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.

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.

'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.

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.