Cloning and Functional Analysis of Glyoxalase I Gene BrGLYI 13 in Brassica rapa L.

Glyoxalase I (GLYI) is a key enzyme that detoxifies methylglyoxal, a toxic byproduct of glycolysis, and is essential for plant pollination. However, the genome-wide identification and functional analysis of <i>GLYI</i> in <i>Brassica rapa</i> L. (<i>B. rapa</i>) remain limited. This study identified 17 <i>BrGLYI</i> genes (<i>BrGLYI1</i>-<i>BrGLYI17</i>) from the <i>B. rapa</i> genome. The self-compatible line 039-1 and the self-incompatible line GAU-28-5 were used as experimental materials, and Real-Time Quantitative Reverse Transcription PCR (RT-qPCR) was performed to examine the effect of <i>BrGLYI</i> genes on self-compatibility in winter <i>B. rapa</i>. Preliminary results showed that <i>BrGLYI13</i> exhibited significant tissue specificity, with higher expression in the flowers of 039-1 compared to GAU-28-5. The open reading frame of <i>BrGLYI13</i> (852 bp) was cloned from both 039-1 and GAU-28-5 cDNA, with no base mutations observed between the two lines. RT-qPCR revealed higher <i>BrGLYI13</i> expression in the stigma of 039-1 compared to GAU-28-5. Based on the functional conservation and sequence homology, <i>BrGLYI13</i> is speculated to play a similar role to that of <i>AtGLYI3</i> in methylglyoxal detoxification and stress response. Furthermore, the knockout of <i>AtGLYI3</i> resulted in reduced silique lengths and seed numbers. These findings suggest that <i>BrGLYI13</i> is involved in the self-compatibility response in <i>B. rapa</i> and promotes the silique length and seed number in the Arabidopsis mutant, providing a basis for further research on the mechanisms of self-compatibility in <i>B. rapa</i>.