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  • Elevated aminopeptidase activities were often detected in

    2023-02-06

    Elevated aminopeptidase activities were often detected in organs of plant with accelerated protein turnover, such as leaves and petals during senescence, fruits maturation or in germinating seeds [22]. Moreover, it was proven that these enzymes take part in defense responses [[23], [24], [25]]. Part of the published research was focused on aminopeptidase activity in plants subjected to various biotic and abiotic factors, during germination and growth processes. In pigeon pea (Cajanas cajan) plants, infected with Fusarium oxysporum and Aspergillus niger var. niger, changes in aminopeptidase activity were observed during 72 h of germination. Two aminopeptidases were identified (AP1 and AP2). AP1 activity was significantly induced by infection with fungi [23]. The most explored and studied enzyme - acidic leucine aminopeptidase (LAP-A) from tomato – was described as being involved in response to different factors, such as water deficiency, salinity stress, plant hormones (abscisic CK 666 or systemin), wounding, heavy metals (e.g. cadmium) or even attack of herbivores (e.g. insects) [[25], [26], [27]]. Leucine aminopeptidase 2 was proven to be a salt-responsive biomarker in leaves of diploid wheat T. monococcum [28]. Changes (mostly increase) in aminopeptidases expression and activity in response to harmful factors were described for many plants, e.g. bean Phaseolus vulgaris [29], triticale barley [30,31], pigeon pea [23,24], black nightshade (Solanum nigrum) [32]. There are only few data about APs from Brassica napus. Recently [33], aminopeptidases from non-germinated seeds of winter oilseed rape (Brassica napus) were characterized. To the best of our knowledge, there are no reports discussing the influence of plant protection agents on the expression and activity of plant aminopeptidases. Since aminopeptidases play crucial role not only in plants but in all living organisms, this study can show the possible impact of such agents on human and other mammals health. Here we present for the first time, the influence of selected fungicides, applied at three different timings, on the expression/activity of aminopeptidases in winter oilseed rape. The activity was determined in different plant organs, namely flowers, leaves, stems and pods. One of the fungicides – Pictor 400 SC contains strobilurin (dimoxystrobin) as an active ingredient, and boscalid which is succinate dehydrogenase inhibitor (SDHI). In Symetra 325 SC, SDHI - isopyrazam serves as an active ingredient, and azoxystrobin as a respiration inhibitor (strobilurin). Propulse 250 SE is two-component fungicide containing SDHI (fluopyram) and sterol biosynthesis inhibitor (prothioconazole). The goal of the paper was to show that studied fungicides, besides their major role, could also be involved in defense and protection mechanisms in winter oilseed rape, by enhancing the expression/activity of specific aminopeptidases. These aminopeptidases catalyse the hydrolysis of storage proteins, especially cleavage of N-terminal amino acids, crucial in the defense and protection processes. The study was performed using partially purified protein extracts from different winter oilseed rape organs. The relationship between applied fungicides, aminopeptidases activity and oilseed rape crop yield as well as thousand kernels weight was also discussed.
    Experimental
    Results and discussion
    Conclusion The influence of widely used fungicides on the expression and activity of aminopeptidases in oilseed rape plants was investigated. Generally, plant proteases participate in many crucial life processes, such as proteins turnover, plant growth, development and defensive activity [46]. Their expression can be affected by many external factors (chemicals, temperature, wounding, etc.). Thus it can be expected, that application of fungicides, can influence the expression and activity of several enzymes in plants, even though they are not molecular targets for those compounds.