Effects of gibberellic acid on sucrose accumulation and sucrose biosynthesizing enzymes activity during banana ripening

During banana ripening there is a massive conversion into sugars, mainly sucrose, which can account for more than 10% of the fresh weight of the fruit. An ethylene burst is the trigger of the banana ripening process but there is evidence that other compounds can act as modulators of some biochemical pathways. As previously demonstrated, gibberellic acid (GA₃) can impair the onset of starch degradation and affect some degradative enzymes, but effects on the sucrose biosynthetic apparatus have not yet been elucidated. Here, the activity and amount of sucrose synthase (SuSy; E.C. 2.4.1.13) and sucrose–phosphate synthase (SPS; E.C. 2.4.1.14), respiration rates, ethylene production, and carbohydrate levels, were evaluated in GA₃-infiltrated and non-infiltrated banana slices. The exogenous supply of gibberellin did not alter the respiration or the ethylene profile but delayed sucrose accumulation by at least 2 days. While SuSy activity was similar in control and treated slices, SPS increase and sucrose accumulation was related in treated slices. Western blotting with specific antiserum showed no apparent effects of GA₃ on the amount of SuSy protein, but impaired the increase in SPS protein during ripening. The overall results indicate that although GA₃ did not block carbohydrate mobilisation in a irreversibly way, it clearly affected the triggering of starch breakdown and sucrose synthesis. Also, the delayed sucrose accumulation in GA₃-infiltrated slices could be explained by the disturbance of SPS activity. In conclusion, gibberellins can play an important role during banana ripening and our results also reinforce the idea of multiple regulatory components in the ripening pathway, as evidenced by the GA₃ effects.     

香蕉果实XET cDNA的克隆及其在成熟软化的果实果皮和果肉中的表达分析

木葡聚糖内糖基转移酶(Xyloglucan endotransglycosylase,XET)通过分解细胞壁半纤维素多糖的主要成分--木葡聚糖而参与果实软化.为了阐明香蕉(Musa acuminata.Colla cv.GrandNain)果实成熟过程中的软化与细胞壁代谢酶XET基因表达模式的关系,采用RT-PCR和RACE-PCR方法,首次从成熟香蕉果实果肉中分离了编码XT基因的全长cDNA(MA-XET1,全长1 095 bp).序列分析表明,MA-XET1的5'端和3'端的非翻译区分别为66 bp和1 89bp,该片段含有一个完整的开放读码框,编码280个氨基酸,推导的MA-XET1蛋白质中存在XET蛋白的催化活性部位DEIDFEFL.Southern杂交表明,MA-XET1在香蕉基因组中由多拷贝基因编码.Northern分析显示,跃变前期的果肉中,不能检测MA-XET1基因的表达,跃变期的果实果肉中MA-XET1表达增加,跃变后期该基因表达略有减弱;在跃变前期的果实果皮中,MA-XET1的积累较低,跃变期的果实果皮中积累大幅增加,而后迅速下降.Propylene(丙烯,乙烯的类似物)处理降低香蕉果实果皮和果肉的硬度,而且propylene促进MA-XET1在果皮和果肉中的积累.这些结果表明,MA-XET1参与香蕉果实成熟过程中的果皮和果肉软化,并且,MA-XET1的表达在转录水平上受乙烯调控.     

Expression of multiple forms of polygalacturonase gene during ripening in banana fruit.

The activity of polygalacturonase (PG, E.C 3.2.1.15) during ripening in climacteric fruits has been positively correlated with softening of the fruit tissue and differential expression of its gene is suspected to be regulated by the plant hormone ethylene. We have cloned four partial cDNAs, MAPG1 (acc. no. AF311881), MAPG2 (acc. no. AF311882), MAPG3 (acc. no. AF542382) and MAPG4 (acc. no. AY603341) for PG genes and studied their differential expression during ripening in banana. MAPG3 and MAPG4 are believed to be ripening related and regulated by ethylene whereas MAPG2 is associated more with senescence. MAPG1 shows constitutive expression and is not significantly expressed in fruit tissue. The genomic clone MAGPG (acc. No. AY603340) includes the complete MAPG3 gene, which consists of four exons and three introns. The structure of the gene has more similarity to tomato abscission PG rather than tomato fruit PG. It is concluded that softening during ripening in banana fruit results from the concerted action of at least four PG genes, which are differentially expressed during ripening.