콜히친 침지처리에 의한 ‘망종화’의 4배체 식물유도
A Tetraploid Induction in Hypericum patulum Thunberg by Colchicine Soaking Treatment
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초록
‘망종화’에서 콜히친 처리에 의한 효율적인 4배체 식물을 유도하고자 적정 식물체 부위, 적정농도 및 침지시간을검토하였다. 종자의 발아율은 콜히친의 농도가 높을수록또한 침지시간이 길수록 저하되었다. 대조구를 제외한 16개의 처리구에서 총 453개체가 발아된 반면 4배체는 유도되지 않았다. 기내 배양 중인 줄기 절편체의 식물체 재생율은 콜히친 처리농도 0.01%에서 최고를 나타내다가 0.1%농도 이상으로 높아질수록 낮았다. 적정식물체 부위는 줄기 절편체로 나타났다. 4배체 식물은 콜히친을 0.05% 이상으로 6시간 침지처리 하였을 때 얻을 수 있었고, 특히0.05%, 12시간 침지처리에서 식물체의 재생수 대비 약42%의 높은 획득율을 보였다. Flowcytometry에 의해DNA함량의 배가여부를 확인한 결과, G1 phase의 DNA 함량 peak가 2배체에서 94.5, 4배체는 192.5로, DNA가 배가됨을 확인할 수 있었다. 또한 공변세포 당 엽록체 수는2배체가 약 10개인 것에 비해 4배체는 17∼19개로 2배체보다 약 1.7∼1.9배 정도 많았다.
Abstract
This study aimed to get the basic data on the breeding of good varieties in Hypericum patulum Thunberg. Theoptimum materials, concentration and soaking time were examined to identify the effective approach to induce tetraploidplant by colchicine treatment to cultivate the varieties. For the seed germination rate of seed by colchicine treatment, thehigher colchicine concentration was and the longer soaking time was, the more the germination rate decreased. Whileindividuals were germinated in 16 test groups except control group (no treatment group), all the plants were diploid and notetraploid was induced. For the plant regeneration rate by colchicine treatment on the explant of Hypericum patulumThunberg that was under in vitro culture, the higher the colchicine concentration increased, the ress the regeneration rate. While total 147 individuals were regenerated in all treatment, when the explant was soaking treatment in more than 0.05%for over 6 hours, tetraploid could be obtained. In the soaking treatment of 0.05% for over 6 hours, tetraploid could beobtained. In particular, for the soaking treatment in 0.05% for 12 hours, 8 tetraploids were induced, which was about 47.1%of the number of plant regenerated. In accordance with the observation on doubling of DNA contents in leaf in order toidentify polyploidy, the peak DNA content of G1 phase was 94.5 for diploid and 192.5 for tetraploid. It confirmed doublingof DNA content. Furthermore, the number of chloroplasts per guard cell depending on polyploid was around 10 in diploidand 17 to 19 in tetraploid, which were around 1.7 to 1.9 times as much as diploid.
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