Induction of Ageing and Apoptosis by Sucrose in Penicillium aurantiogriseum

Induction of Ageing and Apoptosis by Sucrose in Penicillium aurantiogriseum | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Induction of Ageing and Apoptosis by Sucrose in Penicillium aurantiogriseum Assia Bouhoudan, Fatima Chidi, Mustapha Khaddor This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-3876169/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract This work aims to study the physiological and metabolic responses of P. aurantiogriseum to sucrose-induced stress. Comparative analyses on intra-species (within strains of P. aurantiogriseum), and interspecies (between P. aurantiogriseum and P. camembertii ) were conducted to assess the effect of carbon stress on aging phenomena and toxigenesis. Our results revealed a correlation between sucrose concentration and ageing signs. At a sucrose concentration of 500 g/L, the ageing signs of P. aurantiogriseum began to fade, allowing its normal characteristics to resurface. This transformation is believed to be a response to the sucrose and the cells that cannot adapt undergo apoptosis, leaving only the normal cells to thrive. Terrestric acid production was observed during the ageing process and continued even after returning to a normal physiological state, albeit at a reduced level. P. aurantiogriseum Terrestric acid Aging Apoptosis Carbon stress. Toxigenesis Figures Figure 1 Figure 2 1. Introduction Ageing and apoptosis are two highly debated phenomena in moulds (Osiewacz 2002 ; Hamann 2008; Sharon 2009; Scheckhuber 2011; Kamogashira 2017). These natural mechanisms play a critical role in cell development. Apoptosis, a vital process, occurs throughout the development of an organism and is associated with the maintenance of cellular homeostasis, the elimination of damaged cells, the response to infectious agents, as well as the cell's adaptive response to biotic and abiotic stress (Danial and Korsmeyer 2004 ; Green 2005 ). On the other hand, ageing corresponds to a gradual decline in the body's ability to cope with stress and damage (Jazwinski 2002 ). Both of these mechanisms involve the accumulation of reactive oxygen species (ROS) (Elmore 2007 ). In 1956, Harman initially proposed the theory of radical ageing, based on the temporal imbalance between the production and elimination of free radicals (ROS) (Harman 1956 ). These ROS, produced naturally throughout the cell cycle, exert a crucial physiological role, notably in signalling cascades (Ray 2012), and the activated compounds play a pivotal role in inflammation and the balance between growth, apoptosis, and cellular senescence (Wang 2013). Antioxidant systems help maintain redox balance in the cell and significantly mitigate the risk of oxidative damage caused by ROS. Nevertheless, ROS possess varied and significant toxic properties, affecting various cellular components (macromolecules such as DNA, proteins, and lipids) when they evade antioxidant systems (Valko 2007). This process of oxidative alteration leads to mutations and denaturation, causing structural dysfunctions in biological systems. This ultimately leads to the ageing process, according to Harman's theory. An evolved version of the free radical theory of ageing suggests that mitochondria are the primary source of cellular ROS. When these mitochondria, particularly their respiratory chain components, are damaged, an accumulated production of ROS is generated (Harman 1956 , 1981 , 1998 ). Any change in the usual conditions of cell life leads to a reaction called "stress" insofar as the cell has adapted its biological functions to environmental changes. Thus, any environmental or genetic condition, that increases exposure to aggression or decreases the capacity of defence or repair, results in accelerated ageing and, subsequently, apoptosis (Wang 2013). Oxidative stress, caused by an increase in cell ROS, is the most commonly studied. Mycotoxins, as secondary metabolites of moulds, have been detected in several human or livestock foods (Khaddor 2006). The ingestion of mycotoxins represents a real menace to human and animal health (Faid and Tantaoui-Elaraki 1989 ). Penicillium aurantiogriseum is a particular species of the Penicillium genus. Its mycotoxins are of great importance given their largely variable effects between harmful and beneficial to human and animal health (Khaddor 2007). To date, more than 37 mycotoxins have been identified in P. aurantiogriseum . Previous studies had identified some of the mycotoxins such as penicillic acid, aurantiamine, and terrestric acid in P. aurantiogriseum (Khaddor 2007). There are few studies of natural contamination with terrestric acid and its toxicity, and little is known about the factors influencing its production. The offending substance was found to be phytotoxic (Gausman 1991 ) and cardiotoxic (Frisvad 2004). The present work is devoted to studying P. aurantiogriseum physiological response to the stress induced by sucrose and is focused on terrestric acid, a mycotoxin produced by P. aurantiogriseum , exploring its production and toxicity under stress conditions induced by sucrose. Thus, it may help to control the growth conditions of this species and improve mycotoxin production. This study will be a pilot study to control the production of P. aurantiogriseum mycotoxins that have great pharmaceutical interest. 2. Materials and Methods 2.1. Fungal Strains The study was conducted on the P. aurantiogriseum strain that belongs to the collection of the Environmental and Food Biotechnology Research Team (EFBRT), which had been utilized in prior studies (Khaddor 2007; Bouhoudan 2018, 2020). This strain was originally isolated from Pastilla leaves at the Department of Food Microbiology, Agronomic and Veterinary at the Institute Hassan II (IAV) in Rabat and its identification was confirmed by The Scientific Institute of Public Health (ISSP) of Louis Pasteur/Brussels. To initiate the study, the stored strain was cultivated on malt extract agar (MEA) and incubated at 25°C for 7 days. Following incubation, spores were suspended in 0.1% of tween 80 solution, and the suspension’s density was standardized to 10 7 spores/ml. Additionally, two other Penicillium strains, namely P. aurantiogriseum and P. camembertii , were included in our experiments to facilitate conclusive comparisons. These strains, isolated within the laboratory (EFBRT), were authenticated using the Pitt method (Pitt and Hocking 2009 ). 2.2. Growth medium The G25N medium serves as a secondary purification medium conventionally employed for validating strain identification (Zhao 2014; Park 2014). This medium, known for its high glycerol content of 25%, reduces water activity, thus impeding the growth diffusion of P. aurantiogriseum (max 24 mm). conversely, Glycerol facilitates consistent Penicillium colony development (Pitt 1973 ) and stands out as an excellent carbon source conducive to mycotoxin production (Mulè 2004). Hence, we retained this medium’s composition while introducing varied sucrose concentrations, ranging from 0 g/L (G25N) to 700 g/L, to study the impact of stress on the physiology of strains. The strains were inoculated on a G25N medium supplemented with different sucrose concentrations from 0 g/L to 700 g/L, herein referred to as G25N*. the inoculated cultures were then incubated for 10 days at 25°C. Physiological studies included observations on morphology, texture, colour, growth rings, growth status, mycelial weight, and colony diameter of P. aurantiogriseum colonies, as well as the appearance of the hyphae. Radial growth was quantified by measuring the diameter of each colony using a ruler (Zain 2009 ). All the experiments were meticulously conducted in triplicate to ensure accuracy and consistency of results. 2.3. Mycotoxins extraction The mycotoxins extraction process followed the methodology outlined in the work of Bouhoudan et al, (Bouhoudan 2018, 2020). Fluorescence intensity, an indicator of toxigenesis, was represented by a varying number of "+" signs as described by Zain et al, (2009). 2.4. Statistical study Statistical analysis of the results obtained was conducted using the "Duncan's multiple ranges" test at a significance threshold of 5% [Stat Soft]. Nine tests were performed for each medium, and the averages derived from these nine trials (n = 9) were subjected to analysis of variance (ANOVA) with Ducan's Multiple Range Test, also set at a 5% threshold. This statistical approach aimed to precisely define whether the factor (carbon source) had a significant effect on the response, specifically concerning mycotoxin production and fungal growth). 3. Results 3.1. Effect of stress on the physiology of Penicillium strains Under the influence of varying sucrose concentrations, the colony diameter of both P. aurantiogriseum and P. camembertii strains exhibited an increase, as detailed in Table 1 . Table 1 Average colony diameters of Penicillium species as a function of sucrose concentration of the G25N medium. Sucrose concentration (g/L) Colony diameter (mm) (95% IC) PA P7 PC 0 1.98 d 1.70 d 2.28 d 100 45.00 b 38.00 c 43.00 c 400 36.37 a 30.97 a 44 a 500 40.70 c 33.70 c No growth 600 50.36 c 60.41 c 20.90 c 700 62.77 f 57 f 22.15 f In the same column, two results followed by the same letter do not differ significantly at the 5% threshold. For each concentration of YES, nine tests were performed. The averages obtained in the nine trials (n = 9) were compared by analysis of variance (ANOVA) with Ducan's Multiple Range Test at the 5% threshold. PA: Penicillium aurantiogriseum (collection of "EFBRT") at different concentrations of sucrose. P7: Penicillium aurantiogriseum (isolated in the "EFBRT" laboratory) at different concentrations of sucrose. PC: Penicillium camembertii (isolated in the laboratory) at different concentrations of sucrose. The results revealed a consistent response among the three Penicillium strains under sucrose-induced stress, revealing a proportional increase in colony diameter with rising sucrose concentrations. As reported in our previous work by Bouhoudan et al, (Bouhoudan 2020), all strains exhibited a shared critical concentration point at 400 g/L, showcasing an exceptionally aggressive morphological response. At this threshold, the colonies displayed a whitish and notably severe appearance, as depicted in Fig. 1 . Subsequently, signs of ageing began to disappear at a sucrose concentration of 500 g/L, revealing the resurgence of normal Penicillium strain characteristics. However, the disappearance of these signs varied in terms of speed and appearance among the strains. The P. aurantiogriseum strain, from the team's collection (Fig. 1 A), revealed a slower return to normalcy compared to the same strain isolated in the laboratory (Fig. 1 B). additionally, P. camembertii (Fig. 1 C) displayed more significant results compared to the strains of P. aurantiogriseum . At the microscopic level, all three Penicillium strains showed an identical appearance across the various sucrose concentrations added to the G25N* medium. Microscopic observation revealed the presence of two distinguished zones within the colony: the central zone, which contains older cells, and the peripheral zone, which represents younger cells, as illustrated in Fig. 2 . 3.2. Effect of sucrose on the toxigenesis of Penicillium strains The results revealed a significant impact of sucrose concentrations within the G25N* growth medium on the metabolic profile of P. aurantiogriseum strains (Table 2 ). Table 2 Average of Terrestric acid (TA) intensity in Penicillium strains depending on sucrose concentration of G25N* medium. Sucrose concentration (g/L) Intensity of Terrestric Acid PA P7 PC 0 – – No production of TA 100 + + 400 ++++ ++++ 500 ++ ++ 600 ++ ++ 700 ++ ++ (-) No (+) low, (++) medium, (++) strong (+++) very strong PA: Penicillium aurantiogriseum (collection of "EFBRT") at different concentrations of sucrose. P7: Penicillium aurantiogriseum (isolated in the "EFBRT" laboratory) at different concentrations of sucrose. PC: Penicillium camembertii (isolated in the laboratory) at different concentrations of sucrose. The study of toxigenesis revealed a substantial production of terrestric acid coinciding with the appearance of ageing signs. Notably, as the ageing signs became more important, the concentration of terrestric acid produced increased significantly. However, the P. camembertii strain does not exhibit any production of terrestric acid. 4. Discussion The present work serves as a follow-up to prior studies conducted by Bouhoudan et al, (Bouhoudan 2018, 2020) focusing on assessing the effect of carbon sources on both the growth and the toxigenesis of P. aurantiogriseum . Specifically, our emphasis was placed on discussing terrestric acid production given the insufficiency of studies that addressed it. This study acts as a pilot initiative laying the groundwork for future research endeavours that will include a broader spectrum of Penicillium species and their associated mycotoxins. G25N medium is typically a secondary purification medium used to confirm the identification of strain (Zhao 2014; Park 2014). This medium is known for its high content of glycerol (25%) serving to reduce the water activity in the medium which does not allow the growth diffusion for P. aurantiogriseum (max 24 mm). On the other hand, Glycerol facilitates the consistent development of Penicillium colonies (Pitt 1973 ) and it is an excellent carbon source for mycotoxin production (Pitt 1973 ). For this reason, we used this medium by keeping the same composition and added different sucrose concentrations, ranging from 0 g/L (G25N control) to 700 g/L, to study the stress effect related to sucrose concentration on the physiology of P. aurantiogriseum strains. The modified media is named G25N*. In this regard, the phenotypic, morphologic, and metabolic responses identified in this study were caused obviously by the addition of sucrose at different concentrations in the culture medium. Stress caused by the high sucrose concentration on G25N*medium allowed us to observe different phenomena related to the suffering state of P. aurantiogriseum : Although ageing is a normal and essentially biological process in cells and generally leads to cell damage induced by ROS, induction of ageing by external factors leads to a kind of trauma and cellular stress. Cell reactions vary according to stress intensity and timing. In this study, we subjected our Penicillium aurantiogriseum strain to carbon stress (due to the high sucrose concentration). The high sucrose concentration caused intense growth of the strain translated by an increase in colony diameter. The hyphae spread rapidly (seven d of growth gave a very important colony diameter compared to the control). However, as the hypha spreads, the nutrients become progressively restricted for peripheral cells. Nutrient deprivation had induced elevation of the colonies in the central zone showing cells suffering due to oxygen lack. The addition of sucrose induced the acceleration of the growth process and consequently a rapid achievement of the latency phase. The central zone containing the aged cells had a white colour (no sporulation) and the diameter of this zone was larger compared to the peripheral zone which contains the young cells. In the centre of the colony, mitotic cells lose their proliferative capacity and then disappear by apoptosis. The facts discussed so far are the consequences of programmed cell ageing. Various factors can contribute to the phenomenon of radical ageing mentioned in the introduction by Hamann et al, (Osiewacz 2002 ; Hamann 2008; Hameed 2012). We propose that in this study until the concentration of 400 g/L, the cells lost their ability to resist sucrose stress. We have suggested that sucrose indirectly induced several reactions initiated in the mitochondria. Since mitochondria are the main generators of reactive oxygen species (ROS), they were subjected to molecular damage. The time and intensity of exposure of the cells to the stress allowed a significant production of ROS. ROS, therefore, prevented the mitochondria from using scanning systems to repair damaged molecules that lead to early ageing (Osiewacz 2002 ). Besides, as already mentioned in the introduction section, at higher concentrations, ROSs damage cellular compounds and cause cellular dysfunctions (Osiewacz 2002 ). Therefore, oxygen lack in the central zone reflected a disorder in the respiratory chain, which increased ROS levels. Studies on the development of senescent cultures have revealed that the presence of a determinant of senescence seems to accumulate during ageing (Maas 2007). To confirm this fact, we performed a small demonstration in which we took a tip of centre cells and another of peripheral cells and cultivated them in fresh growth media. After growing, we obtained colonies that grow back from the cells of the periphery, whereas cells of the centre could not grow back. This supports Marcou's conclusions that the hyphae were dead in the centre and that the senescence determinant seems to be absent from young cultures and accumulates after the culture passes through a critical point. Marcou demonstrated the hyphae fusion of a senescent culture leading to the transfer of the accumulated senescence determinant to a young culture. This one immediately becomes senescent and dies on the growth front. The determinant of senescence thus appears infectious and cytoplasmic (Maas 2007). This explains the increase in the central zone diameter of stressed colonies. At the metabolic level, the stress induced by the high sucrose concentration in the culture medium affected the toxigenic profile of P. aurantiogriseum . Production of terrestric acid at the time of ageing signs appearance became very important. More aging signs were important more the concentration of terrestric acid produced was high. Sucrose assimilation became more difficult for the centre cells (aged). Also, the oxygen lack caused by cellular outgrowth leads to a decrease in centre cells' antioxidant capacity to maintain the environment in a reduced state due to the increase of (ROS) in the mitochondria. Therefore, the cells respond to stress by dynamic redirection of the glycolysis’s metabolic flux to the pentose phosphate pathway, which is the biosynthetic pathway of terrestric acid. It should be noted that mycotoxins are derived from acetate and shikimate via polyketoacids and the amino acid pathways in the Krebs cycle (which takes place in the mitochondria) (Marcou 1961 ; Pieter S. Steyn 1980 ). This explains the proportional relationship between stress and terrestric acid production. Although the increase in sucrose concentration has shown intense cellular ageing in all colonies, the concentration of 400 g/L, however, presented a special case in which the colony was all white with a very aged look, which supports the radical ageing theory of Harman (Harman 1956 ). Penicillium aurantiogriseum cells have undergone accelerated chronological ageing linked to the long exposure to the high sucrose concentration in which cells still young undergo an early senescence process. Cellular outgrowth has led to nutrient deprivation and oxygen lack. Radical ageing signs, appearing in this study, were accompanied by an overall alteration of a set of physiological functions as well as a higher susceptibility to different alterations. The free radical theory explains these changes by the accumulation of oxidized molecules and the consequences of oxidation as the appearance of mutations. Several arguments are in favour of free radicals’ involvement in ageing mechanisms (Turner and Aldridge 1971 ; Wang 2013; Kamogashira 2017). The situation is more complex when one looks for chronic oxidative stress during which, on the one hand, the ROS increases elevated and, on the other hand, the inductions of the antioxidant and repairing systems are more modest, those are sometimes themselves impaired by oxidation. Wang et al, (Wang 2013) supported this observation by explaining when the repair systems themselves were affected, it is likely to move towards cellular function deterioration that was accompanied by accelerated ageing. If the ROS continues to accumulate, a more consistent adaptation of the cell is necessary with the induction of its antioxidant system. Suppression of systems capable of releasing ROS, particularly the respiratory chain (Osiewacz 2002 ), is observed by reorientation towards the glycolysis pathway mentioned above. This leads to an overproduction of terrestric acid, which reached its maximum level at the concentration of 400 g/L. Several studies have suggested the existence of an "ageing program" that blocks cell protection and accelerates cell death (Madeo 1999; Herker 2004; Hekimi 2011). We took a crop point and re-cultivated it in a fresh growing medium. After growth, we obtained colonies that normally pushed what allowed us to suggest that culture to 400 g/L did not undergo apoptosis. Studies have shown a correlation between oxidative stress and mycotoxin production (Mortimer and Johnston 1959 ; Schmidt-Heydt 2011). Mycotoxin biosynthesis contributed to the adaptation to stressful environments. Interesting in this respect is the fact that these mycotoxins have antioxidant properties that can protect against oxidative stress (Stoll 2014). For this, we suggested the antioxidant effect of terrestric acid vis-à-vis oxidative stress, and, through this; the cells do not have undergone apoptosis. It is now obvious that high sucrose concentration provoked ROS overproduction and therefore mitochondrial dysfunction that causes early ageing. However, in our case, the increase in sucrose concentration above 500 g/L did not result in radical ageing and the cells resisted the stress. Our strain was normal in appearance with good biomass and mycelium production and moderate terrestric acid production. We consider this finding as an escape phenomenon at the high sucrose concentration. Our analysis suggests that between 10g/l and 400g/l of sucrose, P. aurantiogriseum was forced to adapt to the sucrose stress. It used its panoply of intracellular and genetic to maintain cellular growth and survival. This response is similar to the "retrograde response" described for the first time in S. cerevisiae (Sekito 2000; Schmidt-Heydt 2015) Thus, retrograde response occurs during normal aging to offset mitochondrial dysfunction accumulation allowing cells to live as long as they do (Jazwinski 2002 ). The induction of retrograde response increases the replicative lifespan and increases stress resistance (Liao and Butow 1993 ; Kirchman 1999). The retrograde response indicates changes in gene expression (Traven 2001). Studies have clearly shown that the retrograde response results in a wide range of changes in gene expression (Kirchman 1999) that suggest extensive metabolic remodelling of the cell in response to the metabolic constraint associated with mitochondrial dysfunction that ultimately allows for longer cell life (Jazwinski 2002 ). However, below the concentration of 500 g/L, the cell reached a very intense level of stress, which resulted in the accumulation of a very large amount of ROS, leading therefore to dysfunction not only of the mitochondria but also of all cell components and mutations in genes. This necessarily leads to the non-functioning of the retrograde response and the cells undergo apoptosis leaving the other normal cells to live. This explains the existence of a peripheral zone containing young cells. Studies show that the involvement of apoptosis occurs when the molecular and organelle scanning systems can no longer cope with the damage accumulated by oxidative stress (Kamogashira 2017). These studies confirm the link between apoptosis and ageing. They suggested an essential part of an "ageing program" that blocks cell protection and accelerates death, thereby regulating the life span of other cells (Parikh 1987; Fabrizio 2004; Hekimi 2011). On the other hand, terrestric acid production has been moderate compared to high sucrose concentration. This supports our suggestion of apoptosis of damaged cells and therefore fewer cells that have undergone aging and produced terrestric acid. It was essential to understand whether sensitivity to the high concentration of sucrose is a particular characteristic of the strain of P. aurantiogriseum or all strains of this species and likewise for a wide range of Penicillium species. For this reason, we carried out a comparative study of intra-species (between two strains of P. aurantiogriseum ) and inter-species (between two species of Penicillium ). The results showed that the reaction to the high concentration was the same with non-significant differences. We were suggesting, that sensitivity to oxidative stress is not specific to the strain studied. Our preliminary findings will be used to extrapolate the same steps to conduct the study with powerful conclusions. This study showed the relationship between sucrose-induced stress, ageing, apoptosis, and mycotoxin production, in P. aurantiogriseum . Stress, depending on its intensity, could generate a process of programmed or radical ageing and even apoptosis. Besides, the accumulation of ROS due to stress has led to a change in the glycolysis pathway, the activation of certain mechanisms, and the overproduction of terrestric acid that has shown antioxidant properties against ROS to save the cells from apoptosis. The possibility of recovering large quantities of terrestric acid is a key advantage that could later allow studying this mycotoxin well. Insights gained into stress responses pave the way for mycotoxin production, however further studies are needed to validate and extend these preliminary results across diverse Penicillium species. Declarations Funding: This research received no external funding Author(s) contribution declaration: Conceptualization, AB; validation, MK; resources, AB, and FC; writing-original draft preparation, AB; writing-review and editing, MK, and FC; supervision, MK. All authors have read and agreed to the published version of the manuscript. Conflicts of Interest: The authors declare no conflicts of interest. All the experiments undertaken in this study comply with the current laws of the country where they were performed. References Bouhoudan A 1, Chidi F 1, Khaddor M et al (2020) 2 1 E, Effect of Sucrose on the Physiology and Terrestric Acid Production of Penicillium Aurantiogriseum . 179–189. https://doi.org/10.17707/AgricultForest.66.1.17 Bouhoudan A, Chidi F, Tantaoui-Elarak A, Khaddor M (2018) The effect of carbon source concentration on toxigenesis and lipase activity of Penicillium aurantiogriseum. 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Int J Food Microbiol 175:20–29. https://doi.org/10.1016/j.ijfoodmicro.2014.01.010 Traven A, Wong JM, Xu D et al (2001) Interorganellar communication. Altered nuclear gene expression profiles in a yeast mitochondrial dna mutant. J Biol Chem 276:4020–4027. https://doi.org/10.1074/jbc.M006807200 Turner WB, Aldridge DC (1971) Fungal metabolites. Academic Press, London, New York Valko M, Leibfritz D, Moncol J et al (2007) Free radicals and antioxidants in normal physiological functions and human disease. Int J Biochem Cell Biol 39:44–84. https://doi.org/10.1016/j.biocel.2006.07.001 Wang C-H, Wu S-B, Wu Y-T, Wei Y-H (2013) Oxidative stress response elicited by mitochondrial dysfunction: Implication in the pathophysiology of aging. Exp Biol Med 238:450–460. https://doi.org/10.1177/1535370213493069 Zain ME (2009) Effect of olive oil on secondary metabolite and fatty acid profiles of Penicillium expansum, Aspergillus flavus, A. parasiticus and A. ochraceus . Aust J Basic Appl Sci 3:4274–4280 Zhao W-J, An C-H, Han J-R (2014) Wet-plate culture studies of Penicillium sp. PT95 and Q1 for mass production of sclerotia. J Basic Microbiol 54:327–332. https://doi.org/10.1002/jobm.201200469 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-3876169","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":268850337,"identity":"7ea0473f-7000-43a4-abb2-6afa441e296a","order_by":0,"name":"Assia Bouhoudan","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABAklEQVRIiWNgGAWjYJCCA2AkAWTxMDDIka7FmASLoFoSGwip5e8//vDAD4Y7cga3m49JvKm5k77h+OEDDD9qGKL5cWiWuJFjcLCH4ZmxwZ1jaZJzjj3L3XAmLYGx5xhD7owDOKy5wcNwgIfhcOKGGzlm0jxsh3M3HMgxYOBtYMhtwKFF/vzxBwf/wLX8O5xucP79B8a/QC3zcWgxOJBgcBhuC2/b4QSDGzkMzCBbNuDQYgj0y2EZg2fGkjfSki3n9h02nHnjGVDkmETuRhxa5M4ff/zxTcUdOb4byQdvvPl2WJ7vfPLDh29qbHLn4fI+xHlo/AOQaBoFo2AUjIJRQC4AAB79aiWVoLXZAAAAAElFTkSuQmCC","orcid":"","institution":"Faculty of Sciences and Technology","correspondingAuthor":true,"prefix":"","firstName":"Assia","middleName":"","lastName":"Bouhoudan","suffix":""},{"id":268850338,"identity":"91cf2d1b-72b1-4684-8aac-37e8132e3dc9","order_by":1,"name":"Fatima Chidi","email":"","orcid":"","institution":"Abdelmalek Essaadi University","correspondingAuthor":false,"prefix":"","firstName":"Fatima","middleName":"","lastName":"Chidi","suffix":""},{"id":268850339,"identity":"e4513e63-18c2-4986-8eb0-34a22b681097","order_by":2,"name":"Mustapha Khaddor","email":"","orcid":"","institution":"Abdelmalek Essaadi University","correspondingAuthor":false,"prefix":"","firstName":"Mustapha","middleName":"","lastName":"Khaddor","suffix":""}],"badges":[],"createdAt":"2024-01-18 15:14:10","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-3876169/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-3876169/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":50159875,"identity":"ed00d19e-c9c5-4895-a9ec-6a4fe4ffb1dd","added_by":"auto","created_at":"2024-01-25 12:50:20","extension":"jpeg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":41924,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cem\u003eA: Penicillium aurantiogriseum \u003c/em\u003e(collection of \"EFBRT\") at different concentrations of sucrose.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eB: \u003c/strong\u003e\u003cem\u003ePenicillium aurantiogriseum \u003c/em\u003e(isolated in the \"EFBRT\" laboratory) at different concentrations of sucrose.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eC: \u003c/strong\u003e\u003cem\u003ePenicillium camembertii \u003c/em\u003e(isolated in the laboratory) at different concentrations of sucrose.\u003c/p\u003e\n\u003cp\u003eThe microscopic aspect of the three strains was taken at the concentration of 400 g/L where the whole colony presents the same shape of the thallus\u003c/p\u003e","description":"","filename":"floatimage1.jpeg","url":"https://assets-eu.researchsquare.com/files/rs-3876169/v1/910c6c7cd248434550c5bbf6.jpeg"},{"id":50159876,"identity":"fed29f0b-b41f-4aa4-9788-e1dd91c989ed","added_by":"auto","created_at":"2024-01-25 12:50:20","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":35600,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eA: \u003c/strong\u003eColony centre: cells lose branched hyphae and have an abnormal appearance\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eB: \u003c/strong\u003eColony periphery, the strain retains its asexual reproduction type with the presence of conidiophores as well as spores\u003c/p\u003e","description":"","filename":"Onlinefloatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-3876169/v1/c1f344f5a011f94b0d008d6d.png"},{"id":50209596,"identity":"1f952c6b-f841-4762-ba4a-f775f07bb521","added_by":"auto","created_at":"2024-01-26 10:37:49","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":365394,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-3876169/v1/1369aac5-62fb-4195-9384-8e83058466bb.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Induction of Ageing and Apoptosis by Sucrose in Penicillium aurantiogriseum","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eAgeing and apoptosis are two highly debated phenomena in moulds (Osiewacz \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Hamann 2008; Sharon 2009; Scheckhuber 2011; Kamogashira 2017). These natural mechanisms play a critical role in cell development. Apoptosis, a vital process, occurs throughout the development of an organism and is associated with the maintenance of cellular homeostasis, the elimination of damaged cells, the response to infectious agents, as well as the cell's adaptive response to biotic and abiotic stress (Danial and Korsmeyer \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Green \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2005\u003c/span\u003e). On the other hand, ageing corresponds to a gradual decline in the body's ability to cope with stress and damage (Jazwinski \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). Both of these mechanisms involve the accumulation of reactive oxygen species (ROS) (Elmore \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2007\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn 1956, Harman initially proposed the theory of radical ageing, based on the temporal imbalance between the production and elimination of free radicals (ROS) (Harman \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e1956\u003c/span\u003e). These ROS, produced naturally throughout the cell cycle, exert a crucial physiological role, notably in signalling cascades (Ray 2012), and the activated compounds play a pivotal role in inflammation and the balance between growth, apoptosis, and cellular senescence (Wang 2013). Antioxidant systems help maintain redox balance in the cell and significantly mitigate the risk of oxidative damage caused by ROS. Nevertheless, ROS possess varied and significant toxic properties, affecting various cellular components (macromolecules such as DNA, proteins, and lipids) when they evade antioxidant systems (Valko 2007). This process of oxidative alteration leads to mutations and denaturation, causing structural dysfunctions in biological systems. This ultimately leads to the ageing process, according to Harman's theory. An evolved version of the free radical theory of ageing suggests that mitochondria are the primary source of cellular ROS. When these mitochondria, particularly their respiratory chain components, are damaged, an accumulated production of ROS is generated (Harman \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e1956\u003c/span\u003e, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e1981\u003c/span\u003e, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e1998\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAny change in the usual conditions of cell life leads to a reaction called \"stress\" insofar as the cell has adapted its biological functions to environmental changes. Thus, any environmental or genetic condition, that increases exposure to aggression or decreases the capacity of defence or repair, results in accelerated ageing and, subsequently, apoptosis (Wang 2013). Oxidative stress, caused by an increase in cell ROS, is the most commonly studied.\u003c/p\u003e \u003cp\u003eMycotoxins, as secondary metabolites of moulds, have been detected in several human or livestock foods (Khaddor 2006). The ingestion of mycotoxins represents a real menace to human and animal health (Faid and Tantaoui-Elaraki \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e1989\u003c/span\u003e). \u003cem\u003ePenicillium aurantiogriseum\u003c/em\u003e is a particular species of the \u003cem\u003ePenicillium\u003c/em\u003e genus. Its mycotoxins are of great importance given their largely variable effects between harmful and beneficial to human and animal health (Khaddor 2007). To date, more than 37 mycotoxins have been identified in \u003cem\u003eP. aurantiogriseum\u003c/em\u003e. Previous studies had identified some of the mycotoxins such as penicillic acid, aurantiamine, and terrestric acid in \u003cem\u003eP. aurantiogriseum\u003c/em\u003e (Khaddor 2007). There are few studies of natural contamination with terrestric acid and its toxicity, and little is known about the factors influencing its production. The offending substance was found to be phytotoxic (Gausman \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e1991\u003c/span\u003e) and cardiotoxic (Frisvad 2004).\u003c/p\u003e \u003cp\u003eThe present work is devoted to studying \u003cem\u003eP. aurantiogriseum\u003c/em\u003e physiological response to the stress induced by sucrose and is focused on terrestric acid, a mycotoxin produced by \u003cem\u003eP. aurantiogriseum\u003c/em\u003e, exploring its production and toxicity under stress conditions induced by sucrose. Thus, it may help to control the growth conditions of this species and improve mycotoxin production. This study will be a pilot study to control the production of \u003cem\u003eP. aurantiogriseum\u003c/em\u003e mycotoxins that have great pharmaceutical interest.\u003c/p\u003e"},{"header":"2. Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1. Fungal Strains\u003c/h2\u003e \u003cp\u003eThe study was conducted on the \u003cem\u003eP. aurantiogriseum\u003c/em\u003e strain that belongs to the collection of the Environmental and Food Biotechnology Research Team (EFBRT), which had been utilized in prior studies (Khaddor 2007; Bouhoudan 2018, 2020). This strain was originally isolated from Pastilla leaves at the Department of Food Microbiology, Agronomic and Veterinary at the Institute Hassan II (IAV) in Rabat and its identification was confirmed by The Scientific Institute of Public Health (ISSP) of Louis Pasteur/Brussels. To initiate the study, the stored strain was cultivated on malt extract agar (MEA) and incubated at 25\u0026deg;C for 7 days. Following incubation, spores were suspended in 0.1% of tween 80 solution, and the suspension\u0026rsquo;s density was standardized to 10\u003csup\u003e7\u003c/sup\u003e spores/ml.\u003c/p\u003e \u003cp\u003eAdditionally, two other \u003cem\u003ePenicillium\u003c/em\u003e strains, namely \u003cem\u003eP. aurantiogriseum\u003c/em\u003e and \u003cem\u003eP. camembertii\u003c/em\u003e, were included in our experiments to facilitate conclusive comparisons. These strains, isolated within the laboratory (EFBRT), were authenticated using the Pitt method (Pitt and Hocking \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2009\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003e2.2. Growth medium\u003c/h2\u003e \u003cp\u003eThe G25N medium serves as a secondary purification medium conventionally employed for validating strain identification (Zhao 2014; Park 2014). This medium, known for its high glycerol content of 25%, reduces water activity, thus impeding the growth diffusion of \u003cem\u003eP. aurantiogriseum\u003c/em\u003e (max 24 mm). conversely, Glycerol facilitates consistent \u003cem\u003ePenicillium\u003c/em\u003e colony development (Pitt \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e1973\u003c/span\u003e) and stands out as an excellent carbon source conducive to mycotoxin production (Mul\u0026egrave; 2004). Hence, we retained this medium\u0026rsquo;s composition while introducing varied sucrose concentrations, ranging from 0 g/L (G25N) to 700 g/L, to study the impact of stress on the physiology of strains. The strains were inoculated on a G25N medium supplemented with different sucrose concentrations from 0 g/L to 700 g/L, herein referred to as G25N*. the inoculated cultures were then incubated for 10 days at 25\u0026deg;C.\u003c/p\u003e \u003cp\u003ePhysiological studies included observations on morphology, texture, colour, growth rings, growth status, mycelial weight, and colony diameter of \u003cem\u003eP. aurantiogriseum\u003c/em\u003e colonies, as well as the appearance of the hyphae. Radial growth was quantified by measuring the diameter of each colony using a ruler (Zain \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). All the experiments were meticulously conducted in triplicate to ensure accuracy and consistency of results.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003e2.3. Mycotoxins extraction\u003c/h2\u003e \u003cp\u003eThe mycotoxins extraction process followed the methodology outlined in the work of Bouhoudan et al, (Bouhoudan 2018, 2020). Fluorescence intensity, an indicator of toxigenesis, was represented by a varying number of \"+\" signs as described by Zain et al, (2009).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003e2.4. Statistical study\u003c/h2\u003e \u003cp\u003eStatistical analysis of the results obtained was conducted using the \"Duncan's multiple ranges\" test at a significance threshold of 5% [Stat Soft]. Nine tests were performed for each medium, and the averages derived from these nine trials (n\u0026thinsp;=\u0026thinsp;9) were subjected to analysis of variance (ANOVA) with Ducan's Multiple Range Test, also set at a 5% threshold. This statistical approach aimed to precisely define whether the factor (carbon source) had a significant effect on the response, specifically concerning mycotoxin production and fungal growth).\u003c/p\u003e \u003c/div\u003e"},{"header":"3. Results","content":"\u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003e3.1. Effect of stress on the physiology of Penicillium strains\u003c/h2\u003e \u003cp\u003eUnder the influence of varying sucrose concentrations, the colony diameter of both \u003cem\u003eP. aurantiogriseum\u003c/em\u003e and \u003cem\u003eP. camembertii\u003c/em\u003e strains exhibited an increase, as detailed in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eAverage colony diameters of \u003cem\u003ePenicillium\u003c/em\u003e species as a function of sucrose concentration of the G25N medium.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eSucrose concentration\u003c/p\u003e \u003cp\u003e(g/L)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c4\" namest=\"c2\"\u003e \u003cp\u003eColony diameter (mm) (95% IC)\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePA\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eP7\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePC\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e1.98 d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e1.70 d\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2.28 d\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e45.00 b\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e38.00 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e43.00 c\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e400\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e36.37 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e30.97 a\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e44 a\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e40.70 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e33.70 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNo growth\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e600\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e50.36 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e60.41 c\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e20.90 c\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e700\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e62.77 f\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e57 f\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e22.15 f\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"4\"\u003eIn the same column, two results followed by the same letter do not differ significantly at the 5% threshold. For each concentration of YES, nine tests were performed. The averages obtained in the nine trials (n\u0026thinsp;=\u0026thinsp;9) were compared by analysis of variance (ANOVA) with Ducan's Multiple Range Test at the 5% threshold.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e\u003cp\u003e\u003cstrong\u003ePA:\u003c/strong\u003e \u003cem\u003ePenicillium aurantiogriseum\u003c/em\u003e (collection of \u0026quot;EFBRT\u0026quot;) at different concentrations of sucrose.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eP7:\u003c/strong\u003e \u003cem\u003ePenicillium aurantiogriseum\u003c/em\u003e (isolated in the \u0026quot;EFBRT\u0026quot; laboratory) at different concentrations of sucrose.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePC:\u003c/strong\u003e \u003cem\u003ePenicillium camembertii\u003c/em\u003e (isolated in the laboratory) at different concentrations of sucrose.\u003c/p\u003e \u003cp\u003eThe results revealed a consistent response among the three \u003cem\u003ePenicillium\u003c/em\u003e strains under sucrose-induced stress, revealing a proportional increase in colony diameter with rising sucrose concentrations. As reported in our previous work by Bouhoudan et al, (Bouhoudan 2020), all strains exhibited a shared critical concentration point at 400 g/L, showcasing an exceptionally aggressive morphological response. At this threshold, the colonies displayed a whitish and notably severe appearance, as depicted in Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e1\u003c/span\u003e.\u003c/p\u003e \u003cp\u003eSubsequently, signs of ageing began to disappear at a sucrose concentration of 500 g/L, revealing the resurgence of normal \u003cem\u003ePenicillium\u003c/em\u003e strain characteristics. However, the disappearance of these signs varied in terms of speed and appearance among the strains. The \u003cem\u003eP. aurantiogriseum\u003c/em\u003e strain, from the team's collection (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e1\u003c/span\u003eA), revealed a slower return to normalcy compared to the same strain isolated in the laboratory (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e1\u003c/span\u003eB). additionally, \u003cem\u003eP. camembertii\u003c/em\u003e (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e1\u003c/span\u003eC) displayed more significant results compared to the strains of \u003cem\u003eP. aurantiogriseum\u003c/em\u003e.\u003c/p\u003e \u003cp\u003eAt the microscopic level, all three \u003cem\u003ePenicillium\u003c/em\u003e strains showed an identical appearance across the various sucrose concentrations added to the G25N* medium. Microscopic observation revealed the presence of two distinguished zones within the colony: the central zone, which contains older cells, and the peripheral zone, which represents younger cells, as illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e2\u003c/span\u003e.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003e3.2. Effect of sucrose on the toxigenesis of Penicillium strains\u003c/h2\u003e \u003cp\u003eThe results revealed a significant impact of sucrose concentrations within the G25N* growth medium on the metabolic profile of \u003cem\u003eP. aurantiogriseum\u003c/em\u003e strains (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eAverage of Terrestric acid (TA) intensity in \u003cem\u003ePenicillium\u003c/em\u003e strains depending on sucrose concentration of G25N* medium.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eSucrose concentration\u003c/p\u003e \u003cp\u003e(g/L)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c4\" namest=\"c2\"\u003e \u003cp\u003eIntensity of Terrestric Acid\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePA\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eP7\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ePC\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u0026ndash;\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\" morerows=\"5\" rowspan=\"6\"\u003e \u003cp\u003eNo production of TA\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e+\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e400\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e++++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e++++\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e500\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e600\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e700\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e++\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colspan=\"4\" nameend=\"c4\" namest=\"c1\"\u003e \u003cp\u003e(-) No (+) low, (++) medium, (++) strong (+++) very strong\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e\u003cp\u003e\u003cstrong\u003ePA:\u003c/strong\u003e \u003cem\u003ePenicillium aurantiogriseum\u003c/em\u003e (collection of \u0026quot;EFBRT\u0026quot;) at different concentrations of sucrose.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eP7:\u003c/strong\u003e \u003cem\u003ePenicillium aurantiogriseum\u003c/em\u003e (isolated in the \u0026quot;EFBRT\u0026quot; laboratory) at different concentrations of sucrose.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003ePC:\u003c/strong\u003e \u003cem\u003ePenicillium camembertii\u003c/em\u003e (isolated in the laboratory) at different concentrations of sucrose.\u003c/p\u003e \u003cp\u003eThe study of toxigenesis revealed a substantial production of terrestric acid coinciding with the appearance of ageing signs. Notably, as the ageing signs became more important, the concentration of terrestric acid produced increased significantly. However, the \u003cem\u003eP. camembertii\u003c/em\u003e strain does not exhibit any production of terrestric acid.\u003c/p\u003e \u003c/div\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eThe present work serves as a follow-up to prior studies conducted by Bouhoudan et al, (Bouhoudan 2018, 2020) focusing on assessing the effect of carbon sources on both the growth and the toxigenesis of \u003cem\u003eP. aurantiogriseum\u003c/em\u003e. Specifically, our emphasis was placed on discussing terrestric acid production given the insufficiency of studies that addressed it. This study acts as a pilot initiative laying the groundwork for future research endeavours that will include a broader spectrum of \u003cem\u003ePenicillium\u003c/em\u003e species and their associated mycotoxins.\u003c/p\u003e \u003cp\u003eG25N medium is typically a secondary purification medium used to confirm the identification of strain (Zhao 2014; Park 2014). This medium is known for its high content of glycerol (25%) serving to reduce the water activity in the medium which does not allow the growth diffusion for \u003cem\u003eP. aurantiogriseum\u003c/em\u003e (max 24 mm). On the other hand, Glycerol facilitates the consistent development of \u003cem\u003ePenicillium\u003c/em\u003e colonies (Pitt \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e1973\u003c/span\u003e) and it is an excellent carbon source for mycotoxin production (Pitt \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e1973\u003c/span\u003e). For this reason, we used this medium by keeping the same composition and added different sucrose concentrations, ranging from 0 g/L (G25N control) to 700 g/L, to study the stress effect related to sucrose concentration on the physiology of \u003cem\u003eP. aurantiogriseum\u003c/em\u003e strains. The modified media is named G25N*. In this regard, the phenotypic, morphologic, and metabolic responses identified in this study were caused obviously by the addition of sucrose at different concentrations in the culture medium.\u003c/p\u003e \u003cp\u003eStress caused by the high sucrose concentration on G25N*medium allowed us to observe different phenomena related to the suffering state of \u003cem\u003eP. aurantiogriseum\u003c/em\u003e:\u003c/p\u003e \u003cp\u003eAlthough ageing is a normal and essentially biological process in cells and generally leads to cell damage induced by ROS, induction of ageing by external factors leads to a kind of trauma and cellular stress. Cell reactions vary according to stress intensity and timing. In this study, we subjected our \u003cem\u003ePenicillium aurantiogriseum\u003c/em\u003e strain to carbon stress (due to the high sucrose concentration). The high sucrose concentration caused intense growth of the strain translated by an increase in colony diameter. The hyphae spread rapidly (seven d of growth gave a very important colony diameter compared to the control). However, as the hypha spreads, the nutrients become progressively restricted for peripheral cells. Nutrient deprivation had induced elevation of the colonies in the central zone showing cells suffering due to oxygen lack.\u003c/p\u003e \u003cp\u003eThe addition of sucrose induced the acceleration of the growth process and consequently a rapid achievement of the latency phase. The central zone containing the aged cells had a white colour (no sporulation) and the diameter of this zone was larger compared to the peripheral zone which contains the young cells. In the centre of the colony, mitotic cells lose their proliferative capacity and then disappear by apoptosis. The facts discussed so far are the consequences of programmed cell ageing. Various factors can contribute to the phenomenon of radical ageing mentioned in the introduction by Hamann et al, (Osiewacz \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Hamann 2008; Hameed 2012). We propose that in this study until the concentration of 400 g/L, the cells lost their ability to resist sucrose stress. We have suggested that sucrose indirectly induced several reactions initiated in the mitochondria. Since mitochondria are the main generators of reactive oxygen species (ROS), they were subjected to molecular damage. The time and intensity of exposure of the cells to the stress allowed a significant production of ROS. ROS, therefore, prevented the mitochondria from using scanning systems to repair damaged molecules that lead to early ageing (Osiewacz \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). Besides, as already mentioned in the \u003cspan refid=\"Sec1\" class=\"InternalRef\"\u003eintroduction\u003c/span\u003e section, at higher concentrations, ROSs damage cellular compounds and cause cellular dysfunctions (Osiewacz \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). Therefore, oxygen lack in the central zone reflected a disorder in the respiratory chain, which increased ROS levels.\u003c/p\u003e \u003cp\u003eStudies on the development of senescent cultures have revealed that the presence of a determinant of senescence seems to accumulate during ageing (Maas 2007). To confirm this fact, we performed a small demonstration in which we took a tip of centre cells and another of peripheral cells and cultivated them in fresh growth media. After growing, we obtained colonies that grow back from the cells of the periphery, whereas cells of the centre could not grow back. This supports Marcou's conclusions that the hyphae were dead in the centre and that the senescence determinant seems to be absent from young cultures and accumulates after the culture passes through a critical point. Marcou demonstrated the hyphae fusion of a senescent culture leading to the transfer of the accumulated senescence determinant to a young culture. This one immediately becomes senescent and dies on the growth front. The determinant of senescence thus appears infectious and cytoplasmic (Maas 2007). This explains the increase in the central zone diameter of stressed colonies.\u003c/p\u003e \u003cp\u003eAt the metabolic level, the stress induced by the high sucrose concentration in the culture medium affected the toxigenic profile of \u003cem\u003eP. aurantiogriseum\u003c/em\u003e. Production of terrestric acid at the time of ageing signs appearance became very important. More aging signs were important more the concentration of terrestric acid produced was high. Sucrose assimilation became more difficult for the centre cells (aged). Also, the oxygen lack caused by cellular outgrowth leads to a decrease in centre cells' antioxidant capacity to maintain the environment in a reduced state due to the increase of (ROS) in the mitochondria. Therefore, the cells respond to stress by dynamic redirection of the glycolysis\u0026rsquo;s metabolic flux to the pentose phosphate pathway, which is the biosynthetic pathway of terrestric acid. It should be noted that mycotoxins are derived from acetate and shikimate via polyketoacids and the amino acid pathways in the Krebs cycle (which takes place in the mitochondria) (Marcou \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e1961\u003c/span\u003e; Pieter S. Steyn \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e1980\u003c/span\u003e). This explains the proportional relationship between stress and terrestric acid production.\u003c/p\u003e \u003cp\u003eAlthough the increase in sucrose concentration has shown intense cellular ageing in all colonies, the concentration of 400 g/L, however, presented a special case in which the colony was all white with a very aged look, which supports the radical ageing theory of Harman (Harman \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e1956\u003c/span\u003e). \u003cem\u003ePenicillium aurantiogriseum\u003c/em\u003e cells have undergone accelerated chronological ageing linked to the long exposure to the high sucrose concentration in which cells still young undergo an early senescence process. Cellular outgrowth has led to nutrient deprivation and oxygen lack. Radical ageing signs, appearing in this study, were accompanied by an overall alteration of a set of physiological functions as well as a higher susceptibility to different alterations. The free radical theory explains these changes by the accumulation of oxidized molecules and the consequences of oxidation as the appearance of mutations. Several arguments are in favour of free radicals\u0026rsquo; involvement in ageing mechanisms (Turner and Aldridge \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e1971\u003c/span\u003e; Wang 2013; Kamogashira 2017). The situation is more complex when one looks for chronic oxidative stress during which, on the one hand, the ROS increases elevated and, on the other hand, the inductions of the antioxidant and repairing systems are more modest, those are sometimes themselves impaired by oxidation. Wang et al, (Wang 2013) supported this observation by explaining when the repair systems themselves were affected, it is likely to move towards cellular function deterioration that was accompanied by accelerated ageing. If the ROS continues to accumulate, a more consistent adaptation of the cell is necessary with the induction of its antioxidant system. Suppression of systems capable of releasing ROS, particularly the respiratory chain (Osiewacz \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2002\u003c/span\u003e), is observed by reorientation towards the glycolysis pathway mentioned above. This leads to an overproduction of terrestric acid, which reached its maximum level at the concentration of 400 g/L. Several studies have suggested the existence of an \"ageing program\" that blocks cell protection and accelerates cell death (Madeo 1999; Herker 2004; Hekimi 2011). We took a crop point and re-cultivated it in a fresh growing medium. After growth, we obtained colonies that normally pushed what allowed us to suggest that culture to 400 g/L did not undergo apoptosis. Studies have shown a correlation between oxidative stress and mycotoxin production (Mortimer and Johnston \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e1959\u003c/span\u003e; Schmidt-Heydt 2011). Mycotoxin biosynthesis contributed to the adaptation to stressful environments. Interesting in this respect is the fact that these mycotoxins have antioxidant properties that can protect against oxidative stress (Stoll 2014). For this, we suggested the antioxidant effect of terrestric acid vis-\u0026agrave;-vis oxidative stress, and, through this; the cells do not have undergone apoptosis.\u003c/p\u003e \u003cp\u003eIt is now obvious that high sucrose concentration provoked ROS overproduction and therefore mitochondrial dysfunction that causes early ageing. However, in our case, the increase in sucrose concentration above 500 g/L did not result in radical ageing and the cells resisted the stress. Our strain was normal in appearance with good biomass and mycelium production and moderate terrestric acid production. We consider this finding as an escape phenomenon at the high sucrose concentration. Our analysis suggests that between 10g/l and 400g/l of sucrose, \u003cem\u003eP. aurantiogriseum\u003c/em\u003e was forced to adapt to the sucrose stress. It used its panoply of intracellular and genetic to maintain cellular growth and survival. This response is similar to the \"retrograde response\" described for the first time in \u003cem\u003eS. cerevisiae\u003c/em\u003e (Sekito 2000; Schmidt-Heydt 2015) Thus, retrograde response occurs during normal aging to offset mitochondrial dysfunction accumulation allowing cells to live as long as they do (Jazwinski \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). The induction of retrograde response increases the replicative lifespan and increases stress resistance (Liao and Butow \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e1993\u003c/span\u003e; Kirchman 1999). The retrograde response indicates changes in gene expression (Traven 2001). Studies have clearly shown that the retrograde response results in a wide range of changes in gene expression (Kirchman 1999) that suggest extensive metabolic remodelling of the cell in response to the metabolic constraint associated with mitochondrial dysfunction that ultimately allows for longer cell life (Jazwinski \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2002\u003c/span\u003e). However, below the concentration of 500 g/L, the cell reached a very intense level of stress, which resulted in the accumulation of a very large amount of ROS, leading therefore to dysfunction not only of the mitochondria but also of all cell components and mutations in genes. This necessarily leads to the non-functioning of the retrograde response and the cells undergo apoptosis leaving the other normal cells to live. This explains the existence of a peripheral zone containing young cells. Studies show that the involvement of apoptosis occurs when the molecular and organelle scanning systems can no longer cope with the damage accumulated by oxidative stress (Kamogashira 2017). These studies confirm the link between apoptosis and ageing. They suggested an essential part of an \"ageing program\" that blocks cell protection and accelerates death, thereby regulating the life span of other cells (Parikh 1987; Fabrizio 2004; Hekimi 2011). On the other hand, terrestric acid production has been moderate compared to high sucrose concentration. This supports our suggestion of apoptosis of damaged cells and therefore fewer cells that have undergone aging and produced terrestric acid.\u003c/p\u003e \u003cp\u003eIt was essential to understand whether sensitivity to the high concentration of sucrose is a particular characteristic of the strain of \u003cem\u003eP. aurantiogriseum\u003c/em\u003e or all strains of this species and likewise for a wide range of \u003cem\u003ePenicillium\u003c/em\u003e species. For this reason, we carried out a comparative study of intra-species (between two strains of \u003cem\u003eP. aurantiogriseum\u003c/em\u003e) and inter-species (between two species of \u003cem\u003ePenicillium\u003c/em\u003e). The results showed that the reaction to the high concentration was the same with non-significant differences. We were suggesting, that sensitivity to oxidative stress is not specific to the strain studied. Our preliminary findings will be used to extrapolate the same steps to conduct the study with powerful conclusions.\u003c/p\u003e \u003cp\u003eThis study showed the relationship between sucrose-induced stress, ageing, apoptosis, and mycotoxin production, in \u003cem\u003eP. aurantiogriseum\u003c/em\u003e. Stress, depending on its intensity, could generate a process of programmed or radical ageing and even apoptosis. Besides, the accumulation of ROS due to stress has led to a change in the glycolysis pathway, the activation of certain mechanisms, and the overproduction of terrestric acid that has shown antioxidant properties against ROS to save the cells from apoptosis. The possibility of recovering large quantities of terrestric acid is a key advantage that could later allow studying this mycotoxin well. Insights gained into stress responses pave the way for mycotoxin production, however further studies are needed to validate and extend these preliminary results across diverse \u003cem\u003ePenicillium\u003c/em\u003e species.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eFunding:\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis research received no external funding\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthor(s) contribution declaration:\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eConceptualization, AB; validation, MK; resources, AB, and FC; writing-original draft preparation, AB; writing-review and editing, MK, and FC; supervision, MK. All authors have read and agreed to the published version of the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflicts of Interest:\u003c/strong\u003e\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eThe authors declare no conflicts of interest. 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J Basic Microbiol 54:327\u0026ndash;332. \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://doi.org/10.1002/jobm.201200469\u003c/span\u003e\u003cspan address=\"10.1002/jobm.201200469\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"P. aurantiogriseum, Terrestric acid, Aging, Apoptosis, Carbon stress. Toxigenesis","lastPublishedDoi":"10.21203/rs.3.rs-3876169/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-3876169/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThis work aims to study the physiological and metabolic responses of \u003cem\u003eP. aurantiogriseum\u003c/em\u003e to sucrose-induced stress. Comparative analyses on intra-species (within strains of P. aurantiogriseum), and interspecies (between \u003cem\u003eP. aurantiogriseum\u003c/em\u003e and \u003cem\u003eP. camembertii\u003c/em\u003e) were conducted to assess the effect of carbon stress on aging phenomena and toxigenesis. Our results revealed a correlation between sucrose concentration and ageing signs. At a sucrose concentration of 500 g/L, the ageing signs of \u003cem\u003eP. aurantiogriseum\u003c/em\u003e began to fade, allowing its normal characteristics to resurface. This transformation is believed to be a response to the sucrose and the cells that cannot adapt undergo apoptosis, leaving only the normal cells to thrive. Terrestric acid production was observed during the ageing process and continued even after returning to a normal physiological state, albeit at a reduced level.\u003c/p\u003e","manuscriptTitle":"Induction of Ageing and Apoptosis by Sucrose in Penicillium aurantiogriseum","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-01-25 12:50:16","doi":"10.21203/rs.3.rs-3876169/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"21bb40b4-94d4-483d-a42e-5f0187a8b4a5","owner":[],"postedDate":"January 25th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2024-01-26T10:29:42+00:00","versionOfRecord":[],"versionCreatedAt":"2024-01-25 12:50:16","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-3876169","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-3876169","identity":"rs-3876169","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}