Science

#Chlamystress

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#Chlamystress in a nutshell

Green algae are not only one of the best carbon dioxide traps, but also perfect producers of oils and sugars that can be used for biofuel production. The current problem is that these molecules, sugars and oils, are only accumulated when the cells are stressed. This stress is very easy to apply in the lab, but very difficult and expensive at industrial level (we are talking about 2-3000 L of culture).

During my work I have discovered some genes that are activated under stress. It seems that these genes control the response to stress, and in consequence, the accumulation of oils and sugars. I have results that support this hypothesis, but I need to prove this to have no doubt. (this is the aim of this project). This is very important because if this hypothesis is true, we could create Chlamydomonas lines that express these genes constantly. The constant expression of these genes will make possible the accumulation of oils and sugars without needing to stress the algae, reducing costs and increasing productivity.

The extensive use of this technology will replace the use of foods (corn, soybean) for producing biofuels and biomass, avoiding the current "fuel or food" ethical issue. Furthermore algae are more efficient, requiring fewer resources. This is (will be) the third green revolution. But some steps must be walked before. This project could be one of them. I need your help for continue in this research line and free this knowledge.

Background and motivation:
Chlamydomonas reinhardtii is a unicellular freshwater green algae (technically is a chlorophyte). This microalgae is actually a promising alternative to produce biofuels and biomass, acting as a powerful CO2 sink and being a source of renewable energy. This is a novel alternative in biofuels production, which overcomes some plant limitations and the criticism and ethics limitations of using classic biofuel crops (important fractions of the land to grow food are being used to ‘grow’ fuel for the gas tank, increasing the food–crisis, especially in already poor countries).
It is known that stress situations induce the production of hydrogen, lipids (basis of biodiesel) or sugars and starch (basis of bioethanol production) in this species. For example nitrogen starvation induces the formation of lipids and cold induces the accumulation of sugars and starch, but little is known about how Chlamydomonas senses the environment and re-program its metabolism towards the accumulation of this valuable molecules. The knowledge of these mechanisms will represent a stepping stone for designing new cultivation strategies and obtaining more productive strains.

What do we know? What do we want to know? Why do I need your support?
My current project aims to study how the Chlamydomonas cells response and adapt to different kind of stresses (heat, cold, starvation, UV radiation,...). How do these cells adapt to survive in non optimal conditions? What is changing inside them? It is clear that to survive to these stresses the Chlamydomonas cells should adapt. How do the cells adapt? In brief, the first step of adaptation is sensing that the environment has changed and then transmit this information to the nucleus (the "headquarters" of the cell). In the headquarters the stress is analyzed and a response plan is defined. The messages for carrying out the response plan are then written and sent back. These messages have orders for producing new workers (proteins) that will product the necesarry products (metabolites) to survive to the stress.
I started studying the final cellular response, the changes in the metabolites (sugars, lipids, pigments,...), under the different stresses. We have discovered some new compounds that are stress-responsive and also determined the changes in some sugars, lipids and organic acids. Some of these compounds are very interesting for us, like the lipids and the sugars (thinking in biofuel and bioethanol). It would be great if we could manage to produce more of these sugars and lipids by the Chlamydomonas, but before doing that we need to know how the cell is doing for accumulating these molecules. This is why we have also studied changes in the proteins ("the workers") and in the transcripts ("the messages" that the nucleus send to produce new "workers"). We have seen very different responses at protein and transcript levels, most of them were known in other species but we get some specific to Chlamydomonas and some completely new (not described anywhere).
During the development of this project we have discovered a small group of genes that appears to be part of the master metabolic switch under stress conditions. We think that when these genes are "switched on" they produce "internal orders" that organize what should be changed to adapt to the stress. Different stress should be managed by different organizers and get a different adaptative response. We have get promising results in which we saw that the adaptative response is different and specific to each stress, and we have also found different organizers. So at this moment we have strong experimental evidences that supports this hypothesis. But we need more experiments to confirm it. We need your help for making this research possible:
First of all we need to test again when this genes are switched on an off (sending orders or not) and at which speed (number of messages they sent). This is important, because the larger number of copies of the order the larger number of proteins (workers) that will be ready in the cell (technically: we need to test the expression levels of the genes in this family employing RT-qPCR). But getting more "workers" is not a conclusive proof to determine the function of the gene (you can have lots of workers, but you don't know if they are really working). We also need to see what happens if we block these orders. If we block this orders, theoretically, no workers will be available so probably we won't see a complete adaptative response (technically: I plan to use RNAi to block the expression of these genes). Comparing the response to this stress between Chlamydomonas that are using this gene and that are not we will determine the function of these genes. We want to see which sugar, or which lipid or whatever is changed.

Why 5000?
The experiments I have plan are not very expensive, but they are not cheap enought than I can "sneak" enought money from my current project.

With your help I will buy some small equipment, a license for using Geneious, and part of the chemicals and kits that I need for doing this research. This project depends mostly on your support, please contribute.

Why this research is important? Why would you consider supportting this project?
If we can confirm that these organizer-genes are really organizing the adaptation to the stress in Chlamydomonas it will be an extremely important discovery. This would be the first time in which these new organizers are described and characterized in a wide range of stresses. Furthermore once we know the organizers, we will be able to to select the best organizers for our interests (i.e. production of lipids, sugars, both, ...). Then we could easily made these genes hyper-functional for sending orders constantly to the cell for an increased production.

It is very important that this knowledge is free for everybody (not property of one or two big companies) since the confirmation of the function of these genes, and its relation to sugar and lipid accumulation, will open a new door to extend the ongoing second green revolution. Extensive use unicellular green algae to harness the solar power and exploit it in the form of biomass has the further advantages of being cheap, not contributing to the food vs fuel dilemma, in addition to trapping CO2 from the athmosphere and to produce value-added co-products like hydrogen, sugars or lipids for biofuel. All the countries and people should have the right to use this promising technology and this could be one initial step.