Giovannoni J J, DellaPenna D, Bennett A B, Fischer R L
Division of Molecular Plant Biology, University of California, Berkeley 94720.
Plant Cell. 1989 Jan;1(1):53-63. doi: 10.1105/tpc.1.1.53.
Tomato fruit ripening is accompanied by extensive degradation of pectic cell wall components. This is thought to be due to the action of a single enzyme, polygalacturonase, whose activity is controlled, at least in part, at the level of gene expression. At the onset of tomato fruit ripening, polygalacturonase enzyme activity, mRNA levels, and relative rate of gene transcription all increase dramatically. To elucidate the role of polygalacturonase during tomato fruit ripening, we utilized a pleiotropic genetic mutation, rin, that blocks many aspects of ripening, including the activation of polygalacturonase gene transcription. The polygalacturonase structural gene was ligated to a promoter that is inducible in mature rin fruit and inserted into the fruit genome, and plants were regenerated. This allowed expression of the polygalacturonase gene in transgenic rin fruit at a time corresponding to ripening in wild-type fruit. Expression of this gene resulted in the accumulation of active polygalacturonase enzyme and the degradation of cell wall polyuronides in transgenic rin fruit. However, no significant effect on fruit softening, ethylene evolution, or color development was detected. These results indicate that polygalacturonase is the primary determinant of cell wall polyuronide degradation, but suggest that this degradation is not sufficient for the induction of softening, elevated rates of ethylene biosynthesis, or lycopene accumulation in rin fruit.
番茄果实成熟伴随着果胶细胞壁成分的大量降解。这被认为是由于一种单一酶——多聚半乳糖醛酸酶的作用,其活性至少部分地在基因表达水平上受到控制。在番茄果实成熟开始时,多聚半乳糖醛酸酶的酶活性、mRNA水平和基因转录相对速率都急剧增加。为了阐明多聚半乳糖醛酸酶在番茄果实成熟过程中的作用,我们利用了一种多效性基因突变体rin,它阻断了成熟的许多方面,包括多聚半乳糖醛酸酶基因转录的激活。将多聚半乳糖醛酸酶结构基因连接到一个在成熟rin果实中可诱导的启动子上,并插入果实基因组中,然后再生植株。这使得多聚半乳糖醛酸酶基因在转基因rin果实中在与野生型果实成熟相对应的时间表达。该基因的表达导致转基因rin果实中活性多聚半乳糖醛酸酶的积累和细胞壁多聚半乳糖醛酸的降解。然而,未检测到对果实软化、乙烯释放或颜色发育有显著影响。这些结果表明,多聚半乳糖醛酸酶是细胞壁多聚半乳糖醛酸降解的主要决定因素,但表明这种降解不足以诱导rin果实的软化、乙烯生物合成速率升高或番茄红素积累。