• Korean Journal of Plant Tissue Culture
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• v.24 no.5
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• pp.257-262
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• 1997

Intergeneric crosses over 34 cross combinations between genus Brassica and Raphanus were made. Ovules taken out of crossed were cultured on MS media supplemented with 1.0 mg/L GA and BA. Germination of embryo from ovule culture was not influenced by BA and GA in medium but by parental characters in its cross combination. Use of Raphanus sativus cv. Daibyosobudore and Jungkukcheongpi showed low embryo germination when they were used as male part. Cross combination with Brassica juncea as male parent showed slightly increased germination compared to other cross. These results indicated that embryo germination in ovule culture was not much influenced by casein hydrolysate, malt extract, BA, kinetin and glutamine in the medium, but parents in combination were key factor for increasing embryo germination.

• FLOWER RESEARCH JOURNAL
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• v.19 no.4
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• pp.231-237
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• 2011

In this study, we performed ovule culture after reciprocal crosses of two Alstroemeria accessions and investigated genetic contribution of parents by using RAPD markers. The best method was half-ovule culture on MS medium supplemented with $60g{\cdot}L^{-1}$ sucrose and $2.2g{\cdot}L^{-1}$ gelrite at 14 days after pollination. Embryos began to germinate after 6 weeks of culture. The complete plantlets were formed after 4 months of culture. In eight progenies and two parental cultivars, 59 polymorphic bands were obtained out of 89 total bands by RAPD analysis using 7 primers. Eight $F_1$ progenies from the crosses between two accessions using reciprocal crosses showed 1:1 contribution of maternal and paternal parents. It is confirmed that $F_1$ progenies were obtained from parental accessions by using RAPD markers. We conclude this cross combination showed pre-fertilization barriers with incompatibility between stigma or style, and pollen because progeny number was different in each cross combination. Thereby, it warrants overcoming pre-fertilization barrier together with post-fertilization barrier in order to broaden the heterozygosity within progeny populations in Alstroemeria breeding program.

• Horticultural Science & Technology
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• v.19 no.3
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• pp.378-383
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• 2001

Embryo culture, ovule culture and ovary slice culture were tested to find optimum method for overcoming post fertilization barrier in interspecific crosses between L. longiflorum 'Gelria' and L. cernuum. Although reciprocal crosses between the species were carried out by cut-style pollination method, fruits developed only in crosses of L. longiflorum${\times}$L. cernuum. On the 40 days after pollination, ovaries were sliced into 2-4mm thickness and cultured on a hormone-free Murashige-Skoog (MS) medium, supplemented with 2%, 4%, 6%, 8% and 10% sucrose. For the L. longiflorum Gelria'${\times}$L. cernuum cross, ovule development was found to be best at 6% sucrose and a lot of hybrid plant lets established directly from the ovary slice culture and subsequent ovule culture. High concentration of sucrose above 8% made ovules abort or vitrificate from 40 days after culture. In contrast, ovules from the L. cernuum${\times}$L. longiflorum 'Gelria' cross swelled well in ovary slice culture, however, they did not germinated in subsequent ovule culture. On the 60 days after pollination, ovules thicker than 0.6mm was interpreted as one containing embryo. The embryo size ranged from 1.2 mm to 1.7 mm, and in vitro development of the excised embryos was found to be best with the MS medium (pH 5.8), supplemented with $0.1-1 mg{\cdot}L^{-1}$ NAA and 6% sucrose. Thick ovules excised 60 days after pollination germinated about 60% as normal seeds in MS medium supplemented with 6% sucrose and free hormone. The ovule culture 60 days after pollination was concluded to be most recommendable to produce interspecific hybrids in large scale crosses between L. longiflorum 'Gelria' and L. cernuum by the reason of easy procedure.

• Journal of the Korean Society of Tobacco Science
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• v.17 no.2
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• pp.109-113
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• 1995

Nicotiana repanda has resistance to many important tobacco diseases but no tobacco cultivars are currently available that carry risistance genes derived from this species. Numerous attempts to hybridize N, repanda with commercial tobacco cultivars have been largely unsuccessful because of cross incompatibility or the uncovering of lethal genes. In vitro pollination of placenta attached ovules was useful in by passing prezygotic barriers for interspecific hybrid combination between N.tabacum cv. NC82 and N. repanda. Six days after in vitro pollination of N. tabacum cv. NC82×N. repanda, enlarged ovules on plancenta were removed and transferred into ovule culture medium of kitsch and kitsch (1969). Within 15 days of ovule culture, germination occurred. Most of the hybrid seedlings obtained had poor root system and finally died, while few of them had good root system and grew well.

• Plant Resources
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• v.7 no.2
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• pp.87-92
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• 2004

Five different poplar hybrids were tested for rescuing embryo to elongate in vitro plantiets after hybridization. Ovaries and ovules were cultured on Woody Plant Medium (WPM) supplemented with cytokinins, 6-benzylamine (BA) and zeatin. Multiple shoots were initiated from half section of capsule with immature embryos after 21 days from pollination and tiny shoots were formed after the expansion of cotyledons in ovule cultures. Germinating response was better in intraspecific hybrids $(6.53\pm1.66)$ than interspecific crosses $(0.93\pm0.54)$ from half section of capsules on WPM medium. In general, zeatin was better than BA in inducing multiple shoots from isolated ovules. The highest average number $(19.40\pm4.53)$ of shoots was produced from immature ovules of 21 days post-pollination of WPM medium supplemented with 5.0 mg/L zeatin. The highest percentage of germination was 93% from the half section of in vitro ovary cultures. Soil acclimation was successfully conducted in cell tray containing artificially mixed soil with 96% survival rate.

• Korean Journal of Plant Tissue Culture
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• v.22 no.5
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• pp.261-266
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• 1995

This study was carried out to develop new varieties which are dwarf and tolerant to winter cold in the Aurantioideae by intergeneric crossing. to do that, the reciprocal crosses of Hwanggeumyooza and trifoliate orange, yooza and trifoliate orange were done and in vitro immature ovule culture of their hybrid was carried out .The callus formation from immature ovule was good in order of Hwanggeumyooza, Hwanggeumyooza $\times$ tifoliate orange, yooza, and trifoliate orange and best at 1 to 3 mg/L NAA+0.5mg/L zeatin on MT medium. In vitro germination percentage of 20week old hybrid of Hwanggeumyooza $\times$ tifoliate orange and trifoliate orange $\times$ Hwanggeumyooza were 41.3% and 37.7, respectively. The phenotype of hybrid (95%) of Hwanggeumyooza $\times$ trifoliate orange and that (100%) of trifoliate orange $\times$ Hwanggeumyooza were similar to that of trifoliate orange. After Hwanggeumyooza was pollinated by pollens of trifoliate orange, the pollen tubes grew on stigma after 3h of pollination and entered into micropyle after about 24~28 h. One gamete in pollen was fused with polar nuclei after 2 days and other one fused with egg nucleus at 3days after pollination. The fruit set percentage by intergeneric crossing was 14.0% in Hwanggeumyooza $\times$ trtfoliate orange and 17.5% in trifoliate orange $\times$ Hwanggeumyooza. The fruit set percentages of Hwanggeumyooza. and trifoliate orange were 34.2% and 39.5% by artificial self-fertilization, 34.2% and 39.5% by artificial cross fertilization, 3.1% and 1.4% by parthenocarpy and 13.0% and 3.0% by natural fertilization, respectively. The somatic and gametic chromosome numbers of Hwanggeumyooza, yooza, and trifoliate orange were 2n=18 and n=9.

• Proceedings of the Zoological Society Korea Conference
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• 1997.10b
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• pp.313.1-313
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• 1997

No Abstract, See Full Text

• Korean Journal of Plant Tissue Culture
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• v.22 no.6
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• pp.317-322
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• 1995

As a basic research for breeding new varieties, reciprocal -intergeneric crosses between Citrus junos and P.trifoliata were made. F$_1$ hybrid production using in vitro ovule culture, gametogenesis, and fertilization phenomena were investigated. Frequency of fruit set resulting from crossing of Citrus junos and Poncirus Trifoliata was 16.6% while that of Poncirus Trifoliata and Citrus junos was 11.7%. Callus formation occurred well when ovules at the 6th week after pollination were cultured on MT (Murashige and Tucker) medium supplemented with zeatin 0.5 mg/L and NAA 1.0 or 3.0 mg/L. Immature ovules developed into mature embryos of the MT medium supplemented with 2,4-D 0.1 or 3.0 mg/L. Immature ovules developed into mature embryos of the MT medium supplemented with 2,4 D 0.1 or 0.5 mg/L. The invitro germination rates of 20-week-old ovules set C. junos $\times$ P. Trifoliata and P. Trifoliata $\times$ C. junos were 54.5% and 48.6%, respectively. The emergence ratios of trifoliate hybrids obtained by C. junos $\times$ P. Trifoliata and P. Trifoliata $\times$ C. junos were 56.7% and 100%, respectively. The chromosome number of C. junos and P. Trifoliata was n = 9 or 2n = 18, and the sizes of their pollen grain were 33.75 $\mu$ and 25.0 $\mu$. The length and width of embryo sac in C. junos and P. Trifoliata were 69.38~79.23 $\mu$ and 27.50~38.56 $\mu$, and those of egg cells were 17.50~41.50 $\mu$ and 6.25~8.12$\mu$. Fertilization of C. junos and P. trifoliata terminated 72 h after pollination.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.32 no.4
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• pp.455-461
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• 1987

The ovaries or the ovules of grasses were pollinated and cultured in vitro to raise the interspecific or the intergeneric hybrids between tall fescue, meadow fescue, and Italian ryegrass. The isolated and suface-sterili-zed pistils were dusted with compatible pollens on stigma, on stump after removing stigma, or on excised ovule. Furthermore, the fertilized ovaries and ovules were cultured on MS, M6, or White's media and treated with plant growth regulators: IAA, kinetin, BA to promote embryo development and seed maturity. The in vitro fertilization in grass species ranged from 44 to 92% depending on ovary and pollen parents. The stigmatic pollination was resulted in 67.8% fertilization, the stump pollination 89.0%, and the excised ovule pollination 61.0%, repectively. White's medium was the most effective to provide embryo development and seed maturity in grass species. And the combined treatment of IAA 10mg/$\ell$, kinetin 0.2mg/$\ell$, was better than the non-treatment. Only two seedlings, one complete and one abnormal with root formation were obtained from 127 ovaryies cultured. The anatomy of ovules in vitro cultured was revealed the differentiation of vascular system and meristematic tissue, and the formation of sclerenchyma cells inside ovule.

• Journal of Plant Biotechnology
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• v.7 no.4
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• pp.259-265
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• 2005

Indian citrus ringspot virus (ICRSV)-free plants of Kinnow mandarin (Citrus nobilis Lour x C. deliciosa Tenora) were raised from virus-infected plants using unfertilised ovules as explants. Plants were tested by indirect ELISA and RT-PCR before using their explant. An amplified product of 539 bp was obtained by RT- PCR in ICRSV infected plants. Unfertilized ovules were excised from unopened flower buds of plants tested postive for virus and were cultured on Murashige and Skoog's (MS) basal medium supplemented with various concentrations of kinetin (KN) or malt extract (ME). Maximum induction (31.94%) of embryogenic callus was observed on MS medium supplemented with KN ($9.29\;{\mu}M$). Transfer of embryogenic calli to similar media composition resulted in somatic embryogenesis in all cultures, with an average number of 60.36 globular, 17.39 heart and 7.71 cotyledonary-shaped somatic embryos per culture. All cotyledonary shaped embryos developed into complete plantlets within 60 days on transfer to similar medium. Embryogenic callus induction, somatic embryo formation, maturation, germination and plantlet formation were achieved on MS medium supplemented with KN ($9.29\;{\mu}M$) alone. The plantlets derived from somatic embryos were transferred to sterilized soil, sand and vermiculite (3:1:1) mixture. After acclimatization, the plantlets were transferred to screen house and were indexed for ICRSV employing indirect ELISA and RT-PCR and found free of virus. A distinct feature of this study is the induction of somatic embryogenesis from unfertilised ovules to produce virus-free plants.