Soybean is a powerhouse of plant-based protein and oil. It packs a punch with 35-45% protein and 18-22% oil. However, there is a catch. A substance called Kunitz Trypsin Inhibitor (KTI) lurks within, making it tough for our bodies to use the protein effectively. This is where genetic science comes in.

The goal was clear: create soybeans without KTI. This would make the soybeans more nutritious and appealing to a global audience. The process involved a clever mix of plant breeding and genetic screening. Two sets of parent plants were chosen. The first set, known as recipient parents, included AMS-MB-5-18 and AMS-MB-5-19. The second set, the donor parents, were NRC-101 and NRC-127. These pairs were crossed to produce four unique combinations.

The next step was to identify plants carrying the desired null KTI allele. This was done through marker-assisted selection across several backcross generations. A whopping 342 SSR markers were used to check for genetic diversity and to ensure the new plants closely resembled the original parents. This was crucial for maintaining the desirable traits of the parent plants.

The process yielded 60 F1 seeds, with a success rate of 12%. Backcrossing continued, resulting in 259 BC1F2 plants, 45 BC1F3 plants, and finally, 8 BC1F4 plants that carried the null KTI allele. The recovery of the recurrent parent genome content increased with each generation. By the BC1F4 stage, Cross A (AMS MB 5-18 X NRC 101) achieved 84. 15% recovery, while Cross B (AMS MB 5-18 X NRC 127) reached 83. 72%.

The absence of the KTI peptide was confirmed using native polyacrylamide gel electrophoresis. This method is a reliable way to check for the presence of specific proteins. Additionally, the expression of the KTI3 gene was studied during two key seed developmental stages, R5 and R6. The results showed that the KTI3 gene was more active during the R5 stage. This finding is important for understanding how to control the expression of this gene in future soybean varieties.

The null parent lines showed minimal KTI3 expression, which aligns with their genetic makeup. This confirms that the introgression of the null KTI allele was successful and had the desired effect on the trait in advanced backcross populations. The journey to improve soybean nutrition is ongoing, but these findings are a significant step forward.

The Quest to Boost Soybean Nutrition

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