• Plant Resources
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• 제8권2호
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• pp.87-90
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• 2005

In tobacco, in vitro pollination has been successfully applied to overcome interspecific incompatibility. The use of this technique will make it possible to introduce DNA into pollen tubes just before fertilization. In this study, we showed improvement of the efficiency of in vitro self-pollination and introduction of foreign genes into pollen tubes by the method of polycation. A plasmid harbouring the GUS gene was introduced into pollen grains and pollen tubes, which had incubated on pollen germination medium(PGM), by polyornithine method. Transient expression of the GUS in pollen grains and pollen tubes that were treated with 0, 2, 5 and $10{\mu}g/m\ell$ DNA was observed. In results, combination of the techniques of polyornithine and in vitro pollination was efficient new technique for genetic transformation through fertilization processes.

• 한국연초학회지
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• 제17권2호
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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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• 제7권2호
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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.

• 한국작물학회지
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• 제32권4호
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• pp.455-461
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• 1987

Festuca속의 토올페스큐와 메도우페스큐, 그리고 Lolium속의 이탈리안라이그라스들 간에 종속간 잡종을 얻기 위하여 기내수분과 기내배양을 실시하였다. 이들 작물들의 자방과 화분을 채취하여 소독을 하고 주두수분, 주두제거 수분, 배주수분의 3가지 방법으로 기내수분을 실시하였다. 그리고 이들의 배발육상태를 MS, N6 , White's의 배지종류와 IAA Kinetln, BA의 생장조절물질의 처리로서 비교하였다. 1. 토올페스큐, 메도우페스큐, 이탈리안라이그라스의 상호 조합간 기내수분에서 44~92%의 수정율을 나타내었다. 이들의 주두수분은 평균 67.8% 주두제거 수분은 평균 89%의 수정율을 보였으나 배주수분은 평균 61 %로 낮았다. 2. 화본과 수정배주에서 배발육을 촉진하는데는 White's 배지가 가장 효율적이었고 식물생장조절물질의 처리는 IAA(10mg/$\ell$)와 Kinetin(0.2 mg/$\ell$)의 혼합처리가 무처리보다 효과적이었으나 발아율은 극히 저조하였다 3. 기내배양으로 성숙 배주의 배부조직에서 유관책의 형성과 시원체조직의 분화를 확인할 수 있었으며 배주 내부에 후막세포들의 형성이 관찰되었다.

• 한국연초학회지
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• 제14권1호
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• pp.3-11
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• 1992

The present study examined various environmental and cultural media conditions for in vitro "rescue" of cross-fertilized ovules formed through sexual crosses between Nicotiana rustica and N. tabacum cv. BY4. The response ovules to two cultural procedures was compared; ovules were cultured either separately or left attached to the placenta. Total yield of seedlings and percent of normal seedlings were increased by culturing individual ovules separately, rather than on excised placenta. Total yield of seedlings and number of normal seedlings were produced following in vitro culture of individual fertilized ovules of N. rustica X M tabacum cv. BY4 at four days post-pollination on NN medium containing 2% sucrose. In the in vitro culture of fertilized ovules, high sucrose concentration increased the frequency of seedlings of abnormal appearance. Therefore, sucrose should be supplied to developing ovules at gradually decreased concentrations. Culture of fertilized ovules from three to eight days after pollination gave increased number of seedlings, but with delayed cultral time the number of morphologically normal seedling were decreased. Hybrids were uniform in appearance and showed vegetative heterosis but flower characteristics were generally intermediate between those of the parents. All hvbrids evaluated were self-sterile.f-sterile.

• 원예과학기술지
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• 제16권2호
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• pp.236-238
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• 1998

심비디움의 교배에 의한 신품종 육성시 발아율을 증진시키고 발아기간을 단축시켜 육종 효율을 높이고자 자가 또는 타가수분 후 경과일수에 따라 무균발아에 가장 적합한 수분 후 일수 및 배지에 대해 조사한 결과는 다음과 같다. 자방의 길이 생장은 교배조합에 관계없이 수분후 110일경, 자방의 폭은 교배조합에 따라 다소차이가 있으나 대체로 140일경에 생육이 정지되었다. 자방의 수가 많고 교배된 꼬투리의 교배율이 높은 조합일수록 발아상태가 양호하였다. 수분후 150일 부터 180일 경과된 꼬투리를 배양하는 것이 발아 상태가 가장 양호하였고 발아 소요일수도 짧았다. 무균파종 및 발아 후 생육에 적합한 배지는 하이포넥스 배지로 나타났다.

• 한국작물학회:학술대회논문집
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• 한국작물학회 1987년도 추계 학술대회지
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• pp.62-63
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• 1987

• 한국연초학회지
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• 제13권2호
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• pp.52-58
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• 1991

Interspecific cross between Nicotiana trigonophylla and N. tabacum cv. BY4 is highly sterile because of abnormal ovule and embryo development. In vitro culture of excised N. trigonophylla ovules after polination by N. tabacum allows significant numbers of hybrid embryos to develop into mature plants. Total yield of seedlings and number of normal seedlings were produced following in vitro culture of individual fertilized ovules of N. trigonophylla X N. tabacum at four days post-pollination on B5 medium containing 6% sucrose. Hybrids were uniform in morphology and peroxidase isozyme composition and the majority were cytologically stable: flower characteristics were generally intermediate between those of the parents. All hybrids evaluated were self-sterile.

• 원예과학기술지
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• 제19권3호
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• pp.378-383
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• 2001

L. longiflorum 'Gelria'와 우리 나라 자생종인 솔나리(L. cernuum)와의 종간 교잡시 수정 후 장벽을 극복하여 효율적인 식물체 생산을 목적으로 자방절편 배양, 배주배양, 배배양을 수행하였다. 두 종의 정역교잡에서 L. longiflorum 'Gelria'${\times}$L. cernuum의 편방향 교잡에서만 과실이 발달하였다. 수분 40일 경과된 자방의 자방절편 배양에서 sucrose 농도를 2, 4, 6, 8, 10%로 달리하여 배양한 결과 6% sucrose+MS 배지가 배주발달에 가장 좋은 것으로 나타나 많은 잡종 개체를 획득하였다. 자방절편 배양에서 획득된 배주의 배양도 6% sucrose배지에서 제일 좋았다. Sucrose가 발아 및 미숙배의 생육에 미치는 영향은 자방절편 배양 시 8% 이상의 sucrose농도에서는 배양 40일 후부터 비대 발달된 배주가 투명화 되거나 퇴화되는 현상이 많이 관찰되었다. 한편, L. cernuum${\times}$L. longiflorum 조합에서는 30일간 자방절편 배양하였을 때 배주가 자라는 모습을 볼 수 있었으나, 이어진 배주 배양에서 식물체를 획득할 수가 없었다. 수분 60일 후의 배주를 관찰한 결과 0.6mm보다 두꺼운 배주는 배를 함유하는 것으로 간주되었고, 이들의 배는 1.2-1.7mm 정도였다. 이들 배주 NAA를 첨가하지 않은 6% sucrose만 첨가한 MS배지에서 일반 종자처럼 발아하였다. 이들 배주로부터 적취된 배의 배양은 MS+NAA $0.1-1 mg{\cdot}L^{-1}+6%$ sucrose에서 가장 좋은 효과가 나타났다. 작업의 간편성을 고려하면 L. longiflorum 'Gelria'와 솔나리(L. cernuum)와의 종간잡종 육성은 수분 후 60일에 채취된 0.6mm 이상의 배주를 배양하는 방법이 추천될 수 있었다.

• 원예과학기술지
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• 제29권1호
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• pp.74-76
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• 2011

An interspecific hybrid lily cultivar 'Green Star' was bred in 2005 at the National Institute of Horticultural and Herbal Science (NIHHS), Rural Development Administration (RDA), Korea. The crossing and in vitro embryo rescue was conducted between Lilium FA97-2 (L. ${\times}$ formolongi 'Silky White' ${\times}$ L. Asiatic 'Sunray') and L. Asiatic 'Bomi (Byeongga ${\times}$ Connecticut King)' by cut style pollination method (CSM) at Suwon in 2000. The first selection was done and was tentatively named as 'FA03-5' in 2003. After in vitro multiplication and bulbing production of 'FA03-5' line, growth and flowering characteristic tests were conducted from 2003 to 2005. The evaluation of characteristics and consumer preferences were surveyed at a lily flower show of NIHHS in 2005. 'Green Star' flowered in the middle of June and grew more than 120 cm stem in length. Flowers bloomed facing upward, unspotted in petals and greenish yellow (RHS, Y6D). 'Green Star' was male sterile. Year-round flowering can be done by storing the bulb under $-1.5^{\circ}C$ conditions. It was needed to control the Botrytis disease in wet season.