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-<h1> NitroBLAST: Laying the foundation for nitrogen fixation.</h1>
+<h1>NitroBLAST: Laying the foundation for nitrogen fixation.</h1>
 
 <h2>Background</h2>
 
 <p>The Netherlands has been facing a pressing <strong>nitrogen crisis</strong> for several years. This crisis is largely attributed to the <strong>agriculture sector</strong>, with over 80% of ammonia (a nitrogenous compound) emissions coming from manure <a href="#cite1">[1]</a> and chemical fertilizers <a href="#cite2">[2]</a>. The over-use of fertilizers has a detrimental effect on the environment through the deposition of excess nitrogen oxides and ammonia in the ground, excessively enriching the environment with nutrients promoting uncontrolled plant and algal growth, or eutrophication, a form of nutrient imbalance <a href="#cite3">[3]</a> that negatively impacts the local biodiversity. This highlights the need of the hour: a solution for <strong>increasing global food supply while maintaining environmental standards</strong>.</p>
 
-<h2> Nitrogen Crisis in the Netherlands </h2>
+<h2>Nitrogen Crisis in the Netherlands </h2>
 
     <div class="image-text-wrapper">
       <figure>
@@ -19,7 +19,7 @@
         <figcaption>Figure 1: Nitrogen manure production in kilograms/hectare in the Netherlands in 2010 <a href="#cite8">[8]</a>.</figcaption>
       </figure>
     <p>
-      The Nitrogen Action Programme, introduced by the Dutch government in 2015, aimed at reducing nitrogen deposition, was deemed <strong>insufficient</strong> in 2019 by the council of state. This declaration restricted the building of new residential areas, until the nitrogen emissions were compensated for, further augmenting the ongoing housing crisis of the Netherlands <a href="#cite1">[1]</a>. This emphasizes the urgency of addressing this crisis. <br>
+      The Nitrogen Action Programme, introduced by the Dutch government in 2015, aimed at reducing nitrogen deposition, was deemed <strong>insufficient</strong> in 2019 by the council of state. This declaration restricted the building of new residential areas, until the nitrogen emissions were compensated for, further augmenting the ongoing housing crisis of the Netherlands <a href="#cite1">[1]</a>. This emphasizes the urgency of addressing this crisis. <br> <br>
       On the other hand, to combat global hunger, an increase in global food production is of the essence. This is addressed through the increase in crop yield, which is possible due to the Haber-Bosch process of fertilizer production, where elemental nitrogen is converted into ammonia. <strong>Over-fertilization</strong> and its direct and indirect impact on the environment make agriculture the second leading contributor to short-term <strong>increases in global surface temperature</strong> <a href="#cite4">[4]</a>.
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@@ -27,7 +27,7 @@
     <br>
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       <p>
-        In 2022, Dutch agriculture lost 74% (312,000 tons) of the nitrogen it spread as manure and synthetic fertilizer to the air and soil. Synthetic fertilizer production alone is also the cause of nearly <strong>2% of global CO<sub>2</sub> emissions</strong> <a href="#cite5">[5]</a>. In addition to <strong>water pollution</strong> by leakage of nitrate, <strong>air pollution</strong> due to the conversion to N<sub>2</sub>O leads to a global greenhouse effect equivalent to 10% of that caused by the increase in atmospheric CO<sub>2</sub> <a href="#cite6">[6]</a>. For staple crops like cereals and maize, <strong>up to 40% of a farm’s operating cost is spent purchasing fertilizer</strong> <a href="#cite4">[4]</a>. Rising prices for fertilizer have been one of the problems leading to farmers' protests in Europe, and efforts to reduce nitrogen emissions in the Netherlands have been met with its own wave of protests <a href="#cite7">[7]</a>. <br>
+        In 2022, Dutch agriculture lost 74% (312,000 tons) of the nitrogen it spread as manure and synthetic fertilizer to the air and soil. Synthetic fertilizer production alone is also the cause of nearly <strong>2% of global CO<sub>2</sub> emissions</strong> <a href="#cite5">[5]</a>. In addition to <strong>water pollution</strong> by leakage of nitrate, <strong>air pollution</strong> due to the conversion to N<sub>2</sub>O leads to a global greenhouse effect equivalent to 10% of that caused by the increase in atmospheric CO<sub>2</sub> <a href="#cite6">[6]</a>. For staple crops like cereals and maize, <strong>up to 40% of a farm’s operating cost is spent purchasing fertilizer</strong> <a href="#cite4">[4]</a>. Rising prices for fertilizer have been one of the problems leading to farmers' protests in Europe, and efforts to reduce nitrogen emissions in the Netherlands have been met with its own wave of protests <a href="#cite7">[7]</a>. <br> <br>
         Altogether, there is a clear and urgent need for an alternative environmental-friendly solution to the nitrogen problem. This can not only make a huge impact on the Netherlands, but also globally, by enabling a sustainable and food secure future.
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-<h2> Motivation </h2>
+<h2>Motivation </h2>
 
 <p>Being a team from the Netherlands, we have actively followed the <strong>unfolding of the nitrogen crisis</strong> and seen the farmer's protests on the news. While nitrogen deposition is incredibly harmful to the environment, the Dutch agriculture sector is a big driving factor behind its economy, with <strong>agricultural exports</strong> being worth 124 billion euros in 2023 alone <a href="#cite10">[10]</a>. The Netherlands is also considered one of the <strong>front runners in terms of food and agriculture technology</strong>. The world's first lab-grown meat burger was a Dutch invention, introduced in 2013 <a href="#cite11">[11]</a>. Given our leadership in this field, why not leverage synthetic biology to address the nitrogen crisis? We were inspired by previous iGEM teams such as Wageningen 2021 <a href="#cite12">[12]</a> and Stony-Brook 2023 <a href="#cite13">[13]</a> that have tackled similar challenges, alongside a recent publication in Nature in April <a href="#cite14">[14]</a>.</p>
 
+<h2>Solution</h2>
+
+<div class="image-text-wrapper">
+  <p>
+    The Nature publication by Coale <i>et al.</i> examines UCYN-A, a cyanobacterial species capable of converting N<sub>2</sub> into organic nitrogen, and its relationship with the marine algae <i>Bradurosphera bigelowii</i>. It has already been established that UCYN-A and <i>B. bigelowii</i> have a symbiotic relationship, where <i>B. bigelowii</i> functions as a so-called host, and has taken up the UCYN-A bacteria into its cell in a process known as endosymbiosis. <strong>The symbiont, UCYN-A, fixes nitrogen for the host</strong> whereas <i>B. bigelowii</i> supplies organic carbon and a conducive living environment. This paper proved that UCYN-A is not a common symbiont, but has instead evolved into a eukaryotic organelle for nitrogen fixation, termed the <strong>"nitroplast"</strong> <a href="#cite14">[14]</a>. <br> <br>
+    The discovery of the nitroplast captured our interest - we had considered a project on nitrogen fixation before but failed to see a way in which we could innovate or propose new solutions to the problems previous teams faced. All diazotrophs (bacteria and archaea that fix atmospheric N<sub>2</sub>) use the <strong>enzyme nitrogenase</strong> to fix nitrogen, but the expression of this enzyme presents great difficulties: it is <strong>irreversibly damaged by reacting with oxygen</strong>, while at the same time catalyzing an energetically demanding reaction. Due to this, diazotrophs have evolved very complex mechanisms to couple nitrogen fixation with respiration and/or photosynthesis, which so far has been beyond reach in terms of reproduction by synthetic biologists. The <strong>nitroplast solves this problem, acting as a fully contained compartment within a eukaryote where nitrogen fixation takes place</strong>, utilizing millions of years of evolutionary optimization. <br> <br>
+    Replicating endosymbiosis, while more ambitious than root-bacteria symbiosis, <strong>ensures by design that cell and organelle will work tightly together</strong>, preventing the difficulties associated with either root-dependence or nitrogenase expression. Our ideal <strong>long-term goal would be to introduce this organelle into crops</strong>. By doing this, it may be possible to <strong>reduce the reliance on synthetic fertilizers</strong>, thereby lowering the environmental impact of their production and use and enhancing sustainability in agriculture. This potential for positive change inspired our group to explore this innovative solution further. <br> <br>
+    We are motivated by the vision of making <strong>the first step of what could be one of the biggest contributions to sustainable agriculture in the not-so-distant future</strong>. We believe that the use of the nitroplast's capabilities could lead to more eco-friendly farming practices and help address some of the pressing challenges associated with current fertilization techniques, both in the Netherlands where there is a major nitrogen crisis, and globally where a growing demand for feed crops clashes with a need to reduce greenhouse emissions. Our project aims to harness the power of this organelle to create a <strong>more sustainable and efficient approach to crop cultivation</strong>, ultimately benefiting both the environment and the agricultural industry. <br> <br>
+  </p>
+
+  <figure>
+    <img src="https://static.igem.wiki/teams/5054/farmer-protests-in-the-hague-the-netherlands.jpg" alt='Farmer protests in the Netherlands [9].' width="400" height = "300"/>
+    <figcaption>Figure 2: Farmer protests in the Netherlands <a href="#cite9">[9]</a>.</figcaption>
+  </figure>
+</div>
+
+
     <hr>
     <h2>References</h2>
     <ol>