PCR-RFLP 시스템을 이용한 배 및 사과의 속간잡종 검정
PCR-RFLP Screening of Intergeneric Hybrid between
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참고문헌
초록
속간 교배조합 배´사과로부터 137립의 후대종자를 얻었으며, 평균 착과율은 12.0% 이었다. 사과´배 조합에서는 80립의 후대종자를 얻었으며, 6.4%의 착과율을 나타내었다. 사과보다 배를 모본으로 사용하였을 때 더 높은 착과율과 과실당 종자수가 많았다. 또한, 교배조합별 종자 발아율을 조사한 결과, 배´사과 조합은 평균 56.6%의 발아율을 나타냈으며, 사과´ 배 조합은 41.7%를 나타냈다. 온실내에서 발아된 실생의 많은 개체는 발아 후 1개월 내에 고사하였으며, 발아 6개월 후에 배´사과 조합에서 67개체, 사과´배 조합에서 13개체만이 생존하였다. 사과´배 또는 배´사과 속간교배 조합으로부터 S-genotype과 속간잡종 여부를 검정할 수 있는 을 PCR-RFLP방법에 의해 배´사과 조합에서 속간잡종 3계통, 사과´배 조합에서 속간잡종 2계통을 선발하였다. 잡종으로부터 사과 S3-RNase와 S7-RNase를, 배의 S4-RNase를 증폭하였다. 증폭된 3종의 S-RNase의 염기서열을 분석하여 S-genotype을 동정하였다. 또한 사과와 배의 S-RNase 염기간의 79~86%의 높은 상동성은 자가불화합성 유전자의 분화가 속 분화 보다 선행되었음을 시사하였다.
Abstract
Through the intergeneric hybridization of pear ´apple, 137 seeds were successfully obtained. The average fruit setting showed about 12.0%. Eighty seeds were harvested from the combination of apple ´pear intergeneric crossing with the 6.4% of fruit setting. Hybridization using pear as the female parent provide higher ratio of fruit setting and number of seeds than those of apple. Germination rate of harvested seeds in pear ´apple combination also showed higher of 56.6% than that in apple ´pear combination of 41.7%. However, some seedlings died in one month after germination, and 67 seedlings from pear ´apple combination and 13 seedlings from apple ´pear combination survived after 6 months from germination. The PCR-RFLP system was developed which could determine S-genotypes and screen intergeneric hybrids among apple ´pear and pear ´apple hybridization. Based on this system, we selected 3 intergeneric hybrids from progenies among pear ´apple combination and 2 progenies among apple ´pear combination. PCR fragments of S3-RNase and S7-RNase of apple and S4-RNase of pear were amplified in 5 intergeneric hybrids. The nucleotide sequences of amplified 3 S-RNases were determined and confirmed by cloning and sequencing. Alignments of nucleotide sequences also showed high similarity of 79 - 86% between the S3-, S7-RNase of apple and S4-RNase of pear.
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