약용식물의 건전종자 선발 및 발아율 검정을 위한 Tetrazolium Test 소개

이가연,민지윤,문병철,강영민 한약정보연구회 2015 한약정보연구회지 Vol.3 No.1

Propagation technology and evaluation of seed viability is a highly demanded technique in the production of Korean herbal medicine. To produce Raphanus seeds as the examples of Korean herbal medicine, seed germination improvement is very important. In this study, tetrazolium test was used because of rapid and simple method. After the selection of seed groups by the tetrazolium test, healthy seeds were A group and bad seeds were B group. The germination of seeds in A group were significantly different (alpha=0.05) from the germination of seeds in B group the given time. In addition, B seed group could not be cultured and propagated a long time because of tetrazolium chemical damage and had bad embryo physiologically. Also, nondestructive evaluation of viability for individual seeds as X-ray seed analysis or hyperspectral reflectance imaging system was required in future study. These seed analytical studies by tetrazolium test will be improved technology for reproductive medicinal herbs including Raphanus seeds.

민들레와 할미꽃 종자의 정선기술 개발

김석현 한국국제농업개발학회 2004 韓國國際農業開發學會誌 Vol.16 No.2

종자의 크기가 매우 작으면서 무게도 가벼운 민들레와 할미꽃 종자를 효과적으로 정선하는 기술을 개발하기 위하여 실시 한 본 연구에서 다음과 같은 결과를 얻었다. 민들레는 손으로 비빈 후 1.0~1.7㎜체로 쳐서 남는 가운데 부분의 종자를 선별하는 것이 가장 효과적이었다. 할미꽃은 깃털과 함께 부착된 꽃송이를 Seed Scarifier에 넣어 500 rpm으로 60초간 솜털과 종자를 분리한다. 분리된 종자 부분을 2.0㎜ 체로 쳐서 남는 부분을 농황산에 30초간 침지 한 후 물로 100배 희석하여 마대에 종자가 포함된 황산희석 액을 넣어 흐르는 물에 수세하여 종자를 선별하는 것이 가장 효과적이었다. 기계적 장치를 이용하거나 농황산을 사용하여 종자를 정선 할 경우 약간의 파손립이 발생하는데, 이것은 정선되는 종자의 전체량에 비하면 적은 량이므로 이 정도의 파손은 큰 문제가 아니라고 생각한다. 민들레와 할미꽃 종자에서 모두 개량정선 방법으로 종자를 정선할 경우 발아율이 관행정선한 경우에 비하여 약간 저조하였으나 유의적인 차이는 없으므로 보아 이 연구에서 개발된 개량정선방법은 민들레와 할미꽃 종자를 인공적으로 대량번식할 경우 시간과 노동력 및 경비를 줄이는데 효과가 클 것으로 판단된다. The development of efficient refining procedure for the seeds of wild plant species is required to reduce the cost and labor input in artificial propagation of wild plant. This study was carried out to develop methods for collecting and refining tiny seeds from wild plants. For obtaining refined Taraxacum mongolicum seeds, the inflorescence was rub with hands, and subsequently the roughly refined seeds were selected between 1.7~1.0 ㎜ sieve. The seeds of Pulsatilla koreana were refined as follows. Inflorescences were separated using a Seed Scarifier at 500 rpm for 60 sec. The separated seeds were passed 2.0 ㎜ screen and immersed into conc. (95%) sulfuric acid for 30 sec. and collected seeds after dilution of the sulfuric acid solution 100 times with tap water. During seed refining process using mechanical or sulfuric acid treatments, a small portion of damaged seed was evolved, however, the amount was not noticeable as compared to the total amount of collected seeds. Because the differences of germination percentages between hand-refined seeds and seeds refined by above methods were not statistically significant, the developed procedures for refining tiny seed of wild plants are helpful to reduce the cost and labor input in artificial propagation of two species.

History of orchid propagation: a mirror of the history of biotechnology

Tim Wing Yam,Joseph Arditti 한국식물생명공학회 2009 Plant biotechnology reports Vol.3 No.1

Orchid seeds are nearly microscopic in size. Because of that, many fanciful theories were proposed for the origin of orchids. Almost 400 years separate the time when orchid seeds were seen for the first time and the development of a practical asymbiotic method for their germination. The seeds were first observed and drawn during the sixteenth century. Seedlings were first described and illustrated in 1804. The association between orchid and fungi was observed as early as 1824, while the requirement for mycorrhiza for seed germination was established in 1899. An asymbiotic method for orchid seed germination was developed in 1921. After Knudson’s media B and C were formulated, orchids growing and hybridization became widespread. Hybrids which early growers may not have even imagined became possible. Orchid seeds are nearly microscopic in size. Because of that, many fanciful theories were proposed for the origin of orchids. Almost 400 years separate the time when orchid seeds were seen for the first time and the development of a practical asymbiotic method for their germination. The seeds were first observed and drawn during the sixteenth century. Seedlings were first described and illustrated in 1804. The association between orchid and fungi was observed as early as 1824, while the requirement for mycorrhiza for seed germination was established in 1899. An asymbiotic method for orchid seed germination was developed in 1921. After Knudson’s media B and C were formulated, orchids growing and hybridization became widespread. Hybrids which early growers may not have even imagined became possible.

갈대와 띠 종자의 정선기술 개발

김석현 한국잔디학회 2004 한국잔디학회지 Vol.18 No.1

The efficient refinement of seed is required to reduce the cost and labor input in artificial propagation of wild plant. This study was carried out to develop methods for collecting and refining tiny seeds from wild plants. For obtaining Phragmites communis seeds, the inflorescence was cut into small fragments using a Straw Cutter and subsequently detached pappus hairs from seed coat by Hammer Mill. The primary refined seeds were passed 1.0 mm sieve. The screened seeds were subjected to Seed Blower with wind speed of 0.25 mㆍsec-1 to collected intact and well-ripen seeds. The seeds of Imperata cylindrica were refined as follows. Inflorescences were cut using a Straw Cutter first. The pappus was removed from cut fragments using a Hammer Mill and subsequently subjected to Seed Scarifier at 500rpm for 60 sec. for further separation. The separated seeds were passed 1.0 mm screen and collected after blowing with Seed Blower of wind speed of 0.15 mㆍsec-1. When the amount of seed was too little to refine with Seed Scarifier and Blower, the procedure was slightly modified from the procedure described above. The crude seed mixture obtained from Hammer Mill step was hand-refined roughly and then immersed into cone. (95%) sulfuric acid for 2 min. and collected floating portion after dilution of sulfuric acid solution 100 times with tap water. The collected seeds were dried and passed 0.149 mm sieve. During seed refining process using mechanical or sulfuric acid treatments, a small portion of damaged seed were evolved, however, the amount was not noticeable as compared to the total amount of collected seeds. Because the germination percentages between hand-refined seeds and seeds refined by above methods were not statistically different, the developed procedures for refining tiny seed of wild plants are helpful to reduce the cost and labor input in artificial propagation of two species.

Improvement of Seed Germination in Scrophularia takesimensis, Korean Native Plant

Jeong Hee Kang(강정희),Kyoung Ok Choi(최경옥),Sang Yeol Ahn(안상열),Dong Shik Kim(김동식),Young Shin Chon(천영신),Jae Gill Yun(윤재길) 한국원예학회 2009 원예과학기술지 Vol.27 No.4

Scrophularia takesimensis, a Korean native plant, is one of the rare and endangered species designated by the Korea Forest Service (KFS) because seed germination hardly occurs in nature due to of some propagation difficulties and its survival has been threatened by overexploitation. A better understanding of dormancy in S. takasimensis would contribute to successful propagation of this species from seeds. For the purpose of a higher percentage of germination, the seeds collected from the native place (Ullungdo) were stored in a 4℃ refrigerator, sown in petri dishes (90 ㎜) at 10-day intervals for 60 days, and placed in a seed germinator at 20℃. Seed germination was not observed in any duration of low temperature treatments. To improve poor germination, seeds were soaked in 50-400 ㎎ㆍℓ?¹ gibberellic acid (GA₃) or indole 3-butyric acid (IBA) for 24 hours. Soaking in 200 ㎎ㆍℓ?¹ GA₃ remarkably promoted germination up to 80%, but the other concentrations were not effective (>40%). On the other hand, soaking in all concentrations of IBA did not improve germination at all. To select an adequate temperature for seed germination, seeds, previously soaked in a 200 ㎎ㆍℓ?¹ GA₃ for 24 hours, were incubated at 10, 15, 20, or 25℃. Seed germination of over 70% was attained at 15 and 20℃, but only 20% at 25℃. These results indicate that seeds of S. takesimensis are in such deep dormant state that they hardly germinate even by low temperature treatment. Soaking in 200 ㎎ㆍℓ?¹ GA₃ for 24 hours, however, markedly improved seed germination up to 80%. 한국자생식물인 섬현삼(Scrophularia takesimensis)은 자연상태에서 발아가 잘 이루어지지 않고 자생지인 울릉도에서의 과도한 채취로 인해 멸종 위기에 직면해 있다. 이 멸종위기 식물을 보존하고 유지하기 위해서 종자에 의한 대량증식기술이 요구된다. 섬현삼 종자의 휴면여부를 확인하기 위해서 울릉도에서 자생하고 있는 종자를 채종하여 4℃냉장고에 60일 동안 저장하면서 10일간격으로 90㎜ 페트리디쉬에 파종하여 20℃ 항온기에 배치하였다. 그 결과 저온처리하지 않은 대조구와 60일까지 저온처리한 모든 종자들은 발아되지 않았다. 종자발아를 향상시키기 위해서 50-400㎎ㆍℓ?¹ GA₃용액 또는 IBA(indole 3-butyric acid)용액에 24시간 침지시킨 후 90㎜ 페트리디쉬에 파종하여 항온기에 배치 하였다. GA용액에 침지처리로 발아율이 현저하게 향상되었는데, 특히 200㎎ㆍℓ?¹ GA₃용액에 24시간 침지시켰을 때, 발아율이 80%까지 향상되었다. 하지만 다른 모든 농도와 IBA용액 처리에서는 발아가 되지 않았다. 섬현삼의 종자를 200㎎ㆍℓ?¹ GA₃용액에 24시간 침지시킨 후 10, 15, 20, 25℃ 항온기에 각각 파종한 결과, 15℃와 20℃에서 70% 발아율을 나타내었고 25℃에서는 20% 발아율을 나타내었다. 결론적으로 섬현삼(Scrophularia takesimensis)종자는 저온처리에 의해서도 휴면이 타파되지 않을 정도로 깊은 휴면을 가지고 있다는 것이 확인되었다. 그렇지만 200㎎ㆍℓ?¹ GA₃ 용액에 24시간 침지시킨 섬현삼 종자는 휴면이 타파되어 80%까지 종자발아율이 향상되었다.

Factors That Affect Seed Germination and Changes in Endogenous Gibberellins and Abscisic acid Concentrations in Mukdenia rossii (Oliv.) Koidz

Ju Sung Cho,Jeong Hak Jeong,Seok Min Lee,In Jung Lee,Cheol Hee Lee 한국원예학회 2018 원예과학기술지 Vol.36 No.4

The present study was conducted to establish storage conditions that accelerate germination and maintain the germination rate of Mukdenia rossii (Oliv.) Koidz. seeds. Fresh seeds of M. rossii exhibited an excellent germination rate of over 95.5%, and cotyledon development was confirmed by the presence of a sufficiently developed embryo. Therefore, the seeds of M. rossii were classified as nondormant type. Seeds with weak germination potential due to long-term storage were used for the experiment to determine the temperature and light condition and improve germination rate. To evaluate how changes in storage conditions affect the relationship between germination characteristics and hormone content, fresh seeds were stored at different temperatures for 2, 4, and 6 months and germinated at 20°C in the presence of light. Treatment with 100 mg·L<SUP>-1</SUP> gibberellic acid (GA) improved the germination rate of M. rossii seeds by 94.8% whose germination rate had decreased due to long-term storage. Seeds stored at different temperatures and for different periods of time exhibited high germination rates between 88.8%-99.5% under all storage conditions; however, storage period had negative effects on mean germination time, germination energy, and time taken for the germination rate to reach 50% that were statistically significant (p < 0.001). Endogenous GA content in seeds that had been stored for 6 months at 25°C changed marginally, therefore, it can be predicted that the relative proportion of ABA in the seed is increased. On the other hand, the relative concentration of abscisic acid (ABA) increased significantly. In contrast, the M. rossii seeds stored at -20°C had a decrease in activated GA (GA4) levels and increased levels of precursor GAs, such as GA24 and GA9. Furthermore, endogenous ABA content decreased significantly, which increased the relative GA concentration in seeds. Therefore, -20°C is an effective storage temperature for the propagation of M. rossii seeds.

지리산바위솔과 제주연화바위솔 종자의 형태특성 및 저온과 GA에 대한 발아 반응

강정희(Jeong Hee Kang),정경진(Kyeong Jin Jeong),최경옥(Kyoung Ok Choi),천영신(Young Shin Chon),윤재길(Jae Gill Yun) 한국원예학회 2010 원예과학기술지 Vol.28 No.6

우리나라 자생식물의 하나인 지리산바위솔(O. japonicus A. Berger)과 제주연화바위솔[O. iwarenge (Mak.) Hara]의 종자 형태를 관찰하고 종자발아생리를 구명하기 위해 저온과 GA를 처리하였다. 종자의 형태를 실체현미경으로 관찰한 결과 2가지 바위솔 모두 세로로 주름진 쌀알 모양이었으며, 크기는 길이 0.77-1.00㎜, 폭 0.25-0.37㎜로 미세 종자임이 확인되었다. 휴면유무와 발아최적 온도를 구명하기 위해, Petri-dish에 파종후 온도가 다른 파종상에서 발아 시켰을 때, 지리산바위솔은 모든 온도에서 20% 이하의 발아율을 보였고, 제주연화바위솔은 10℃에서만 80% 정도의 발아율을 보였다. 4℃ 냉장고에 건식저장을 하면서 10일 간격으로 온도별로 파종한 결과, 지리산바위솔은 무처리에서는 발아가 전혀 보이지 않았다. 10일간 저온처리 후 10℃ 파종상에 두었을 때 발아율 44%로 많이 향상되었으나, 그 이상의 저온 처리에서는 발아율이 다시 감소하였다. 제주연화바위솔의 경우 저온 처리 30일까지는 발아율이 크게 향상되어 10℃에서 95% 발아율을 보였다. 그러나 그 이상의 저온처리(40일)에서는 발아율이 급격히 저하되었다. GA₃ 농도를 50-400㎎?L<SUP>-1</SUP>으로 하고, 침지시간은 3-24시간 범위로 하였을 때, 두가지 바위솔 모두 발아 속도가 현저하게 빨라지고 발아율이 획기적으로 높아지는 결과를 얻었다. 지리산바위솔은 모든 침지 시간에서 80-100% 발아율을 보였다. 제주 연화바위솔도 GA₃ 농도와 시간에 상관 없이 파종 후 6일만에 90-100%의 발아율을 나타내었다. This study was conducted to improve the seed germination by low temperature and GA₃ in Orostachys japonicus A. Berger (Jirisan) and O. iwarenge (Mak.) Hara (Jejuyeonhwa), Korean native plants. Observation of the seeds using a stereo microscope showed that all seeds of the two species have wrinkled surface and oblong shape. Seed size ranged 0.77-1.00/0.25-0.37 ㎜ (length/width), indicating that the seeds are minute seeds. When the seeds of two Orostachsis species were sown into petri-dish and placed in a plant growth chamber of 10, 15, 20, or 25℃,’Jirisan’ showed seed germination below 20% at all temperatures and ‘Jejuyeonhwa’ 80% at only 10℃. Seed germination of ‘Jirisan’ increased up to 44% at 10℃ by low temperature (4℃) storage for 10 days, but decreased again at storage for more than 20 days. The seeds of ‘Jejuyeonhwa’ showed a large increase in seed germination by low temperature for 20-30 days, which was 95% at 10℃, but low temperature for more than 40 days significantly decreased seed germination. Dipping treatment in GA₃ solution of 50-400 ㎎?L<SUP>-1</SUP> for different periods (3, 6, 12, and 24 hrs) remarkably improved germination rate and speed in both species, 80-100% in ‘Jirisan’ and 90-100% in ‘Jejuyeonhwa’ at all concentrations and dipping times used in this study.

등수국의 종자발아에 미치는 몇 가지 요인

조주성,정정학,김수영,이주영,이철희 한국자원식물학회 2014 한국자원식물학회지 Vol.27 No.5

This study was carried out to establish seed propagation method of Hydrangea petiolaris Siebold & Zucc., one of the specific and protected plant species for the floral region. Seed size ranged 1.36 × 0.84 ㎜, weight of thousand seeds was 2.3 ± 0.02 ㎎. Observation of seed morphology characters showed wrinkled surface and elliptical shape, indicating amber color and fine seeds. Moisture contents of seeds, increased rapidly by water-soaking treatment, recording maximum moisture contents (31.9%) after 48 hours. Percent germination of seeds was higher under the light than dark condition. Seed germination was the best at 25℃ under the light condition (78.0%) among temperature and light conditions treated. Percent germination, germination energy and T 50 was greatly improved by soaking in GA 3 solutions for 48 hours. Especially, 100∼ 500 ㎎·L -1 GA 3 treatment resulted in the highest germination rate as 90.0% and above. 본 연구는 식물구계학적 특정종 III 등급이며, 국외반출 시 승인이 필요한 보호종인 등수국의 종자 번식법을 개발하기 위해수행되었다. 종자의 크기는 1.36×0.84 ㎜이며, 1,000립 중은2.3 ± 0.02 ㎎이었다. 종자는 타원형으로 표면에 주름이 있었으며, 황색의 미립종자였다. 침지처리에 의해 종자의 함수율은 48시간 만에 3.9%에서 31.9%로 급속히 증가하여 최대 함수율을보였다. 온도 및 광조건에 따른 발아율은 25°C, 명조건에서 가장 높았으며(78.0%), GA3 용액에 48시간 처리함으로써 발아율,발아세 및 T50이 매우 향상되었다. 특히 GA3 처리구는 100∼500㎎·L⁻¹의 농도에서 발아율이 90.0% 이상으로 매우 우수였으며발아세와 T50을 무처리에 비해 향상시킬 수 있었으므로, GA3는등수국 종자의 발아력을 효과적으로 향상시킬 수 있는 적절한화학적 처리방법으로 판단되었다

Germination of West African Ebony (Diospyros mespiliformis Hochst) Seeds: Effects of Dehydration and Different Pre-sowing Treatments

Kanmegne, Gabriel,Mbakop, Christelle Nya,Fonkou, Theophile Institute of Forest Science 2020 Journal of Forest Science Vol.36 No.4

Diospyros mespiliformis is a highly valued and threatened tree species within the Sahelo-Sudanian zone of Africa, but its seed germination requirements under cultivation are not well researched. In a first experiment which aimed at determining germination response of seeds to dehydration, fresh seeds were dried at room temperature for 26 days during which their moisture content, their germinability, and their viability were monitored at two-day intervals. In the second experiment, 14 pre-germination treatments were tested for their effect on the germination of dried seeds. Results showed that fresh seeds had 52.7% moisture and achieved 97.7% germination. As seeds were dried, percentage germination gradually decreased with decreasing moisture content and reached 0% when moisture content had dropped to 18%. Meanwhile, seed viability remained at 100% over drying duration. Seeds that were not germinated after air dry also recorded 100% viability. The most effective treatment for inducing germination of dried seeds was scarification using 98% sulfuric acid for 30 min which resulted in 96.6% germination. This study reports for the first time in D. mespiliformis seeds a desiccation-induced dormancy which can be efficiently alleviated by acid scarification. This study provides useful information that will contribute to efficient management of D. mespiliformis seed resources for propagation.

Effect of germination and water absorption on scarification and stratification of kousa dogwood seed

조주성,이철희 한국원예학회 2018 Horticulture, Environment, and Biotechnology Vol.59 No.3

Cornus kousa (kousa dogwood), used as a landscape tree and a source of food and medicine, is difficult to propagate from seeds owing to dormancy. The present study was conducted to develop an effective method of seed propagation and identify the seed dormancy type. Kousa dogwood seeds were classified as presenting physical dormancy (PY) and physiological dormancy (PD) from an immature embryo. Seeds were contained within an impermeable endocarp that conferred PY and had an undifferentiated embryo that grew and differentiated after scarification and cold moist stratification, leading to improved germination. To break PY + PD, four replicates of 50 seeds (3200 in total) for each treatment were prepared by scarification and cold moist stratification. These methods combined improved moisture absorption and germination more than either treatment alone. In particular, the addition of cold moist stratification treatment to scarified seeds could effectively induce embryo development. By contrast, untreated seeds showed low germination rate of less than 1% at 30 °C with continuous light. Sulfuric acid (H2SO4) at different concentrations and durations was used to uniformly wound the endocarp of seeds. Treatment with 80% H2SO4 for 10 min resulted in the highest germination rate (68.6%) and moisture content (65.7%), which were higher than those following polishing with a brushing machine for 5–40 min. Longer treatment with high concentrations of H2SO4 (80 and 100%) caused the acid to infiltrate the seed and inhibit germination by damaging internal tissues, while low concentrations (40 and 60%) did not affect germination rates even after treatment for more than 10 min. To obtain maximum germination, concentration and treatment duration of sulfuric acid should be carefully controlled.