Cloning and expression of phenylalanine ammonia-lyase (<i>PAL</i>) gene in Chinese water chestnut

SONG Mubo; SHUAI Liang; DUAN Zhenhua; QIN Li; LI Lin; CHEN Zhenlin; FANG Fang

doi:10.7671/j.issn.1001-411X.201804020

Journal of South China Agricultural University > 2019 > 40(1): 77-83. > DOI: 10.7671/j.issn.1001-411X.201804020

SONG Mubo, SHUAI Liang, DUAN Zhenhua, et al. Cloning and expression of phenylalanine ammonia-lyase (PAL) gene in Chinese water chestnut[J]. Journal of South China Agricultural University, 2019, 40(1): 77-83. DOI: 10.7671/j.issn.1001-411X.201804020

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Cloning and expression of phenylalanine ammonia-lyase (PAL) gene in Chinese water chestnut

SONG Mubo^{1, 2,},
SHUAI Liang^{1, 2},
DUAN Zhenhua^{1, 2},
QIN Li¹,
LI Lin¹,
CHEN Zhenlin¹,
FANG Fang^{1, 2, ,}

1.
Institute of Food Science and Engineering Technology, Hezhou University, Hezhou 542899, China
2.
Guangxi Talent Highland of Preservation and Deep Processing Research in Fruit and Vegetables, Hezhou 542899, China

More Information

Received Date: April 08, 2018
Available Online: May 17, 2023

Graphical Abstract

Abstract

Abstract

Objective
To clone full-length cDNA of phenylalanin ammonia-lyase (PAL) in Chinese water chestnut (Eleocharis tuberosa), investigate its sequence characteristics, analyze its expressions in different organs and during storage of fresh-cut E. tuberosa, and provide theoretical references for revealing the mechanism of E. tuberosa etiolation.

Method
The full sequence of PAL gene was cloned from E. tuberosa by RT-PCR and RACE techniques. Bioinformatics method was used to analyze the cDNA sequence and the encoded amino acid sequence. Real-time PCR was used to analyze the expression profile of PAL gene in different organs and in fresh-cut E. tuberosa during storage.

Result
The full-length cDNA of PAL gene in E. tuberosa was cloned and named CwPAL. The sequence consists of 2 485 bp with an intact open reading frame of 2 142 bp, encoding a polypeptide of 713 amino acids. The formula of CwPAL protein is C₃₄₃₇H₅₅₁₄N₉₄₄O₁₀₅₈S₃₄, the relative molecular weight is 78 079, pI is 5.97, and the total number of atoms is 10 987. CwPAL protein contains two functional domains including PAL-HAL and PLN02457, and the typical PAL active site sequence (GTITASGDLVPLSYIAG). The main structural element in CwPAL secondary structure is α-Helix, the three-dimension structure of CwPAL protein shows a typical “sea horse” shape. The phylogenetic analysis showed that CwPAL was very closely related to PAL proteins of Ananas comosus and Phoenix dactylifera. The Q-PCR analysis showed that the expression level of CwPAL gene was the highest in peel, CwPAL gene expression quickly increased during storage of fresh-cut E. tuberosa, and salicylic acid treatment significantly inhibited CwPAL gene expression.

Conclusion
The cloned CwPAL gene is a typical member of PAL family, and CwPAL gene may affect E. tuberosa etiolation by regulating the phenylpropanoid pathway.
- Eleocharis tuberosa,
- phenylalanine ammonia-lyase,
- gene cloning,
- expression analysis,
- etiolation,
- salicylic acid

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References (35)

References

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Cloning and expression of phenylalanine ammonia-lyase (PAL) gene in Chinese water chestnut

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