Zhang Mei, Yuan Bing, Leng Ping
College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, PR China.
J Exp Bot. 2009;60(6):1579-88. doi: 10.1093/jxb/erp026. Epub 2009 Feb 26.
In order to understand more details about the role of abscisic acid (ABA) in fruit ripening and senescence of tomato, two cDNAs (LeNCED1 and LeNCED2) which encode 9-cis-epoxycarotenoid dioxygenase (NCED) as a key enzyme in ABA biosynthesis, two cDNAs (LeACS2 and LeACS4) which encode 1-aminocyclopropane-1-carboxylic acid (ACC) synthase, and one cDNA (LeACO1) which encodes ACC oxidase involved in ethylene biosynthesis were cloned from tomato fruit using a reverse transcription-PCR (RT-PCR) approach. The relationship between ABA and ethylene during ripening was also investigated. Among six sampling times in tomato fruits, the LeNCED1 gene was highly expressed only at the breaker stage when the ABA content becomes high. After this, the LeACS2, LeACS4, and LeACO1 genes were expressed with some delay. The change in pattern of ACO activity was in accordance with ethylene production reaching its peak at the pink stage. The maximum ABA content preceded ethylene production in both the seeds and the flesh. The peak value of ABA, ACC, and ACC oxidase activity, and ethylene production all started to increase earlier in seeds than in flesh tissues, although they occurred at different ripening stages. Exogenous ABA treatment increased the ABA content in both flesh and seed, inducing the expression of both ACS and ACO genes, and promoting ethylene synthesis and fruit ripening, while treatment with fluridone or nordihydroguaiaretic acid (NDGA) inhibited them, delaying fruit ripening and softening. Based on the results obtained in this study, it was concluded that LeNCED1 initiates ABA biosynthesis at the onset of fruit ripening, and might act as an original inducer, and ABA accumulation might play a key role in the regulation of ripeness and senescence of tomato fruit.
为了更详细地了解脱落酸(ABA)在番茄果实成熟和衰老过程中的作用,采用逆转录-聚合酶链反应(RT-PCR)方法从番茄果实中克隆了两个编码9-顺式环氧类胡萝卜素双加氧酶(NCED)(ABA生物合成中的关键酶)的cDNA(LeNCED1和LeNCED2)、两个编码1-氨基环丙烷-1-羧酸(ACC)合酶的cDNA(LeACS2和LeACS4)以及一个编码参与乙烯生物合成的ACC氧化酶的cDNA(LeACO1)。同时还研究了成熟过程中ABA与乙烯之间的关系。在番茄果实的六个采样时间点中,LeNCED1基因仅在转色期ABA含量升高时高表达。此后,LeACS2、LeACS4和LeACO1基因的表达出现一定延迟。ACO活性的变化模式与乙烯产量在粉红期达到峰值一致。种子和果肉中ABA含量的最大值均先于乙烯产量出现。ABA、ACC和ACC氧化酶活性的峰值以及乙烯产量在种子中均比果肉组织更早开始增加,尽管它们出现在不同的成熟阶段。外源ABA处理增加了果肉和种子中的ABA含量,诱导了ACS和ACO基因的表达,促进了乙烯合成和果实成熟,而氟啶酮或去甲二氢愈创木酸(NDGA)处理则抑制了这些过程,延迟了果实成熟和软化。基于本研究获得的结果,得出结论:LeNCED1在果实成熟开始时启动ABA生物合成,可能作为原始诱导物,ABA积累可能在番茄果实成熟和衰老的调控中起关键作用。