Genetically Stable Somatic Embryos of F1 Cucumber (Cucumis sativus) Hybrids (original) (raw)
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Genetically stable cell lines of cucumber for the large-scale production of diploid somatic embryos
Physiologia Plantarum, 1996
Raaijmakers, N. and van Hoist, G.-J. 1996. Genetically stable celi lines of cucumber for the large-scale production of diploid somatic embryos.-Physiol. Plant. 97: 303-310. For the initiation of embr\'ogenic cucumber (Cucumis sativus L.) cell lines, from excised radicles, directly in liquid medium, the culture regime, explant density' and type and concentration of hormones were adjusted so that pro-embryogenic masses (PEMs) were formed within about 8 weeks. The established cucumber cell lines were maintained for severai years without loss of embryogenic and genetic stability. The ploidy level of somatic embryos from different cucumber eel! lines was either diploid or tetraploid and depended on the ploidy level of Ihe cell line. Cucumber cell lines that produced only diploid embryos were obtained by selecting completely diploid explant material and growing 11 in the dark during the initiation phase, Mixoploid explants could lead to tetraploid or mixoploid cell lines. Isolation and additional selection and subculturing of single PEMs resulted in either completely diploid or tetraploid eel! lines, indicating that all ceils of individual PEMs are either diploid or tetraploid. The embryogenic cucumber cell lines, differing only in ploidy level, were indistinguishable in growth rate and embrj'ogenic potential and were genetically stable over several years.
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.
Somaclonal variation in cucumber (Cucumis sativusL.) plants regenerated via embryogenesis
Acta Botanica Neerlandica, 1990
Plants were regenerated via embryogenesis from leaf explant-derived callus of cucumber (Cucumis sativus L. cv. Hokus). Somaclonal variation was evaluated in the regenerated plants (R) generation) and in the selfed progeny (R z). Considerable variation was found in the young R) plants, but it was of a transient nature. Mature R) plants showed slight abnormalities which were not directly transmitted to the progeny. Seed production of the R1s was variable and very poor, as was seed germination. This is probably caused by dysfunctioning of generative tissue and cells and might have a genetic base. Of eight R z lines analysed, one was tetraploid and five expressed a low extent of variability, probably of a genetic nature. Most of the R, plants were phenotypically normal. In a yield analysis, genetic variation useful for improvement of crop performance has not been detected. It is suggested that in cucumber the generative phase acts as a sieve for genetically based somaclonal variation.
Somatic Embryos Derived from Cotyledons of Cucumber
Journal of the American Society for Horticultural Science, 1990
Two studies were conducted to test the effects of various tissue culture media on somatic embryogenesis from cotyledon tissue of cucumber (Cucumis sativus L.). The two best media for embryo initiation were Murashige and Skoog (MS) salts and vitamins containing either 1 or 2 mg 2,4-D/liter and 0.5 mg kinetin/liter. In the second study, embryos developed more normally. More plantlets developed when tissue was removed from the initiation medium after 3 weeks and transferred to MS containing 1 mg NAA/liter and 0.5 mg kinetin/liter for 3 weeks, rather than leaving the embryos on a medium containing 2,4-D. Histological evidence indicated that the embryos were multicellular in origin. Charcoal in the maturation medium inhibited embryo development. Chemical names used: (2,4-dichlorophenoxy) -acetic acid (2,4-D); N-(2-furanylmethyl)-lH-purine-6-amine (kinetin); 1-naphthaleneacetic acid (NAA).
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.
Somatic Embryogenesis and Shoot Regeneration Induced in Cucumber Leaves
Pakistan Journal of Botany
Commercial cucumber cultivars were explored for embryogenesis and plant regeneration induced in somatic tissues on plant growth regulators (PGRs). Maximum callus induction 94.16% and 76% was observed in leaf disc explants on MS medium supplemented with 2,4-D (2 mgL -1), NAA and BAP (1.5 mgL -1 , each), respectively. Seed cotyledon explants induced maximum calli (77%) on 4.0 + 0.75 mgL -1 (BAP + NAA, respectively). Calli induced in leaf disc on the highest level of 2,4-D (5 mgL -1) yielded the highest embryo formation (23%) whereas calli induced on BAP and BAP + NAA (5 + 1 mgL -1) regenerated into 14% and 12%, shoots respectively. These shoots were excised and rooted on MSO medium. The plantlets were transplanted in pots and transferred to field after acclimatization. The developed plant material will be morphologically and genetically characterized for homozygosity.
During indirect regeneration of plants, chances of somaclonal variations may arise. These variations should be identified to produce true to type plants. Inter-simple sequence repeat (ISSR) and sodium dodecyl sulphate - poly acrylamide gel electrophoresis (SDS-PAGE) were used to assess the somaclonal variation among regenerated plantlets, an embryogenic calli produced from leaf explants compared to in vivo plants of three cucumber (Cucumis sativus L.) cultivars (Waffir F1, Beith Alpha and Fares). Five ISSR primers yielded 34 distinct and reproducible band patterns with an average of 6.8 bands per primer. Jaccard's similarity coefficients revealed presence of a relatively high genetic similarity (75%) among regenerated plantlets and in vivo plants of Beith Alpha cultivar compared to other cultivars. However, relatively high genetic variability have been recorded among calli and in vivo plants of Waffir F1 (62%) and Fares (45%) cultivars. Protein profile derived by SDS-PAGE analysis revealed that there are variation among in vivo plants, calli and regenerated plantlets of three cucumber cultivars. The molecular weights of detected bands for all samples ranged from 23 to 107 KDa. A hundred percent (100 %) similarity between in vivo plants of Waffir and Beith Alpha cultivars as well as between their calli has been noted. The regenerated plantlets of Beith Alpha showed a relatively higher similarity of 78% with in vivo plants. Variations in protein pattern were recorded between calli and regenerated plantlets of Waffir (70%) and Beith Alpha (57%) cultivars from one side and between calli and in vivo plants from another side (67% in both cultivars). We think that leaf explant culturing influenced by the cucumber cultivar genotype and some genotypes are more susceptible to somaclonal variation, or that the in vitro instability is actually a consequence of a genotype versus culture medium interaction. Our results suggested that rapid propagation of cucumber Beith Alpha cultivar via leaf organogenesis is an acceptable method with a lower risk of somaclonal variation. [Mamdouh D, Marghany HA and Ewais EA. Assessment of Somaclonal
Viability assessment of in-vitro produced synthetic seeds of cucumber
AFRICAN JOURNAL OF BIOTECHNOLOGY
Friable, embryogenic calli of F1 cucumber (Cucumis sativus) cultivar, Royal, were induced from the hypocotyl pieces cultured on solidified MS-basal media supplemented with 2,4-dichlorophenoxy acetic acid (2,4-D) and benzyl amino purine (BAP). Embryogenic calli were transferred to liquid Murashige and Skoog (MS)-basal media supplemented with 5 µM naphthaleneacetic acid (NAA) and 1 µM BAP. The mature somatic embryos were encapsulated in sodium alginate mixture in synthetic seeds. The encapsulation mixture containing 3% sodium alginate, 100 mM calcium chloride and one-fourth volume of the cell suspension nutrient mixture containing 5 × 10-4 somatic embryos per ml was found the best. Synthetic seeds remain viable up to 14 weeks when stored at 4°C. Germination efficiency of synthetic seeds was decreased to 57% after 10 weeks of storage followed by rapid decrease in survival rate to 0% after 15 weeks. Genetic diversity between mother plants and in vitro produced synthetic seeds showed res...
Induction of somatic embryogenesis in absence of exogenous auxin in cucumber (Cucumis sativus L.)
International Journal of Plant Production, 2012
Induction of somatic embryogenesis in absence of exogenous auxin can clarify some basic concepts of somatic embryogenesis. Explants from leaf and petiole of Cucumber (Cucumis sativus) were used in this experiment for somatic embryogenesis using three different modified B5 basal media; boron and 2,4-D but no IAA as control (I), boron and IAA but no 2,4-D (II) and no boron, 2,4- D and IAA (III). Leaf explants grown on III produced somatic embryos indicating an induction potential for somatic embryogenesis with an incomplete medium. The endogenous auxin of the leaf explants might have a potential for triggering induction of somatic embryogenesis.
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 (