• 한국산림과학회지
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• 제109권4호
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• pp.413-420
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• 2020

본 연구는 체세포배 발아 식물체의 세포 라인별, 순화용 기질별 생육 특성을 분석하여, 일본잎갈나무 조직 배양묘의 순화 중 생존율을 높이려는 방안을 마련하고자 실시하였다. 체세포배 발아 식물체는 라인 L14-66, L16-18, L17-B4를 사용하였으며, 순화용 기질은 낙엽송 전용 상토, 원예용 상토, 엘리포트, 피트플러그를 사용하였다. 평균 초기 신초와 뿌리 길이는 L14-66 라인이 가장 짧았고, L17-B4 라인이 가장 길었다. 세포 라인별 평균 생존율은 L17-B4 라인이 87.0%로 가장 높았고, L14-66 라인이 64.3%로 가장 낮았다. 순화용 기질별 생존율은 엘리포트와 피트플러그에서 각각 88.5%, 88.9%로 가장 높았다. L14-66 라인의 초기 신초 길이는 낙엽송 전용 상토(r = 0.852), 원예용 상토(r = 0.692), 엘리포트(r = 0.867)에서 생존율과 상관이 높았으나, 전 질소 함량이 높은 피트플러그에서는 상관이 없었다. L17-B4 라인의 초기 신초 길이는 모든 순화용 기질에서 생존율과 상관이 없었다. L14-66 라인의 초기 뿌리 길이는 낙엽송 전용 상토(r = 0.986), 엘리포트(r = 0.846), 피트플러그(r = 0.802)에서 생존율과 상관이 높았으며, 유식물체의 생존율은 초기 뿌리 길이가 길수록 높았다. L17-B4 라인의 초기 뿌리 길이는 낙엽송 전용 상토(r = 0.896)와 엘리포트(r = 0.696)에서만 생존율과 상관을 보였다. 결론적으로, 유식물체의 생존율을 높이기 위해서 신초 길이보다 뿌리 길이를 우선 고려해야 하며, 피트플러그와 같이 기질 내 질소 함량이 높은 재료를 사용하거나 순화 과정 중에 질소 시비를 높여 주는 것이 좋다. 또한, 생존율을 높이기 위해 초기 생장이 빠른 라인 개발도 함께 이루어져야 한다.

• 한국식물학회:학술대회논문집
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• 한국식물학회 1987년도 식물생명공학 심포지움 논문집 Proceedings of Symposia on Plant Biotechnology
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• pp.213-237
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• 1987

In vitro flowering system may minimize the confounded influence of non-floral meristem parts of plants in studying the relationship of a given treatment and flowering responses. We have induced flower buds from plantlets regenerated from zygotic embryo-derived somatic embryos of ginseng, which circumvented the normal 2-year juvenile period before flowering. The result suggests that the adulthood of ginseng root explants in the experiment previously conducted by Chang and Hsing (1980; Nature 284: 341-342) is not prerequired to flowering of plantlets regenerated through somatic embryogenesis. We have also induced flower buds from elongated axillary brandches from cotyledonary nodes by culturing ginseng zygotic embryos, seedlings, and excised cotyledonary nodes. It was found that 6-benzyladenine (BA) supplemented to the medium was essential for flowering, whereas abscisic acid (ABA) was inhibitory. Gibberellic acid(GA3) was also required for flowering when ABA was present with BA in the medium. The results suggest that cytokinins, gibberellins, and inhibitors play primary, permissive, and preventive roles, respective-ly, in the induction of flowering of ginseng. Tran Thanh Van (1980; Int. Rev. Cytol., Suppl. IIA: 175-194) has developed the "thin cell layer system" in which the induction of shoots, roots, or flower buds from epidermal layer explants were controlled by culture conditions and exogenous growth regulators in the medium, Utilizing the thin cell layer system, Meeks-Wagner et al. (1989; The Plant Cell 1: 25-35) have cloned genes specifically expressed during floral evocation. However, the system is too tedious for obtaining a sufficient amount of plant materials for biochmical and molecular biological studies of flowering. We have developed a garlic callus culture system and one obvious advantaging over the thin cell layer system is that an abundant cells committed to develope into flower buds proliferate. When the above cells were compared by two-dimensional gel electrophoresis with those which have just lost the competence for developing into flower buds, a few putative proteins specific to floral evocation were detected. The garlic callus culture system can be further explored for elucidation of the molecular biological mechanism of floral evocation and morphogenesis.hogenesis.

• 식물조직배양학회지
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• 제28권2호
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• pp.103-108
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• 2001

카사바 (Manihot esculenta Crantz cv. MColl 22)의 마디절편을 제아틴 0.01%가 첨가된 MS 기본배지에서 2주간 현탁배양하여 경엽부 길이가 1.5∼2.5 cm로 생장된 유식물의 부정근을(1) 1∼l.5 cm길이만 남기고 짧게 전정, (2) 1∼2 mm 이내로 제거, (3)뿌리 발생부 전체를 절단한 후 마사토를 담은 유리병에 이식하여 비무균조건에서 순화생장시켰다. 그 결과 이식 7∼10일경부터 각 조건에서 새 뿌리가 발생되기 시작하여 6주 후에는 조건에 따라 73∼93%의 생존율을 나타냈다. 기내 발생 뿌리 가운데 손상 없이 토양에 이식된 것은 그대로 생존을 계속하여 유식물 생장에 도움을 주었으며, 제거된 부정근은 이식 후 신장이 중단되고 새로 뿌리가 발생되었다. 그러나 간혹 전정되고 남은 부분에 측근이 발생된 것이 관찰되었다. 각 조건에서 새로 발생된 기외 뿌리의 수와 길이 및 경엽부 길이와 마디 수에서는 서로 유의성 있는 차이를 나타내지 않았으나, 생체중에서는 전정>제거>절단의 순으로 약간의 차이를 보였다. 미세증식된 카사바의 기내발생 부정근을 적절히 전정 또는 제거한 후 이식하여 순화시키는 것은, 작업을 간편하게 할 뿐만 아니라 유식물의 생존율이 높고 생장에도 지장이 거의 없으므로 경제적인 미세증식 방법으로 유용하게 이용될 수 있다.

• Journal of Plant Biotechnology
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• 제6권1호
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• pp.39-43
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• 2004

A successful, efficient system for multiple shoot induction and plant regeneration of soybean (Glycine max) was established. Four soybean genotypes were compared for organogenic responses on various media cultured under light conditions. The adventitious shoots (98%, 2.6 shoots/cotyledon) directly from one-day-old cotyledon after germination induced by the hormone treatment and its efficiency was higher than any other conditions. The optimal medium for the induction of multiple shoots from cotyledon in Pungsannamulkong(shoot formation rate, 98%), Lx 16 (83%) and IIpumgeomjeongkong(63%) was MS medium supplemented with 2 mg/L BAP, but for Alchankong(75%), MS medium supplemented with 1mg/L zeatin and 1mg/L IAA, 3% sucrose, 4% Phytagel. Higher root induction (88%) was observed from the shoots placed on rooting medium (hormone-free MS basal). Plantlets were transferred onto the same medium supplemented with 1% activated charcoal for further development. With this treatment, regenerated plantlets were obtained within 7-8 weeks (shoot induction for 4 weeks, rooting and shoot elongation for 3-4 weeks).

• Current Research on Agriculture and Life Sciences
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• 제31권2호
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• pp.94-100
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• 2013

An efficient plant regeneration protocol is developed for a standard-type chrysanthemum. When petal segments derived from flower buds (4 or 8cm in diameter) were used as the culture material, the highest shoot regeneration frequency (96%) was obtained on a Murashige and Skoog (MS) medium supplemented with 0.5 mg/L IAA, 2 mg/L BA, 3% sucrose, and a 0.8% agar. Pre-culturing the explants under dark conditions for 14 days produced better results for the shoot regeneration frequency than the explants cultured under a continuous 16 h photoperiod ($40{\mu}molm^{-2}s^{-1}$). The shoot regeneration frequency ranged from 19.0% for the Shinmato cultivar to 89.1% for the Baeksun cultivar. Activated charcoal (0.2%) enhanced the root formation of the regenerated shoots in a hormone-free MS medium. The rooted plantlets were acclimatized and successfully established in a greenhouse.

• 식물조직배양학회지
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• 제22권2호
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• pp.115-120
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• 1995

국내품종인 동진벼와 태백벼의 미숙접합자배 유래 배발생 현탁배양 세포의 초저온 보존 시스템을 개발하였다. 동결/해동 후 캘러스 재생률은 동진벼의 경우 2 M DMSO와 0.4 M sucrose를 태백벼의 경우 0.64 M DMSO와 0.4 M sucrose를 혼용처리하였을 때 캘러스 재생률이 각가 88%와 90%로 가장 높았다. 또한 고농도의 삼투용액에서 배양세포 의 전처리 과정은 필요하지 않았다. 재생된 캘러스를 1 mg/L NAA와 5 mg/L kinetin이 첨가 된 $N_{6 }$, 배지로 이식하여 명배양하였을 때 체세포배발생을 통하여 다수의 유식물체가 발달하였다. 약 100여개의 식물체가 재분화되었으며, 이중 25%는 albino이었다.다.

• Plant Resources
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• 제6권2호
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• pp.108-113
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• 2003

This study was carried out to obtain basic information for possibility of oral vaccine in carrot using Agrobacteruim -mediated transformation system. The epitope region of porcine epidemic diarrhea virus (PEDV) spike gene which is classified as a member of the Coronaviridae and causes an acute enteritis in pigs was successfully expressed in carrot (Daucus carota) using the Agrobacterium-mediated transformation system. Hypocotyl segments of in vitro germinated plantlets were infected with Agrobacteriun tumefaciens LBA 4404 harboring PEDV spike gene. Embryogenic callus (EC) was induced on MS selection medium with 1 mg/L 2,4-D, 50 mg/L kanamycin and 300 mg/L cefotaxime after 45 days of culture. Subcultured ECs on MS selection medium without 2,4-D were converted to somatic embryos (SE) of various stage; globular, heart and torpedo stage. Putative transgenic embryos were selected on MS medium with 50 mg/L kanamycin and 300 mg/L cefotaxime. Regenerated plantlets from transformed SE were induced on MS medium containing 50 mg/L kanamycin after 30 days of culture. Genomic PCR confirmed the integration of PEDV spike gene into nuclear genome of carrot and northern blot analysis demonstrated the expression of PEDV spike gene in transgenic carrot.

• 한국자원식물학회지
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• 제22권6호
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• pp.506-512
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• 2009

To study the effect of genotype of donor plants on anther culture, anthers of nine hybrid cultivars (Derby, Special, Bossanova, Minipaprika, Fiesta, Boogie, Phenlene, Kufrah, and Clarity) of sweet pepper (Capsicum annuum L.) were cultured in a petridish containing C medium (Sibi, Dumas De Vaulx medium) supplemented with 0.1 mg/L 2,4-D and 0.1 mg/L kinetin, 3% sucrose and 0.32% Phytagel. The cultures were incubated in the dark at $35^{\circ}C$ for seven days, and then cultured at $25^{\circ}C$ with a photoperiod of 16 hr. daylength for 40 days. Frequency of callus formation and plant regeneration was varied among cultivars. Callus formation was ranged from 6% in Phenlene to 69.8% in Kufrah. The highest percentage of regenerated plantlets was obtained in cv. Phenlene (2.67%) followed by Bossanova (2.41%). Result of ploidy analysis; chromosome number observation and flowcytometry analysis, showed that haploid plants could be developed from all of these hybrid cultivars except cv. Fiesta, where highest percentage of haploid plants were obtained in Minipaprika (40%) followed by cv. Bossanova (36.1%). Haploid plants derived from these hybrid cultivars contained single set of chromosome (12 in numbers), higher stomata density (numbers), and smaller sized stomata as compare to diploid plants. The mean length of stomata was 26.9 ${\mu}m$ in haploid plants and 35.7 ${\mu}m$ in diploids.

• 한국약용작물학회지
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• 제17권3호
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• pp.192-197
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• 2009

Camptotheca acuminata, a native of South China is a well known natural source of monoterpene-indole alkaloid camptothecin(CPT), one of the most promising anti-tumoural compounds. This study was conducted to optimize plant growth regulators and culture conditions on plantlets regeneration through organogenesis from callus of Camptotheca acuminta. Callus were induced from various explants of in vitro germinated plantlets of C. acuminta using WPM medium containing 0.2 ㎎/L 2,4-D. Hypocotyl segments were exhibited higher embryogenic callus than the other explants. Shoot buds formation from embryogenic callus was affected by plant growth regulators, pre-treated dark condition and liquid culture. Organogenesis was optimal in WPM liquid medium containing 0.5 ㎎/L BA. The dark pre-treatment for 2 weeks before the solid culture was effective for organogenesis. The regenerated shoots were rooted in WPM medium with 0.2 ㎎/L NAA and successfully acclimated in green-house conditions.

• 한국육종학회지
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• 제42권5호
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• pp.431-438
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• 2010

Anthers of sweet pepper $F_1$ cultivar 'Special' were cultured on Dumas De Vaulx (C medium), supplemented with $0.1mgL^{-1}$ 2, 4-D and $0.1mg{\cdot}L^{-1}$ kinetin with 3% sucrose, and 0.32% phytagel. The calluses obtained were further sub-cultured on Murashige and Skoog (MS) medium without growth regulators for regeneration. Regenerated plantlets were grown in plastic pots under plastic house and characterized their cytological and morphological characters in spring, 2008. Twenty percent plantlets were identified as haploid plants after chromosome and ploidy analysis. Haploid plants contained 12 chromosomes, high stomatal density with small stomatal length as compared to diploid plants. Stomatal length in haploids was 23.3% smaller than diploids. Haploid plants were characterized as small leaf and petiole size, poor vigor, thin stem and short plant height, short internodes and small flower buds, fruit size and fruit weight as compared to diploid plants and most of the haploid fruits were seedless. SP55, SP62, SP68, SP72 and SP77 are found high yielding double haploids with high total soluble content (8.6, 8.7, 9.2, 9.1 and $9.8^{\circ}Brix$, respectively) and desirable fruit shape, and recommended them to exploit as inbred lines for heterosis breeding.