Histological comparison between wheat embryos developing in vitro from isolated zygotes and those developing in vivo

Bakos, Ferenc; Szabó, László; Olmedilla, Adela; Barnabás, Beáta

doi:10.1007/s00497-008-0087-7

Cited by 7 publications

(5 citation statements)

References 77 publications

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“…The similar observation for abundant starch and protein accumulation was found in the scutellum of hexaploid wheat embryos developing in vitro from isolated zygotes [52]. In agreement with this, the “dormant” structures formed in the presence of DAM showed visual similarity with scutellum of zygotic embryos being hard, white and non translucent.…”

Section: Discussionsupporting

confidence: 81%

Protocol for efficient regulation of in vitro morphogenesis in einkorn (Triticum monococcum L.), a recalcitrant diploid wheat species

et al. 2017

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Einkorn (Triticum monococcum L.) is A-genome diploid wheat that has a potential to become a useful model for understanding the biology and genomics in Triticeae. Unfortunately, the application of modern technologies such as genetic engineering, RNAi-based gene silencing and genome editing is not available for einkorn as there is no efficient in vitro tissue culture and plant regeneration system. In the present study an efficient and simple protocol for plant regeneration via direct or indirect somatic embryogenesis and organogenesis has been developed. Various auxins used as sole inductors in einkorn displayed low effect for morphogenesis (0–8%) and plant regeneration (1–2 shoots per explant). The addition of Daminozide, the inhibitor of biosynthesis of gibberellins, together with auxin significantly improved the formation of morphogenic structures, especially when Dicamba (51.4%) and Picloram (56.6%) were used for combination; furthermore, the simultaneous addition of cytokinin into induction medium significantly promoted in vitro performance. Among the tested cytokinins, the urea-type substances, such as TDZ and CPPU were more effective than the adenine type ones, BA and Zeatin, for the regulation of morphogenesis; especially, TDZ was more effective than CPPU for shoot formation (11.73 vs. 7.04 per regenerating callus). The highest morphogenic response of 90.2% with the production of more than 10 shoots per initial explant was observed when 3.0 mg/L Dicamba, 50.0 mg/L Daminozide and 0.25 mg/L TDZ were combined together. Along with the identification of appropriate induction medium, the optimal developmental stage for einkorn was found as partially transparent immature embryo in size of around 1.0 mm. Although in the present study the critical balance between plant growth regulators was established for einkorn only, we assume that further the proposed strategy could be successfully applied to other recalcitrant cereal species and genotypes.

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Section: Discussionsupporting

confidence: 81%

Protocol for efficient regulation of in vitro morphogenesis in einkorn (Triticum monococcum L.), a recalcitrant diploid wheat species

et al. 2017

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“…A slight cellular disorganization, limited mitotic activity, intercellular spaces, and a moderate degree of vacuolization can be observed. Bakos et al (2009), who have also observed similar characteristics in embryos of Triticum aestivum obtained in vitro, have considered these characteristics as a discrete dysfunction typical of the in vitro environment that does not compromise the conversion of plants, as found in P. aduncum. On the other hand, somatic embryos with partially organized stem meristem, with differentiated cells with large vacuoles and small nuclei, as reported in Vitis vinifera (Goebel-Tourand et al 1993), or with limited or absent meristematic activity and presence of vacuolated cells, as reported in Ipomoea potatoes (Padmanabhan et al 1998), presented problematic conversion.…”

Section: Germination Of Somatic Embryos and Acclimatization Ofmentioning

confidence: 83%

“…Considerable amounts of starch have also been seen in in vitro embryos of Dendrocalamus hamiltonii (Godbole et al 2004) and T. aestivum (Bakos et al 2009). In contrast, several other authors have not identified significant amounts of starch grains in somatic embryos at different stages of development, and their consumption was attributed to the formation of embryogenic structures.…”

Section: Germination Of Somatic Embryos and Acclimatization Ofmentioning

confidence: 97%

“…The integrity of the meristems, mainly of the shoot apical meristem, is closely related to the ability of somatic plant regeneration (Yeung et al 1996;Bakos et al 2009). Specifically in this system, meristems developed normally, generating whole plants, despite the characteristics of the shoot meristem cells mentioned above and the smaller differentiation of the root meristem.…”

Section: Germination Of Somatic Embryos and Acclimatization Ofmentioning

confidence: 99%

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Somatic embryogenesis and plant regeneration in Piper aduncum L

Sousa

Meira

et al. 2020

In Vitro Cell.Dev.Biol.-Plant

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An efficient protocol is reported for in vitro plant regeneration through somatic embryogenesis in Piper aduncum, a Brazilian Amazon species with high economic potential. The species is important due to a variety of components found in its essential oil, with emphasis on dillapiole. Leaf explants from five accessions identified for high oil yield and levels of dillapiole were evaluated for their embryogenic potential. To induce embryogenic calli, the explants were cultivated in MS medium supplemented with 5 mg L −1 of 1-naphthaleneacetic acid (NAA) and 2.5 mg L −1 of N6-benzylaminopurine (BAP) for 80 d. For somatic embryogenesis, the embryogenic calli were transferred to MS medium with 10 mg L −1 of NAA and 2.5 mg L 1 of BAP and incubated for 45 d. The obtained somatic embryos were germinated in MS medium without regulators by 45 d and the obtained plantlets were subjected to acclimatization. Somatic embryos and calli from this process were subjected to anatomical and histochemical analyses. Biochemical analyses (total soluble sugars, starch, total amino acids, and proteins) were also performed to identify markers for embryogenic competence acquisition. In addition, the germination of somatic embryos was evaluated in a semi-solid and liquid system (R.I.T.A.® temporary immersion bioreactors). The obtained plants were evaluated for genetic fidelity using ISSR markers. The present study indicate that the accessions did not differ in embryogenic potential, with a mean percentage of calli with somatic embryos of 82.4%. Anatomical analyses confirmed the occurrence of the embryogenic route and the histochemical analyses identified starch grains in somatic embryos at different developmental stages. The biochemical analyses showed high total soluble sugars and total amino acids in embryogenic calli, marks of the acquisition of the embryogenic competence of P. aducum. The R.I.T.A.® temporary immersion bioreactors were highly efficient in the regeneration of somatic plants, with 100% germination. The plants regenerated in the semi-solid and liquid systems showed high genetic homogeneity. The survival rate of the acclimatized plants was 100%.

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“…The in vitro culture of ovules a few hours after pollination [400] or isolated zygotes [401] provides an excellent system to study embryogenesis, as this process is more complex to analyze in plants. Therefore, in vitro investigations during embryo development have already been established in maize [402,403], barley [404], wheat [401,[404][405][406][407][408], rice [409][410][411][412], tobacco [413], and Setaria viridis [414]. The use of histological and other microscopic tools is simplified during in vitro culture.…”

Section: Analysis Of Embryogenesis With Embryo Rescue Technical Holdermentioning

confidence: 99%

Embryo Rescue in Plant Breeding

Rogo,

Fambrini,

Pugliesi

2023

Plants

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Embryo rescue (ER) techniques are among the oldest and most successful in vitro tissue culture protocols used with plant species. ER refers to a series of methods that promote the development of an immature or lethal embryo into a viable plant. Intraspecific, interspecific, or intergeneric crosses allow the introgression of important alleles of agricultural interest from wild species, such as resistance or tolerance to abiotic and biotic stresses or morphological traits in crops. However, pre-zygotic and post-zygotic reproductive barriers often present challenges in achieving successful hybridization. Pre-zygotic barriers manifest as incompatibility reactions that hinder pollen germination, pollen tube growth, or penetration into the ovule occurring in various tissues, such as the stigma, style, or ovary. To overcome these barriers, several strategies are employed, including cut-style or graft-on-style techniques, the utilization of mixed pollen from distinct species, placenta pollination, and in vitro ovule pollination. On the other hand, post-zygotic barriers act at different tissues and stages ranging from early embryo development to the subsequent growth and reproduction of the offspring. Many crosses among different genera result in embryo abortion due to the failure of endosperm development. In such cases, ER techniques are needed to rescue these hybrids. ER holds great promise for not only facilitating successful crosses but also for obtaining haploids, doubled haploids, and manipulating the ploidy levels for chromosome engineering by monosomic and disomic addition as well substitution lines. Furthermore, ER can be used to shorten the reproductive cycle and for the propagation of rare plants. Additionally, it has been repeatedly used to study the stages of embryonic development, especially in embryo-lethal mutants. The most widely used ER procedure is the culture of immature embryos taken and placed directly on culture media. In certain cases, the in vitro culture of ovule, ovaries or placentas enables the successful development of young embryos from the zygote stage to maturity.

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Histological comparison between wheat embryos developing in vitro from isolated zygotes and those developing in vivo

Cited by 7 publications

References 77 publications

Protocol for efficient regulation of in vitro morphogenesis in einkorn (Triticum monococcum L.), a recalcitrant diploid wheat species

Protocol for efficient regulation of in vitro morphogenesis in einkorn (Triticum monococcum L.), a recalcitrant diploid wheat species

Somatic embryogenesis and plant regeneration in Piper aduncum L

Embryo Rescue in Plant Breeding

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