A Bacterial Indole-3-acetyl-L-aspartic Acid Hydrolase Inhibits Mung Bean (Vigna radiata L.) Seed Germination Through Arginine-rich Intracellular Delivery |
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Authors: | Kevin Liu Han-Jung Lee Sio San Leong Chen-Lun Liu Jyh-Ching Chou |
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Institution: | (1) Department of Life Science, National Dong Hwa University, Shou-feng, Hualien, 97401, Taiwan |
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Abstract: | Indole-3-acetyl-L-aspartic acid (IAA-Asp) is a natural product in many plant species and plays many important roles in auxin
metabolism and plant physiology. IAA-Asp hydrolysis activity is, therefore, believed to affect plant physiology through changes
in IAA metabolism in plants. We applied a newly discovered technique, arginine-rich intracellular delivery (AID), to deliver
a bacterial IAA-Asp hydrolase into cells of mung bean (Vigna radiata) seeds and measured its effects on mung bean seed germination. IAA-Asp hydrolase inhibited seed germination about 12 h after
the enzyme was delivered into cells of mung bean seeds both covalently and noncovalently. Mung bean seed germination was delayed
by 36 h when the enzyme protein was noncovalently attached to the AID peptide and longer than 60 h when the enzyme protein
was covalently attached to the AID peptide. Root elongation of mung bean plants was inhibited as much as 90% or 80%, respectively,
when the IAA-Asp hydrolase was delivered with the AID peptide by covalent or noncovalent association. Further thin-layer chromatography
analysis of plant extracts indicated that the levels of IAA increased about 12 h after treatment and reached their peak at
24 h. This result suggests that IAA-Asp hydrolase may increase IAA levels and inhibit seed germination of mung bean plants
and that the AID peptide is a new, rapid, and efficient experimental tool to study the in vivo activity of enzymes of interest in plant cells. |
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Keywords: | Hydrolase IAA IAA conjugate Intracellular delivery Mung bean Seed germination Vigna radiata |
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