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J.m. Van Tuyl - One of the best experts on this subject based on the ideXlab platform.
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THE EFFECT OF Ovule AGE ON OVARY-SLICE Culture AND Ovule Culture IN INTRASPECIFIC AND INTERSPECIFIC CROSSES WITH TULIPA GESNERIANA L.
2016Co-Authors: M. G. M, Van Creij, J.m. Van TuylAbstract:The effect of Ovule age on ovary-slice Culture and Ovule Culture in intraspecific and interspecific crosses with Tulipa gesneriana L
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The effect of medium composition on ovary-slice Culture and Ovule Culture in intraspecific Tulipa gesneriana crosses
Plant Cell Tissue and Organ Culture, 2000Co-Authors: M.g.m. Van Creij, D.m.f.j. Kerckhoffs, S.e.m. De Bruijn, Dick Vreugdenhil, J.m. Van TuylAbstract:The effect of several media components on the germination percentage of Ovules in intraspecific T. gesneriana L. crosses was studied by using two embryo rescue techniques, viz. ovary-slice Culture followed by Ovule Culture and direct Ovule Culture. The addition of 9% sucrose to medium for ovary-slice Culture, started at 3 or at 5 weeks after pollination (WAP), significantly improved the germination percentage as compared to 5% sucrose. The germination percentage did not differ between both sucrose concentrations (3% and 5%) used in Ovule Culture started 4 weeks later with Ovules excised from the ovary-slices (at 9 WAP). Similar germination percentages were obtained with media containing the full or half of the concentrations of micronutrients and macronutrients of the MS-medium during ovary-slice Culture and Ovule Culture. For direct Ovule Culture, started at 4, at 6, and at 8 WAP, the germination percentages did not differ between Ovules Cultured on media with 3%, 6% or 9% sucrose. The addition of the cytokinin BAP (0.01 or 0.1 mg l-1) had no effect on the germination percentage. The use of liquid-shaken Culture resulted in germination percentages which were similar to those on agar-solidified media. Analysis of the carbohydrate concentration of the media revealed that, in both media for ovary-slice Culture and for Ovule Culture, ultimately all sucrose is converted into glucose and fructose. The total concentration of carbohydrates decreased with 19%–48% in the media for ovary-slice Culture, whereas the total concentration of carbohydrates did not decrease remarkably in media for Ovule Culture.
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Application of in vitro pollination, ovary Culture, Ovule Culture and embryo rescue for overcoming incongruity barriers in interspecific Lilium crosses
Plant Science, 1991Co-Authors: J.m. Van Tuyl, M.g.m. Van Creij, M.p. Van Dien, T.c.m. Van Kleinwee, J. Franken, Raoul J. BinoAbstract:The use of a complete and integrated in vitro pollination, fertilization and embryo rescue system in lily was examined. By combining pollination techniques to overcome pre-fertilization barriers with in vitro methods to overcome post-fertilization barriers, interspecific lily crosses could be made more efficiently. In vitro cut-style pollination and in vitro grafted style technique were developed and applied on various interspecific crosses using Lilium longiflorum, L. dauricum, L. henryi, and both Asiatic and Oriental hybrids as the parents. In addition, methods for ovary Culture, ovary-slice Culture and Ovule Culture were generated. Ovule swelling score in ovary Culture was used to evaluate media effects on Ovule development. Using the integrated in vitro pollination and fertilization protocol it was not only possible to raise the total number of hybrid plantlets in a single interspecific cross, but also the number of successful interspecific combinations.
Nobuhiro Kudo - One of the best experts on this subject based on the ideXlab platform.
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A novel interspecific hybrid plant between Hydrangea scandens ssp. chinensis and H. macrophylla via Ovule Culture
Plant Biotechnology, 2008Co-Authors: Nobuhiro Kudo, Tadashi Matsui, Tomoyuki OkadaAbstract:In the genus Hydrangea, H. macrophylla is the most popular species. For this species, numerous cultivars with showy colorful flowers have been bred since the early 1900s through selection of natural mutants and intraspecific crosses among a limited number of early ancestral varieties. Although breeding of H. macrophylla has been successful, further improvements in flower shape, flower color, and growth habit are desirable. H. scandens ssp. chinensis is a small shrub that is native to South and Southeast Asia and valued for its evergreen foliage, winter flowering and broad adaptability in mild climates. Cross-pollination between H. scandens ssp. chinensis and H. macrophylla, and subsequent Ovule Culture, resulted in the production of an interspecific hybrid plant. The hybridity of this plant was confirmed by RAPD analysis. The hybrid plant had flower and leaf morphologies intermediate between the two parental species. Since the hybrid showed more vigorous growth than both parents, had evergreen foliage, and flowered in winter, it has sufficient horticultural merit for commercialization and may be suitable for greenhouse Culture.
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Efficient Production of interspecific Hybrids between Spiraea thunbergii Sieb. ex Blume and S. japonica L. fil. through Ovule Culture
Engei Gakkai zasshi, 2003Co-Authors: Masahide Iizuka, Nobuhiro Kudo, Yasuo Kimura, Isao OgiwaraAbstract:To establish an efficient means of regenerating plantlets via Ovule Culture from an interspecific cross between Spiraea thunbergii Sieb. ex Blume and S. japonica L. fil., the effects of concentrations of macro-elements in the MS medium and sucrose and the sampling time of Ovules were investigated. Survival rates of Ovules and normal plants, on the MS medium in which macro-elements were diluted to 1/2 or 1/4 were better than those Cultured on standard, undiluted MS medium. The high concentrations of macro-elements increased the rate of blighted plants. Hybrid plantets developed on 1/4 MS medium supplemented with 3% sucrose. Although the frequency of albino plant was not influenced by the concentration of macro-elements in the medium, their rate increased as Ovules were Cultured in media high in sucrose. When Ovules were excised 5 to 22 days after pollination and Cultured in 1/4 MS medium supplemented with 3% sucrose, the percentages of normal plants that developed ranged from 1 to 31. When Ovules in their differentiation and enlargement stages were excised from 14 to 19 days after pollination and Culture, the frequency of normal plants ranged from 25 to 31%. Thus, the success rate of obtaining interspecific hybrids between Spiraea thunbergii Sieb. ex Blume and S. japonica L. fil., was optimum when hybrid Ovules were excised from 14 to 19 days after pollination and Culture on 1/4 MS medium supplemented with 3% sucrose.
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Interspecific Hybrids between Spiraea thunbergii Sieb. ex Blume. and S. japonica L. fil. via Ovule Culture
Engei Gakkai zasshi, 2001Co-Authors: Masahide Iizuka, Nobuhiro Kudo, Yasuo Kimura, Isao OgiwaraAbstract:Reciprocal crosses between Spiraea thunbergii Sieb. ex Blume. and S. japonica L. fil. were carried out, to incorporate red color to S. thunbergii. In vitro germination rate of the stored pollen, germination of pollen grains on the stigma, growth of the pollen tubes, and the development of Ovule were investigated, in addition Ovule Culture was tried to obtain interspecific hybrids. 1. Since the flowering time of both species differed, pollen grains of both species were stored dry at -30°C, 5°C, and 20°C. Only the pollen of both species stored at -30°C for 360 days remained viable for crossing. 2. Pollen grains of S. japonica, stored at -30°C, germinated well on the stigma of S. thunbergii. The pollen tubes elongated rapidly to the ovary as the developing Ovules were observed 10 days after pollination. Stored pollens of S. thunbergii germinated well on the stigma of S. japonica, but whether the pollen tubes reached the ovary it was not confirmed. 3. In the interspecific crosses between S. thunbergii (♀) and S. japonica (♂), enlarged Ovules excised 16 days after pollination were Cultured in 1/2MS medium containing 30g·liter-1 sucrose and 8g·lite-1 agar. The resulting plantlets seem to be true hybrids between S. thunbergii and S. japonica., based on several morphological characteristics and the RAPD banding patterns which exhibited the same bands as the parents.
Yoshihito Takahata - One of the best experts on this subject based on the ideXlab platform.
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Original Research Production of Haploids and Doubled Haploids from Unfertilized Ovule Culture of Various Wild Species of Gentians (Gentiana spp.)
2021Co-Authors: Yutaro Takamura, Takashi Hikage, Ryo Takahashi, Katsunori Hatakeyama, Yoshihito TakahataAbstract:Abstract 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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Efficient haploid and doubled haploid production from unfertilized Ovule Culture of gentians (Gentiana spp.)
Breeding science, 2013Co-Authors: Hisako Doi, Nobue Hoshi, Eri Yamada, Shuji Yokoi, Masahiro Nishihara, Takashi Hikage, Yoshihito TakahataAbstract:Factors affecting reliable plant regeneration from unfertilized Ovule Culture of gentians (Gentiana spp.) were examined. Cold pretreatment (4°C) of flower buds enhanced or maintained production of embryo-like structure (ELS). When 43 genotypes were surveyed in two different labs, 40 of them produced ELSs ranging from 0.01 to 26.5 ELSs per flower bud. No ELSs could be obtained in three genotypes. A significant correlation (r = 0.64) was observed between the number of ELS per flower and the frequency of responding flower buds. Eight genotypes of G. triflora, which were used as common materials in two different labs, produced ELSs in both labs. The ploidy levels of a total of 1,515 regenerated plantlets were determined, revealing that the majority of these plants consisted of haploids (57.9%) and diploids (34.3%). However, the frequency of haploids and diploids was different between G. triflora and G. scabra, and G. triflora showed higher frequencies of haploids than G. scabra. When haploids were treated with oryzalin for chromosome doubling, diploids and tetraploids were obtained. These results demonstrate that the unfertilized Ovule Culture technique of gentians is a powerful tool for obtaining haploids and DHs because of its reproducible and reliable nature and application to a wide range of genotypes.
M.g.m. Van Creij - One of the best experts on this subject based on the ideXlab platform.
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The effect of medium composition on ovary-slice Culture and Ovule Culture in intraspecific Tulipa gesneriana crosses
Plant Cell Tissue and Organ Culture, 2000Co-Authors: M.g.m. Van Creij, D.m.f.j. Kerckhoffs, S.e.m. De Bruijn, Dick Vreugdenhil, J.m. Van TuylAbstract:The effect of several media components on the germination percentage of Ovules in intraspecific T. gesneriana L. crosses was studied by using two embryo rescue techniques, viz. ovary-slice Culture followed by Ovule Culture and direct Ovule Culture. The addition of 9% sucrose to medium for ovary-slice Culture, started at 3 or at 5 weeks after pollination (WAP), significantly improved the germination percentage as compared to 5% sucrose. The germination percentage did not differ between both sucrose concentrations (3% and 5%) used in Ovule Culture started 4 weeks later with Ovules excised from the ovary-slices (at 9 WAP). Similar germination percentages were obtained with media containing the full or half of the concentrations of micronutrients and macronutrients of the MS-medium during ovary-slice Culture and Ovule Culture. For direct Ovule Culture, started at 4, at 6, and at 8 WAP, the germination percentages did not differ between Ovules Cultured on media with 3%, 6% or 9% sucrose. The addition of the cytokinin BAP (0.01 or 0.1 mg l-1) had no effect on the germination percentage. The use of liquid-shaken Culture resulted in germination percentages which were similar to those on agar-solidified media. Analysis of the carbohydrate concentration of the media revealed that, in both media for ovary-slice Culture and for Ovule Culture, ultimately all sucrose is converted into glucose and fructose. The total concentration of carbohydrates decreased with 19%–48% in the media for ovary-slice Culture, whereas the total concentration of carbohydrates did not decrease remarkably in media for Ovule Culture.
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Interspecific hybridization in the genus Tulipa L.
1997Co-Authors: M.g.m. Van CreijAbstract:The genus Tulipa L. comprises about 55 species. The tulip species are classified in two subgenera, Tulipa and Eriostemones, which are subdivided into five and three sections respectively. Commercial tulips are mainly cultivars of T. gesneriana L . and of Darwin hybrids, the latter of which are obtained by interspecific hybridization between T. gesneriana and T. fostefiana Hoog ex . Irving. The present-day tulip assortment could be improved considerably by introducing traits from other tulip species, such as resistance for diseases and short forcing period. With conventional breeding methods, hybrids can only be obtained from a limited number of interspecific crosses. Most interspecific crosses are hampered by crossing barriers. The main goal of the research on crossing barriers in tulip was to bypass crossing barriers between cultivars of the present-day assortment and the other tulip species. This main goal was divided into two subgoals: (1) the identification of crossing barriers in crosses between T. gesneriana and a number of representative tulip species and (2) the development of techniques to bypass these barriers. Pre-fertilization development and post-fertilization development have both been studied in a number of crosses to identify crossing barriers. Pre-fertilization development was analyzed in crosses between cultivars of T.gesneriana and 13 tulip species from all eight sections of the genus Tulipa (Chapter 2). Pollen tube growth in the pistil and pollen tube penetration in the Ovules showed much variation between the crosses in progamic development. Depending on the cross, pollen tubes grew as far as the stigma or the style or continued growing down into the ovary. This resulted in percentages of Ovules with pollen tube penetration varying between 0%-79%. The percentages Ovules with pollen tube penetration differed between the flowers of a specific cross, between the different years and between the maternal genotypes used. The progamic phase and embryo and endosperm. development were studied in the incongruent cross T.gesneriana x T. agenensis DC. (former name T. oculus-solis St. Amans) (Chapter 3) and in a compatible T. gesneriana cross. Less pollen tubes penetrated the Ovules in the interspecific cross in comparison with the compatible cross. In the interspecific cross, the embryo development was more often retarded and higher percentages of Ovules with aberrant embryogenesis (from 32 days after pollination) were found than in the compatible cross. In most Ovules with aberrations in embryo development, also the endosperm showed abnormalities. Endosperm degeneration seems to be the major cause of starvation of the embryo before seed maturity in the cross T. gesneriana x T. agenensis. Several techniques have been studied for bypassing pre-fertilization barriers in interspecific tulip crosses: the cut-style method, the grafted-ovary method and placental pollination (Chapter 4). The application of the cut-style method or the grafted-ovary method in the interspecific crosses did not improve the pollen tube growth and pollen tube penetration in the Ovules as compared to ordinary stigmatic pollination. The percentage of Ovules with pollen tube penetration was also not increased after placental pollination. However, the pollination procedure applied for placental pollination might still be optimized to improve the percentage of Ovules penetrated by pollen tubes. The effect of hormone treatments and embryo rescue techniques on the ability of bypassing post-fertilization barriers was studied. Ovaries were treated with the cytokinin BAP (0. 1 % or I %) or the auxin NAA (1 %) at 12 days after pollination (Chapter 4). Seeds of the cross T . gesneriana x T. agenensis were obtained on the plant after treating ovaries with 0. 1 % BAR Seed production on the plant from this cross has not been reported previously. Additional research is needed before definite conclusions can be drawn about the effect of hormone treatments for bypassing crossing barriers in a wider range of interspecific tulip crosses. Two embryo rescue techniques were studied: ovary-slice Culture and Ovule Culture. Compatible T. gesneriana crosses were used as model system. The efficiency of direct Ovule Culture and ovary-slice Culture followed by Ovule Culture was compared for Cultures started at 2-9 weeks after pollination (Chapter 5). The influence of media composition on the percentage of Ovules showing germination was also studied (Chapter 6). In most cases, the percentage of embryos that germinated increased significantly with a more advanced developmental stage of the embryos at the start of the Culture. The lower efficiency at early Culture dates is due to embryo abortion and retarded embryo development. The germination percentages for ovary- slice Culture followed by Ovule Culture, started at various dates, were for some Culture dates comparable to direct Ovule Culture, but for other Culture dates significantly higher. The length of the period of ovary-slice Culture prior to Ovule Culture mostly did not affect the germination percentage. Media composition influenced the percentage of Ovules showing germination after embryo rescue. The germination percentage was influenced by the sucrose concentration used for ovary-slice Culture. 9% Sucrose resulted in higher germination percentages as compared to 5 % sucrose. The sucrose concentrations in media used for subsequent Ovule Culture (3 %, 5%) or in media used for direct Ovule Culture (3%, 6%, 9%) did not influence the germination percentages. Analysis of the carbohydrates concentrations revealed that the total concentration of carbohydrates decreased with 22%-50% in media for ovary-slice Culture (9% sucrose), whereas the total concentration of carbohydrates remained rather constant in Ovule Culture (3% sucrose) applied after ovary-slice Culture. Comparable germination percentages were obtained by using media with the full or half of the concentrations micronutrients and macronutrients of the MS-medium during ovary-slice Culture and subsequent Ovule Culture. For direct Ovule Culture, started at 4, 6 and 8 weeks after pollination, the germination was not improved by the addition of the cytokinin BAP (0.01 and 0. 1 mg/1), nor by the use of liquid shaken Culture. An in vitro pollination procedure has been developed in order to perform an integrated system of pollination, fertilization and embryo rescue techniques under optimal controlled environmental conditions. Once an in vitro pollination procedure has been developed, it can also be used for the post-fertilization Culture of whole ovaries. By using compatible intraspecific T. gesneriana crosses as model, the effect of the following media components on seed set and seed germination could be studied (Chapter 7): concentration of macronutrients and micronutrients of MS-medium, concentration sucrose, fructose and glucose, the auxin NAA, the cytokinin BAP, the gibberellin GA 3 , spermine, casein hydrolysate, the buffer MES and the use and type of agar. The application of MS-medium at full strength, with 5%-7% sucrose and 1 mg/l NAA proved to be suitable. Analysis of carbohydrate uptake revealed that on liquid media significantly more carbohydrates were absorbed by the ovaries than on agar solidified media. However, the numbers of seeds produced were higher on media with agar than on liquid media. The A of Daichin agar improved the seed set as compared to bacteriological agar. The addition of the buffer MES to liquid media, to control the pH, affected the seed set positively. The effect of the use of casein hydrolysate needs additional research. Other components tested had no or negative effects on the seed set and/or seed germination. Unique hybrids have been obtained from the crosses T. gesneriana x T. praestans Hoog and T.gesneriana x T. agenensis by using hormone treatments and/or ovary-slice Culture and Ovule Culture. These techniques proved to be suitable for rescuing embryos of incongruent interspecific tulip crosses. Improvement of Culture conditions and Culture media can still increase the efficiency, especially at early Culture dates. Bulblet formation in vitro after embryo germination and the transfer of the bulblets grown in vitro into the soil were not the subject of our research, but proved to be problematic. For hybrid plant production, it is therefore of great importance to improve the methods of hybrid plant recovery after embryo germination in vitro. For the introduction of genes for resistance in the cultivar assortment of tulip by means of the in this study developed methods, it is of importance to locate possible resistance genitors in the tulip species. Bypassing F1-sterility due to the formation of triploids and sterile diploids in interspecific tulip hybridization, also needs additional research. Traits from a number of tulip species such as forcing time, flower colours, flower shapes and other morphological characteristics can already be introduced in the present-day assortment with the aid of the described embryo rescue techniques. This can give a new impulse to e breeding of tulips
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Application of in vitro pollination, ovary Culture, Ovule Culture and embryo rescue for overcoming incongruity barriers in interspecific Lilium crosses
Plant Science, 1991Co-Authors: J.m. Van Tuyl, M.g.m. Van Creij, M.p. Van Dien, T.c.m. Van Kleinwee, J. Franken, Raoul J. BinoAbstract:The use of a complete and integrated in vitro pollination, fertilization and embryo rescue system in lily was examined. By combining pollination techniques to overcome pre-fertilization barriers with in vitro methods to overcome post-fertilization barriers, interspecific lily crosses could be made more efficiently. In vitro cut-style pollination and in vitro grafted style technique were developed and applied on various interspecific crosses using Lilium longiflorum, L. dauricum, L. henryi, and both Asiatic and Oriental hybrids as the parents. In addition, methods for ovary Culture, ovary-slice Culture and Ovule Culture were generated. Ovule swelling score in ovary Culture was used to evaluate media effects on Ovule development. Using the integrated in vitro pollination and fertilization protocol it was not only possible to raise the total number of hybrid plantlets in a single interspecific cross, but also the number of successful interspecific combinations.
Kere George Mbira - One of the best experts on this subject based on the ideXlab platform.
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In vitro haploid and dihaploid production via unfertilized Ovule Culture.
Plant Cell Tissue and Organ Culture (PCTOC), 2010Co-Authors: Jinfeng Chen, Li Cui, Ahmed Abbas Malik, Kere George MbiraAbstract:Haploids and doubled haploids are very important in plant breeding, enabling the time needed to produce homozygous lines to be shortened compared with conventional breeding. In the present review, emphasis is given to haploid induction through unfertilized Ovule/ovary Culture. Attention is given to induction of haploid plants from female gametophyte Culture through analysis of factors in the processes of gynogenesis, including genotype selection, stage of Ovule development, pretreatment, and Culture media containing nutritional components and phytohormones. The gynogenetic approach may be of great value in discovering novel genetic recombinations. Application of double haploids in genetics and plant breeding is also highlighted. This review also identifies some existing knowledge gaps where work may increase the efficiency of this process in different plant species.
