Limitations of Current Agricultural Facilities:

Agriculture is a body of work that depends heavily upon the use of pesticides. No matter how efficient pesticides may be, the chemicals that are permeated through its application can cause severe issues to both humans and the environment. Chemicals present in pesticides, such as paraquat, have shown a direct negative impact upon the consumer when directly ingested. These chemicals can cause a variety of diseases including heart failure, kidney failure, and liver scarring. Alongside these unintended side effects on humans, these chemicals also have the capacity to pollute the surrounding environment, wreaking havoc upon the ecosystem of soils, causing a drop in soil efficiency, subsequently lowering crop yield. Thailand, being a country that depends on an agricultural based economy, relies heavily upon the usage of pesticides. Especially with the rapid economic development Thailand has undergone in the past few decades, our agricultural industry continues to grow swiftly. This growth is hastened by pesticide utilization, as the usage has increased significantly over these couple of decades. This startling increase is a cause of concern seeing as more farmers and workers are exposed to the harmful chemicals. Thus, alternatives to these pesticides must quickly be found before further damage is done to the agricultural community and the environment that surrounds the farmlands.

Pictures taken from: says, Bruce, et al. “The Truth about Pesticide Use in Thailand.” The Thailand Life | Award-Winning Thai Blog, 11 Apr. 2018, https://www.thethailandlife.com/truth-about-pesticides-thailand.

Farmland Nemesis:

On the topic of pesticides, the majority of the general public think of pesticide as anti-insect, which is only correct to a certain extent. Insects and pests aren’t the only two problems faced when trying to keep crops healthy. Alongside these two hindrances, farmers also have to deal with bacteria that is capable of causing infectious diseases on plants. Ralstonia solanacearum, is an example of these disease-causing bacterias. Bacterial wilt, a disease caused by R. solanacearum, is a fatal infection that affects water uptake of the plant, causing malnutrition and eventually death within the crop. Alongside this malignant effect, bacterial wilt is easily spreadable throughout entire fields, making it extremely detrimental towards farmlands if not contained properly and immediately. Bacterial wilt is an example of why pesticides are so crucial. Without the help of pesticides, containment of plant infections would be exceedingly difficult which could result in large losses of crop yield. On one hand, this may highlight the significance of pesticides, but it also shows how overly dependent and reliant we have to be on the usage of these chemicals. Knowing this, we must find a more sustainable and safer alternative for pesticides that can yield the same results.

Picture taken from: “Ralstonia Solanacearum.” Wikipedia, Wikimedia Foundation, 30 Sept. 2022, https://en.wikipedia.org/wiki/Ralstonia_solanacearum.

Significance of Bombolitin:

Most cells are protected and regulated through a phospholipid bilayer. This bilayer acts as the cell’s shield and fence, inhibiting certain objects from entering, while also holding the shape of the cell together. Bombolitin holds the ability to interact with this phospholipid bilayer, allowing for disruption within the cell and eventually cell lysis. This holds especially true for cell membranes with more negative charges, specifically Ralstonia. Since Ralstonia contains a negatively charged cell membrane, the positively charged bombolitin will be attracted to the membrane of the Ralstonia, allowing the peptide to interact with the lipid bilayer of the bacteria. These interactions will result in the aforementioned cell membrane disruption and lysis, making bombolitin capable of inhibiting Ralstonia activity. This capability allows bombolitin to be a potential safer alternative to pesticides when targeting R. Solanacearum.

Our Goal:

Having said that, one main issue is the production of bombolitin. Prior techniques of production entails harvesting the bombolitin from bee venom itself, however, this is extremely costly and inefficient as it doesn’t yield the necessary amount of peptide. Hence, our goal is to find a way to directly express the bombolitin genome sequence via synthetic biology. Through various steps of digestion, ligation, and transformation, we’ll use competent cells to explicitly produce bombolitin enabling more efficient production of the peptide itself.

Pictures taken from:“Bl21 (DE3) Chemically Competent E. Coli Cells.” GoldBio, https://www.goldbio.com/product/14440/bl21-de3-chemically-competent-e-coli-cells.