Haploid and Doubled Haploid Plant Production in Gentian (Gentiana spp.)

Doi, Hisako; Takahata, Yoshihito

doi:10.1007/978-3-642-54102-5_8

Cited by 6 publications

(6 citation statements)

References 28 publications

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“…Haploid cultures in gentian family are restricted to G. triflora Pall., G. scabra Bunge, and many of their F1 hybrids and clonal cultivars (Doi and Takahata, 2015). These two species are widely distributed in the alpine zone of Japan and cultivated commercially as cut flowers and pot plants, making the gentian an important ornamental plant all over the world.…”

Section: Embryogenesis In Haploid Culturesmentioning

confidence: 99%

“…The haploid culture was not able to produce the plants. Since then, the list of published works and successful protocols developed for the generation of haploid or doubled haploid plants from developing microspores and ovules has lengthened (Doi et al, 2010, 2011, 2013; Doi and Takahata, 2015).…”

Section: Embryogenesis In Haploid Culturesmentioning

confidence: 99%

“…The embryos from the cotyledonary to torpedo stages easily regenerated to plantlets in comparison to the earlier stage embryos. Abnormal embryos sometimes regenerated to plantlets as well, when transferred to the 0.5MS medium supplemented with 1.0 mg l -1 GA 3 (Doi and Takahata, 2015).…”

Section: Embryogenesis In Haploid Culturesmentioning

confidence: 99%

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Somatic Embryogenesis in the Family Gentianaceae and Its Biotechnological Application

et al. 2019

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The family Gentianaceae consists of 1736 species, which play an important role in human being existence due to their pharmacological and horticultural values. Many species accumulate bitter iridoid substances used medicinally and in flavorings, while others are cultivated because of beauty of their flowers showing a wide range of colors and patterns. Out of 99 genera belonging to the gentian family, process of somatic embryogenesis (SE) was reported for 5. The first reports, aimed at micropropagation of ornamental cultivars and production of secondary metabolites, concerned Centaurium erythraea Rafn., Eustoma russellianum Grieseb. and Exacum affine Balf. Somatic embryos were induced on different explants cultured in the liquid Murashige and Skoog medium supplemented with auxins and cytokinins. In the 1990s of the last century, significant progress in the exploration of the phenomenon of SE and its biotechnological application was made for the genus Gentiana . The process was induced on various explants and studied at the structural and ultrastructural levels. Regenerated plants were screened for genetic stability using flow cytometry, chromosome counting, and molecular markers. Besides typical indirect SE, the use of leaf fragments enabled to obtain single-cell origin of somatic embryos. On the other hand, proliferation of embryogenic callus in liquid medium resulted in the establishment of long-term embryogenic cell suspension cultures, paving the way not only to study the formation of somatic embryos and the development of regenerants but also to preserve the morphogenic potential of cell aggregates by cryopreservation. Cell suspensions re-established after storage in liquid nitrogen maintained their embryogenic character and allowed to obtain somatic embryo-derived regenerants that were true-to-type at both genetic and epigenetic levels. Another application of SE was related to genetic manipulation purposes. Efficient protocols of plant regeneration from callus-, cell suspension-, or leaf mesophyll-derived protoplasts allowed engaging procedures of somatic hybridization or protoplast electroporation for gentian genome modifications. Also, high embryogenic potential existing in the numerous gentian species enabled successful Agrobacterium -mediated transformation of G. cruciata L. and G. dahurica Fisch.

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Section: Embryogenesis In Haploid Culturesmentioning

confidence: 99%

Section: Embryogenesis In Haploid Culturesmentioning

confidence: 99%

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Somatic Embryogenesis in the Family Gentianaceae and Its Biotechnological Application

et al. 2019

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“…In cultivated gentians, uniform inbred lines have been difficult to obtain through recurrent selfing of conventional methods because of severe inbreeding depression, however doubled haploid (DH) technology has been developed using in vitro culture of anther (androgenesis) and unfertilized ovule (gynogenesis) to address this issue (Doi et al 2010, 2013, Pathirana et al 2011. Further, the latter has more advantages than the former in terms of the high frequencies of embryogenesis and haploid/DH production and small influence on genotypes (Doi and Takahata 2015). Although unfertilized ovule culture is an effective tool for obtaining DHs of cultivated gentians (G. scabra and G. triflora), it is not yet known whether this technique can be used with wild gentians.…”

Section: Introductionmentioning

confidence: 99%

Original Research Production of Haploids and Doubled Haploids from Unfertilized Ovule Culture of Various Wild Species of Gentians (Gentiana spp.)

Takamura¹,

Takahashi²,

Hikage³

et al. 2021

Preprint

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The production of haploids and doubled haploids (DHs) on unfertilized ovule culture was examined in 19 wild species of gentians ( Gentiana ssp.) classified into eight sections. Of the 19 species including 35 strains, embryo-like structures (ELSs) were obtained in 15 species, and regenerated plants were produced in 11 species. ELS production has varied greatly among the15 species, i.e., 0.5%-79.2% frequency of responding flower buds and 0.01-1.99 ELSs per flower bud. Of the ELS-producing species, almost all were classified into the sects. Pneumonanthe or Cruciata . Species in sect. Pneumonanthe showed higher responses than those in sect. Cruciata . In examining the effect of flower bud stage on ELS production, more than twice as many ELSs were observed at the anther-dehiscent stage than that at the anther-indehiscent stage. Ploidy level was determined in 117 randomly selected regenerated plantlets, which suggests that most were haploid (32.5%) and diploid (46.2%). When 12 diploid plants were examined using simple sequence repeat (SSR) markers, 8 (66.7%) were DH. This study revealed that unfertilized ovule culture can be applied not only on cultivated gentian species but also on a number of wild species. The production of haploids and DHs in wild gentians provides novel prospects for ornamental and/or medicinal gentian breeding.

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“…To promote gentian breeding, in addition to conventional crossbreeding, several methods have been developed, i.e., mutation, polyploidy, protoplast culture, doubled haploid production, genetic transformation, marker assisted selection, etc. (Doi and Takahata 2015, Hikage 2016, Nishihara et al 2015, Takahata et al 1995. On the other hand, though the genus Gentiana is comprised of 15 sections and about 360 species (Ho and Liu 2001), breeding using wide hybridization was only slightly carried out.…”

Section: Introductionmentioning

confidence: 99%

Production of interspecific hybrids between Japanese gentians and wild species of <i>Gentiana</i>

Takamura

Asano

Hikage³

et al. 2019

Breed. Sci.

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Wide hybridization, which is a powerful tool to broaden genetic variation, has been used in breeding of many crops. However, in ornamental gentian few wide hybridizations have been reported. Interspecific hybridizations between two gentian cultivated species (Gentiana scabra and G. triflora) and 11 wild species, which were classified in five sections, were carried out using ovule culture. When G. scabra was used as a female parent, normal seedlings and hybrid plants were obtained from eight and five interspecific combinations, respectively. The yield of seedling produced from ovule culture depended on interspecific combinations, ranging from 0.3 to 427.7 normal seedling per flower. In the hybridization of G. triflora with five wild species, normal seedlings and plants were produced in five and four interspecific combinations, respectively. The yield of normal seedling ranging from 0.4 to 228.3 was different between not only interspecific combinations but also reciprocal crosses. Two cultivated species are classified in sect. Pneumonanthe, and successful production of hybrids was obtained from the hybridization with species classified in sections Pneumonanthe or Cruciata. The hybrid nature of the produced plants was confirmed by molecular marker and morphology. The production of interspecific hybrids opens a novel prospect in ornamental gentian breeding.

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Haploid and Doubled Haploid Plant Production in Gentian (Gentiana spp.)

Cited by 6 publications

References 28 publications

Somatic Embryogenesis in the Family Gentianaceae and Its Biotechnological Application

Somatic Embryogenesis in the Family Gentianaceae and Its Biotechnological Application

Original Research Production of Haploids and Doubled Haploids from Unfertilized Ovule Culture of Various Wild Species of Gentians (Gentiana spp.)

Production of interspecific hybrids between Japanese gentians and wild species of <i>Gentiana</i>

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Haploid and Doubled Haploid Plant Production in Gentian (Gentiana spp.)

Cited by 6 publications

References 28 publications

Somatic Embryogenesis in the Family Gentianaceae and Its Biotechnological Application

Somatic Embryogenesis in the Family Gentianaceae and Its Biotechnological Application

Original Research Production of Haploids and Doubled Haploids from Unfertilized Ovule Culture of Various Wild Species of Gentians (Gentiana&nbsp;spp.)

Production of interspecific hybrids between Japanese gentians and wild species of <i>Gentiana</i>

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Original Research Production of Haploids and Doubled Haploids from Unfertilized Ovule Culture of Various Wild Species of Gentians (Gentiana spp.)