초록 ▼

1. 김의 유전적 순계 개발
엽상체 1개체를 잘게 잘라 15℃, 80 μmol·m^-2·s^-1의 단일조건(10L:14D)에서 단포자 방출을 유
도한 후, 단포자 1개체만을 분리시켜 하나의 엽상체를 얻어냈다. 이와 같은 과정을 3회 반복하여 얻어진 엽상체를 통기배양시켜 20℃, 30 μmol·m^-2·s^-1의 장일조건(14L:10D)에서 자가수정 유래의 사상체를 얻어내어 순계사상체를 확보하였다. 순계사상체에서 형성된 각포자낭의 각포자로부터 유래된 엽

1. 김의 유전적 순계 개발
엽상체 1개체를 잘게 잘라 15℃, 80 μmol·m^-2·s^-1의 단일조건(10L:14D)에서 단포자 방출을 유
도한 후, 단포자 1개체만을 분리시켜 하나의 엽상체를 얻어냈다. 이와 같은 과정을 3회 반복하여 얻어진 엽상체를 통기배양시켜 20℃, 30 μmol·m^-2·s^-1의 장일조건(14L:10D)에서 자가수정 유래의 사상체를 얻어내어 순계사상체를 확보하였다. 순계사상체에서 형성된 각포자낭의 각포자로부터 유래된 엽상체를 순계엽상체(순계주)라고 하였다.
2. 적정 DNA 마커 탐색
CAPS (Cleaved Amplified Polymorphic Sequence) 분석은 교잡실험에 이용한 한국산 참김(진도 회동 순계주), 한국산 방사무늬김(완도 청산 순계주), 일본산 방사무늬김(JPY-TKR 10-1 순계주)을 이용하였다. CAPS 분석을 위한 3 set의 프라이머는 방사무늬김 핵 유전자 부위 염기서열(EF-1α, TOP2, V-ATPase)을 바탕으로 설계하였다. 세 개의 유전자좌를 PCR한 후, 총 20 종류의 제한효소로 처리하고 전기영동을 통해 CAPS 밴드 유형을 비교․분석하였다.

Abstract ▼

In the present study, we tried to develop new variety of seaweed against the "Plant Variety Protection" which just started in 2012. The main research items of this study are 1) isolation of the pure lines in Porphyra, 2) screening of DNA markers, 3) inducement of heterozygotes using the pure lines,

In the present study, we tried to develop new variety of seaweed against the "Plant Variety Protection" which just started in 2012. The main research items of this study are 1) isolation of the pure lines in Porphyra, 2) screening of DNA markers, 3) inducement of heterozygotes using the pure lines, 4) confirmation of cross-fertilization and maternal line using CAPS markers, 5) growth of the pure lines, 6) growth of the heterozygotes, and 7) primary study for cross-breeding of Undaria pinnatifida.
A total of 14 strains of Korean and Japanese Porphyra were successfully established as pure lines. The codominant CAPS (cleaved amplified polymorphic sequence) markers in several loci were found to become a useful tool for screening of heterozygotes and their maternal line produced by inter- or intraspecific cross between different lineages of Porphyra having genetic variation. The crossing with Korean and Japanese pure lines in Porphyra strains were attempted to make up for the weak point of Korean ones. A total of 1,476 single conchocelis colonies were obtained from four crossing combinations applying three methods to rase the rate of cross-fertilization. Heterozygotic sporophytes and their maternal line from the cross-fertilization were determined and separated by the discriminate three CAPS markers (EF-1α/Mse I, TOP2/Mse I, car A/ApaL I). A total of 218 heterozygotes were obtained from 107 (25.8%) out of 414 colonies in [Korean Porphyra yezoensis Wando Cheong-san (♀·♂) × Japanese Porphyra yezoensis JPY-TKR 10-1(♀·♂)] and 111 (28.1%) out of 394 colonies in [Korean Porphyra tenera Jindo Hoe-dong (♀·♂) × Japanese Porphyra yezoensis JPY-TKR 10-1(♀·♂)] using the nuclear CAPS markers. DNA fragments of cross-fertilized heterozygotes showed both CAPS patterns of the parents. Nuclear CAPS marker would be an extremely useful method for preliminary screening for rapid identification of heterozygotes. Among 107 heterozygotes obtrained from [Korean Porphyra yezoensis Wando Cheong-san (♀·♂) × Japanese Porphyra yezoensis JPY-TKR 10-1(♀·♂)] crossing, the colonies originated from maternal line of Korean P. yezoensis and Japanes P. yezoensis were 95 (88.8%) and 12 (11.2%), respectively. Among 111 heterozygotes obtained from [Korean Porphyra tenera Jindo Hoe-dong (♀·♂) × Japanese Porphyra yezoensis JPY-TKR 10-1(♀·♂)] crossing, the colonies originated from maternal line of Korean P. tenera and Japanes P. yezoensis were 105 (94.6%) and 6 (5.4%), respectively. Organelle CAPS marker would be an useful and reliable tool for determining the origine of their maternal line. The growth characteristics of the two pure lines(Korean Porphyra yezoensis Wando Cheong-san, Japanese Porphyra yezoensis JPY-TKR 10-1) and one selected strain as superior variety(Korean Porphyra yezoensis Heanam Oe-ran) were examined by indoor culture under the different temperature (5, 10, 15, 20℃). The three pure lines were maintained in active growth under 10 and 15℃. In addition, the growth characteristics of the randomly selected 10 heterozygoes were examined to screening the candidates for developing superior varieties by indoor culture under the different temperature (5, 10, 15, 20℃). It is worth to secreening suitable conditions for growth and selecting the superior varieties before cultivation test. In the primary study in cross-breeding of Undaria pinnatifida, four local varieties were collected and investigated their morphological characteristics. Among them, seven superior strains were selected and separated into the male and female gametophytes. In this study, the procedure for cross-breeding in Porphyra was established by applying simple diagnostic markers for rapid screening of heterozygotes and their maternal lines in conchocelis phase. Therefore, the procedure for cross-breeding in Porphyra were revolutionary improved against the time consuming cross-breeding using pigmentation mutants as genetic markers for confirmation of cross-fertilization and their maternal lines. And success in obtaining inter- and intraspecific heterozygotes of Korean and Japanes Porphyra, male and female gametophytes of Undaria pinnatifida will serve to develop several superior varieties of them hereafter.