Articles
CLONING AND EXPRESSION ANALYSIS OF PAL GENE IN LITCHI (LITCHI CHINENSIS SONN.) PERICARP
Article number
894_10
Pages
105 – 116
Language
English
Abstract
To elucidate molecular mechanisms of phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) gene in the pigmentation of litchi pericarp, a full-length cDNA sequence of PAL gene (named Lc-pal, GenBank No.
FJ944018.1) was isolated from the pericarp of Litchi chinensis Sonn. Feizixiao. The full length cDNA sequence of PAL gene is 2444 bp, contains a long open reading frame (ORF) of 2172 bp), a 120 bp 5 untranslated region (5-UTR), and a 152 bp 3-UTR with an 18 nt poly (A+). Sequence analysis of Lc-pal showed 76-87% similarities at both nucleotide and amino acid levels to previously recorded plant pal genes and shared the highest (87%) sequence identity with Citrus clementine (CAB42793.1) pal1. High homology confirmed the identity of the cloned pal gene and suggested that pal is relatively conserved in systematic evolution.
Results from RT-PCR and Real-Time PCR showed that Lc-pal was constitutively expressed in the pericarp of different litchi cultivars, different developmental stages of Nuomici and different color phenotypes of Feizixiao. Significant differences were detected in the expression of Lc-pal in the pericarp from five litchi cultivars with different color phenotypes.
Highest expression levels of Lc-pal were detected in the pericarp of Wupili compared to the lower levels in the pericarp of Sanyuehong. For different color phenotypes of Feizixiao, highest Lc-pal expression levels were detected in yellow pericarp, followed by red pericarp as compared to the lower levels in green phenotypes.
For different developmental stages of Nuomici, the highest expression levels of Lc-pal were detected at the mature stage, followed by the color break stage (turning to yellow) as compared to the low levels at the green stage.
FJ944018.1) was isolated from the pericarp of Litchi chinensis Sonn. Feizixiao. The full length cDNA sequence of PAL gene is 2444 bp, contains a long open reading frame (ORF) of 2172 bp), a 120 bp 5 untranslated region (5-UTR), and a 152 bp 3-UTR with an 18 nt poly (A+). Sequence analysis of Lc-pal showed 76-87% similarities at both nucleotide and amino acid levels to previously recorded plant pal genes and shared the highest (87%) sequence identity with Citrus clementine (CAB42793.1) pal1. High homology confirmed the identity of the cloned pal gene and suggested that pal is relatively conserved in systematic evolution.
Results from RT-PCR and Real-Time PCR showed that Lc-pal was constitutively expressed in the pericarp of different litchi cultivars, different developmental stages of Nuomici and different color phenotypes of Feizixiao. Significant differences were detected in the expression of Lc-pal in the pericarp from five litchi cultivars with different color phenotypes.
Highest expression levels of Lc-pal were detected in the pericarp of Wupili compared to the lower levels in the pericarp of Sanyuehong. For different color phenotypes of Feizixiao, highest Lc-pal expression levels were detected in yellow pericarp, followed by red pericarp as compared to the lower levels in green phenotypes.
For different developmental stages of Nuomici, the highest expression levels of Lc-pal were detected at the mature stage, followed by the color break stage (turning to yellow) as compared to the low levels at the green stage.
Authors
G.B. Hu, F.C. Hu, Y.H. Qin, Y.Z. Wei, X.J. Li, Z.C. Zhao, H.C. Wang, H.B. Chen, X.M. Huang
Keywords
Litchi chinensis Sonn., phenylalanine ammonia lyase (PAL), gene cloning, sequence analysis, expression analysis
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