나무이끼(Climacium japonicum)의 기내배양에 영향을 미치는 몇 가지 요인

Giush Uddin Ahmed,Cheol Hee Lee 한국화훼산업육성협회 2010 화훼연구 Vol.18 No.1

본 연구는 나무이끼(Climacium japonicum) 배우체의 기내증식에 영향을 미치는 몇 종류의 화학적 및 물리적 환경을 표준화하기 위하여 실시하였다. 초기 배양재료는 멸균한 배우체의 정단을 Knop 배지(1865)의 다량요소와 Nitsch와 Nitsch배지 (1956)의 미량요소를 첨가한 고체배 지에 배양하여 획득하였다. 배우체 생산을 위해 좋은 배양재료를 얻기 위하여 다져진 배우체 및 shoot의 정 단부와 기저부를 평가하였다. 배우체의 기내생산에 미 치는 영향을 검정하기 위해 배지 7종류, 총질소 4농도, sucrose 5농도, 광도와 온도 등의 요인들을 분석하였다. Shoot의 정단부가 배우체 번식을 위해 사용한 3종류의 절편체들 중 가장 우수하였으며, Knop 배지(1865)의 다량요소와 Nitsch와 Nitsch배지 (1956)의 미량요소를 첨가한 배지가 다른 종류의 배지들에 비해 배우체의 증식에 효과적 이었다. Sucrose의 농도가 높아질수록 배우체의 신장 및 증식에 좋은 영향을 미쳤으며, 질소의 결핍 및 과다는 배우체의 생장을 억제하였다. 광도의 강약에 따라 배수체의 수, 길이 및 생체중에 큰 변화를 보였으며, 배우체 생장을 위해 최적의 광도는 3000- 4000lx. 인 것으로 생각되었다. 고온 또는 저온 조 건은 배우체의 증식과 생산에 부정적인 영향을 미쳤다. 본 연구에 의해 개발된 생산과정의 단순화, 대량화 및 영 양체의 고품질화는 효과적인 이끼의 기내배양 방법으로 생각되었다. These investigations were conducted to standardize several chemical and physical environments affecting in vitro propagation of gametophytes of Climacium japonicum. Propagation of this moss species was established on basal medium containing Knop macro salts and Nitsch and Nitsch trace elements. Primary cultures were initiated from apical shoots of gametophytes. Gametophyte production was accessed using chopped gametophytes, apical shoots and basal shoots. Seven ty pes of culture media and four concentrations of total nitrogen and five strengths of sucrose were tested for in vitro gametophyte production. Light and temperature factors were also evaluated. Apical shoots were the greatest among three types explants used for gametophyte propagation. Medium containing Knop macro salts and Nitsch and Nitsch trace elements was more effective than other types of media. Higher sucrose concentrations showed a positive effect on the elongation and multiplication of gametophytes. Both nitrogen deficiency and excessiveness inhibited gametophyte growth. Light intensity variation showed highly significant changes in numbers, length and fresh weight of gametophytes. Optimum light intensity for gametophyte growth seemed to be around 3000- 4000 lx. Both lower and higher temperature had a negative effect on gametophyte propagation and production. This study will provide large scale and high quality propagules, and effective moss propagation system.

다시마(Saccharina japonica) 배우체의 미동정 진균증

정하나 ( Ha-na Jeong ),오명주 ( Myung-joo Oh ),최성제 ( Sung-je Choi ),서정수 ( Jung-soo Seo ),박명애 ( Myoung-ae Park ),김위식 ( Wi-sik Kim ) 한국수산과학회 2017 한국수산과학회지 Vol.50 No.2

In 2015, white cottony tufts were observed on gametophytes of the kelp Saccharina japonica. Wet mount and histopathology examination revealed numerous fungal hyphae and mycelium around the gametophytes. The gametophytes surrounded by fungal hyphae were generally round and empty. A specific 610-bp fragment of the internal transcribed spacer (ITS)-5.8S rDNA-ITS gene of fungi was amplified by polymerase chain reaction and the nucleotide sequence showed 100% identity with those of Acremonium sclerotigenum, Acremonium sp. and Ascomycota sp. When fungus-infected gametophytes were mixed with healthy gametophytes, a high transmission rate (100%) resulted. This is the first report of mycosis of gametophytes in Korea.

꼬시래기의 사분포자체와 배우체의 초기 생장에 대한 온도와 광도의 영향

이상용,최한길 한국해양과학기술원 2020 Ocean and Polar Research Vol.42 No.2

The aim of this study is to examine the physiological characteristics of an agarophyte Agarophyton vermiculophyllum (Ohmi) Gurgel, J.N. Norris et Fredericq in the early life stage of tetrasporophytes (2n) and gametophytes (n) to select appropriate seedlings for mariculture. Growth experiments were carried out at the combinations of four temperatures (20, 25, 30, and 35°C) and three light intensity levels (20, 60, and 100 μmol photons m-2 s-1) in the two ontogenetic stages: discoid holdfasts and erect sporelings. Holdfast areas and sporeling lengths of tetrasporophytes and gametophytes were estimated after 14 days in culture. Relative growth rates (RGRs) for holdfast areas were 7.08–28.38% day-1 for tetrasporophytes and 11.58–23.67% day-1 for gametophytes. At 35°C, holdfasts of tetrasporophytes survived with RGRs of 7.08–23.28% day-1 but those of gametophytes died. Maximal holdfast growth of tetrasporophytes occurred at 30°C and 100 μmol photons m-2 s-1, which were different from gametophytes (25°C and 100 μmol photons m-2 s-1). RGRs of tetrasporophytic sporelings were 2.93–11.11% day-1 and were between 0.78–10.82% day-1 for gametophytes. Maximal growth of A. vermiculophyllum sporelings occurred at 25°C and 60 μmol photons m-2 s-1 for tetrasporophytes, and at 20°C and 100 μmol photons m-2 s-1 for gametophytes. In conclusion, the present results indicate that carpospores could be used as resources of spore-seedling methods having genetic diversity for mass field cultivation because tetrasporophytes showed higher-temperature tolerance and faster-growing ability than gametophytes of A. vermiculophyllum in the discoid holdfast and sporeling stages.

Propagation methods for gametophyte proliferation and sporophyte formation in silver cloak fern (Cheilanthes argentea)

장보국,조주성,이철희 한국원예학회 2019 Horticulture, Environment, and Biotechnology Vol.60 No.3

Cheilanthes argentea (S.G. Gmel.) Kunze is a highly valued indigenous/endemic Korean bracken species used for ornamentalpurposes. In this study, we developed a practical proliferation method for mass production of C . argentea plants using tissueculture. The gametophyte proliferation using the chopping method produced gametophytes that were morphologicallyidentical to those obtained from in vitro spore germination. This new method increased plant fresh weight by more than40-fold, from 300 mg to 12.7 g, in Murashige and Skoog medium. A blending method was used to produce 74.3 sporophytesin 7.5-cm 2 plastic pots using 1 g of gametophytes. Furthermore, addition of exogenous gibberellin promoted sporophytedevelopment and growth, suggesting the possibility of controlling the number of sporophytes formed. However, furtherstudies are needed to explain the eff ect of exogenous gibberellin, which is closely related to the sex-determining hormoneantheridiogen, on the number of sporophytes and its mechanism of action. Our new tissue culture system is capable of massproliferation of C. argentea gametophytes in vitro and the formation of sporophytes by ex vitro preparation using the generatedgametophytes.

내부공생 켈프 배우체의 숙주 선택

김광용,최태섭,이영호,Kim, Gwang-Yong,Choe, Tae-Seob,Lee, Yeong-Ho 한국조류학회(藻類) 2004 ALGAE Vol.19 No.1

Farmed kelp gametophytes were previously observed to be living endophytically in filamentous red algae. The interactions of two farmed kelp species and six red algae were examined in laboratory culture. Undaria pinnatifida (Harvey) Suringar and Laminaria religiosa Miyabe demonstrated the differing abilities of zoospores to become endophytic in four host red algae and neither kelp became endophytic in two non-filamentous red algae. There was a strong seasonal component regarding infectiousness that is associated with the changes in fron erosion in U. pinnatifida from April to June. At the same time, L. religiosa showed no significant changes in frond erosion, and there were no apparent changes in infection levels in the two species they were able infect. This study indicated clear differences between two keip species with regard to their symbiotic relationship to red algae in terms of host specificity and preference of kelp gametophytes.

Gametophyte life-history dominance of Chondrus crispus (Gigartinaceae, Rhodophyta) along the Atlantic coast of Nova Scotia, Canada

Mclachlan, Jack L.,Blanchard, Wade,Field, Christopher,Lewis, Nancy I. The Korean Society of Phycology 2011 ALGAE Vol.26 No.1

Similar to other species of Gigartinaceae Chondrus crispus has an alternation of perennial, isomorphic gametophytic and sporophytic generations. As these two generations co-exist independently within populations and obtain their resources in a similar manner, intraspecific competition is expected. In populations within the southern Gulf of St. Lawrence, fronds of both generations of C. crispus occur in similar numbers. This equivalency can be related to substratum instability, where the population is dynamic with a high turn-over rate of genets. These observations support a stochastic hypothesis to account for distribution of gametophytes and sporophytes in this area. Along the Atlantic coast of Nova Scotia, where the substratum is stable, gametophytes are overwhelmingly predominant. Gametophytic predominance is greatest in the lower littoral zone where C. crispus is abundant and space is limited. Under the fucoid canopy where "free-space" exists, the gametophyte to sporophyte ratio is lower. Gametophytic and sporophytic fronds are distributed equally among different size-classes and size-distribution is not considered a competitive factor. Previous studies have shown that sporophytic fronds of C. crispus are more susceptible to infections by endophytic algae and other pathogens, and are more heavily grazed by herbivores than are gametophytic fronds. Thus, mechanistic factors are strongly implied in the selection of gametophytes in the Atlantic population.

Gametophyte life-history dominance of Chondrus crispus (Gigartinaceae, Rhodophyta) along the Atlantic coast of Nova Scotia, Canada

Jack L. McLachlan,Wade Blanchard,Christopher Field,Nancy I. Lewis 한국조류학회I 2011 ALGAE Vol.26 No.1

Similar to other species of Gigartinaceae Chondrus crispus has an alternation of perennial, isomorphic gametophytic and sporophytic generations. As these two generations co-exist independently within populations and obtain their resources in a similar manner, intraspecific competition is expected. In populations within the southern Gulf of St. Lawrence, fronds of both generations of C. crispus occur in similar numbers. This equivalency can be related to substratum instability, where the population is dynamic with a high turn-over rate of genets. These observations support a stochastic hypothesis to account for distribution of gametophytes and sporophytes in this area. Along the Atlantic coast of Nova Scotia, where the substratum is stable, gametophytes are overwhelmingly predominant. Gametophytic predominance is greatest in the lower littoral zone where C. crispus is abundant and space is limited. Under the fucoid canopy where “free-space” exists, the gametophyte to sporophyte ratio is lower. Gametophytic and sporophytic fronds are distributed equally among different size-classes and size-distribution is not considered a competitive factor. Previous studies have shown that sporophytic fronds of C. crispus are more susceptible to infections by endophytic algae and other pathogens,and are more heavily grazed by herbivores than are gametophytic fronds. Thus, mechanistic factors are strongly implied in the selection of gametophytes in the Atlantic population.

Drynaria bonii spore culture: optimization of culture conditions and analysis of gametophyte and sporophyte development

Quyen Van Nguyen,Hang Thi Khuat,Yen‑Ngoc Thi Nguyen,Dung Thi Vu,Thu‑Ha Bui,부경환 한국식물생명공학회 2020 Plant biotechnology reports Vol.14 No.5

Drynaria bonii H. Christ is a medicinal plant that has traditionally been used for the treatment of chronic diseases and tuberculosis. However, D. bonii has not yet been cultivated or propagated. In this study, we optimized the conditions for the spore culture of D. bonii to produce sporophytes. First, we evaluated the efects of various factors, such as temperature, light intensity, potting material, and pH, on spore germination and early gametophyte development. Under optimal culture conditions, 95–99% of the spores germinated within 2 weeks of culture. Next, based on these results, we established a culture system for gametophyte development and sporophyte production. Mature gametophytes frst appeared 5 months post-germination (mpg), and at 12 mpg, the rate of production of heart-shaped gametophytes reached 56.6%. Sporophytes at the early frst-leaf stage frst appeared at 6 mpg, and at 12 mpg, the rate of formation of sporophytes reached 15.4%. Interestingly, 6.3% of all gametophytes produced multiple sporophytes. Additionally, 42–62% of the gametophytes also produced multiple secondary gametophytes, indicating a high potential of D. bonii gametophytes to generate new gametophytes and, subsequently, sporophytes. When transferred to new trays or pots, sporophytes grew well and showed 100% survival. Overall, we conclude that this spore culture system can be successfully used for the propagation of D. bonii sporophytes.

Thermal and light impacts on the early growth stages of the kelp Saccharina angustissima (Laminariales, Phaeophyceae)

Augyte, Simona,Yarish, Charles,Neefus, Christopher D. The Korean Society of Phycology 2019 ALGAE Vol.34 No.2

Anthropogenic disturbances, including coastal habitat modification and climate change are threatening the stability of kelp beds, one of the most diverse and productive marine ecosystems. To test the effect of temperature and irradiance on the microscopic gametophyte and juvenile sporophyte stages of the rare kelp, Saccharina angustissima, from Casco Bay, Maine, USA, we carried out two sets of experiments using a temperature gradient table. The first set of experiments combined temperatures between $7-18^{\circ}C$ with irradiance at 20, 40, and $80{\mu}mol\;photons\;m^{-2}\;s^{-1}$. The second set combined temperatures of $3-13^{\circ}C$ with irradiance of 10, 100, and $200{\mu}mol\;photons\;m^{-2}\;s^{-1}$. Over two separate 4-week trials, in 2014 and again in 2015, we monitored gametogenesis, the early growth stages of the gametophytes, and early sporophyte development of this kelp. Gametophytes grew best at temperatures of $8-13^{\circ}C$ at the lowest irradiance of $10-{\mu}mol\;photons\;m^{-2}\;s^{-1}$. Light had a significant effect on both male and female gametophyte growth only at the higher temperatures. Temperatures of $8-15^{\circ}C$ and irradiance levels of $10-100{\mu}mol\;photons\;m^{-2}\;s^{-1}$ were conditions for the highest sporophyte growth. Sporophyte and male gametophyte growth was reduced at both temperature extremes-the hottest and coldest temperatures tested. S. angustissima is a unique kelp species known only from a very narrow geographic region along the coast of Maine, USA. The coupling of global warming with high light intensity effects might pose stress on the early life-history stages of this kelp, although, as an intertidal species, it could also be better adapted to temperature and light extremes than its subtidal counterpart, Saccharina latissima.

Thermal and light impacts on the early growth stages of the kelp Saccharina angustissima (Laminariales, Phaeophyceae)

Simona Augyte,Charles Yarish,Christopher D. Neefus 한국조류학회I 2019 ALGAE Vol.34 No.2

Anthropogenic disturbances, including coastal habitat modification and climate change are threatening the stabilityof kelp beds, one of the most diverse and productive marine ecosystems. To test the effect of temperature and irradianceon the microscopic gametophyte and juvenile sporophyte stages of the rare kelp, Saccharina angustissima, from CascoBay, Maine, USA, we carried out two sets of experiments using a temperature gradient table. The first set of experimentscombined temperatures between 7-18°C with irradiance at 20, 40, and 80 μmol photons m-2 s-1. The second set combinedtemperatures of 3-13°C with irradiance of 10, 100, and 200 μmol photons m-2 s-1. Over two separate 4-week trials, in 2014and again in 2015, we monitored gametogenesis, the early growth stages of the gametophytes, and early sporophytedevelopment of this kelp. Gametophytes grew best at temperatures of 8-13°C at the lowest irradiance of 10-μmol photonsm-2 s-1. Light had a significant effect on both male and female gametophyte growth only at the higher temperatures. Temperatures of 8-15°C and irradiance levels of 10-100 μmol photons m-2 s-1 were conditions for the highest sporophytegrowth. Sporophyte and male gametophyte growth was reduced at both temperature extremes—the hottest and coldesttemperatures tested. S. angustissima is a unique kelp species known only from a very narrow geographic region alongthe coast of Maine, USA. The coupling of global warming with high light intensity effects might pose stress on the earlylife-history stages of this kelp, although, as an intertidal species, it could also be better adapted to temperature and lightextremes than its subtidal counterpart, Saccharina latissima.