Differential Display PCR을 이용한 사과 자가적과성 연관 유전자 탐색
Identifying Genes Related with Self-thinning Characteristics in Apple by Differential Display PCR
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사과의 경우 한 과총에 5개의 꽃이 피는데 그 중 한 가운데의 중심화가 먼저 개화하여 과일로 발달하고 주위의 측과 4개는 스스로 낙과되는 현상을 자가적과성이라고 한다. 적과는인위적으로 과실의 숫자를 줄여 잎 수와 과실 수의 균형을 맞추는 작업으로 과실의 크기를 증가시키고, 수세, 수형을 유지시켜 안정적인 생산에 도움을 준다. 노동력 절감을 위해 인간에게 유용하게 사용될 수 있는 특성이 자가적과성인데, 자가적과성 품종의 사과에서 측과는 만개 후 30일 이내에 떨어지고 중심과만 남아서 성숙하게 된다. 51개의 사과 품종으로부터 비자가적과성 그룹 20종, 6월 생리적 낙과 그룹 16종, 자가적과성 그룹 15종을 분류하였다. 대표적인 자가적과성 품종인 Aori #9 로부터 중심과와 측과에서 다르게 발현이 되는유전자들을 DD-PCR 방법으로 확인하였다. 중심과에서 30개의 clones 과 측과에서 24개의 clones을 선발하여 염기서열을분석하였다. 주로 측과에서 발현되는 유전자들은 pathogenesis,senescence, temperature stress, protein degradation, fruit browning, sorbitol metabolism에 관여하는 유전자들과 높은상동성을 나타내고, 중심과에서 발현되는 유전자들의 염기서열을 분석해 보면 anthocyanin의 up-regulation이나 flavonol생합성, ethylene 생합성에 관여하는 유전자들이 분포한다. Cytochrome P450 유전자의 발현양상을 보기 위해 Real time PCR 분석을 한 결과 중심과보다 측과에서 발현량이 높게 나타났다. 중심과와 측과에서 다르게 발현되는 유전자들의 실제발현양상을 분석하기 위해 Real time PCR을 이용해서 상대정량을 분석할 계획이며 분자수준에서의 자가적과를 조절하는 기작에 대한 앞으로의 연구는 생력 재배가 가능한 품종 육성의 육종 소재 개발에 활용될 수 있을 것이다.
Abstract
Thinning of apple fruitlets is one of the most laborious and important works for the improvement of fruit quality and for the promotion of sufficient flower bud formation to prevent alternate bearing in commercial cultivars. Lateral fruits of self-thinning apple cultivars fall naturally within 30 days after full bloom and only central fruit remains to mature. Differences of gene expression between central fruit and lateral fruit were investigated by differential display (DD) PCR. Partial cDNAs of 30 clones from the central fruit and 24 clones from the lateral fruit were selected for nucleotide sequence determination and homology searches. The levels of transcripts coding for proteins involved in pathogenesis related proteins, senescence,temperature stress, protein degradation, fruit browning, sorbitol metabolism were significantly higher in pedicels of lateral fruit than in pedicels of central fruit. On the other hand, the up-regulation of proteins involved in anthocyanin and flavanol biosynthesis and ethylene synthesis were observed in pedicels of central fruit. In Real time PCR analysis, cytochrome P450 gene was confirmed as showing a higher expression level in lateral fruit than in central fruit. The results of this study indicate that differentially expressed genes are related to self-thinning characteristics in apple tree.
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