양파의 성숙배 배양을 통한 체세포배발생 캘러스 유기 및 식물체 재분화
Induction of Embryogenic Callus and Plant Regeneration by Mature Embryo Culture of Onion (Allium cepa L.)
1
조회
0
다운로드
1
인용
0
참고문헌
초록
양파의 재분화 체계 확립을 위해 성숙된 양파의 종자로부터 성숙배를 적출한 후 2,4-D와 BA의 다양한 농도를 이용하여 체세포배발생 캘러스의 형성율을 조사하였다. 체세포배발생 캘러스는 쉽게 부서지는 특성을 가지고 있었으며 비체세포배발생 캘러스는 물기를 많이 포함하고 있어 쉽게 구분이 가능하였다. 체세포배발생 캘러스는 picloram 2.5 mg/L 또는 5 mg/L가 단독으로 포함된 MS배지를 이용하여 25℃ 암배양 4주 후 가장 높은 유도율을 나타내었다. 그러나 유도된 캘러스를 shoot로 재분화 시킨 결과 2,4-D와 BA 1 mg/L가 혼합된 처리구에서 가장 높은 shoot 유기율을 나타내었다. 따라서 다른 처리에서 유도된 캘러스를 2,4-D와 BA 1 mg/L가 포함된 액체배지에 현탁배양을 시도하였다. 현탁배양은 캘러스의 증가 뿐만 아니라 캘러스의 cell cycle을 synchronized 함으로서 이후 형질전환이 용이할 것으로 판단되었다. 현탁 배양된 캘러스는 Kinetin 1 mg/L가 포함된 1/2MS 배지에서 60일간 광배양 하여 shoot 유기하였으며 유도된 shoot는 NAA 0.5 mg/L가 포함된 발근배지에서 발근 후 순화하여 완전한 식물체로 재생할 수 있었다
Abstract
To obtain regeneration system of onion, we analyzed the effects of 2,4-D and BA concentration on the embryogenic callus induction from mature embryos. The highest embryogenic callus induction ratio was shown on MS medium (Murashie and Skoog 1962) containing 2.5 mg/L or 5 mg/L picloram after mature embryos were placed on medium. When induced callus were cultured on half strength of MS medium containing 1 mg/L Kinetin, the highest shoot formation ratio was observed on MS medium containing 1 mg/L 2,4-D and 1 mg/L BA. Embryogenic callus were cultured in MS liquid medium containing 1 mg/L of 2,4-D and 1 mg/L BA. The suspension cultured cell clumps could be mass propagated. Embryogenic callus were friable, but non-embryogenic callus included a lot of moisture, hence the identification between embryogenic and non-embryogenic callus as easily achieved. When embryogenic callus as cultured on half strength of MS medium containing 1 mg/L Kinetin, shoots were induced. The whole plantlet was obtained on rooting medium containing 0.5 mg/ of NAA.
참고문헌 (21)
- Smith RH, "Regeneration in cereal tissue culture", pp.1328~1336 (1990)
- Bohanec B, "Variation in gynogenic response among long-day onion accessions", 18, pp.737~742 (1999)
- Campion B, "Advances in haploid plants induction in onion through in vitro gynogenesis", pp.97~104 (1992)
- Dommisse EM, "Onion is a monocotyledonous host for Agrobacterium", 69, pp.249~257 (1990)
- Dunstan DI, "Shoot production from the flower head of Allium cepa L", 10, pp.345~365 (1977)
- Eady CC, "Transient expression of uidA constructs in in vitro onion culture following particle bombardment and Agrobacterium- mediated DNA delivery", 15, pp.958~962 (1996)
- Hansen EE, "Regeneration of shoots from cell suspension-derived protoplasts of Allium cepa", 15, pp.8~11 (1995)
- Jeong HB, "Plant redifferentiation and in vitro multiplication of onion by shoot primodium culture", 38, pp.123~128 (1997)
- Karim MA, "isolation and culture of protoplasts of Allium cepa", 51, pp.43~47 (1997)
- Keller J, "Culture of unpollinated ovules and flower buds in some species of the genus Allium and haploid induction via gynogenesis in onion", pp.241~47 247 (1990)
- Kim MD, "Plant regeneration from protoplasts of suspension cultured cells in Arabidopsis thaliana", 27, pp.125~131 (2000)
- Kunneman BPAM, "Effects of tissue culture and acclimatization conditions on the survival and growth of rooted and unrooted Mallus and Pyrus microcutting", 314, pp.147~154 (1992)
- Kwon EJ, "Plant regeneration of major cultivars of sweetpotato in Korea via somatic embryogenesis", 29
- Ministry of Agri. and Forestry, Korea, "Crop's statistics" (2003)
- Murashige T, "A revised medium for rapid growth and bioassays with tabacco tissue cultures", 15, pp.473~479 (1962)
- van Rooyen A, "Plant regeneration via organogenesis and somatic embryogenesis in callus cultured derived from mature zygotic embryos of leek", 91, pp.261~91 270 (1996)
- Ojima M, "Factors influencing Agrobacterium-mediated transformation of Brassica rapa L", 47, pp.127~134 (1997)
- Van der Valk P, "High frequency somatic mebryogenesis and plnat regeneration frm zygotic embyo-derived callus culture of three Allium species"
- Yoon S, "Effects of excising in vitro-formed roots on acclimatization of micropropagated cassava plantlets", 28, pp.103~108 (2001)
- Zheng SJ, "propagation and regeneration of mature zygotic embryo-derived callus from Allium cepa L", 53, pp.99~105 (1998)
- Zheng SJ, "Effect of cytokinins and lines on plant regeneration from long-term callus and suspension culture of Allium cepa L", 108, pp.83~108 90 (1999)