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Identification and functional characterization of a new flavonoid synthase gene MdFLS1 from apple

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Abstract

Main conclusion

The flavonoid synthase gene MdFLS1 from apple, which possibly plays an important role in anthocyanin synthesis, accumulates in the purple-red branches of Malus ‘Pink spire’.

Abstract

Flavonoid metabolism serves an important function in plant growth and development. In this study, we selected 20 varieties of apple lines, 10 green and ten red branches, from the plant nursery of Qingdao Agriculture Academy. Metabolite analysis revealed that large amounts of anthocyanins accumulated in the purple-red branches of M. ‘Pink spire’. Real-time polymerase chain reaction showed that the expression of the flavonol synthase gene MdFLS1 was over 1500-fold higher in M. ‘Pink spire’ than in the other varieties. A single base A was inserted at the first three bases of the active binding site of MdFLS1 to prove that the purple-red colour of apple leaves and stems in M. ‘Pink spire’ may be caused by the inactivation of MdFLS1 protein. The results of in vitro enzymatic reaction revealed that the MdFLS1 protein lost its activity. MdFLS1 was expressed in Arabidopsis thaliana to explore further its functions. High-expression wild-type strains (OE1 and OE2) and high-expression strains of A-base insertion (A-OE1 and A-OE2) were obtained. Compared with the wild-type strains, the overexpression lines showed lighter tissue colour and less accumulation of anthocyanins. However, A-OE1 and A-OE2 showed no difference in colouration. In conclusion, we speculated that the MdFLS1 gene in M. ‘Pink spire’ cannot bind flavonoids, triggering the synthesis of anthocyanins in another branch of the flavonoid metabolic pathway and resulting in the purple-red colouration of apple leaves and stems. These results suggest that MdLS1 is a potential genetic target for breeding high-flavonoid apples in future cultivar development.

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Abbreviations

CHS:

Chalcone synthase

DFR:

Dihydroflavonol-4-reductase

DHQ:

Dihydroquercetin

FLS:

Flavonol synthase

F3H:

Flavanone 3-hydroxylase

PAL:

Phenylalanine ammonia lyase

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Acknowledgements

This project was supported by grants from the National Natural Science Foundation of China (Grant No. 32001982, 31701827, 31772254, 31972357), the Shandong Provincial Natural Science Foundation of China (Grant No. ZR2019PC041, ZR2017CM009) and the China Postdoctoral Science Foundation (Grant No. 2020M671984). Supported by the Open Project Program (2020KF09) of State Key Laboratory of Crop Biology, SDAU, Tianan, Shandong.

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  1. Pan Li and Kang Lei contributed equally to this work.

Authors and Affiliations

  1. School of Pharmacy, Liaocheng University, Liaocheng, Shandong, 250000, People’s Republic of China

    Pan Li, Kang Lei & Lusha Ji

  2. State Key Laboratory of Crop Biology, College of Life Sciences, Shandong Agricultural University, Tai’an, Shandong, 271018, People’s Republic of China

    Lin Liu, Chengchao Zheng, Huairui Shu & Shizhong Zhang

  3. School of Pharmacy, Linyi University, Linyi, Shandong, 276000, People’s Republic of China

    Guizhi Zhang

  4. Qingdao Agriculture Academy, Qingdao, Shandong, 266100, People’s Republic of China

    Hongjuan Ge

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  1. Pan Li
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Li, P., Lei, K., Liu, L. et al. Identification and functional characterization of a new flavonoid synthase gene MdFLS1 from apple. Planta 253, 105 (2021). https://doi.org/10.1007/s00425-021-03615-2

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