cucumis melo L.), watermelon (Citrullus lanatus (Thunb.) Matsum. & Nakai), winter squash (Cucurbita maxima Duch. ex Lam.), summer squash (Cucurbita pepo L.) and other cucurbits. This report is focused on studies that were carried out during the last 20 years.
TURKISH JOURNAL OF BIOLOGY, 2016
Introduction Cucumber, a plant belonging to the family Cucurbitaceae, is among the top ten vegetables produced globally (Plader et al., 2007). This species is believed to have been domesticated in India for 3000 years and in eastern Iran and China for probably 2000 years (Aydemir, 2009). In cucumber, the development of homozygous parental lines using the traditional self-pollination method takes from 6 to 8 years (Gémes-Juhász et al., 2002). Therefore, the development of an efficient production system of doubled haploids and its further application in breeding programs could reduce the time required for cultivar development. In the family Cucurbitaceae, haploid plants have recently been obtained by different methods such as in vitro gynogenesis in summer squash (Cucurbita pepo L.) (Shalaby, 2007), in situ induction of haploid embryos via irradiated pollen in winter squash (Cucurbita maxima Duchesne ex Lam.) (Kurtar and Balkaya, 2010), parthenogenesis method using gamma-irradiated pollen in snap melon (Cucumis melo L.) (Godbole and Murthy, 2012), and anther culture method in watermelon (Citrullus lanatus L.) (Abdollahi et al., 2015). Several methods including spontaneous parthenogenesis (
Ploidy instability of embryogenic cucumber (Cucumis sativus L.) callus culture
Biologia Plantarum, 1996
Embryogenic callus cultures were established from immature cucumber (Cucumis sativus L.) embryos on E20A or MS (Murashige and Skoog 1962) media supplemented with 6-benzylaminopurine (BAP), ct-naphthylacetic acid (NAA) and/or 2,4-dichlorophenoxyacetic acid (2,4-D). Regeneration of plants was observed after a transfer to culture media either without growth regulators or supplemented with kinetin and NAA. Flow cytometry was employed to estimate DNA ploidy levels. Most of cell nuclei in young leaf tissues were found in G 1 phase with 2C DNA content. Callus cultures were mixoploid with DNA content ranging from 2C to 32C. The frequency of polyploid cells was increasing with the age of culture and the polyploidization was accompanied by a gradual loss of regeneration ability. Plants regenerated from callus cultures were classified as diploid (57 %), tetraploid (18 %), octoploid (4 %) and mixoploid (2n/4n, 4 %) and (4n/8n, 17 %). The results of this study confirmed a close link between the polyploidization and the loss of totipotency in vitro. Tetraploid plants obtained in this study have a potential to be used in interspecific crosses where their tetraploid status could help in overcoming existing breeding barriers due to differences in chromosome number.
Long-term suspension cultures of cucumber (Cucumis sativus L.) with high embryogenic potential
Acta Physiologiae Plantarum, 2009
Cucumber (Cucumis sativus L.) cytokininindependent embryogenic cell suspension cultures were derived and maintained for more than 3.5 years without losing the embryogenic potential. The preparation and the characteristics of the cucumber embryogenic cell suspension possess many similarities to that of carrot. The cultures were induced from hypocotyl explants of in vitro grown cucumber plants in liquid MS media containing 2,4dichlorophenoxyacetic acid as the sole growth regulator during 6 weeks and they contained a heterogeneous array of several different types of single cells and cell clusters (PEMs). The established cell suspensions were subcultured in 1-week interval, while the inoculation density was optimized to 2.0 9 10 5 cells ml-1 using cell viability as a marker. Somatic embryos were obtained after the transfer of the proembryogenic masses to a hormone-free semisolid MS medium with a frequency of 388 ± 57 somatic embryos per 1 ml of packed cell volume of the established cucumber embryogenic culture within 7 days. The frequency of normal somatic embryos with two cotyledons was found to be 78%. Such embryos possessed the potential of spontaneous maturation and the embryo conversion rates were 87%. The yield of normally growing plants was much higher compared with that previously described for cucumber systems. Somatic embryo-derived plants were successfully transferred to the greenhouse where they flowered and fruited. Keywords 2,4-D Á Cell lines Á Somatic embryo Á Embryo conversion Á Cucumber Communicated by E. Lojkowska.
Shoots formation from gynosinesis Cucumis sativus l
Engineering & Technology, 2020
CBM, Cucumis sativus L., haploid, gynogenesis, TDZ Haploid plants achieve through androgenesis or gynogenesis. In the gynogenesis method, the ovary or ovule are used as explants induct haploid plants. The female flower one day before the flowering of Cucumis sativus L. are collected. Cold pretreatment of ovaries at 4°C up to 24 hours and culture under dark conditions. Significantly enhanced callus induction response is compared with cultures under 4-week cultured on CBM medium supplemented with various concentrations of TDZ 0.01 0.04 mg/L. After 4 weeks, ovaries are transferred to medium with kinetin 0.05 0.20 mg/L. Then, ovaries were transferred to medium supplemented with BA: IAA 3:1. Finally, green ovaries were transferred to BA 1.5 mg/L and GA3 1.5 mg/L. The results showed that ovary induction has best affected on CBM with TDZ 0.03 mg/L with 11 callus/sample. Ovaries developed on kinetin 0.1 mg/L with 7.4 callus/sample. Ovaries become green and had leaves and roots formation on ...
Reproductive biology in monoecious and gynoecious cucumber cultivars as a result of IBA application
Horticultura Brasileira, 2008
I n the Brazilian vegetable growing sector, the economic value of cucumber (Cucumis sativus) is quite relevant for the continuation and development of the farms devoted to its exploitation. The average yield is approximately 10 t ha -1 for the pickle type, and 21 t ha -1 for the fresh type . Yield potential is not fully achieved due to several reasons, among DIOLA V; ORTH AI; GUERRA MP. 2008. Reproductive biology in monoecious and gynoecious cucumber cultivars as a result of IBA application. Horticultura Brasileira 26: 030-034.
In vitro culture of Cucumis sativus L
Theoretical and Applied Genetics, 1989
The ability to regenerate plants from leaf explants has been tested for three highly inbred cucumber lines (B, G, S), their reciprocal hybrids, F 2 and BC 1 generations. The lines differed from each other in their regenerating ability, which was expressed by the percentage of explants regenerating embryoidal callus and mean number of plantlets per plant. Thus, the lines could be classified as frequently (B), intermediately (G) or occasionally regenerating ones (S). There were no reciprocal cross differences in the regeneration. It was found that the intermediately and intensively regenerating lines contain two pairs of dominant genes responsible for plant regeneration, characterized by complementary and probably additive interaction. The frequently regenerating line differed from the intermediately regenerating in the effect of one gene. It is supposed that the above-mentioned genes belong to three different loci. The ability to regenerate plants from leafexplants had high heritability.
Embryogenesis and plant regeneration from anther cultures of Cucumis sativus L
Scientia Horticulturae, 2003
The response of anthers to in vitro culture and effect of growth regulators, temperature pretreatment of anthers have been studied in Calypso and Green Long cultivars of cucumber (Cucumis sativus L.). Direct and callus mediated embryogenesis (embryogenic calli with embryos) was induced in Calypso and Green Long cultivars respectively on B5 medium (Gamborg's medium) supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) and a-naphthaleneacetic acid (NAA) alone as well as 2,4-D in combination with N 6-benzylaminopurine/kinetin/thidiazuron (BAP/KN/TDZ). Optimal embryogenic calli/embryos were induced on B5 medium supplemented with 2.0 mM 2,4-D and 1.0 mM BAP. Temperature pretreatment of flower buds at 4 8C for 0-10 days and also at 32 8C for 1 day were tested and best response was obtained at 4 8C for 2 days. Embryo differentiation was achieved on B5 medium containing 0.09 M sucrose, 0.25 mM NAA and 0.25 mM KN. Embryo maturation was on B5 medium supplemented with 5.0 mM abscisic acid (ABA). Embryos germinated into plantlets on B5 medium containing 0.09 M sucrose. Plantlets were acclimatized in the controlled environment. In each cultivars, the root tips of 24 regenerated plantlets were analyzed for ploidy level, of which 21 and 17 have been haploids in Calypso and Green Long, respectively.
Genetically Stable Somatic Embryos of F1 Cucumber (Cucumis sativus) Hybrids
Acta Hort, 2007
Explants were taken from hypocotyls, cotyledons and leaves of five cucumber cultivars. The maximum callus weight in the induction medium was 1215.4 mg with the cv. Natica using hypocotyl explants and the minimum was 487.6 mg with cv. Mascot using leaf explants. When auxin was available in the induction phase somatic embryos were obtained; the absence of cytokinin caused a reduction in the number of embryos. The number of somatic embryos increased with sucrose concentration to a maximum of 9% sucrose. Three weeks on induction medium was needed for production of somatic embryos; prolonged incubation (6 weeks) gave poor somatic embryo numbers. As auxin concentration decreased in maturation medium somatic embryo yield increased; the highest number was obtained in the absence of auxin. Generally, the culture of callus on maturation medium in darkness was superior to 16 h light for somatic embryogenesis. Leaf explants of the cultivar Profito gave 37.3 embryos per replicate which was the highest number recorded. In addition, this cultivar yielded embryos in the shortest time period (6 weeks). Satisfactory somatic embryo numbers were obtained after three weeks on induction medium, plus three weeks on maturation medium, or four weeks on induction medium, plus two weeks on maturation medium, or five weeks on induction medium, plus one week on maturation medium. Comparison of characteristics of cucumber plants derived from somatic embryogenesis with those obtained from seed showed no significant difference in phenotype of plants, or fruits, or genotype (using RAPD DNA tests).