Towards a sustainable control of wheat dwarf virus disease in China and Norway by application of CRISPR and Next Generation Sequencing

Cereals (wheat, barley and rye) are dominant staple food crop grown worldwide and are among the most important crops in the world. In Norway and China, wheat has significant importance as a food source and also has a socio-economic impact on the development of both countries. These cereals are affected by pathogens, insect pests, and weeds. Among them, viruses are the most difficult to control since, unlike diseases caused by fungi, viral diseases cannot be controlled with the use of fungicides. Once a viral disease has established on a field, there are no means by which it can be eradicated. To date, 200 different viral species are known to infect cereals worldwide. Most of the cereal viruses are transmitted by a winged-vector. These vectors, due to climate change, are believed to increase their distribution. Therefore, it of utmost importance to understand the biology of viruses and device methods to control viral diseases that might propose a risk for sustainable agriculture in the near future. In this project, The Norwegian Institute of Bioeconomy Research (NIBIO) and The Chinese Academy of Agricultural Sciences (CAAS) have cooperated to work towards a sustainable control of the wheat dwarf virus disease. We have studied the molecular interactions between the virus, the vector and the host using cutting-edge molecular techniques and discovered various plant proteins that might have an important role in the transmission of the virus by the vector. In addition, utilizing next generation sequencing, this project has determined various plant genes which seem to be involved in the ability of the virus to infect the plant. Using this data, we have developed plants, gene edited by CRISPR, which are resistant to WDV infection. These plants can be used to reduce yield losses due to viral infection. However, due to the current GMO regulations, even CRISPR modified plant are considered genetically manipulated. We have also developed new systems for Cas9 mediated gene editing in plants such as a base-editing-mediated gene evolution (BEMGE). This will facilitate the generation of gene edited plants and has direct application in the breeding programs. Moreover, we have initiated novel applications of the CRISPR-Cas9 and CRIPSR-Cas13 system focusing on viral replication and viral localization in cells (protoplasts). This is an initial step for the development of system that theoretically allows the elimination of viruses in plant cells, which has great potential in the generation of clean (virus-free) planting material, which is highly demanded by plants producers, breeding companies and farmer.

The results from this project are significantly important for the industry for the following reasons: (1) We have developed plant which are resistant to WDV infection. These plants can be used to reduce yield losses due to viral infection. (2) We have developed new systems for Cas9 mediated gene editing in plants. This will facilitate the generation of gene edited plants and has direct application in the breeding programs. (3) We have initiated novel application of the CRISPR-Cas9 and CRIPSR-Cas13 system focusing on viral replication and viral localization in cells. This is an initial step for the development of system that theoretically allows the elimination of viruses in plant cells, which has great potential in the generation of clean (virus-free) planting material; which is highly demanded by plants producers, breeding companies and farmer. Laboratories in Greece, Czech Republic and the United States will use our developed methods and constructs in their plant related research. Some of the results which we have obtained have provided enough scientific evidence to continue more novel and risky research on the CRIPSR-Cas system, giving us the opportunity to apply not only for applied research funds but also for basic research funds. Further joint application with CAAS, Hebei Plant Protection Institute- Hebei and Southwest University - Chongqing, are in plan. In addition , we're exploring possible joint application with the university of Kentucky for highly competitive NSF grants and joint applications with Greece and Czech Republic to EU grants.

The Chinese Academy of Agricultural Sciences (CAAS) and the Norwegian Institute of Bioeconomy Research (NIBIO) will cooperate to work towards a sustainable control of the wheat dwarf virus disease. Wheat has significant importance as a food source, and also an important socio-economic impact on the development of the two countries. Viruses are the most difficult pathogen to control since viral diseases cannot be controlled by the use of fungicides. Wheat dwarf virus is the most damaging wheat virus in China, and one of the most important wheat-infecting viruses in Norway. The positive outcome of the project will: (1) Contribute towards sustainable agriculture of wheat, the second most grown crop in the world. (2) Demonstrate the value and importance of applying novel, cutting-edge techniques (CRISPR-Cas9) next generation sequencing to plant health and plant protection. (3) In an climate change framework, handle a disease transmitted by a winged-vector, which due to climate change is believed to become an increasing threat to sustainable agriculture all over the world including, China and Norway. (4) Develop resistant varieties of wheat and thus contributing to the establishment of an environmentally friendly agriculture of wheat. The project will result in knowledge that can be directly used to control viral diseases in wheat. In this context, the project will promote information and result sharing to important farmer associations that would largely benefit from project results. The project will also focus on dissemination and transfer of scientific knowledge on sustainable agriculture by education of MSc and PhD students in both countries and thus securing adequate scientific follow up of the project. Finally, it is important to underline that NIBIO and CAAS have ongoing bilateral projects and that this project will pursue and enhance the current fruitful ongoing research cooperation between Norway and China.