Inhibitory effect of colchicine on growth and gravitropic responses in Arabidopsis root was explored to find whether there was an involvement of ethylene production. It has been known that cytoskeleton components are implicated in sedimentation of statoliths to respond to gravitropism and growth. The root growth was inhibited by 25% and 40% over control for 8 hr treatment of colchicine at a concentration of 10-5 M and 10-7 M, respectively. The roots treated with colchicine at the concentration of 10-7 M showed the same pattern as control in 3 hr, however, gravitropic response was decreased in the next 5 hr. The colchicine treatment at the concentration of 10-5 M inhibited the gravitropic response resulting in 60° of curvature. In order to better understand the role of colchicine, the production of ethylene was measured with and without the treatment of colchicine. Colchicine increased the ethylene production by 20% when compared to control via the activation of ACC oxidase and ACC synthase activity. These results suggest that the inhibition of the growth and gravitropic responses of Arabidopsis roots by the treatment of colchicine could be attributed to the rearrangement of microtubule, and increase of ethylene production.

Background: For stable induction of tetraploidy in Fallopia multiflora Haraldson, colchicine was treated to establish the condition of induction and investigated the morphological and cytogenetic traits of the tetraploid plants obtained compared to those of diploid ones. Methods and Results: For the induction of tetraploidy, F. multiflora plants were soaked in aqueous solutions of colchicine at various concentration (0.1, 0.5, and 1.0%). After this, 2% dimethyl sulfoxide (DMSO) was added at room temperature on a shaker set at 150 rpm for periods of 12, 24, and 48 h. The induction rate of tetraploids appeared to be the highest in plants treated with 0.5% colchicine for 24 h. As the colchicine concentration and soaking time increased above these levels, the growing tip of the roots did not develop and they began to rot. When compared to diploid plants, tetraploids differed greatly in various characteristics, including the sizes and shapes of the leaves, fruits, flowers and roots. The induced tetraploid F. multiflora had larger guard cells, and chloroplasts, increased number of chloroplast in the guard cells and decreased stomatal densities. Conclusions: When colchicine induced plants for tetraploid, it can be distinguished from diploids, in various characteristics such as morphological changes as stomatal size, number of chloroplasts per guard cell, number of chromosomes and flow cytometry. Therefore, it proved that these methods are suitable, quick and easy methods for the identification of the ploidy level of F. multiflora.

This was carried out to develop a chromosome-doubled (12x) persimmon that will be used as a crossing parent to select seedless persimmon cultivars with the change of the consumption trend recently. To obtain a chromosome-doubled (12x) persimmon, colchicine was applied at the meristem of seedlings in vitro derived from cross among hexaploid persimmon (Diopyros kaki Thunb.). These were treated with 0.03%, 0.05% and 0.1% colchicine respectively for doubling chromosome, and it was most effective at the concentration of 0.05% colchicine. After colchicine treatment, we conducted tests to elucidate conditions for inducing shoot and root development. As the result, the shoots grew best when cultivated at 1/2MS media plus 10 and 30 μM zeatin respectively, and the roots grew best when cultivated at 1/2MS media after dipping for 5 seconds at 10 mM NAA+5% DMSO. We also compared seedlings that have chromosome (6x) do not doubled and crossing parents (6x) and chromosome-doubled seedlings (12x). As the result, these chromosome-doubled seedlings (12x) showed lower stomatal density and larger stomatal size.

Background : This study was performed to investigate the morphological and growth characteristics and metabolomics approach for the diploid and tetraploid Polygonum multiflorum Thunb (PMT). Methods and Results : The present study was performed to investigate the effective induction method of tetraploid plants using colchicine treatment. To know the morphological and growth characteristics of PMT overground growing was investigated about the stem diameter, leaf length and width, a scape, inflorescence length, chlorophyll, seed length and seed width. Metabolite profiles from P. multiflorum were analyzed by an ultra performance liquid chromatography–quadrupole - time of flight (UPLC-Q-TOF) mass spectrometry (MS). The number of stomata and chlorophyll content was also higher in tetraploid. However, the physiological and morphological characteristics of the newly developed tetraploid showed noticeable results compared to the diploid plant. Also, the inherent characteristics of a newly acquired tetraploid also exhibited better results compared to diploid. Twenty one compounds including catechins from leaf and thirty compounds including stilbenes from roots were identified as major metabolites of PMT. The levels of most root metabolites except dethiobiotin, epigallocatechin gallate, procyanidin B1, epiafzelechin 3-O-gallate, and moupinamide from PMT were higher than those of diploid. The levels of 2,3-digalloylglucose, fallacinol, tricin, physcion-8-O-β-D-glucoside, torachrysone-8-O-β-D-glucopyranoside, questin, aloe-emodin 8-O-(6′-O-acetyl)-glucoside, polydatin, 2,5-dimethyl-7-hydroxychromone, 3,5,4'-trihydroxystilbene, aloe-emodin, myrciacitrin II and ω-hydroxyemodin from teraploid PMT were 342, 32, 31, 21, 11, 11, 9, 8, 6, 5, 4, and 2 times, respectively, higher. Conclusion : From above results, overground growing of tetraploid P. multiflorum is fairly better than that of native P. multiflorum.

Background : This study was performed to investigate the morphological and growth characteristics of tetraploid Polygonum multiflorum L. with colchicine treatment, and to clarify its effective induction and time for the increase of rhizome hypertrophy and effective components. Methods and Results : The induction of tetraploid P. multiflorum were done with colchicine treatment of 0.1, 0.5, and 1.0% conc. and the time was treated with 12, 24, and 48 hrs, respectively. DMSO 1% is treated for the effective penetration of colchicine with constant-temperature oven at 24C. Leaf explant was added the HR-A solution stained with HR-B solution, and then determined whether DNA contents of the doubles or not. Seed stand rate showed lower tendency as higher treatment concentration and as delayed the treatment time. When treated in DMSO 1% with 48 hr, seed stand was so small rate with 2%, and treated in 0.5% with 12 hr, seed stand was recorded higher rate with 33%. When treated in 0.5% colchicine with 24 hr, chimera and tetraploid induction rates were highest and obtained 29 individual chimera and 5 individual tetraploid. On overground growing, mean stem diameter of tetraploid (11.3 ㎜) is two times thicker than that of diploid (6.4 ㎜), with the vigorous growth. leaf length and width of tetraploid was 9.1 and 6.2 ㎝, respectively, and that of diploid was 6.8, and 5.1 ㎝, respectively. Stem diameter of a scape was also more thicker tendency in tetraploid than in diploid, chlorophyll contents over 29% existed in tetraploid. Investigated result from stoma no. and size of leaf with 400 magnification, stoma no. of diploid (26 individual) was four times higher than that of tetraploid (7 individuals), the size of tetraploid (38 ㎛) is twice longer in size of diploid (18 ㎛) and stoma area are broaden in four times. Seed length of tetraploid (3 ㎜) is longer than that of diploid (22 ㎜); but, seed width of tetraploid (22 ㎜) is widen than that of diploid (14 ㎜) in seed size comparison. That is, seed length is bigger in 1.4 times and seed width is bigger in 1.6 times, and seed surface area is broaden in two times. Conclusion : From above results, overground growing of tetraploid P. multiflorum is fairly better than that of native P. multiflorum. In the future, this tetraploid variety will promote to register when recognize the excellence than native species after tested the disease tolerance, underground growing characteristics, and key index component.

The present study was performed to investigate the effects of the colchicine concentrations on chromosome doubling for producing of tetraploid plants of Codonopsis lanceolata, and its effect on plant morphology. A total of 180 individuals germinated from 16 treatment groups, were exposed to various concentrations (0.05-1.0% w/v) of colchicine for different soaking duration (3-24 hour). The highest numbers of tetraploid plants (3) were observed from the lowest concentration of colchicine (0.05%), and one (1) tetraploid plant was obtained from the 0.5% concentration group with a 6 hour treatment. However, no tetraploid individual was observed in any other treatment groups. The plant height of the diploid (18.1 ㎝) was slightly shorter than that of the tetraploid (13.4 ㎝). The fresh weight of the main root in the diploid (0.5 g) was four-fold higher than the tetraploid (2.2 g). The colchicine-treated plant regeneration rate in C. lanceolata was decreased when the plants were subjected to high concentration of colchicine. In particular, the highest number of tetraploid plants (5 and 3) was obtained from the lower concentration (0.05% and 0.1%) of colchicine for 6-hour treatment, which were a higher rate (29.4% and 30%) of regenerated tetraploid plants than other regenerated plants. As in the seed treatment result, the plant height of the diploid was significantly higher (10.4 ㎝) than tetraploid. The higher morphological changes were observed comparatively from tetraploid plants than the diploid.

Background : Recently new breeding goal setting is required to develop new boxthorn variety with superior treats of bigger fruits, higher yield in fruits and sprout, pest tolerance, higher medicinal functionality, and self-compatiblity. Various genetic resources also should be retained for this goal. This study was carried out to obtain boxthorn tetraploid for breeding resource. Methods and Results : Colchicine concentration and treatment time for polyploid induction were 0.1, 0.2, 0.4, 0.6% and 12, 24, 36, 48hours respectively. Polyploid was identified by chromosome numbering of individuals by aceto-carmine stain which were examined for polyploid by flow cytomertry(Partec, PA-1) in advance. Conclusion : When cuttings of Cheongyang-jaerae were treated by colchicine to induce tetraploid, survival rate decreased as colchicine concentration and treatment time increased. Rate of individuals having over 30 chloroplast number of stomata guard cell was highest as 37.5% in colchine 0.1% and 48 hours treatment. 8 of them were identified as polyploid by flow cytometry and tetraploid induction efficiency was highest as 6.0% in colchine 0.1% and 48 hours treatment. According to chromosome number examination, 3 of 8 individuals were aneuploid and 5 were tetraploid.

This study was conducted to find out the effective induction method of tetraploid plants to obtain potential data for cultivating superior varieties by colchicine treatment. The seed germination were decreased by the higher concentration of colchicine treatment and longer soaking time. A total of 907 individuals were germinated in 16 treated plots except control (untreated plot) and 28 tetraploids were induced which was about 3.1% of the number of seed germinated. The plant regeneration rate by colchicine treatment on explant of Prunella vulgaris for. albiflora Nakai under in vitro culture was decreased with the higher concentration of colchicine. While a total of 312 individuals were regenerated in all treatments, the explant was soaked in more than 0.05% for over 1 hour, tetraploid could be obtained. In particular, for the soaking treatment in 0.05% for 6 hours and 12 hours, 37 tetraploids were induced, which was about 57.8% of the number of plant regenerated. In accordance with the observation on doubling of DNA contents in leaf in order to identify polyploid, the peak DNA content of G1 phase was 101.3 for diploid and 197.2 for tetraploid. The result confirmed the doubling of DNA content. Furthermore, the number of chloroplasts per guard cell depending on polyploid was around 10 in diploid and 19.3 in tetraploid, which was around 1.9 times as much as diploid.

본 연구는 심비디움의 배수성 육종의 기초자료를 얻기 위하여 실시하였다. 서양란 심비디움 품종인 C. Mystery Island ‘Silk Road’를 자가수분하여 얻은 선발계통(Silk Road-4)의 생장점배양을 통하여 생산된 PLBs(Plant like bodies)에 뾰족한 핀셋으로 10회 찔러 상처를 준 후 oryzalin을 3, 5, 10, 20mg L⁻¹를 첨가한 액체배지에서 3, 6. 9일간 진탕처리하였다. 모든 처리농도에서 처리시간이 길어질수록 생존율이 낮아지는 경향을 보였으며 배수화의 정도 는 4배체 뿐 아니라 2배체와 4배체, 4배체와 8배체의 혼수체, 기타 염색체키메라도 처리에 따라 나타났다. 그중 10mg L⁻¹ 농도로 3주간 처리했을 때와 20mg L⁻¹ 농도로 1주간 처리했을 때 4배체 출현빈도가 12.5%로 가장 높았으 나 생존율이 상대적으로 낮아 효율은 0.27로 나타났고 3mg L⁻¹ 농도로 2주간 처리했을 때는 4배체 출현빈도가 4.5%였지만 상대적으로 생존율이 높아 효율이 0.83으로 가장 높게 나타났다. 콜히친의 경우 50, 100, 300, 500mg L⁻ ¹ 농도로 1, 2, 3주 처리하였다. 그중 300mg L⁻¹ 농도로 3주간 처리했을 때 4배체 출현빈도가 25%로 가장 높았으나 생존율이 상대적으로 낮아 효율은 0.13로 나타났고 50mg L⁻¹ 농도로 1주간 처리했을 때는 4배체 출현빈도가 13.8% 였지만 상대적으로 생존율이 높아 효율이 3.87로 가장 높게 나타났다. 배수화가 된 식물은 2배체 식물보다 생육이 더뎠고 잎의 길이는 짧은데 비해 넓이가 넓고 두께가 두껍게 나타났으며 기공의 크기는 2배체 식물에 비해 커지고 면적당 기공의 수는 감소하였다

본 연구는 심비디움의 배수성 육종의 기초자료를 얻기 위하여 실시하였다. 서양란 심비디움 품종인 C. Show Girl ‘Silky’의 생장점배양을 통하여 생산된 PLBs(Plant like bodies)에 뾰족한 핀셋으로 10회 찔러 상처를 준 후 oryzalin 을 3, 5, 10, 20mg L⁻¹를 첨가한 액체배지에서 1, 2, 3주간 진탕처리하였다. 모든 처리농도에서 처리시간이 길어질수 록 생존율이 낮아지는 경향을 보였으며 배수화의 정도는 4배체 뿐 아니라 2배체와 4배체, 4배체와 8배체의 혼수체, 기타 염색체키메라도 처리에 따라 나타났다. 그중 20mg L⁻¹ 농도로 2주간 처리했을 때 4배체 출현빈도가 46.7%로 가장 높았으나 생존율이 상대적으로 낮아 효율은 34.21로 나타났고 5mg L⁻¹ 농도로 2주간 처리했을 때는 4배체 출 현빈도가 44.4%였지만 상대적으로 생존율이 높아 효율이 40.0으로 가장 높게 나타났다. 콜히친의 경우 50, 100, 300, 500mg L⁻¹ 농도로 1, 2, 3주 처리하였다. 모든 처리시간에서 100%의 생존율을 보인 50mg L⁻¹ 농도를 제외하고 oryzalin 처리와 마찬가지로 처리시간이 길어질수록 생존율이 낮아지는 경향을 보였으며 배수화 정도의 종류도 oryzalin 처리와 마찬가지로 나타났다. 그중 500mg L⁻¹ 농도로 2주간 처리했을 때 4배체 출현빈도가 63.6%로 가장 높았으나 생존율이 상대적으로 낮아 효율은 46.67로 나타났고 100mg L⁻¹ 농도로 2주간 처리했을 때는 4배체 출현 빈도가 63%였지만 상대적으로 생존율이 높아 효율이 56.67로 가장 높게 나타났다. 배수화가 된 식물은 2배체 식물 보다 생육이 더뎠고 잎의 길이는 짧은데 비해 넓이가 넓고 두께가 두껍게 나타났으며 기공의 크기는 2배체 식물에 비해 커지고 면적당 기공의 수는 감소하였다.

'망종화'에서 콜히친 처리에 의한 효율적인 4배체 식물을 유도하고자 적정 식물체 부위, 적정농도 및 침지시간을 검토하였다. 종자의 발아율은 콜히친의 농도가 높을수록 또한 침지시간이 길수록 저하되었다. 대조구를 제외한 16개의 처리구에서 총 453개체가 발아된 반면 4배체는 유도되지 않았다. 기내 배양 중인 줄기 절편체의 식물체 재생율은 콜히친 처리농도 0.01%에서 최고를 나타내다가 0.1% 농도 이상으로 높아질수록 낮았다. 적정식물체 부위는 줄기 절편체로 나타났다. 4배체 식물은 콜히친을 0.05% 이상으로 6시간 침지처리 하였을 때 얻을 수 있었고, 특히 0.05%, 12시간 침지처리에서 식물체의 재생수 대비 약 42%의 높은 획득율을 보였다. Flowcytometry에 의해 DNA함량의 배가여부를 확인한 결과, G1 phase의 DNA 함량 peak가 2배체에서 94.5, 4배체는 192.5로, DNA가 배가됨을 확인할 수 있었다. 또한 공변세포 당 엽록체 수는 2배체가 약 10개인 것에 비해 4배체는 17~19개로 2배체보다 약 1.7~1.9배 정도 많았다.

이 연구는 수량이 많은 4배체 호밀 품종을 육성하기 위한 일환으로 기존의 2배체 호밀에 0.05% 농도의 콜히친을 처리하여 염색체가 배가된 4배체 식물체를 만드는 과정에서 배수체 작성효율, 처리시기, 4배체의 작물학적 특성 등 몇 가지 기초자료를 얻었다. 1. Flow cytometry(FC) 방법으로 호밀에서 배수성 여부를 구분하였는데, 2배체는 DNA 양(DAP1)이 50 전후, 4배체는 80~110 범위이었다. 2. 처리시기에 따른 2배체 호밀의 콜히친 처리후 생존율은 최아종자 처리가 18.5%, 2엽기 처리가 78%로 나타났지만, 생존한 개체 중에서 4배체 배가비율은 각각 50.9%, 1.1%로써, 처리시기별 총 4배체 작성효율은 최아종자 처리(9.42%)가 2엽기 처리(0.86%)보다 월등히 높았다. 3. 최아종자에 콜히친 처리 시 처리종자의 생존율은 1차 년도에 평균 18.5%(곡우호밀 33%, 조그린 4%), 2년차에 평균 42%(곡우호밀 56%, 조그린 21%, 참그린 49%)를 보였다. 생존한 개체들 중에서 4배체 비율은 1년차에 평균 50.9%(곡우호밀 53.7%, 조그린 32.4%), 2년차에 평균 60.5%(곡우호밀 64.1%, 조그린 51.5%, 참그린 60%)를 보였다. 4. 최아종자 처리 시 4배체 작성 효율(생존율 × 4배체 유기율)은 1차년도에 곡우호밀 17.7%, 조그린 1.3%, 2차년도에 곡우호밀 35.9%, 조그린 10.8%, 참그린 29.4%로 품종간 차이를 보였다. 5. 2배체 호밀에 콜히친 0.05%를 12시간 처리하여 35개의 4배체 식물체에서 148개 이삭과 2,673립의 종자를 얻었다. 식물체당 이삭수는 곡우호밀 4.4개, 조그린 3.3개, 수당 립수는 곡우호밀 18.3개, 조그린 15.6개, 천립중은 곡우호밀 37.6g, 조그린 46.8g 이었으며, 14개 이삭을 갖는 식물체도 2개나 되었다.

The effect of colchicine treatment on the agronomic performance and polyploid formation of Bupleurum falcatum using flow cytometry technique was investigated. The roots of 4-leaf stage plants were treated with colchicine (0.5%) for 3, 6, 12, and 24 hours and then transplanted in the field. Agronomic characters (survival rate, plant height, chlorophyll content, bolting rate) were recorded at 4 weeks and 6 months after transplanting while flow cytometry technique was conducted for determination of polyploid formation. Flow cytometry technique revealed polyploid nuclear DNA formation in colchicine treated plants. The highest number of polyploids was obtained at the shortest colchicine treatment time indicating an inverse relationship between colchicine treatment time and polyploid formation. Results also showed that survival and bolting rates were inversely related with the treatment time while plant height and chlorophyll were not significantly affected by the treatment. This study showed a convenient method for determination of colchicine-induced polyploid in B. falcatum and its superior agronomic performance at shorter treatment time.

Present studies were carried out to produce tetraploid plants by colchicine treatment using seeds, seedlings and shoot tips of Codonopsis lanceolata. Three tetraploid plants of C. lanceolata were produced from seeds which absorbed 0.1 % colchicine solution for 12 hours, and 0.5% colchicine solution for 1 and 6 hours from seedlings, respectively. But tetraploid was not produced from shoot tips treated by colchicine solution. Compared to diploid, tetraploid plants had larger stomata, but less number of stomata. Fresh weight of tetraploid plants was 1.4∼3.6 times heavier than diploid plants.

Present studies were carried out to produce tetraploid plants by colchicine treatment using seeds, seedlings and shoot tips of Platycodon grandiflorum in Campanulaceae. The most successful colchicine treatment for tetraploid production in P. grandiflorum was soaking treatment using 0.01 and 0.5% colchicine solution for 1 hour and 12 hours, respectively. Morphological characteristics of both diploid and tetraploid were similar, but tetraploid plants had more leaves. Compared to diploid, tetraploid had the larger stomata, but less number of stomata. Fresh weight of tetraploids was 20∼40% heavier than that of diploid.

고추냉이의 배수체를 유도하기 위하여 EMS와 colchicine을 처리하였을 때 변화되는 단백질과 과산화효소 활성은 다음과 같이 나타났다. 발아율은 무주산이 울릉도산 보다 높았으며, GA3 100 ppm+ EMS 10 ppm 혼용처리시 무처리구에 비하여 모두 6배 이상 증가되었다. Mutagen 처리후 생존율도 울릉도산 보다 무주산에서 더 높게 나타났으며, 과산화효소 활성과 와사비의 초장은 감소되어 나타난 반면, 줄기와 뿌리는 비대되는 경향을 보였다. 무주산과 울릉도산의 단백질 양상이 비슷하고, mutagen을 처리했을 때 polypeptide spot수가 증가되었으나 단백질 pattern은 다르게 나타났고, 특히 EMS 처리는 무처리구에 비하여 현저하게 다른 양상을 보였다.