Itai A, Kawata T, Tanabe K, Tamura F, Uchiyama M, Tomomitsu M, Shiraiwa N
Laboratory of Horticultural Science, Faculty of Agriculture, Tottori University, Japan.
Mol Gen Genet. 1999 Feb;261(1):42-9. doi: 10.1007/s004380050939.
The shelf life of Japanese pear fruit is determined by its level of ethylene production. Relatively high levels of ethylene reduce storage potential and fruit quality. We have identified RFLP markers tightly linked to the locus that determines the rate of ethylene evolution in ripening fruit of the Japanese pear. The study was carried out using sequences of two types of 1-aminocyclopropane-1-carboxylic acid (ACC) synthase genes (PPACS1 and pPPACS2) and a ACC oxidase gene (PPAOX1) as probes on 35 Japanese pear cultivars expressing different levels of ethylene (0.0 to approximately 300 microl/kg fresh weight/h) in ripening fruit. When total DNA was digested with HindIII and probed with pPPACS1, we identified a band of 2.8 kb which was specific to cultivars having very high ethylene levels (> or = 10 microl/kg f.w./h) during fruit ripening. The probe pPPACS2 identified a band of 0.8 kb specific to cultivars with moderate ethylene levels (0.5 microl/kg f.w./h-10 microl/kg f.w./h) during fruit ripening. The cultivars that produce high levels of ethylene possess at least one additional copy of pPPACS1 and those producing moderate levels of ethylene have at least one additional copy of pPPACS2. These results suggest that RFLP analysis with different ACC synthase genes could be useful for predicting the maximum ethylene level during fruit ripening in Japanese pear.
日本梨果实的货架期由其乙烯生成水平决定。相对较高的乙烯水平会降低贮藏潜力和果实品质。我们已经鉴定出与决定日本梨成熟果实中乙烯释放速率的基因座紧密连锁的RFLP标记。该研究使用两种1-氨基环丙烷-1-羧酸(ACC)合成酶基因(PPACS1和pPPACS2)以及一个ACC氧化酶基因(PPAOX1)的序列作为探针,对35个在成熟果实中表现出不同乙烯水平(0.0至约300微升/千克鲜重/小时)的日本梨品种进行检测。当用HindIII消化总DNA并用pPPACS1进行探针杂交时,我们鉴定出一条2.8 kb的条带,该条带是果实成熟期间乙烯水平非常高(≥10微升/千克鲜重/小时)的品种所特有的。探针pPPACS2鉴定出一条0.8 kb的条带,该条带是果实成熟期间乙烯水平中等(0.5微升/千克鲜重/小时 - 10微升/千克鲜重/小时)的品种所特有的。产生高水平乙烯的品种至少有一个额外的pPPACS1拷贝,而产生中等水平乙烯的品种至少有一个额外的pPPACS2拷贝。这些结果表明,用不同的ACC合成酶基因进行RFLP分析可用于预测日本梨果实成熟期间的最大乙烯水平。