نوع مقاله : مقاله پژوهشی
نویسندگان
1 بخش تحقیقات علوم زراعی و باغی، مرکز تحقیقات و آموزش کشاورزی و منابع طبیعی استان مرکزی، سازمان تحقیقات، آموزش و ترویج کشاورزی، اراک، ایران
2 گروه علوم کشاورزی، دانشگاه پیام نور، تهران، ایران
چکیده
کلیدواژهها
موضوعات
عنوان مقاله [English]
نویسندگان [English]
This study aimed to investigate the changes in content of secondary metabolite and activity of phenylalanine amonialyase (PAL) enzyme under heat stress and normal conditions (probable and improbable conditions for the occurrence of aril paleness, respectively) in pomegranate. To this end, two pomegranate orchards with the highest and lowest percentages of aril paleness were selected based on our previous study. Fruits were harvested at four stages of growth and development, including stage 1 (fruit set), stage 2 (hazelnut size), stage 3 (fast growth), and stage 4 (full ripening), to measure total phenolic, total flavonoid and anthocyanin contents, and PAL enzyme activity.The results showed that under probable conditions, the most sensitive stages of fruit growth and development were from the hazelnut size toward the full ripening stage, synchronized with high changes in the content of secondary metabolites. Meanwhile, activity of PAL enzymes was observed in all stages of fruit growth and development. Therefore, during establishing a pomegranate orchard, alleviating the adverse effect of heat stress is of utmost importance. At the first three stages of fruit growth, a simultaneous change in PAL activity and anthocyanin content suggests the role of this enzyme in the biosynthesis of anthocyanin. On the other hand, decreasing and increasing trends of PAL activity and anthocyanin content, respectively, in improbable condition for aril paleness, as well as a reduction in PAL activity, under both conditions, was found to be occurred from fast growth stage to fruit ripening stage. Such issues suggest that the role of other enzymes involved in flavonoids biosynthesis pathways, besides the PAL enzyme, in the synthesis and accumulation of anthocyanin in the pomegranate fruit should be considered.
کلیدواژهها [English]
Extended Abstract
Introduction
Secondary metabolites in some plants such as pomegranate generally influence fruit quality (taste, smell, and color), marketability, and therapeutic properties. Pomegranate (Punica granatum L.) plays a remarkable role in food and pharmaceutical industries, due to its substantial antioxidant activity and being rich of secondary metabolites (especially anthocyanin). In the last decade, the emerging of aril-paleness disorder has widely been reported as a novel and prevalent disorder, leading to a reduction in the quality of pomegranate fruit. Therefore, in order to diminish the delirious effect of aril-paleness disorder on pomegranate, this study aimed to identify the most sensitive growth stage (s) of pomegranate fruit to heat stress at which the aril-paleness disorder emerge as well as to assess the substantial changes occurred simultaneously in the content of secondary metabolites and activity of phenylalanine amonialyase (PAL) enzyme during subjecting the plants to two probable and improbable conditions for the occurrence of aril paleness.
Materials and Methods
This study was carried out during two years (2022 and 2023). Based on the factors affecting the emergence of pomegranate aril paleness, such as temperature, irrigation water, and soil salinity, two orchards named 19 and 17 with high and low magnitude of aril paleness, respectively, were selected in the first year. Regarding the value of average temperature during fruit maturation (25-27 oC for the orchard 19 and > 38 oC for the orchard 17), 30 trees were selected and labeled carefully. At the end of September (2022), the percentage of fruit paleness in each tree was calculated. In the second year (i.e., 2023), ten trees were chosen out of 30 trees selected in the first year, and their fruit (three fruits per tree) was harvested at four stages of growth as follows: stage 1 (fruit set), stage 2 (Hazelnut size), stage 3 (fast fruit growth), and stage 4 (full ripening), and then some phytochemical characteristics were measured, including the content of total phenolic compounds (TP), total flavonoids (TF), and anthocyanins (Cyd) , as well as PAL activity.
Results and Discussion
The results showed a regularly downward status in the changes of TP content after the hazelnut size until the end of the fruit growth under both conditions, although this trend was remarkably severe in probable conditions rather than improbable conditions for the aril paleness occurrence. The change in the content of total flavonoid (TF) in improbable conditions was different, in a way that it initially decreased (from stage 1 to 3) followed by an increasing in TF (from stage 3 to 4), while in probable conditions its change was regularly downward from fruit set to the end of fruit growth and development. Overall, the lowest TF was recorded at stage 4. The content of cyanidin (Cyd) increased from stage 1 to stage 3 in both conditions. However, after the fast fruit growth stage (stage 3), a significant change in Cyd content was observed in both improbable (increasing trend) and probable (decreasing trend) conditions for the occurrence of paleness disorder. Under both condition and up to stage 3, the PAL activity and Cyd content were mounted simultaneously, although the activity of PAL decreased after the fast fruit growth stage.
Conclusion
Our findings showed that under improbable conditions for the occurrence of aril paleness, the highest changes in secondary metabolites and most sensitive stage of fruit growth to aril-paleness disorder occurred at the stage of fruit hazelnut size to full ripening. In order to mitigate the harmful effect of heat stress on pomegranate, some issues should be taken into account during establishing its orchard: choosing land with an appropriate height (> 1000 m above sea), possessing a suitable slope for obtaining maximum shade (Northern and Eastern slopes), choosing a suitable tree-planting pattern, especially in tropical and subtropical areas (square planting pattern), using light mulches, maintaining annual weeds, and using light-colored shades. Additionally, the simultaneous changes in PAL activity and anthocyanin content at the first three stages of fruit growth suggest the role of PAL in the synthesis of anthocyanin. However, regarding separately changes in PAL activity and anthocyanin content (under improbable conditions) as well as a reduction in PAL activity from stage 3 to the end of fruit growth under both probable and improbable conditions substantiate the role of other flavonoid-synthesizing enzymes, beside PAL enzyme, in the synthesis of anthocyanin under heat stress.