국내 딸기 재배품종의 잎절편 배양으로부터 고빈도 식물체 재생
High Frequency Plant Regeneration from Leaf Explant Cultures of Domestic Cultivated Strawberry (Fragaria x ananassa Duch)
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딸기 기내배양으로부터 고빈도 식물체재생 방법을 개발하기 위하여 여봉딸기의 잎과 엽병절편을 0.5 mg/L IBA와 3.2 mg/L kinetin, BAP, zeatin을 각각 조합 첨가한 MS배지에서, 그리고 1주, 2주, 4주동안 암 전처리한 잎 절편을 0.5 mg/L IBA와 3.2 mg/L zeatin이 첨가된 배지에서 6주동안 16시간 광주기 조건으로 배양하였다. 신초 발생은 잎을 adaxial상태로 치상하였을때 절단부위로부터 안토시아닌 색소를 함유한 녹색 캘러스로부터 유도되었고 (57.0%), 엽 병의 경우 절단부위로부터 직접 발생되었다 (6.3%). 캘러스와 신초발생율은 잎 절편 크기가 클수록 (1.0～1.5 cm2) 높았으며, 암 전처리는 그 발생율을 점차 증가시켜 4주동안 암 전처리한 후 16시간 광조건으로 옮겼을 때 가장 높은 발생율 (87.0%)을 얻을 수 있었다. 이러한 shoot발생율 (87%)은 암 전처리를 하지 않은 대조군 (27%) 에 비하여 현저하게 증가된 것이다. 신초를 분리하여 ½ MS기본배지에 옮겨 20동안 배양하였을 때 뿌리가 발생되었으며, 이때 소식물체를 토양에 옮겨 순화시켰다.
Abstract
To develop a high efficiency plant regeneration system from in vitro cultures of strawberry, cv.Yeobong, petiole and leaf explants were cultured on MS basal medium containing a combination of 0.5 mg/L IBA and 3.2 mg/L kinetin or zeatin or benzyl amino purine (BAP) for 6 weeks, and leaf explants with dark pretreatment for a week (T1), 2 weeks (T2), and 4 weeks (T3) were cultured on medium suplemented with 0.5 mg/L IBA and 3.2 mg/L zeatin under 16 hr photoperiod for 6 weeks. Shoot organogenesis was observed from the greenish cali containing minimal anthocyanin formed at proximal cuting edges of the leaf explant (57%) when cultured adaxial side on the medium, whereas was directly formed from a cuting edges of petiole explant (6.3%). Frequency of calus formation and shoot organogenesis at large size of leaf explant (1.0～1.5 cm2) was higher than small size (0.5~1.0 cm2that produced calli and shoots. The maximum frequency (87%) for shoot organogenesis was obtained from the leaf explants that transferred to a 16 hr photoperiod condition after the initial 4 weeks dark period. The improved frequency (87%) in comparision with control without dark pretreatment (27%). When the shoots were transferred to MS basal medium, formed roots with 20 d of culture. The rooted plants were subsequently transferred to the pots and to the field.
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