Introduction

Feeding all population and doing it sustainably, is a major global issue. Biotechnology provides us with promising strategies to solve those challenges. However, agricultural necessities to efficiently manage crops industry, change under different geographical, social and other context-depending conditions. From a social perspective, living in a globalised world implies that one way or another, we are all connected. It is our responsibility to positively use those connexions.

In HYPE-it, we think that having solutions to problems affecting worldwide makes it mandatory to let these solutions arrive wherever they are asked. This is the reason why our aim is to improve accessibility to CRISPR/Cas9, delegating the improvement of crops to local institutions which are aware of their context-dependent circumstances and necessities.

Why do we study the impact assessments of our project on society?
Impact assessment is a process and analysis that makes possible to know the positive and negative consequences of a proposed project on society, economy and environment. The aim is to predict the impacts at an early stage in project design, allowing to reduce the found adverse impacts, shape the project to suit local environment, society and economy and take into account the opinions and preferences of relevant stakeholders.

HYPE-IT was created in order to overcome some problems that significantly make difficult the local accessibility of precision genetic engineering techniques, particularly CRISPR/Cas9. For us, there is a direct connection between society problems and project goals. We not only decided to choose this project to solve a real problem in society but we also made an impact assessment to fully understand the problems. This allows us to follow a strategy that will make our project to benefit society, economy and environment to the greatest extent.

Types of Impact assessment.

Nutritional Impact Assessment.

According to our purpose, it was mandatory to study the influence that a project has in a healthy and correct nutrition. Improvements should be translated in better data regarding child development, aging and incidence rates of diseases directly related to proper nutrition. Nevertheless, diet issues differ relying on the country or even between regions. This is the reason why it is important to identify nutritional necessities before analyzing the impact, as there is not a concrete global solution for all contexts.

Environmental Impact Assessment.

Resources required and environmental consequences of a project are technical issues which determine its success. The goal is using natural resources in a safe and optimum manner, without compromising autochthonous flora, fauna and health of population. The identification, prediction and evaluation of environmental consequences linked to a project, must conceal environmental expertise, project stakeholders and other representative affected nucleus. Thus, analyzing the sustainability and acceptation of proper practices will condition the project viability, as both characteristics will delimit tasks execution.

Socioeconomic Impact Assessment.

Activities performed because of a project, are expected to affect the economic and social context wherever they are located. The scale at which the economic study is carried through, can be from families to small and middle enterprises, or even to the national stock market. Microeconomy will play a role in the macroeconomics evolution, and vice versa, making it necessary to analyze both dimensions.

Three indicators widely used to evaluate this impact are:

• Direct impact. It corresponds to the production (gross added product) and employment generated in those fields which are direct beneficiaries of the project.
• Indirect impact. It involves providers of sectors who are direct beneficiaries of the project.
• Induced impact. It is related to changes affecting the society as a collective, due to the new services and products offered.
• Economic context affects health, education and expansion expectations of a society. The study of shifts induced in social trends, can help to reinforce or redesign the strategy and mission of a project, trying to contribute to individual and collective development.

Scenarios

Recovering crops

Powdery mildew (PM) is a fungal disease caused by a species of fungi in the order Erysiphales, that affects many crops such as cucumber, melon, apple, tomato or cereals. In Europe it is particularly problematic in grapevine, where the parasite covers the leafs and stem of the plant, resulting in damage of the plant and consequently a lower quality of the grape. It becomes acid and contains less anthocyanins and sugar.

In France, Spain, Italy and Germany, between 1992 and 2003, 73% of the fungicides were used only for grapevine protection (1). These fungicides for treating powdery mildew are sulfur-based and can damage the environment. Genetically resistant varieties can be the solution for this problem. These varieties exist but are not accepted by consumers and producers, as the grape has a lower quality and are not the traditional varieties of the region. It has been found that resistance to PM can be achieved by knocking out genes residing in the loci MLO (Mildew Locus O) (2). Using the HYPE-IT technology, it is possible to easily knock out several genes in a grapevine variety which is affected by powdery mildew, without changing the characteristic of the variety.

Nutritional impact:
Grapes and wine have a high content in polyphenols -including anthocyanins-, which have antioxidant properties. These compounds have positive effects in human health in the prevention of diseases as cancer, diabetes and atherosclerosis (3). Grapevines resistant to mildew would avoid losses of this highly valuable product, as well as preventing reduction of the anthocyanin content of the grape. This might reduce the price of grapes and wine, making more accessible for the consumers these healthy products.

Environmental impact:
In 2003, around 60000 tons of sulfur-based fungicides were used, and most of it is applied on vineyards (1). Currently, powdery mildew is controlled using mainly these type of fungicide, which is dispersed into the environment. The fungicide used in vineyards can enter the soil surface and persist for a period of type, damaging the soil and the organisms living in it. It can also migrate due to leaching to groundwater our nearby waterways, contaminating water and being detrimental to aquatic life ( 4). If PM resistant varieties of grape are created, it will not be necessary the use of tons of fungicides that can damage the environment and the ecosystem. Other advantage of the use of HYPE-IT for grapevine improvement is the possibility to conserve biodiversity. It is not necessary for the farmer to change his variety to obtain resistance to PM, as he can improve his own local variety.

Socio-economic impact:
Powdery mildew affecting grapevines can cost annually up to 369$/acre, accounting for loses and expenses in fungicides and its application over the crops. PM can imply around 20% of total grapevine production expenses (5). By using PM resistant varieties, these costs can be reduced, which means lower expenses for the farmer and consequently, cheaper products for the consumer. In 2013, 14 million tons of wine were produced in the European Union, and it is the 3rd most produced processed crop. The same year, it exported 6 million tons (6). These indicators serve to illustrate the economic relevance of grapevine crops and the benefits that would bring resistance to one of its most prevalent diseases, powdery mildew.

Nutritional qualities improvements

Nowadays bad proteo-energetic nutrition is a huge concern in plethora of countries. The consumable items do not provide enough energetic and nutritional values required for a proper development. The consequences also affect the infants causing growth deceleration, energy diminution for activities, more difficulties for learning and even a lower resistance against infections. With HYPE-IT improved crops could provide a better hedge against the previous odds named. Food whose assimilation and processing in the organism will be the best possible, maximizing their nutritive properties and contributing to the improvement of life quality of the people.

As an example for a scenario where crop nutritional improvement is needed, we imagine a farmer from South America, for example Brazil. He wants to grow his own tomato variety (Solanum lycopersicum) but with an increased concentration of b-carotene and lycopene. The amount of cases of deficit in the intake of these two compounds is high in these regions.

e-CYC), carotenoid synthesis is up-regulated in seeds. The higher total carotenoid content resulting from reduction of this protein expression suggests that this gene is a rate-limiting step in the carotenoid biosynthesis pathway. (7) Employing the user-friendly strategy of HYPE-IT this gene can be silenced by knocking it out in order to obtain the desired phenotype. @rhttps://static.igem.org/mediawiki/2016/8/8e/T--Valencia_UPV--humancarotenoid.jp

Nutritional impact:
Carotenoids are essential for the organism as they are antioxidants that contribute to lower the risk of developing cardiovascular illnesses or some types of cancer. Additionally, b-carotenes are precursors of vitamin A, needed for a correct development of the organism. Vitamin A deficiency represents an important problem. It is calculated that 251 million children less than 5 years old have vitamin A deficit. In fact, an estimated 250 000 to 500 000 vitamin A-deficient children become blind every year, half of them dying within 12 months of losing their sight. (8) Moreover, between 1 and 2.5 million of deaths are related to enteric and respiratory diseases caused by this deficiency every year (9)(10)

Environmental impact:
Improving quality of the tomatoes will require a higher demand, and consequently, the production. As a consequence of the overexploitation of the terrain, the farmer will need to increase the expenses in water, fertilizer, pesticides and other investments. The consumption of these natural resources causes environmental problems that need to be taken into consideration. Furthermore, carbon emissions due to fuel and energy expenses will be increased.

Socio-economic impact:
A better tomato nutritional value will have a higher demand in the global market because the farmer will increase his yield, rising his income and possibly, creating new job positions. In a competitive market where fresh and healthy products are highly reckoned these tomatoes could make the difference and becomes an advantage for the seller and the consumer. Besides, the accessibility for this technology allows an increased food quality without being a big issue for the familiar economy.

Increasing productivity

11). One of the biggest problems regarding this issue is the low productivity of some crops. Crops that suffer from this problem need more resources, higher land extension and are prone to be more expensive, compared to High Yielding Varieties of the same crop. @rhttps://static.igem.org/mediawiki/2016/b/b3/T--Valencia_UPV--humancrop.jp

High Yielding Varieties of maize, rice and wheat were developed in the 1960’s (12). These varieties had the common characteristic of being semi-dwarf. This meant that photosynthetic investment in the stem is reduced, and the assimilates are redirected to grain production, increasing the yield of the crop. Later studies found that semi-dwarf varieties had a mutation in the pathway of gibberellin synthesis, making the plant shorter. In particular, semi-dwarf rice has a mutation in gibberellin-20-oxidase gene (Ga20ox) (13). With HYPE-IT, any rice variety which does not have the semi-dwarf phenotype can be edited to obtain the same crop with higher productivity.

This is the case of the well-known Bomba rice variety from Valencia, Spain. It is commonly used for paella, and its organoleptic properties make it very valuable for the consumers. However, this rice is expensive and needs large land extensions due to its low yielding. Bomba rice has a high stem, it lacks the semi-dwarf characteristic. Using HYPE-IT, the Ga20ox gene of this variety could be edited, obtaining a semi-dwarf rice that would maintain the same organoleptic properties with a higher production.

Nutritional impact:
Rice provides 20% of the world’s dietary energy supply (14). It contains vitamins, minerals and a high carbohydrate content. Bomba rice is more appreciated due to its organoleptic properties acquired during cooking. There are many varieties of semi-dwarf rice crops which are cultivated around the world. Hence, this particular variety is not a necessity from the nutritional point of view.

Environmental impact:
Higher Yielding Varieties have the advantage of producing higher amount of grain in the same land extension. The outcome is that less land extension is needed to feed the same amount of population. If higher yielding biotech crops had not been available from 1996 to 2011 thanks to the Green Revolution, an additional 108.7 million hectares of farmland would have been needed to maintain global production levels (15). Land extension is a very valuable resource for the environment. On the other hand, these varieties need more chemical fertilizers, that can damage the soil. As they are more efficient, up to three rounds of cultivation and harvest can be made, deteriorating micronutrient content of the soil.

Socio-economic impact:
Bomba rice is very popular in Spain for its use in the famous dish paella. Nowadays, this variety is expensive compared to others, but it is still considered the rice that should be used to properly cook paella. The increase of productivity while using less land and water can lower its price and spread its use, allowing more population to enjoy its organoleptic properties. This turns out to be a benefit for the farmer and for the consumer, who can find a quality product at a lower price. Additionally, a higher production makes it possible to export traditional and local Valencia rice around the world, extending a cultural dish. As a negative consequence, it is possible that less employees are needed for the cultivation of a crop that consumes less land extension. This can be counteracted by the jobs that can be created as a result of the increasing production.

Organoleptic Properties Improvement

Organoleptic properties constitute the overall appearance of a product: smell, touch, flavor... Undoubtedly, a better appearance means higher consumer acceptance and a significant reduction of food loss and waste due to slower ripening, for example (16). A 2015 report published by FAO estimated that a 40-50% of global fruit is yearly wasted or lost (17).

The following scenario is set in China, the major world fuzzy kiwi (Actinidia deliciosa) producer, with 1.765.847 tons produced in 2013 ( 18). The gene encoding ACS3 is related to the harvesting of several fruits, including kiwis. Using HYPE-IT, they could easily find this information in our software and knockout the gene, slowing kiwi ripening.

Nutritional impact:

Kiwi organoleptic properties are definitely worth to be improved. It is a well-known source of dietary fiber, vitamin E, K, C and carotenoids. Those are needed nutrients in some isolated places where foreign fruits cannot arrive because of their early harvesting during their transport. Improving their organoleptic properties could let their preservation until they reached those destinations. Kiwi seed oil is rich in α-linolenic and Ω-3 fatty acids. These unsaturated acids are helpful in the prevention of cardiovascular diseases, which have significantly increased in China during the last half century. The positive impact of those nutrients could be higher by the improvement of kiwi organoleptic properties, as it may incentivize its intake.

Environmental impact:
In terms of production, a slower fruit ripening is a logistic priority. Concretely in China, the loss rate of fruit is a 20% and barriers imposed to outer food, compromising the fresh fruit transport. Although there may be strict control policy for an optimum layout, there is always a stock being produced. Fruit storage means additional expenses in the control of conditions that affect organoleptic properties, and in 2007, Chinese storage facilities only could accommodate the 15% of production (19)(20). This makes especially difficult the maintenance of small and middle fruit enterprises, which cannot cope a scale in the production with the lack of conservation technology.

Storage costs could be much minor in case of modified kiwis with slower ripening, reducing the risk of losing product, augmenting the stock quality and the competitiveness of final product prices. From the consumer perspective, better organoleptic properties enrich the gastronomic experience, facilitating the intake of positive nutrients mentioned above.

Socio-economic impact:
Closely related with logistic in production, the use of natural resources to reach the expected demand could benefit as well by slowing the harvest losses. A stock with good and lasting quality would lead to a decrease of the fruit amount to be newly produced. Reducing the water and soil used to grow fruit is key for environmentaly friendly strategies. In China, the availability of arable land is also a compromising fact due to the high urbanization and population increase. By slower fruit ripening, resources would be exploited in a more sustainable way, as a higher rate of the product could finally reach the market.

Nutrition is a big part of our life, the gear that allows us to achieve our personal goals every day. If we improve the characteristics of the food that provides that energy, we will increase our life quality.
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References

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