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Australian Journal of Botany Australian Journal of Botany Society
Southern hemisphere botanical ecosystems
RESEARCH ARTICLE

Molecular cloning, expression and sequence analysis of a phenylalanine ammonia-lyase gene from Poncirus trifoliata under iron deficiency

Jian-Fu Li A , Wen-Jun Zheng A , Li Zeng A , Jian-Fu Liu A and Ming-Yuan Wang A B C
+ Author Affiliations
- Author Affiliations

A Department of Horticulture, Huaqiao University, Xiamen, 361021, PR China.

B Engineering Research Center for Biomass Resource Utilization and Modification of Sichuan Province, 621010, PR China.

C Corresponding author. Email: w_mingyuan@163.com

Australian Journal of Botany 62(8) 698-704 https://doi.org/10.1071/BT14251
Submitted: 7 September 2014  Accepted: 5 January 2015   Published: 26 March 2015

Abstract

Phenylalanine ammonia-lyase (PAL) is a specific branch point enzyme of primary and secondary metabolism. It is deemed to play a key role in plant development and defence. Homology cloning of the cDNA sequence of PAL gene, Pt-PAL1, from Poncirus trifoliata found a complete open reading frame (ORF) of 2166 bp, with 721 encoded amino acids. The sequence alignment indicated that the amino acid sequence of Pt-PAL1 shared a high identity with PAL genes found in other plants. Both the dominant and catalytic active sites of Pt-PAL1 were similar to PAL proteins observed in Petroselinum crispum. Phylogenetic tree analysis indicated that Pt-PAL1 was more closely related to PALs in Citrus clementina × C. reticulata than to those from other plants. Real-time polymerase chain reaction showed that the expression of Pt-PAL1 gene in roots under iron (Fe) deficiency (0 μM o,o-FeEDDHA) was significantly higher than that under Fe sufficiency (50 μM o,o-FeEDDHA). The same result was noted for total phenolic content. Phenolic compounds play an important role in response to iron deficiency in Strategy I plants. In the present study, root exudates of Poncirus trifoliata strongly promoted the reutilisation of apoplastic Fe in roots. Furthermore, more Fe was desorbed from the cell wall under Fe deficiency than during Fe sufficiency, indicating a relationship between Fe and total phenolics in Strategy I plants under Fe deficiency.

Additional keywords: citrus, clone, phenolic compounds, real-time PCR.


References

Abadía J, Morales F, Abadía A (1999) Photosystem II efficiency in low chlorophyll, iron-deficient leaves. Plant and Soil 215, 183–192.
Photosystem II efficiency in low chlorophyll, iron-deficient leaves.Crossref | GoogleScholarGoogle Scholar |

Ahmad T, Sablok G, Tatarinova TV, Xu Q, Deng XX, Guo WW (2013) Evaluation of codon biology in citrus and Poncirus trifoliata based on genomic features and frame corrected expressed sequence tags. DNA Research 20, 135–150.
Evaluation of codon biology in citrus and Poncirus trifoliata based on genomic features and frame corrected expressed sequence tags.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3sXmt1Ogs7Y%3D&md5=90fe7996b25ec7ac415dfe44418ca5b5CAS | 23315666PubMed |

Álvarez-Fernández A, Melgar JC, Abadia J, Abadia A (2011) Effects of moderate and severe iron deficiency chlorosis on fruit yield, appearance and composition in pear (Pyrus communis L.) and peach (Prunus persica (L.) Batsch). Environmental and Experimental Botany 71, 280–286.
Effects of moderate and severe iron deficiency chlorosis on fruit yield, appearance and composition in pear (Pyrus communis L.) and peach (Prunus persica (L.) Batsch).Crossref | GoogleScholarGoogle Scholar |

Appert C, Logemann E, Hahlbrock K, Schmid J, Amrhein N (1994) Structural and catalytic properties of the four phenylalanine ammonia-lyase isoenzymes from parsley (Petroselinum crispum Nym.). European Journal of Biochemistry 225, 491–499.
Structural and catalytic properties of the four phenylalanine ammonia-lyase isoenzymes from parsley (Petroselinum crispum Nym.).Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2MXhtVaqtb4%3D&md5=d5c06d8b8e90126b868b069f069bdc77CAS | 7925471PubMed |

Bienfait HF, van den Briel W, Mesland-Mul NT (1985) Free space iron pools in roots generation and mobilization. Plant Physiology 78, 596–600.
Free space iron pools in roots generation and mobilization.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaL2MXlsF2ks78%3D&md5=b449a174522394352d7e6f1655a1329bCAS | 16664289PubMed |

Cesco S, Neumann G, Tomasi N, Pinton R, Weisskopf L (2010) Release of plant-borne flavonoids into the rhizosphere and their role in plant nutrition. Plant and Soil 329, 1–25.
Release of plant-borne flavonoids into the rhizosphere and their role in plant nutrition.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXjtFGks7o%3D&md5=0d55a162a13812435fe393d8e3f24714CAS |

Cochrane FC, Davin LB, Lewis NG (2004) The Arabidopsis phenylalanine ammonia lyase gene family: kinetic characterization of the four PAL isoforms. Phytochemistry 65, 1557–1564.
The Arabidopsis phenylalanine ammonia lyase gene family: kinetic characterization of the four PAL isoforms.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2cXmtVCjsrk%3D&md5=64192c19a09196e829f95a11ee336442CAS | 15276452PubMed |

Durner J, Shah J, Klessig DF (1997) Salicylic acid and disease resistance in plants. Trends in Plant Science 2, 266–274.
Salicylic acid and disease resistance in plants.Crossref | GoogleScholarGoogle Scholar |

Gong XQ, Liu JH (2013) Genetic transformation and genes for resistance to abiotic and biotic stresses in citrus and its related genera. Plant Cell, Tissue and Organ Culture 113, 137–147.
Genetic transformation and genes for resistance to abiotic and biotic stresses in citrus and its related genera.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3sXmt1ykt7w%3D&md5=f7f7aa2b3febc8b5795038de01e1387eCAS |

Guex N, Peitsch MC (1997) SWISS-MODEL and the Swiss-PdbViewer: an environment for comparative protein modeling. Electrophoresis 18, 2714–2723.
SWISS-MODEL and the Swiss-PdbViewer: an environment for comparative protein modeling.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1cXhvFaks70%3D&md5=63347391e1685e0d300557ae98d9ccccCAS | 9504803PubMed |

Hell R, Stephan UW (2003) Iron uptake, trafficking and homeostasis in plants. Planta 216, 541–551.

Hisaminato H, Murata M, Homma S (2001) Relationship between the enzymatic browning and phenylalanine ammonia-lyase activity of cut lettuce, and the prevention of browning by inhibitors of polyphenol biosynthesis. Bioscience, Biotechnology, and Biochemistry 65, 1016–1021.
Relationship between the enzymatic browning and phenylalanine ammonia-lyase activity of cut lettuce, and the prevention of browning by inhibitors of polyphenol biosynthesis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3MXktFais74%3D&md5=d5be04389e391a3592419796838e0b81CAS | 11440111PubMed |

Hsieh LS, Hsieh YL, Yeh CS, Cheng CY, Yang CC, Lee PD (2011) Molecular characterization of a phenylalanine ammonia-lyase gene (BoPAL1) from Bambusa oldhamii. Molecular Biology Reports 38, 283–290.
Molecular characterization of a phenylalanine ammonia-lyase gene (BoPAL1) from Bambusa oldhamii.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXhsFSmt7fK&md5=ba1184055fb14984155a55a68749f60bCAS | 20354908PubMed |

Jin CW, You GY, He YF, Tang C, Wu P, Zheng SJ (2007) Iron deficiency-induced secretion of phenolics facilitates the reutilization of root apoplastic iron in red clover. Plant Physiology 144, 278–285.
Iron deficiency-induced secretion of phenolics facilitates the reutilization of root apoplastic iron in red clover.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXls1KjtLc%3D&md5=da5e189034dc5ed48c6e0105ea715e11CAS | 17369430PubMed |

Joos HJ, Hahlbrock K (1992) Phenylalanine ammonia-lyase in potato (Solanum tuberosum L.). European Journal of Biochemistry 204, 621–629.
Phenylalanine ammonia-lyase in potato (Solanum tuberosum L.).Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK3sXhs1ajtLw%3D&md5=8e9405c25ff56d7ac7655ecaaad3e660CAS | 1541277PubMed |

Kaçar YA, Simsek O, Donmez D, Boncuk M, Yesiloglu T, Ollitrault P (2014) Genetic relationships of some citrus genotypes based on the candidate iron chlorosis genes. Turkish Journal of Agriculture and Forestry 38, 340–347.
Genetic relationships of some citrus genotypes based on the candidate iron chlorosis genes.Crossref | GoogleScholarGoogle Scholar |

Kobayashi T, Nishizawa NK (2012) Iron uptake, translocation, and regulation in higher plants. Annual Review of Plant Biology 63, 131–152.
Iron uptake, translocation, and regulation in higher plants.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38Xos1ams78%3D&md5=e386d6e8c6cf094bb23031e09cc8270dCAS | 22404471PubMed |

Kumar A, Ellis BE (2001) The phenylalanine ammonia-lyase gene family in raspberry. Structure, expression, and evolution. Plant Physiology 127, 230–239.
The phenylalanine ammonia-lyase gene family in raspberry. Structure, expression, and evolution.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3MXmvFCrtL0%3D&md5=6d0d7df523c6f5519f7f8eea110c17b7CAS | 11553751PubMed |

Liu R, Xu S, Li J, Hu Y, Lin Z (2006) Expression profile of a PAL gene from Astragalus membranaceus var. Mongholicus and its crucial role in flux into flavonoid biosynthesis. Plant Cell Reports 25, 705–710.
Expression profile of a PAL gene from Astragalus membranaceus var. Mongholicus and its crucial role in flux into flavonoid biosynthesis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28XlvVSrurw%3D&md5=845e2e51f348d660e23c8a810b713908CAS | 16456646PubMed |

Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(T)(-Delta Delta C) method. Methods (San Diego, Calif.) 25, 402–408.
Analysis of relative gene expression data using real-time quantitative PCR and the 2(T)(-Delta Delta C) method.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD38XhtFelt7s%3D&md5=211b4aef8e2790758d48e76af1428fd6CAS |

Long G, Song J, Deng Z, Liu J, Rao L (2012) Ptcorp gene induced by cold stress was identified by proteomic analysis in leaves of Poncirus trifoliata (L.) Raf. Molecular Biology Reports 39, 5859–5866.
Ptcorp gene induced by cold stress was identified by proteomic analysis in leaves of Poncirus trifoliata (L.) Raf.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XksVChtL0%3D&md5=164b32132564a484a919d5a72e86f799CAS | 22205537PubMed |

MacDonald MJ, D’Cunha GB (2007) A modern view of phenylalanine ammonia lyase. Biochemistry and Cell Biology 85, 273–282.
A modern view of phenylalanine ammonia lyase.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXnvVaqsro%3D&md5=6a49ff12ce8f95501c4fcc4c5793dd3eCAS | 17612622PubMed |

Mandal SM, Chakraborty D, Dey S (2010) Phenolic acids act as signaling molecules in plant–microbe symbioses. Plant Signaling & Behavior 5, 359–368.
Phenolic acids act as signaling molecules in plant–microbe symbioses.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXhsFemu7zF&md5=fb14ecc65e076f7101c29e6d052647e2CAS |

Marschner H (1995) ‘Nutritional physiology: mineral nutrition of higher plants.’ (Academic Press: London)

Mellisho CD, González-Barrio R, Ferreres F, Ortuño MF, Conejero W, Torrecillas A, García-Mina JM, Medina S, Gil-Izquierdo A (2011) Iron deficiency enhances bioactive phenolics in lemon juice. Journal of the Science of Food and Agriculture 91, 2132–2139.

Minami EI, Ozeki Y, Matsuoka M, Koizuka N, Tanaka Y (1989) Structure and some characterization of the gene for phenylalanine ammonia-lyase from rice plants. European Journal of Biochemistry 185, 19–25.
Structure and some characterization of the gene for phenylalanine ammonia-lyase from rice plants.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK3cXhslegtrc%3D&md5=2f66873450aead17b36311a2fefa6039CAS |

Pellegrini L, Rohfritsch O, Fritig B, Legrand M (1994) Phenylalanine ammonia-lyase in tobacco. Molecular cloning and gene expression during the hypersensitive reaction to tobacco mosaic virus and the response to a fungal elicitor. Plant Physiology 106, 877–886.
Phenylalanine ammonia-lyase in tobacco. Molecular cloning and gene expression during the hypersensitive reaction to tobacco mosaic virus and the response to a fungal elicitor.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2MXitlensL4%3D&md5=eb2da8bcd528980d336126ddb9dc3c93CAS | 7824656PubMed |

Ritter H, Schulz GE (2004) Structural basis for the entrance into the phenylpropanoid metabolism catalyzed by phenylalanine ammonia-lyase. The Plant Cell 16, 3426–3436.
Structural basis for the entrance into the phenylpropanoid metabolism catalyzed by phenylalanine ammonia-lyase.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2MXhtVKkuw%3D%3D&md5=fdf1e953c9d1b4e0325bedd329ae4953CAS | 15548745PubMed |

Sarma AD, Sreelakshmi Y, Sharma R (1998) Differential expression and properties of phenylalanine ammonia-lyase isoforms in tomato leaves. Phytochemistry 49, 2233–2243.
Differential expression and properties of phenylalanine ammonia-lyase isoforms in tomato leaves.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK1MXjslOhuw%3D%3D&md5=34679b5dd8a8f1cede0d206272cd598bCAS | 9887524PubMed |

Schieber A, Keller P, Carle R (2001) Determination of phenolic acids and flavonoids of apple and pear by high-performance liquid chromatography. Journal of Chromatography. A 910, 265–273.
Determination of phenolic acids and flavonoids of apple and pear by high-performance liquid chromatography.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3MXpslyrtg%3D%3D&md5=5d0862acc436f3e3200c6bcdf444a678CAS | 11261721PubMed |

Song J, Wang Z (2009) Molecular cloning, expression and characterization of a phenylalanine ammonia-lyase gene (SmPAL1) from Salvia miltiorrhiza. Molecular Biology Reports 36, 939–952.
Molecular cloning, expression and characterization of a phenylalanine ammonia-lyase gene (SmPAL1) from Salvia miltiorrhiza.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXjvFaqurs%3D&md5=1a8dd9955bd368079cdfa1318f6fcd5fCAS | 18454352PubMed |

Vinson JA, Cai YX (2012) Nuts, especially walnuts, have both antioxidant quantity and efficacy and exhibit significant potential health benefits. Food & Function 3, 134–140.
Nuts, especially walnuts, have both antioxidant quantity and efficacy and exhibit significant potential health benefits.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XhslaltLw%3D&md5=eb231d2ad3a66c56f94acdfacaa89a11CAS |

Wasternack C, Parthier B (1997) Jasmonate-signalled plant gene expression. Trends in Plant Science 2, 302–307.
Jasmonate-signalled plant gene expression.Crossref | GoogleScholarGoogle Scholar |

Waterman PG, Mole S (1994) ‘Analysis of phenolic plant metabolites.’ (Wiley-Blackwell: Oxford, UK)

Wulandari C, Muraki S, Hisamura A, Ono H, Honda K, Kashima T, Subandiyah S, Masaoka Y (2014) Effect of iron deficiency on root ferric chelate reductase, proton extursion, biomass production and mineral absorption of cirtus root stock orange jasmine (Murraya exotica L.). Journal of Plant Nutrition 37, 50–64.
Effect of iron deficiency on root ferric chelate reductase, proton extursion, biomass production and mineral absorption of cirtus root stock orange jasmine (Murraya exotica L.).Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3sXhsVShtrbF&md5=44152eeb02db33efbf2345dd0b5d0972CAS |

Xu WH, Liu H, Ma QF, Xiong ZT (2007) Root exudates, rhizosphere Zn fractions, and Zn accumulation of ryegrass at different soil Zn levels. Pedosphere 17, 389–396.
Root exudates, rhizosphere Zn fractions, and Zn accumulation of ryegrass at different soil Zn levels.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXmvVOrurw%3D&md5=e8bc4430b9d37a412e5336fb17f392b1CAS |

Zhang FS, Römheld V, Marschner H (1991) Role of the root apoplasm for iron acquisition by wheat plants. Plant Physiology 97, 1302–1305.
Role of the root apoplasm for iron acquisition by wheat plants.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK38XlsFKgtA%3D%3D&md5=9c57c3da382c8aba83eca636a923d431CAS | 16668547PubMed |

Zhang RQ, Zhu HH, Zhao HQ, Yao Q (2013) Arbuscular mycorrhizal fungal inoculation increases phenolic synthesis in clover roots via hydrogen peroxide, salicylic acid and nitric oxide signaling pathways. Journal of Plant Physiology 170, 74–79.
Arbuscular mycorrhizal fungal inoculation increases phenolic synthesis in clover roots via hydrogen peroxide, salicylic acid and nitric oxide signaling pathways.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38Xhs1ShtLvL&md5=bd3ec47cebec2a4eff12744122b00365CAS | 23122788PubMed |

Zhong H, Läuchli A (1993) Changes of cell wall composition and polymer size in primary roots of cotton seedlings under high salinity. Journal of Experimental Botany 44, 773–778.
Changes of cell wall composition and polymer size in primary roots of cotton seedlings under high salinity.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK3sXkvFWmsrs%3D&md5=1a92f4fbb271b7650accd35879c05869CAS |