CRISPR/Cas9 technology implementation for improved resistance to Abiotic Stress in cereals

Project Information

CRISP-4-CROPS

Grant agreement ID: 739778

DOI

10.3030/739778

Project closed

EC signature date 1 March 2017

Start date 15 September 2017

End date 14 September 2018

Funded under

• INDUSTRIAL LEADERSHIP - Innovation In SMEs

Total cost

€ 72 500,00

EU contribution

€ 72 500,00

Coordinated by

IDEN BIOTECHNOLOGY SL
Spain

Periodic Reporting for period 1 - CRISP-4-CROPS (CRISPR/Cas9 technology implementation for improved resistance to Abiotic Stress in cereals)

Reporting period: 2017-09-15 to 2018-09-14

Summary of the context and overall objectives of the project

Rapidly changing global climate is affecting the crop productivity thus food availability in the world. Global warming, heat stress, drought, salinity etc., affect crop productivity drastically and threaten global food security. Under this situation, there is a great necessity of multidisciplinary research to select/breed high yielding crop cultivars for resistance to these abiotic stresses which can sustain/enhance crop productivity in the farmers’ fields. Cereal crops are the major source of food material and nutritional components for human health and feed for livestock throughout the world. It needs to be mentioned that 1/5th of arable land of the world is arid and semiarid and 2/3 of it is saline thereby affecting crop productivity and nutritional profile severely. Under this situation, the selection of drought and salt tolerant crop cultivars is proposed as a feasible alternative to sustain the productivity of the crops in these regions.
The use of traditional breeding is effective but very slow: around 10 years of research and field experiments.
The implementation of the use of the gene editing techniques requires highly skilled staff. The overall proposal objective is to overcome the barriers to the recruitment of highly qualified PHD or equivalent specialists in gene editing techniques for agriculture applications, in particular the CRISPR-Cas 9 system.
Iden Biotechnology S.L. an agribiotech company focused on the development of products for the agri-food industry aims at exploiting CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats), a newly developed technology for targeted genome modification, to produce new cereal varieties that over-accumulate healthy compounds, improving their nutritional profile. CRISPR/Cas9, gives scientists the ability to more precisely modify DNA by turning genes on or off or editing DNA. There are three main advantages of managing to use this technique for crop improvement: technical, legislative and competitive:
• The results obtained are similar to what could be obtained through natural mutations and conventional breeding, though it is directed, more precise and quick.
• Besides the technical advantages mentioned above, crops modified with this technique may open up new market opportunities, including the European market
• Competitive advantage.

Work performed from the beginning of the project to the end of the period covered by the report and main results achieved so far

Among cereals, maize (Zea mays) is the most produced one (almost 900 million of metric tons per year) and the third most economically valuable cereal, after rice and wheat, according to FAO (FAOSTAT, 2016). Therefore, in CRISPR-4-CROPS framework, IDEN has performed the process for generation of a new maize variety using CRISPR technology.
The 1-year long project was proposed as an initial step for maize improvement. The project was structured in three milestones.
Target gene selection. Initially it was designed to improve one single characteristic of maize. For this purpose, a bibliographic review and a patent analysis for target genes of interest were performed.
In this procedure, other genes of interest related to the production of healthy compounds were also identified. Finally, the set objective was to develop maize lines that over-accumulate healthy compounds, improving the nutritional profile (target gene information is kept confidential to avoid future patentability issues). It is expected that removal of the expression of those genes using CRISPR technology can contribute to develop new maize varieties with the mentioned characteristics.
Gene editing. Two plant transformation methods, Agrobacterium-mediated or biolistic, were used for CRISPR technology implementation but different success ratios depending on this selection were obtained. For this reason, the project was divided in two branches, each including a gene of interest and a transformation methodology.
The project included only the first stages of the maize varieties development as at least three years are needed for the generation of bulk seeds of mutant plants which will be ready to assay for their improved performance. Thereafter, field trials must be performed, which could take between two or three years. Altogether, upon the completion of CRISPR-4-CROPS project, about six years are needed to get a new maize variety.

Progress beyond the state of the art and expected potential impact (including the socio-economic impact and the wider societal implications of the project so far)

Crop improvement during last century has contributed to successfully feed the world population. This success relies not only in a higher yield, but also in a better food quality and a more efficient use of resources.
Gene modification has been widely used along the 20th century to increase the productivity of plants used for agriculture purposes. The fine edition of plant genomes by new technologies such as CRISPR (Clustered Regulatory Interspaced Palindromic Repeats) can contribute to achieve in a more efficient manner the aims of modern agriculture. This technology can be applied to many plant species, including cereals as has been commented before.
Before CRISPR technology, these projects consisted on the random mutagenesis of plant genomes and the detection of a desired characteristic (i.e. a plant with a better performance) among a population of more than 23 k lines, each containing at least 10 plants. In other words, about 230.000 plants had to be sown, grow and analyzed.
The implementation of CRISPR for gene editing has an enormous potential since it is possible to specifically modify a character of an organism. IDEN’s focus was directed to modify a characteristic of maize that would increase its agronomic value (i.e. more nutrient, lower water use) and therefore will increase the market value of the crop. Although the final product in the market would arrive in several years, the first step consists on the optimization of the system, and the edition of the genes in order to validate the interest of their use to obtain better varieties.
With CRISPR technology the investment needed to obtain a mutant plant has been dramatically reduced and makes it feasible for an SME such as IDEN, opening the door for new product development and commercialization.
For these reasons, the objective of CRISPR-4-CROPS project is the generation of a new variant of maize with an increased market value. On one hand, to generate a maize variant that over accumulates nutritional compounds (biofortification). As a parallel project, to generate a maize variant that is more tolerant to abiotic stress as a feasible alternative to sustain the productivity under adverse climate conditions.
Technical feasibility, high specificity, low cost and short generation time compared to other methods derive on an important economic impact.
It is important to consider that the timeline of the project consists mainly in the implementation of CRISPR technology in maize, but after completion there is still a long process to get a commercial product into the market. Even though this is a very initial step in the crop improvement process, the implementation of this technology makes IDEN able to use it for the modification of other characteristics (other target genes), other plant species, even other organisms, in the future. The ability to use CRISPR technology positions IDEN as an innovative SME on the worldwide gene editing scenario and opens new product development opportunities.

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