Molecular cloning, functional characterization and expression of a drought inducible phenylalanine ammonia-lyase gene (ObPAL) from Ocimum basilicum L.

Khakdan, Fatemeh; Alizadeh, Houshang; Ranjbar, Mahdy

doi:10.1016/j.plaphy.2018.07.026

Cited by 43 publications

(24 citation statements)

References 38 publications

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“…Studies have shown that PAL is an entry-point enzyme which can convert L -Phe into trans-cinnamic acid and that it plays a vital role in channeling carbon flux from primary metabolism into the phenylpropanoid pathway [78]. So far, PAL gene has been cloned from many medicinal plants, such as Ocimum basilicum [79], Ginkgo biloba [80], Salvia miltiorrhiza [81], and A. annua [82]. In G. biloba , the highest expression of GbPAL gene was found in leaves, followed by stems, and the lowest expression was in roots; transcription levels of GbPAL were closely related to flavonoid accumulation [80].…”

Section: Discussionmentioning

confidence: 99%

Analysis of Centranthera grandiflora Benth Transcriptome Explores Genes of Catalpol, Acteoside and Azafrin Biosynthesis

Zhang

Wang

et al. 2019

IJMS

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Cardiovascular diseases (CVDs) are a major cause of health loss in the world. Prevention and treatment of this disease by traditional Chinese medicine is a promising method. Centranthera grandiflora Benth is a high-value medicinal herb in the prevention and treatment of CVDs; its main medicinal components include iridoid glycosides, phenylethanoid glycosides, and azafrin in roots. However, biosynthetic pathways of these components and their regulatory mechanisms are unknown. Furthermore, there are no genomic resources of this herb. In this article, we provide sequence and transcript abundance data for the root, stem, and leaf transcriptome of C. grandiflora Benth obtained by the Illumina Hiseq2000. More than 438 million clean reads were obtained from root, stem, and leaf libraries, which produced 153,198 unigenes. Based on databases annotation, a total of 557, 213, and 161 unigenes were annotated to catalpol, acteoside, and azafrin biosynthetic pathways, respectively. Differentially expressed gene analysis identified 14,875 unigenes differentially enriched between leaf and root with 8,054 upregulated genes and 6,821 downregulated genes. Candidate MYB transcription factors involved in catalpol, acteoside, and azafrin biosynthesis were also predicated. This work is the first transcriptome analysis in C. grandiflora Benth which will aid the deciphering of biosynthesis pathways and regulatory mechanisms of active components.

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Section: Discussionmentioning

confidence: 99%

Analysis of Centranthera grandiflora Benth Transcriptome Explores Genes of Catalpol, Acteoside and Azafrin Biosynthesis

Zhang

Wang

et al. 2019

IJMS

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“…The tertiary structure of PaPAL was homology modeled using Petroselinum crispum PAL as a template, and four 'seahorse shape' subunits were also observed. Thus far, the seahorse shape has been reported for Melissa officinalis [56], Solenostemon scutellarioides [37] and Ocimum basilicum [54]. The results suggest that PaPAL might have catalytic functions similar to those of other PAL proteins.…”

Section: Discussionmentioning

confidence: 76%

“…These sites were also found in the PaPAL protein, suggesting that PaPAL has a similar function to that of other PALs. PaPAL contains five types of modification sites, i.e., N-myristoylation, casein kinase II phosphorylation, protein kinase C phosphorylation, N-glycosylation and tyrosine kinase phosphorylation sites, which are also reported for ObPAL [54]. The first tertiary structure of PAL was determined for that from Rhodosporidium toruloides [55], and the first crystal structure was characterized for that from Petroselinum crispum [41].…”

Section: Discussionmentioning

confidence: 83%

“…The enzyme activity of recombinant PaPAL was determined to be 3224.10 U/mg at pH 8.6. Moreover, the optimal temperature of recombinant PaPAL was 60 • C, which was the same as that of PALs from Salvia miltiorrhiza [60], Jatropha curcas [52], Solenostemon scutellarioides [37] and Ocimum basilicum [54] and close to that of PALs from Petroselinum crispum (58 • C) [70] and Zea mays (55-60 • C) [71] but higher than that of PALs from Bambusa oldhamii (50 • C) [50], Rhus chinensis (45 • C) [67] and Arabidopsis thaliana (31-48 • C) [69]. The enzyme activity of recombinant PaPAL at 60 • C was determined to be 4536.14 U/mg.…”

Section: Discussionmentioning

confidence: 85%

“…When expression of recombinant PaPAL was induced at 16 • C, we found that the amount of soluble protein increased slightly in the supernatant. ObPAL from Ocimum basilicum was expressed in E. coli using the pET-28a(+) vector, and the result showed that the content of recombinant protein was also low [54]. In contrast, the content of recombinant RcPAL was increased in the supernatant, also expressed in E. coli using the pET-28a(+) vector [67].…”

Section: Discussionmentioning

confidence: 99%

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Cloning, Characterization and Expression of the Phenylalanine Ammonia-Lyase Gene (PaPAL) from Spruce Picea asperata

Liu¹,

Lou²,

Yang³

et al. 2019

Forests

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Phenylalanine ammonia-lyase (PAL) is the crucial enzyme of the phenylpropanoid pathway, which plays an important role in plant disease resistance. To understand the function of PAL in Picea asperata, in this study, the full-length cDNA sequence of the PAL gene from this species was isolated and named PaPAL. The gene contains a 2160-bp open reading frame (ORF) encoding 720 amino acids with a calculated molecular weight of 78.7 kDa and a theoretical isoelectric point of 5.88. The deduced PaPAL protein possesses the specific signature motif (GTITASGDLVPLSYIA) of phenylalanine ammonia-lyases. Multiple alignment analysis revealed that PaPAL has high identity with other plant PALs. The tertiary structure of PaPAL was predicted using PcPAL from Petroselinum crispum as a template, and the results suggested that PaPAL may have a similar function to that of PcPAL. Furthermore, phylogenetic analysis indicated that PaPAL has a close relationship with other PALs from the Pinaceae species. The optimal expression condition of recombinant PaPAL in Escherichia coli BL21 (DE3) was 0.2 mM IPTG (isopropyl β-D-thiogalactoside) at 16 • C for 4 h, and the molecular weight of recombinant PaPAL was found to be approximately 82 kDa. Recombinant PaPAL was purified and exhibited high PAL activity at optimal conditions of pH 8.6 and 60 • C. Quantitative real-time PCR (qRT-PCR) showed the PaPAL gene to be expressed in all tissues of P. asperata tested, with the highest expression level in the needles. The PaPAL gene was induced by the pathogen (Lophodermium piceae), which caused needle cast disease, indicating that it might be involved in defense against needle cast disease. These results provide a basis for understanding the molecular mechanisms of the PAL gene in the process of P. asperata disease resistance.

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Identification and Expression Analysis of Two Genes Involved in the Biosynthesis of t-Anethole in Fennel (Foeniculum vulgare Mill.) and Their Up-Regulation in Leaves in Response to Methyl Jasmonate Treatments

Abdollahi

Kianersi

Moosavi

et al. 2022

J Plant Growth Regul

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Molecular cloning, functional characterization and expression of a drought inducible phenylalanine ammonia-lyase gene (ObPAL) from Ocimum basilicum L.

Cited by 43 publications

References 38 publications

Analysis of Centranthera grandiflora Benth Transcriptome Explores Genes of Catalpol, Acteoside and Azafrin Biosynthesis

Analysis of Centranthera grandiflora Benth Transcriptome Explores Genes of Catalpol, Acteoside and Azafrin Biosynthesis

Cloning, Characterization and Expression of the Phenylalanine Ammonia-Lyase Gene (PaPAL) from Spruce Picea asperata

Identification and Expression Analysis of Two Genes Involved in the Biosynthesis of t-Anethole in Fennel (Foeniculum vulgare Mill.) and Their Up-Regulation in Leaves in Response to Methyl Jasmonate Treatments

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