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Microbiome analysis to white truffle cultivation – science fiction or just science?

The NATURALLY project team had an eventful summer! Our days were packed with analyses, publications, reports, and fieldwork. We also juggled applying for new projects and forming collaborations, keeping every moment occupied over the past few months. Amidst all this, there was continuous learning, both officially with exams and unofficially, which we absolutely enjoy! We also spent time preparing for conferences and gatherings because, let’s be honest, we have a passion for travelling and presenting our work! All of these activities were intertwined with precious moments of socializing with our colleagues and friends,
something we absolutely adore! However, as a result of our busy schedule, we’ve been less active in the virtual world, writing fewer posts and taking fewer photos. Instead, we’ve added more tasks to our ‘to-do’ list. But there are so many beautiful experiences we’d love to share with you!

One of these beautiful new stories is the visit of the PhD student Mara Rondolini from the University of Perugia in Italy to our laboratory. At the age of 27, Mara possesses a deep passion for merging laboratory work with nature, with her ultimate goal being the cultivation of white truffles. Her ambition is no small feat; it requires substantial efforts, but it’s also a fascinating journey of scientific discovery. We immediately liked her! Both cool Mara and cool science 😉

Mara and her mentors, in collaboration with various institutions in Italy, Serbia, and Croatia, aim to investigate the conditions and techniques required for cultivating truffles in natural ecosystems and agricultural settings to promote coastal biodiversity. But what does this precisely entail?

The white truffle (Tuber magnatum Picco) is the rarest and most highly prized underground mushroom globally, primarily found in natural environments across Europe, where it forms symbiotic relationships with various woody plant species like oak, willow, poplar, and others. The cultivation of white truffles is significantly more intricate and less successful than other truffle varieties, such as the black truffle. The exact reasons for this disparity remain a
mystery. Part of the explanation undoubtedly lies in the highly specific ecological conditions required for the growth of this species, and these unique requirements have yet to be fully
characterized and understood. Mara, along with numerous researchers worldwide, continues the quest for a successful
formula for cultivating white truffles—an intricate combination of soil conditions, climate, vegetation, and other crucial factors that impact this delicate truffle. Multiple studies highlight the significant influence and active role of microorganisms in the white truffle’s life cycle, making microbiome analysis an essential component of this research. Our laboratory focused on analyzing the soil microbiome, a pivotal aspect of the study. Mara collected soil samples from various locations characterized by different environmental conditions, intending to employ diverse analytical methods to compare soil conducive to truffle growth
with less favourable soil. All this aims to uncover that elusive, essential factor crucial for the truffle’s existence! Isn’t that fascinating?

In our laboratory, we isolated DNA from different soil types, subjecting it to sequencing to identify the organisms that, alongside various soil and environmental parameters, either
facilitate or hinder the truffle’s life. These analyses are still ongoing, and upon concluding the research, we hope to unravel at least part of this intricate puzzle. We extend our best wishes
to Mara, who will return to our team at the end of the year. For those interested in this topic and the continual assembly of new pieces in the mosaic of the white truffle’s secret life, you can follow our progress on our social media channels.
Lastly, you might be wondering about the connection between invasive plants and white truffles. Well, there is indeed a link! Since everything in nature is interconnected within the magical cycle of life, our story shares a common thread in this chain of existence. To elaborate, new theories suggest that a significant portion of plant invasiveness is attributed to the microbiome. That’s why we plan to apply the knowledge and demanding laboratory protocols we’ve gained from soil analysis to our study of invasive plants.

We participated in three international meetings abroad

The results of the NATURALLY project were presented this fall at three international meetings. At the 8th Edition of Global Conference on Plant Science and Molecular Biology (GPMB 2023) held in Valencia (Spain) on 11-13 September 2023, Dr. Danijela Poljuha held a lecture entitled “Dr Jekyll and Mr Hyde: Two-Faced Invasive Plants” in which she presented the results of the phytochemical analysis of extracts of four invasive alien species in Istria, emphasizing the unexplored potential of invasive species as a source of phytopharmaceuticals.

Dr. Barbara Sladonja presented an assessment protocol of the ecological impact (positive and negative) of invasive species with a poster presentation entitled “Impact Assessment of Four Invasive Alien Plant Species in Croatia” in Pisa (Italy), at the 118th Congresso della Società Botanica Italiana onlus (IX International Plant Science Conference) 13.-16. September 2023

Mirela Uzelac Božac, PhD student, participated in the 4th International Conference on Community Ecology held in Trieste on September 20, 2023, where she presented her research as part of her doctoral thesis. In the presentation entitled “Invasive Alien Plants as Ecosystem Service Provider”, she presented the results of research into the antioxidant activity of extracts of four foreign invasive species in Istria, as well as individual phenols obtained by LC-MS analysis, and presented the next steps in the research.

Presenting the results of our research to the international scientific public is an important part of our work, which enables us to connect with colleagues from related fields, new collaborations and the opening of new questions, as well as the upgrading of existing research.

The results of the Naturally project were presented in a new scientific paper

In the international scientific journal Plants (IF 4.5) dedicated to plant research, our scientists published a paper entitled “Invasive Alien Species as a Potential Source of Phytopharmaceuticals: Phenolic Composition and Antimicrobial and Cytotoxic Activity of Robinia pseudoacacia L. Leaf and Flower Extracts”. The paper is a part of the NATURALLY project (HRZZ-IP-2020-02-6899), which studies the phytopharmaceutical potential of extracts of four foreign invasive plant species.

In this paper, an international team of researchers present for the first time the profile of phenolic compounds of Black locust populations, an invasive species from the area of the Istrian peninsula. The paper also presents the first data on the antimicrobial and cytotoxic activity of black locust leaf and flower extracts from this area tested on in vitro cell cultures. This is one of the first steps towards the final goal – the proposal of a model for exploiting invasive species for beneficial purposes, and thus contributing to the sustainable management of invasive species, which is one of the global priorities caused by climate change.

Uzelac, M., Sladonja, B., Šola, I., Dudaš, S., Bilić, J., Famuyide, I.M., McGaw, L.J., Eloff, J.N., Mikulic-Petkovsek, M., Poljuha, D. Invasive Alien Species as a Potential Source of Phytopharmaceuticals: Phenolic Composition and Antimicrobial and Cytotoxic Activity of Robinia pseudoacacia L. Leaf and Flower Extracts. Plants 2023, 12, 2715. 

The paper is available at the following link: https://www.mdpi.com/2223-7747/12/14/2715.

Interim meeting of the Naturally project was held at the Institute

At the Institute of Agriculture and Tourism on 6.-7. 7. In 2023, an interim meeting of the project NATURALLY was held in a hybrid form. Project partners from the University of Zagreb, Ljubljana and Pretoria (Republic of South Africa) and from the Polytechnic of Istria and the Polytechnic in Rijeka during the two-day meeting presented the results obtained so far on the project and agreed on further steps in the implementation of the project and potential guidelines for new collaborations.

The project “NATURE as an ALLY: Foreign plant invasive species as a source of pharmaceuticals – NATURALLY” was financed by the Croatian Science Foundation, and its goal is to investigate the phytopharmaceutical potential of natural extracts of four alien invasive plant species in Istria as a basis for new possible ecosystem services.

The participants of the meeting also had the opportunity to hear public lectures on the topic of phytochemistry of invasive species and the indirect influence of global warming on the physiological parameters of mammals by eating plants grown at high temperatures. A tour of the Institute’s laboratory and olive oil tasting were organized for the guests.

Public lecture announcement

On Friday, 7.7. at 9:00 a.m. at the Institute for Agriculture and Tourism, Prof. Ph.D. Maja Mikulič Petkovšek from the Biotechnical Faculty of the University of Ljubljana will hold a public lecture entitled “Phytochemistry of invasive species” as part of the activities of the HRZZ project NATURALLY – NATURE AS AN ALLY: FOREIGN PLANT INVASIVE SPECIES AS A SOURCE OF PHARMACEUTICALS (HRZZ-IP-2020-02-6899).

We invite all those interested to join us via the Zoom link:
https://us06web.zoom.us/j/82325177776?pwd=TUl3YUM0d21Wckc4aytjcFVpKzVPUT09

Results of the NATURALLY project presented at the international conference ICNPU-2023 in Bulgaria

The 5th International Conference on Natural Products Utilization: From Plants to Pharmacy Shelf (ICNPU-2023) was held in Varna, Bulgaria, on May 30 – June 2, 2023. At the conference, Dr Danijela Poljuha presented the results of research conducted as part of the NATURALLY project funded by the Croatian Science Foundation. Through the poster presentation, she presented the results of the analysis of the phenolic composition and antioxidant capacity of leaf extracts of four invasive alien plant species in Istria. This study is part of research aimed at evaluating the potential of invasive plant species as a source of phytopharmaceuticals.

Presentation of the NATURALLY project results at the University of the North

On the occasion of the Fascination of Plants Day, on Thursday, May 18, 2023, Dr. Danijela Poljuha and PhD student Mirela Uzelac, mag. oecol. visited the Department of Food Technology of the University of the North in Koprivnica.

They presented the results of the NATURALLY project (IP-2020-02-6899) and results of the doctoral dissertation, which is carried out as part of the program “Project for the development of careers of young researchers – training of new doctors of science” (DOK -2021-02-3094). A new collaboration was also agreed between three projects: TEMPHYS project (IP-2020-02-7585) led by Dr. Ivana Šola, GinkoBiFlav (UIP-2019-04-1018) lead by Dr. Dunja Šamec and our project NATURALLY, all three financed by the Croatian Science Foundation.

Dr. Dunja Šamec welcomed everyone with opening speech and presented the future Agreement on Cooperation on Strengthening the Research Network of Projects, after which Dr. Ivana Šola from the Faculty of Science, University of Zagreb held a lecture entitled “How does global warming affect the nutritional value of plants?“. She presented the research results of the TEMPHYS project, which deals with the indirect impact of global warming on the physiological parameters of mammals by feeding them plants grown at high temperatures.

Dr. sc. Danijela Poljuha held the lecture entitled “Small and large projects fascinated by plants” in the frame of project NATURALLY and several smaller projects that are carried out at the Institute, and PhD student Mirela Uzelac, mag. oecol., presented the topic and results of her doctoral dissertation entitled “Phytopharmaceutical potential of leaf and flower extracts of alien invasive plant species”.

We are looking forward to future cooperation!

Results of the NATURALLY project presented at the 28HSKIKI international meeting in Rovinj

The 28th Croatian Meeting of Chemists and Chemical Engineers (28HSKIKI) is organized by the Croatian Society of Chemical Engineers and the Croatian Chemical Society in Rovinj from March 28-31, 2023. At the meeting, Dr Danijela Poljuha presented the results of research conducted as part of the NATURALLY project
financed by the Croatian Science Foundation.

Through the poster presentation, she presented the results of the analysis of the phenolic composition and antioxidant capacity of leaf and flower extracts of Robinia pseudoacacia L., an invasive plant species in Istria. This study is part of research aimed at evaluating the potential of invasive plant species as a source of phytopharmaceuticals.

A paper on invasive alien (animal) species was published

In addition to plants, our Naturally team also investigates animals.

A scientific paper on the topic of agricultural and forest pests and newly introduced species in Istria entitled “The Impact of Beehive Proximity, Human Activity and Agricultural Intensity on Diptera Diversity in a Mediterranean Mosaic of Agroecosystems, with a Focus on Pest Species” was published in a special issue of the scientific journal Animals dedicated to interactions between animals in different habitats (Q1; IF: 3,231). The work was created as a result of international cooperation and teamwork of scientists from the Institute, dr. Barbara Sladonja, dr. Danijela Poljuha and Mirela Uzelac, mag. with entomologists from Italy and the Czech Republic.

A statistical data analysis workshop held at the Institute of Agriculture and Tourism

On Monday, February 6th, at the Institute for Agriculture and Tourism, the workshop “Statistical Data Analysis (IBM SPSS Statistic)” was held for members of the project team and associates. Workshop leader Slavica Dudaš, PhD, Assoc. professor at the Polytechnic in Rijeka, explained the methods of data analysis and interpretation of the results in the context of the project’s goals, using practical examples of the analysis results obtained as part of the project.

The participants had the opportunity to go through the entire analysis process and discuss the obtained results. The project “NATURE as an ALLY: Alien Invasive Plants as Phytopharmaceuticals – NATURALLY” is funded by the Croatian Science Foundation. The project’s main goal is to investigate the
phytopharmaceutical potential of natural extracts of four invasive alien species (three of heaven, black locust, Jerusalem artichoke, and Canada goldenrod) in Istria (Croatia) as a basis for new
ecosystem services.