achieving sustainable agriculture - The BioClub Project - Universidade ...

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organism is an island; all the phenotypes observed are the ... genotypes in very dependent and regulated ways.' .... Ano
ACHIEVING SUSTAINABLE AGRICULTURE

www.bioclub.fc.ul.pt 

A groundbreaking biofertiliser Transdisciplinary research underway at the University of Lisbon, Portugal, is seeking to contribute to the long-term sustainability of food production via the creation of a new, innovative biofertiliser

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s the global population increases, meeting demand for crops in order to ensure food security is becoming more and more challenging; particularly doing so in a sustainable way with minimal impact on the environment. There is a high global demand for biofertilisers – a compost which contains living microorganisms able to colonise the interior of a plant – as a potential solution, but there are issues relating to their use and design, such as they tend to be specific to crop, soil type and weather conditions; those with higher nitrogen use efficiency (NUE) are expensive; and most are designed without considering rhizospheric socialisation.

transdisciplinary collaboration. The BioClub researchers believe the efficiency of biofertiliser can be significantly improved with a strong symbiotic relationship between the partners involved in biofertiliser formulation. ‘The BioClub team believes the next steps towards a sustainable agriculture require understanding the networks, which regulate the distinct components of the agrosystem,’ explains project leader Cristina Cruz. ‘No organism is an island; all the phenotypes observed are the product of the interplay of millions of organisms that express their genotypes in very dependent and regulated ways.’

A three year project underway at the Faculty of Sciences of the University of Lisbon (Faculdade de Ciências da Universidade de Lisboa - FCUL), Portugal – ‘Designing biofertilizers by mimicking plants’ recruitment of rhizospheric partners’ (BioClub) – has been working since 2016 to produce an efficient and economically accessible biofertiliser prototype through

EXTENSIVE EXPERTISE Cruz is working with a multidisciplinary team comprising experts in crop productivity, plant and soil ecology, plant physiology, microbiology, the environmental impacts of agriculture, molecular biology and plant physiology, and next-generation sequencing (NGS) techniques. The project also relies on a ‘BioClub society’ to assist with cohesion, as Cruz explains: ‘BioClub is a large project with a diverse number of different experts that needs to create a cohesion, take profit and trigger the effectiveness of this transdisciplinary team,’ she states. ‘To give the cohesion of this BioClub society a board of experts was formed, putting together the task coordinators and the representatives of each of the companies (AMC Chemical, Soilvitae, ADP-Fertilizantes) and end-users (Cimmyt, Fertiprado and EPP).’

Plant

Fungus

Fungus (Glomus intraradices, Piriformospora indica) Azospirillum brasilense (associative)

Bacteria

Autoinducers

Rhizosphere

Exudates

Feedback

Plant

Root

Azospirillum brasilense (endophytic)

Root cells interacting with the microbiota: bacteria and fungi, which regulates plant defence, metabolism and growth. (design by Florian Ulm)

The role of the society is four-fold: to create the spirit of a BioClub society within the team members involved; facilitate the creation of bridges among these members;

define working rules and prototypes; and promote the BioClub society letter. The team will focus on the interplay between microbial agents, for example fungi and bacteria, and the plant. The idea is this will produce an efficient and affordable biofertiliser prototype. The project will study four wheat varieties: two of Triticum durum and two of Triticum aestivum, supplied as grain by Estação Agrária de Elvas, ADP-fertilizantes and LusoSem. As a starting point, the team will characterise the composition of the root exudates (REs) produced by each of these varieties during the first week after germination and how these exudates interfere with the recruitment of the bacteria. CLEAR STEPS The project is divided into different work packages (WPs), the first of which involves obtaining information on the chemical composition of REs of several wheat varieties and how they are or are not affected by biotic and abiotic factors. This will be followed by an exploration of whether the lifestyle of the symbiont interferes with the composition of RE, and an evaluation of the possibility of using the RE of an old wheat variety as a component of the BioClub biofertiliser. WP2 and WP3 will involve evaluating the beneficial effects of the two BioClub versions for plant development, with WP2 comparing the performance of plants grown in pots inoculated with four potential Bioclub.pt, all receiving 70 per cent of the recommended mineral fertiliser dose (RMFD), or noninoculated plants receiving 100 per cent RMFD. ‘The final proof of concept will be performed in field trials testing the effects of the BioClub biofertiliser in three fields representing three distinct edaphoclimatic conditions suitable

We are firmly convinced that the future of agriculture relies on the general use of biofertilisers more than chemical fertilisers

for wheat production (WP four),’ highlights Cruz. ‘In the field trials, apart from the impact of the biofertiliser (both BioClub formulations) on grain yield, the impact on grain quality and soil dynamic properties will also be assessed.’ To date, the researchers boast a number of significant achievements. These include the fact the main wheat varieties and microbial strains have been selected, the REs of each variety are ready for analyses, the REs of colonised plants are in preparation, the biofilm formation of the microbial strains has been characterised, and the interactions between fungal and bacteria partners are being assessed in vitro. ‘Until now our main achievement was the establishment of a hypothesis relating the production of oxide nitric and phytopathogenecity. We inspected for the presence in bacteria of similar sequences to the NO-producing nitrite reductase nirS gene of Thermus thermophilus, an aerobe of the Thermus/ Deinococcus group, which constitutes an ancient lineage related to Cyanobacteria,’ Cruz reveals. ‘The in silico analysis revealed the relationship between the presence of nirS genes and phytopathogenicity in gram-negative bacteria. This is an important achievement since it may be used as a tool to discriminate phytopathogenic from potential PCPB producing biofilm and having very good effects on plant phenotype. This has already been published in Frontiers in Microbiology.’ Another significant finding is that of the importance of the priming effect in plant

response to microorganisms, as Cruz explains: ‘The impact of a microorganism (plant growth-promoting rhizobacteriaPGPR) on plant phenotype is much higher if the plant is not yet colonised by other microorganisms. This is interesting because it opens doors to improve certain plant traits by inoculating plants with certain PGPM during early phases of plant development.’ DRIVING THE POINT HOME Looking ahead, the team will continue to strive to help reduce the strain placed on agriculture by the expanding population. The researchers have identified the need to improve nutrient use efficiency (NUE) and have found that one way to do so is through a partial substitution of the RMFD by plant growth promoting microorganisms (PGPMs). ‘PGPM are able to promote root system development, increase acquisition of nutrients and improve plant defence in exchange for RE,’ Cruz highlights. ‘Based on previous project results we demonstrated the advantages of using biofertilisers on plant development and NUE. Thus, we are firmly convinced that the future of agriculture relies on the general use of biofertilisers more than chemical fertilisers.’ Cruz believes the team requires time and solid results to drive this point home and plans to accumulate evidences and data, and incorporate and interpret drawbacks with a view to improving and facilitating the use of biofertilisers.

Project Insights FUNDING Science and Technology Foundation (PTDC/AGR-PRO/1852/2014) PROJECT TEAM Juliana Melo da Conceição Luís Carvalho Manuela Carolino Amarilis de Varennes Maria Amélia Martins-Loução Rogérioo Tenreiro Patrícia Correia Florian Ulm Francisco Dionísio CONTACT Cristina Cruz Project Leader W: www.bioclub.fc.ul.pt  F: https://www.facebook.com/ BioClubNet/ Twitter: https://twitter.com/bioclubnet BIO Cristina Cruz is a lecturer in Plant Biology at the Faculty of Sciences of the University of Lisbon (Faculdade de Ciências da Universidade de Lisboa FCUL), Portugal, where she has taught and carried out research since 1994. Within the University she teaches in the Department of Plant Biology (Departamento de Biologia Vegetal - DBV), and carries out her research in the Plant-Soil Ecology Group of the Centre for Ecology, Evolution and Environmental Changes (cE3c).

Impact Objectives • Develop an affordable biofertiliser which will not only ensure food security, but will do so without harming the environment • Ultimately reduce the strain placed on agriculture by the expanding world population

Achieving sustainable agriculture Cristina Cruz introduces the BioClub project, which is producing an efficient and affordable biofertiliser prototype that she hopes will contribute to meeting the challenge of feeding a growing population, while minimising the impact on the environment Can you introduce the ‘Designing biofertilisers by mimicking plants’ recruitment of rhizospheric partners’ (BioClub) project? The BioClub biofertiliser will focus on: the interplay between the microbial agents (fungi and bacteria) and the plant:plant exudome (wheat varieties); biofilm production; and lifestyle (associative versus endophytic), in order to produce an efficient and affordable biofertiliser prototype. In what ways will the prototype be beneficial? BioClub is a project that intends to produce an efficient and economically accessible biofertiliser prototype. The goal is that it will greatly benefit plant production and reduce the addition of mineral fertiliser by 30 per cent. It will do so by: improving plant tolerance to pathogens; improve plant productivity and nutrient use efficiency independently of soil types and atmospheric conditions, namely temperature and water availability; and be compatible with European directives in reducing the addition of mineral fertilisation. What are the downfalls of existing biofertilisers? Available biofertilisers present variable benefits and certain problems associated with their use and design. For example, they are generally specific for crop, soil type

and weather conditions and those with higher nitrogen use efficiency are expensive. Most are also generally designed without considering rhizospheric socialisation. Who are the key participants that make up BioClub’s team? BioClub has a transdisciplinary team with specialists in crop productivity, plant and soil ecology, plant physiology, microbiology, environmental impacts of agriculture, molecular biology and plant physiology and next generation sequencing (NGS) techniques. BioClub also involves companies that produce synthetic fertilisers and biofertilisers, as well as end-users including Cimmyt, Fertiprado and EPP. Who are the expected end users of your work and how will they benefit? The end users will be the farmers, as they are the ones that will use the product. In this respect, the participation of Cimmyt, Fertiprado and EPP is very important because it enables us to receive direct feedback from the farmers with whom we directly work. But in order to produce BioClub.pt (Bioclub protype) in sufficient amounts and to achieve its commercialisation, we also need companies able to perform the upscale and companies capable of transforming the prototype into a product. We also have contacts with this type of end-user (Trichodex) and BioClub makes an effort in the formation of the technical advisers in order to improve their understanding

surrounding the differences between a fertiliser and a biofertiliser. Can you discuss your hopes for the future of agriculture and the environment? The increased use of fertilisers based mainly on nitrogen has been critical for increased crop yields and protein production to keep pace with the growing world population. We are now facing a similar problem with the use of fossil burning carbon. Increased industrial fixation of nitrogen can have adverse environmental consequences at local, regional and global scales. Due to all these demands, our use of both fossil carbon and synthetically fixed nitrogen has grown exponentially in the past 150 years. However, with the growing population we need to increase crop production, but increasing crop productivity while ensuring food security and decreasing the environmental impacts of agriculture is a major challenge for the coming decades. Increased productivity was previously mainly achieved through improvement of crop varieties and increased use of inorganic fertilisers and pesticides. Although successful, the long-term sustainability of the strategy is debatable, mainly due to its environmental impacts. The question we face is: how can the Earth provide food security for the growing human population, and continue to be a safe place for humanity? This is BioClub’s challenge: to use a better product that is able to increment crop production needed to feed the population while minimising environmental impact.