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The 3-ketoacyl-CoA synthase WFL is involved in lateral organ development and cuticular wax synthesis in Medicago truncatula

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A 3-ketoacyl-CoA synthase involved in biosynthesis of very long chain fatty acids and cuticular wax plays a vital role in aerial organ development in M. truncatula.

Abstract

Cuticular wax is composed of very long chain fatty acids and their derivatives. Defects in cuticular wax often result in organ fusion, but little is known about the role of cuticular wax in compound leaf and flower development in Medicago truncatula. In this study, through an extensive screen of a Tnt1 retrotransposon insertion population in M. truncatula, we identified four mutant lines, named wrinkled flower and leaf (wfl) for their phenotype. The phenotype of the wfl mutants is caused by a Tnt1 insertion in Medtr3g105550, encoding 3-ketoacyl-CoA synthase (KCS), which functions as a rate-limiting enzyme in very long chain fatty acid elongation. Reverse transcription-quantitative PCR showed that WFL was broadly expressed in aerial organs of the wild type, such as leaves, floral organs, and the shoot apical meristem, but was expressed at lower levels in roots. In situ hybridization showed a similar expression pattern, mainly detecting the WFL transcript in epidermal cells of the shoot apical meristem, leaf primordia, and floral organs. The wfl mutant leaves showed sparser epicuticular wax crystals on the surface and increased water permeability compared with wild type. Further analysis showed that in wfl leaves, the percentage of C20:0, C22:0, and C24:0 fatty acids was significantly increased, the amount of cuticular wax was markedly reduced, and wax constituents were altered compared to the wild type. The reduced formation of cuticular wax and wax composition changes on the leaf surface might lead to the developmental defects observed in the wfl mutants. These findings suggest that WFL plays a key role in cuticular wax formation and in the late stage of leaf and flower development in M. truncatula.

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Acknowledgements

We are grateful to Dr. Xianpeng Yang (Shandong Normal University) and Dr. Fei Li (Service Center for Experimental Biotechnology) for their assistance in wax constituent analysis. The project was jointly funded by the National Natural Science Foundation of China (Grant No. U1702234 and 31700285), Yunnan Applied Basic Research Projects (2018FB037), and the CAS “Light of West China” Program.

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  1. Tianquan Yang and Youhan Li have contributed equally to this work.

Authors and Affiliations

  1. CAS Key Laboratory of Tropical Plant Resources and Sustainable Use, CAS Center for Excellence in Molecular Plant Sciences, Xishuangbanna Tropical Botanical Garden, Chinese Academy of Sciences, Kunming, 650223, China

    Tianquan Yang, Youhan Li, Yu Liu, Liangliang He & Jianghua Chen

  2. Germplasm Bank of Wild Species, Kunming Institute of Botany, Chinese Academy of Sciences, Kunming, 650204, China

    Tianquan Yang

  3. Key Laboratory for Forest Resource Conservation and Utilization in the Southwest Mountains of China, Ministry of Education, Southwest Forestry University, Kunming, 650224, China

    Aizhong Liu

  4. Noble Research Institute, LLC, 2510 Sam Noble Parkway, Ardmore, OK, 73401, USA

    Jiangqi Wen

  5. Department of Plant and Soil Sciences, Institute for Agricultural Biosciences, Oklahoma State University, 3210 Sam Noble Parkway, Ardmore, OK, 73401, USA

    Kirankumar S. Mysore & Million Tadege

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  1. Tianquan Yang
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Contributions

JC, AL, and TY conceived and designed the experiments; TY, YHL, YL, and LH performed the experiments and statistical analysis; JW, KSM, and MT provided plant material and sequence analysis. JC, TY, and YHL wrote the paper. All authors read the manuscript and agree with its content.

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Correspondence to Jianghua Chen.

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Yang, T., Li, Y., Liu, Y. et al. The 3-ketoacyl-CoA synthase WFL is involved in lateral organ development and cuticular wax synthesis in Medicago truncatula. Plant Mol Biol 105, 193–204 (2021). https://doi.org/10.1007/s11103-020-01080-1

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