• Plant Biotechnology Reports
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• 제5권3호
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• pp.265-271
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• 2011

The cultivated potato as well as its tuber-bearing relatives are considered model plants for cell and tissue culture, and therefore for exploiting the genetic variation induced by in vitro culture. The association between molecular stability and tissue culture in different genetic backgrounds and ploidy levels has already been explored. However, it still remains to be ascertained whether somaclonal variation differs between callus-derived chromosome-doubled and undoubled regenerants. Our research aimed at investigating, through amplified fragment length polymorphism (AFLP) markers, the genetic changes in marker-banding patterns of diploid and tetraploid regenerants obtained from one clone each of Solanum bulbocastanum Dunal and S. cardiophyllum Lindl (both 2n = 2x = 24) and tetraploids from cultivated S. tuberosum L. (2n = 4x = 48). Pairwise comparisons between the banding patterns of regenerants and parents allowed detecting considerable changes associated to in vitro culture both at diploid and tetraploid level. The percentages of polymorphic bands between diploid and tetraploid regenerants were, respectively, 57 and 69% in S. bulbocastanum and 58 and 63% in S. cardiophyllum. On average, the frequencies of lost parental fragments in regenerants were significantly higher than novel bands both in S. bulbocastanum (48 vs. 22%) and S. tuberosum (36 vs. 18%) regenerants. By contrast, in S. cardiophyllum, a similar incidence of the two events was detected (32 vs. 29%). Our results revealed that structural changes after tissue culture process strongly affected the genome of the species studied, but diploid and tetraploids regenerated plants responded equally.

• 한국자원식물학회지
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• 제36권6호
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• pp.588-596
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• 2023

Long-term culture of cell lines is an important issue in in vitro culture and in plant science. In this study, the regeneration ability and ex vitro acclimatization of regenerants were evaluated. The ploidy level of regenerants derived from long-term cultured cell lines was measured in Imperata cylindrica 'Rubra', Poaceae. Adventitious buds (shoots) were successfully induced from five-year-cultured calli on MS medium containing 0.1 mg/L BA or 2.0 mg/L TDZ, combined with 0.01 mg/L auxins (IAA, IBA, NAA and 2,4-D), respectively. Adventitious roots were also induced on MS medium containing 0.01 mg/L auxins (IBA, NAA and 2,4-D), respectively. Interestingly, regenerants with both red and green leaf were successfully obtained when regenerants were cultured on MS medium with 9% sucrose. Regenerants derived from long-term cultured calli were transferred to pots using an optimal acclimatization process and successfully adapted to both pot and soil conditions. Moreover, the ploidy level was measured using calli and regenerants that had been kept on MS medium containing various kinds of plant growth regulators (PGRs).

• 한국자원식물학회지
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• 제36권5호
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• pp.496-507
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• 2023

화본과 식물 홍띠(Imperata cylindrica 'Rubra')의 기관분화 관련 유전자의 동태와 기내재생체의 유전적 안정성을 조사하고자 기관분화 단계별 재분화체를 작성하여 기관분화 관련 유전자 발현과 ISSR 마커 기반 변이성을 조사하였다. 5종류 총 15개체의 기관분화 단계별 재분화체에서 캘러스 발생 유전자인 FIE는 모식물체 1번을 제외한 14개의 식물체에서 모두 발현되었으며, 뿌리 발생 유전자인 WOX11도 15개의 모든 단계별 재분화체에서 발현하였다. 체세포 발생 유전자인 LEC1B는 15개 식물체에서 모두 발현하였으나 비교적 약하게 발현하였다. 7종류 총 21개체의 기관분화 단계별 재분화체 및 재분화식물체에 대하여 ISSR 분석한 결과, 유전적 다형성은 기관분화 단계별 재분화체 및 순화 재분화체(실내포트 재배식물체 4.1%, 노지 재배식물체 4.3%, 기내배양 홍띠(적색)식물체 4.2%, 녹색신초 발생 캘러스 5.6%, 캘러스 1.4%)에서 대조구인 모식물체(1.4%)와 같거나 높게 나타났다. 또한, 유전적 유사도 지수는 0.747~1.0 사이에 분포하며, 평균 0.868로 나타났다. 군집분석 결과 유전적 유사도 지수 0.809에서 기외 식물체(모식물체, 실내재배 및 노지재배 재분화 녹색 식물체)와 기내식물체 및 재분화 과정상의 분화체(기내배양 중인 홍띠 식물체, 녹색 신초, 적색신초 발생 캘러스, 캘러스)는 독립적인 2개 그룹으로 유집되었다. 이상의 결과는 화본과 식물의 기내배양 시 재분화 과정에서 일어나는 일련의 유전학적 기초자료를 제공해 준다.

• 한국약용작물학회지
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• 제3권3호
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• pp.207-216
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• 1995

시호의 조직 배양묘와 배양식물의 종자를 이용하여 플러그묘를 생산하기 위한 실험을 수행한 결과를 요약하면 다음과 같다. 1. 재분화식물의 순화를 위해 영양배지없이 증류수, GA(0.1mg/l), putrescine(0.1mg/l)을 소식물체에 흡수시킨 결과 GA처리가 신초 및 뿌리생육에 가장 효과적이었다. 2. 무기염류의 농도를 1/2로 줄인 MS배지와 NAA 0.1ng/l +BA 0.5mg/l 를 조합한 호르몬처리에서 배양묘의 생장과 건물중이 가장 양호하였다. 0.4%의 활성탄물의 관수와 $30^{\circ}C$의 온도에서 생장이 촉진되었다. 3, 배양식물의 종자를 파종하여 육성한 플러그묘는 peatmoss상토와 마사토을 혼합한 상토와 원더그로 1, 2, 3호의 배합비를 1. 3-0. 9-0. 3로 하였을 때 생육이 가장 양호하였다. 4. 4%의 보광처리 및 $30^{\circ}C$의 온도에서 묘령별로는 90일묘가 가장 생육이 양호하였다.

• Journal of Plant Biotechnology
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• 제6권3호
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• pp.157-163
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• 2004

An efficient system for the regeneration of adventitious shoots from in vitro cultured leaf sections of Lycium chinense Miller was developed and silent somaclones from the regenerants detected with RAPD method. Among the eight media tested (B5, SH, N&N, 1/2MS, MS, 3/2MS, GD and WPM), and four cytokinins (BA, kinetin, 2ip and zeatin) with different concentrations (1, 5, 10, 20, 30 and 40 $\mu{M}$), 1/2 MS medium supplemented with 20 and 30 $\mu{M}$ zeatin showed the best regeneration frequency (100% and 93.7%) and higher average number of shoots (9.0 and 9.4). All regenerants easily elongated after subculturing on 1/4MS without growth stimulants and produced spontaneous adventitious roots from their basal parts. With phenotypically normal 40 regenerants, RAPD analysis with 15 different random primers was performed to examine the cryptic somaclonal variants. No substantial differences in banding patterns were found in the amplified polymorphic DNAs implying no DNA changes during dedifferentiation into adventitious shoots. However, one (OPF-4) of the 15 primers detected silent somaclonal variation in one regenerant in which two different polymorphic bands did not appear when compared with the rest regenerants. The results indicate that regenerantion via intervening callus phase can be used to establish true-to-type planting stocks for homogeneous population.

• Plant Breeding and Biotechnology
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• 제6권4호
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• pp.426-433
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• 2018

Plant tissue culture is a technique that has invariably been used for various purposes such as obtaining transgenic plants for crop improvement or functional analysis of genes. However, this process can be associated with a variety of genetic and epigenetic instabilities in regenerated plants, termed as somaclonal variation. In this study, we investigated mutation spectrum, chromosomal distributions of nucleotide substitution types of single-nucleotide polymorphisms (SNPs) and insertions/deletions (InDels) by whole genome re-sequencing between Dongjin and Nipponbare along with regenerated plants of Dongjin from different induction periods. Results indicated that molecular spectrum of mutations in regenerated rice against Dongjin genome ranged from $9.14{\times}10^{-5}$ to $1.37{\times}10^{-4}$ during one- to three-month callus inductions, while natural mutation rate between Dongjin and Nipponbare genomes was $6.97{\times}10^{-4}$. Non-random chromosome distribution of SNP and InDel was observed in both regenerants and Dongjin genomes, with the highest densities on chromosome 11. The transition to transversion ratio was 2.25 in common SNPs of regenerants against Dongjin genome with the highest C/T transition frequency, which was similar to that of Dongjin against Nipponbare genome.

• 한국자원식물학회지
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• 제26권2호
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• pp.320-327
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• 2013

활용가치가 높은 부존식물자원인 갈대의 기내 번식을 통한 배양체계를 확립하고 재분화 식물체들의 유전적 다양성을 검토한 결과, 성숙종자 유래의 캘러스를 통한 기내 식물체 재분화는 N6배지에서 MS배지보다 양호하였고, 0.25~0.5 mg/L의 BA를 포함한 N6배지에서 가장 높았다. ISSR 마커를 이용하여 재분화 식물체의 유전적 안정성을 분석한 결과, 검출된 총 94 유전좌중 유전적 다형성은 17%였고, 평균 유전자다양도 값(h)은 0.03, BA 5 mg/L를 포함한 N6배지에서 0.008, NAA 0.1 mg/L와 kinetin 2 mg/L를 포함한 MS 배지에서 0.040으로 나타났다. 이것은 재분화된 갈대식물체 개체간에 유전적으로 구조가 매우 단순하고 균일하며, 유전적 다양성 진단에 ISSR 마커가 효과적임을 시사한다.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.176-176
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• 2017

Hydropriming had positive effects on the time for germination to reach 50%, the germination index, the time to final germination percentage, and the number of uniform seedlings with enlarged cotyledons in in vitro germination of small watermelon. In addition, the highest shoot initiation was obtained from hydroprimed cotyledonary nodes ($95{\pm}6%$), followed by non-primed cotyledonary nodes ($78{\pm}6%$), hydroprimed cotyledons ($72{\pm}4%$), and non-primed cotyledons ($48{\pm}4%$). Meanwhile, no shoots were initiated from hypocotyls. The total number of shoots that initiated from cotyledonary nodes and cotyledon explants was insignificant, indicating that both cotyledons and cotyledonary node were good sources for the in vitro culture. Choosing explant sources that favor tetraploidy should be the key for producing higher polyploidy plants; a total of 10.5% of tetraploid regenerants were entirely identified from cotyledon explants. Cotyledons with highly differentiated cells might show higher variations than cotyledonary nodes with more preexisting meristematic cells. Cells of cotyledon tissue might undergo changes in ploidy level during differentiation of the culture, or it might be that some of the variations were already present in the tissues of the donor plant. Morphological changes in fruit length of tetraploid regenerants are genotype-dependent.

• 식물조직배양학회지
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• 제27권2호
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• pp.125-131
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• 2000

원형질체는 현탁배양된 세포괴로부터 1% cellulase R-10, 0.25% pectolyase Y-23, 그리고 0.5% driselase가 포함된 CPD 효소용액에서 4시간의 진탕 (40 rpm)조건에서 쉽게 분리되었다. 분리된 원형질체는 1 mg/L 2,4-D, 0.5 mg/L kinetin , 200 rng/L spermidine 그리고 68 g/L glucose가 포함된 KM 액체배지에서 배양되었으며, 또한 현탁배양중인 동종의 feeder cell이 있는 CP배지 위에 액체배양배지를 1 mL균일하게 퍼트린 후,$25^{\circ}C$, 암조건으로 4주간 양육배양하였다. 원형질체 유래 SF 캘러스는 0.05 mg/L IAA, 7 mg/L 2lip와 30mg/L sucrose가 첨가된 MS 재분화 배지에서 광조건으로 4주동안 배양되었을 때, 60% 이상의 shoot를 형성하였다. 이후, 식물의생장조절물질이 첨가되지 않은 MS 배지로 옮겨 뿌리를 유도하였으며, 재분화된 식물체는 토양으로 옮겨 배양 8주후에 종자를 형성하였다.

• Journal of Plant Biotechnology
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• 제5권1호
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• pp.25-31
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• 2003

As a consequence of tissue culture of rice, RAPD analysis was peformed to determine whether extended culture periods as undifferentiated calli affected the subsequent genetic constancy, and whether any resulting DNA rearrangements could be detected between sibling plants produced from the same callus. Somaclonal variation was induced at the initial stage of tissue culture and it increased with the length of culture maintenance. Of the 192 total bands, the number of polymorphic bands was 22 (11.5%), 33 (17.2%), and 49 (25.5%) in the callus of 1,3, and 6 months culture, respectively. A significantly higher level of genotypic polymorphisms between regenerants from two different somaclones was also detected, although all the regenerants were derived from a single genotype. In comparison of DNA polymorphisms between regenerants from non-irradiated and from irradiated calli, a scope of variation spectrum by gamma ray irradiation was larger than that by tissue culture. Consideration must be given to this genomic variation where attempts are to be made to use desirable somaclonal variants for plant breeding purpose and in genetic engineering program.