Vitamin deficiencies plague practically every civilization in the world. Even in countries that are well-off, members of the population still have to watch their food intake and ensure that they are consuming an array of foods in order to meet their needs. Governments and food producers often supplement this by adding necessary vitamins into most foods so that at least some amount is eaten no matter what diet a person has.
And for places that don’t have that cornucopia of options and government assistance and regulation of vitamins? Things often do not turn out well.
The Circumstances of the World
In poor nations around the world, the choices that people have on what food to eat is usually limited and, even for farmers, there is only so much variety that can be maintained. Not all crops grow in every environment, depending on temperature, weather conditions, soil consistency, and many other factors. All in all, these largely limit crop availability to those that were already found in antiquity to grow in those regions.
Additionally, there is a cultural aspect to consider, whereby it is difficult to have a population accept radically new crop staples in lieu of the ones they are already used to eating. Thus, even if different crops with missing vitamin requirements could be grown in such places, their actual consumption by the populace couldn’t be compelled and past evidence has shown that intake of such diversity in poor areas doesn’t occur easily.
It is thanks to all of this that lack of food and malnutrition is rampant throughout the world, with over 800 million people being unable to acquire even the necessary amount of calories needed daily to sustain themselves and over 3 billion people lacking proper access to the vitamins and minerals they require to keep themselves healthy.
The majority of these people live in regions like sub-Saharan Africa and Southeast Asian countries. Even with a sharp decline in poverty rates worldwide thanks to the efforts of governmental bodies like the UN and charities like the Gates Foundation, an increased ability to feed oneself and one’s family has not truly coincided with a decrease in malnutrition rates.
Add to this that subsistence farming have been catastrophically hit in recent years with droughts, flooding, and rapid temperature shifts due to the effects of climate change and it adds up to a world population that is only marginally getting out of the grip of food insecurity.
With the world’s population expected to reach 10 billion by 2050, even with it believed to eventually stabilize at 12 billion by the end of the century, there is a distinct need to rapidly increase food production all around the world and source a more vitamin-rich food supply to those same populations.
A similar event occurred in the middle of the past century, where food security was threatened due to rising human growth. This was dealt with by what is now known as the Green Revolution, where high yielding hybrid crops were introduced into struggling nations, along with fertilizers and pesticides to help protect and improve those yields. The creation of proper irrigation systems that were systematically spread out to maximize individual farmer production also lent a hand to this boom in food creation.
But the revolution has been short-lived. We now sit over just half a century later with a similar encroaching issue that must be figured out and new solutions are needed. Thankfully, we are not without our technological innovations, as much has happened in that half century in the multi-faceted fields of science.
Biotechnology, as one might expect, will be taking the forefront in dealing with the issue of food production and malnutrition.
A Vitamin For Sight
Out of all the deficiencies in the world, those suffering from Vitamin A Deficiency (VAD) have received the most focus. And not without good reason. Hundreds of millions of people suffer from VAD in pockets across the planet and nearly 200 million of those are children. Thanks to a lack of this vitamin in their diet, almost 7 million children die annually from VAD before reaching the age of five.
This crucial ingested compound is not just for the eyes and proper sight formation, but also for general growth and development. Those that survive the condition often do so with some amount of impaired vision, if not outright blindness, a severely weakened immune system, and improper development of bone structure and other features.
So, treating VAD has been a huge focus of the medical and agricultural community for decades and research on biofortified foods with Vitamin A has been ongoing via multiple avenues.
The Golden Rice Project
Within biotechnology, there is none so well known as the Golden Rice project, aimed at helping provide Vitamin A within the most common staple of Southeast Asian countries: rice. Bananas as well have been a focus for Africa and other nations.
Traditional artificial breeding methods have been employed, resulting in the Vitamin A enriched sweet potato 2016 World Food Prize and it has already seen great success in the 10 African countries it has been distributed to. Thanks to all these efforts, along with pill supplementation and other options, 69% of children in all the affected regions have received some sort of Vitamin A in their diet by 2014. But there is still a lot of work to do. Especially since easy breeding of such changes takes decades to accomplish and was only possible with the sweet potato due to its already high amount of Vitamin A.
That’s where biotechnology steps in as another, faster method for making Vitamin A-enriched crops. However, a huge setback for the genetic modification effort has been anti-science actions against biotech, resulting in the holdup of Golden Rice deployment for over a decade at this point. Even though research on the project has been conducted since the early 1990’s and the creation of Golden Rice Version 2 was finished in 2005 (with a 23-fold increase in Vitamin A production), finally meeting 100% Vitamin A incorporation and uptake goals, there has still been no usage in any country around the world. Other than ongoing test plots, such as the one in the Philippines that was vandalized by eco-terrorism group Greenpeace in 2014, there has been no actual usage of Golden Rice for populations afflicted with VAD.
This has been especially idiotic even from the science side of the topic, as even claimed safety statements from activist groups are not meaningful toward how Golden Rice was made. The rice itself already possesses a beta-carotene pathway expressed in the leaves.
The only change made was to add 3 biosynthesis genes to move this production into the endosperm of the rice grain, along with later in the second version adding an enzyme from maize capable of producing higher amount of carotenoids. Those have been the only changes made and they are largely minimal in the scheme of things, especially in comparison to other biotech ventures.
The Australian Banana Project
The other major Vitamin A project that has seen a huge push toward active cultivation is the Vitamin A enriched banana, developed at the Queensland University of Technology in Australia.
The countries that deal in primarily staples involving bananas are usually vitamin rich in a number of ways. This has worked out well for the health of the populations, except in two areas where bananas and the other crops grown in the regions fall flat. Namely, Vitamin A and iron production.
This can be seen to be especially true in East Africa, where the rates of infant mortality and blindness are obscenely high thanks to these malnutritions. Thus, a goal was set. Develop a Cavendish banana with a Vitamin A concentration exceeding 20 micrograms per gram of dry weight. Two different methods were employed by the Australian scientists.
The first uses a synthase gene for the creation of the phytoene carotenoid, obtained from another cultivar of banana from a Fe’i line called Asupina. The Fe’i bananas are a group of disparate cultivars found in the Pacific islands region and are distinct from common bananas and plantains found elsewhere. The Asupina banana produces a high amount of Vitamin A, but due to its other characteristics cannot be grown well outside of its island region.
The second method used the same old phytoene synthase gene obtained from maize and which was originally used in the Golden Rice project. This option also produced high Vitamin A levels in the transformed Cavendish bananas, but suffered from other undesirable traits that made it not as great of an option as the Asupina gene.
The former was able to make a line of Cavendish with 55 micrograms per gram of Vitamin A, far above the desired minimum of 20. Another important result found from the experiment was that early activation of the Vitamin A gene in the banana’s life cycle and an extended length of time given for it to mature was necessary for the bananas to properly accumulate enough Vitamin A in their tissues.
With this success, the Queensland University has teamed up with the Bill and Melinda Gates Foundation in order to begin efforts of getting the bananas approved in East African countries and distributing them to farmers. Since the science has been largely completed in this area, much like with Golden Rice, all that’s left is to get through the politics involved in order to start saving lives. Hopefully Greenpeace won’t stall the proceedings and cause thousands more children to die in the process.
Other Vitamin A Research
Other ongoing past programs for providing Vitamin A have been conducted, even to countries that don’t necessarily need the supplementation. This is in order to avoid any possible complications due to having an unbalanced diet. These programs have focused on inducing such fortification in wheat and maize.
Out of these two, wheat is the more difficult to accomplish, due to the main breeds used not having proper pathways for Vitamin A carotenoid production. Durum wheat, as an example, uses lutein as its primary carotenoid and this does not lend to making Vitamin A. Bread wheat has done slightly better and with a maize gene and some bacterial enzymes of various kinds from multiple independent sets of researchers, they have individually managed to increase the Vitamin A concentration by 60-80 times its original rate.
With all of this talk of using maize genes, one can expect it to have at least some amount of Vitamin A production. And it certainly does, though the amount remains modest. However, since corn is a subsistence crop for millions around the world, it is a great candidate for vitamin biofortification. There is one hitch though.
Vitamin A production only occurs within the yellow breed of corn, causing its tinted hue. White corn, which is often the more common corn eaten by humans and not just given to livestock, lacks this. Thus, having cultures make the transition to using an enriched yellow corn will be the ultimate challenge, likely even more so than the scientific effort involved in making it.
On the topic of that effort, the enhanced corn has been termed Carolight and has been undergoing a field trial in Spain since late 2014. Results published in 2016 indicated that the corn, even while producing high amounts of Vitamin A, was able to grow and have a yield performance matching its non-enriched cousin. This identical feature is a boon to the developers, as it shows that their addition to the corn genome did not impair its performance in other areas.
Additionally, there are examples of beta-carotene being used not as a dietary supplement, but also as a marker for other biotech investigations. A recent study in Camelina, an oilseed crop, found that formation of Vitamin A products can be used as a visual measure of the activation and acquisition of other genes tied to that creation gene during the transgenic transfer.
In short, if the resulting crop turns a yellow color, the scientists and farmers know that it is properly expressing the other trait as well that has been applied to it. This would remove the need for using antibiotic resistance markers, controversial in their own right and difficult to use properly. Furthermore, for those desiring an increased protein content, that appears to also be an effect of adding the genetic material for beta-carotene, while only slightly reducing overall oil content.
A Note On China
A final thing to note before moving on to other topics and vitamins and minerals is the looming shadow of China over this and many other biotech projects. The powerhouse country already is an agricultural monolith as well and it has been reaching its arm into the field of biotechnology for many years now. With its centralized government system, it can and has been producing dozens of specialized projects for its scientists to work on to improve crops in a number of ways. Biofortification being one of them.
Rice, as one might expect, is the staple of staples in Chinese cuisine and many Chinese citizens, especially those in poorer outlying regions, suffer from VAD just as much as those in much poorer Southeast Asian countries. When almost 20% of your entire dietary consumption is rice, that isn’t all that surprising of an outcome. But China appears to be determined to stop this, to support its 1 billion person population with both an ample amount of food and a proper amount of nutrition. So it’s had its eye on the Golden Rice project for some time.
There are also economic considerations to make as well. Since China produces more than 30% of the world’s rice, if it is able to add vitamin-enriched rice to that menu, it can likely charge a premium for it as exports.
As a side note, the only other country that produces more rice is India at 43% and, what do you know, it is also quite keen to properly deploy the enhanced crop. But recent political upheaval and extreme efforts on the parts of anti-science organizations have stalled that for now. In the meantime, they have been moving ahead with the release of modified mustard plants, so maybe the fate of golden rice and other enriched crops will change sometime in the near future.
But back to China. There has been a strong focus of introducing the same gene modifications used in Golden Rice into local rice varieties already designed to grow well in Chinese soil. This includes the two most common cultivars of the region, Indica and Japonica rice, with the former being grown in the central and southern regions and the latter in the northern and eastern areas.
Hybridization methods also allowed the traits to be back-crossed and transferred to a number of specific cultivars used after the gene transfers were made. At the same time, however, they also appear to be working on their own options in-house for Vitamin A rice. Thanks to past controversies, such as a small group of Golden Rice scientists not receiving written consent and approval before conducting a consumption test of the rice with children, the official brand of Golden Rice has become somewhat of a persona non grata within Chinese politics.
Creating their own version of the rice might be more politically advantageous and workable with the public, along with enabling them to be in control over its distribution and use. This has largely been their philosophy toward biotech projects for some time, with China moving toward holding something of a monopoly on various types of crop innovations if they can complete their creations before scientists elsewhere in the world do.
A Micronutrient Extravaganza
Beyond just Vitamin A, though it remains one of the most highly researched areas of vitamin biofortification, there are a number of other essential micronutrients that have been studied. The focus has been on applying these gene enhancements to major staple crops such as wheat, rice, barley, millet, and other such choices.
Iron Collecting
Iron is another nutrient found deprived in multiple cuisines thanks to the local crop availability.
A lack of iron can lead to anemia and eventually death, accounting for about a million deaths annually from iron deficiency alone. Negative health effects in general from the anemia and other symptoms affects about 2 billion people worldwide. The desired intake per day ranges from 10 to 20 milligrams depending on sex, body weight, metabolism, and nutritional needs.
Different scientific endeavors have resulted in a multitude of achievements with iron fortification through very diverse gene choices. One study looked through the iron oxidation pathway within the model organism Arabidopsis, finding which one had the highest iron collecting capability. However, getting this gene to work within the consumable portion of other plants has been difficult. Rice with the gene only saw limited root uptake, while soybeans saw a five-fold increase in iron collection, but only in the leaves and pod walls, not the beans themselves.
Another alternative that has seen more success is a different metal-chelating gene that synthases nicotianamine. A study used this gene from soybeans and inserted it into rice, finding that it was able to work with the similar nicotianamine gene in rice already to produce even more iron. Another study used that rice gene itself and inserted it into wheat, finding that it increased uptake of iron and zinc into the wheat tissues. Bean ferritin genes are also a favorite for transgenically adding to other crops.
Folic Acid Accumulation
Folate is a vital vitamin for good health. Also known as vitamin B9, it is responsible for contributing to cellular division and DNA replication. Proper intake has been known to reduce risk of heart disease and stroke, along with a chronic insufficiency of folate resulting in increased cancer risk due to improper cell reproduction. Dietary intake requirements center around 400 micrograms a day.
The first attempts at metabolic fortification of folate involved expressing a bacterial enzyme involved in folate synthesis within Arabidopsis plants. This prevailed in increasing folate amounts four-fold, with a similar increase shown when the gene was used in tomatoes, maize, lettuce, and the Mexican common bean. But these gains only ever went up to a 9-fold rise, indicating that there was some sort of genetic bottleneck stopping further growth. And 9-fold was not nearly enough for what was needed as a daily intake.
It is believed that this stoppage point was due to a lack of enough precursors for folate creation, as giving the plants more of these precursors did improve folate production. This meant, however, that no matter which folate genes were added and how strong they worked, they would still be stuck with the limiting precursor factor.
More recent research using a multi-gene complex was able to increase folate amount by another 50% over the prior best result, but still short of what was desired. Depending on the crop though, some were found to be able to even go up to a 100-fold increase from their norm, meeting the wanted range.
It may just be that certain crops are more suitable for folate enhancement and future work may need to be focused on maximizing those, at least until another method or gene combination is found that can improve the rest even further.
Building The Building Blocks of Proteins
A last fundamental area to note within prime biofortification research is the improvement of certain essential amino acid production. These are referred to as essential because they are necessary for proper function of the human body. The ones termed non-essential are already processed and created within the body normally, but others cannot be made naturally by the body and must be obtained through food sources.
Out of the 9 essential amino acids, there are 4 that are difficult for plants to create in large amounts and often cause problems within people’s diets around the world. They are Lysine, Methionine, Threonine, and Tryptophan. Sadly, the quantities of these found in most plants are far below the necessary levels for human health. And cultures that are often deprived of a variety of plants suffer even more with that lack.
Thus, enrichment of primary crops has been a focus for some time, with a special focus on lysine, the most limited of the four within critical crops like cereals and legumes. The first place to start for this and the rest of the research has been to understand how the biosynthesis pathways work for each, largely through studying them in Arabidopsis. Understanding each part of their synthesis allows biotech researchers to make genetic changes and improvements to any part of those pathways, as necessary.
The feedback loop for lysine production has been exploited with a bacterial synthase gene and a Arabidopsis gene of a similar style. The latter was required because just using the bacterial gene resulted in other negative growth traits for the plants. A recent study from China using these and other methods was able to create nine transgenic lines of lysine-enriched rice, which saw a 48 to 60-fold increase in lysine amount. In addition to rice, victory had been achieved in maize for lysine enrichment, though no other crops thus far.
Methionine is the next most limited in plants in general and similar Arabidopsis genes have been tried, this time on potatoes, tobacco, and alfalfa, seeing increases from a modest 6-fold to a more robust 32-fold jump. Thus far though, that’s all that has been accomplished largely with this amino acid.
Threonine and tryptophan have an analogous lack of information to report. A five-fold increase of threonine was seen in a tobacco plant experiment and a ten-fold in the same for tryptophan. For now, the amount of research into these topics is minimal, but hopefully more effort will be put into them in the future as more achievements are notched in science’s vest for other biofortification undertakings.
A World To Discover
Even with the myriad of vitamins, minerals, and amino acids discussed in this article, there are still plenty others that aren’t mentioned and that are being pursued by small teams of researchers around the world. Too many to really cover in a single article.
Just know that their struggles are not in vain and progress is being made everywhere all the time. We have already attained so much from existing research. It is somewhat depressing to note though that, even with that done, there has yet to be a single biofortified crop approved for public use anywhere, even though many are ready and waiting to be used.
But that’s all left to the political side of things. Science communicators, including myself, along with scientists, scientific organizations, and many more, will continue our efforts to educate the public, fight back against anti-science fearmongering, and try to make this planet a better place for us and everything else.
If you can do your best to help inform those around you about scientific research such as that discussed above, your small effort may do a lot of good toward finally seeing these crops approved. Thank you for your time.
References
1. Lee, H. (2017). Transgenic Pro-Vitamin A Biofortified Crops for Improving Vitamin A Deficiency and Their Challenges. The Open Agriculture Journal, 11, 11-23. doi:10.2174/1874331501711010011
2. Giuliano, G. (2017). Provitamin A biofortification of crop plants: a gold rush with many miners. Current Opinion in Biotechnology, 44, 169-180. doi:10.1016/j.copbio.2017.02.001
3. Dash, S. K., Sahoo, K. K., Chattopadhyay, K., Behera, L., Pradhan, S. K., Molla, K. A., & Mohapatra, S. D. (2016). Biofortification of Food Crops. Chapter: High Beta-Carotene Rice in Asia: Techniques and Implications. Retrieved June 25, 2017, from https://link.springer.com/chapter/10.1007/978-81-322-2716-8_26
4. Wu, Y., Xu, Y., Du, Y., Zhao, X., Hu, R., Fan, X., . . . Zhao, K. (2017). Dietary safety assessment of genetically modified rice EH rich in β-carotene. Regulatory Toxicology and Pharmacology, 88, 66-71. doi:10.1016/j.yrtph.2017.05.019
5. Hefferon, K. L. (2016). Can Biofortified Crops Help Attain Food Security? Current Molecular Biology Reports, 2(4), 180-185. doi:10.1007/s40610-016-0048-0
6. Steur, H. D., Mehta, S., Gellynck, X., & Finkelstein, J. L. (2017). GM biofortified crops: potential effects on targeting the micronutrient intake gap in human populations. Current Opinion in Biotechnology, 44, 181-188. Retrieved June 25, 2017, from https://biblio.ugent.be/publication/8521462/file/8521465.pdf.
7. Wang, G., Xu, M., Wang, W., & Galili, G. (2017). Fortifying Horticultural Crops with Essential Amino Acids: A Review. International Journal of Molecular Sciences, 18(6), 1306. doi:10.3390/ijms18061306
8. Blancquaert, D., Steur, H. D., Gellynck, X., & Straeten, D. V. (2017). Metabolic engineering of micronutrients in crop plants. Annals of The New York Academy of Sciences, 1390, 59-73. doi:10.1111/nyas.13274
9. Vinoth, A., & Ravindhran, R. (2017). Biofortification in Millets: A Sustainable Approach for Nutritional Security. Frontiers In Plant Science, 8(29). doi:10.3389/fpls.2017.00029
10. Strobbe, S., & Straeten, D. V. (2017). Folate biofortification in food crops. Current Opinion in Biotechnology, 44, 202-211. Retrieved June 25, 2017, from https://biblio.ugent.be/publication/8521453/file/8521454.pdf.
11. Ghag, S. B., & Ganapathi, T. R. (2017). Genetically modified bananas: To mitigate food security concerns. Scientia Horticulturae, 214, 91-98. doi:10.1016/j.scienta.2016.11.023
12. Paul, J., Khanna, H., Kleidon, J., Hoang, P., Geijskes, J., Daniells, J., . . . Dale, J. (2017). Golden bananas in the field: elevated fruit pro-vitamin A from the expression of a single banana transgene. Plant Biotechnology Journal, 15(4), 520-532. doi:10.1111/pbi.12650
13. Dutta, S. S., Das, S., Pale, G., Iangrai, B., Aochen, C., Rai, M., & Pattanayak, A. (2016). Current status and future prospects of research on genetically modified rice: A review. Agricultural Reviews, 37(1), 10-18. doi:10.18805/ar.v37i1.9259
14. Zanga, D., Capell, T., Zhu, C., Christou, P., & Thangaraj, H. (2016). Freedom-to-operate analysis of a transgenic multivitamin corn variety. Plant Biotechnology Journal, 14(5), 1225-1240. doi:10.1111/pbi.12488
15. Yang, Q., Wu, H., Li, Q., Duan, R., Zhang, C., Sun, S. S., & Liu, Q. (2017). Characterization of Agronomy, Grain Physicochemical Quality, and Nutritional Property of High-Lysine 35R Transgenic Rice with Simultaneous Modification of Lysine Biosynthesis and Catabolism. Journal of Agricultural and Food Chemistry, 65(21), 4296-4304. doi:10.1021/acs.jafc.7b00621
16. Schmidt, M. A., & Pendarvis, K. (2017). Proteome rebalancing in transgenic Camelina occurs within the enlarged proteome induced by β-carotene accumulation and storage protein suppression. Transgenic Research, 26(2), 171-186. doi:10.1007/s11248-016-9992-y
17. Singh, S. P., Keller, B., Gruissem, W., & Bhullar, N. K. (2017). Rice NICOTIANAMINE SYNTHASE 2 expression improves dietary iron and zinc levels in wheat. Theoretical and Applied Genetics, 130(2), 283-292. doi:10.1007/s00122-016-2808-x
Photo CCs: Golden Rice from Wikimedia Commons
