杉木苯丙氨酸解氨酶基因ClPAL的克隆与表达分析

doi:10.11707/j.1001-7488.20131210

Abstract

Abstract:

In this study, three full-length cDNAs encoding phenylalanine ammonia lyase (PAL) were isolated from Chinese Fir (Cunninghamia lanceolata) using RACE. They encode 709, 710 and 709 amino acids, respectively, and their sequence lengths are 2 455, 2 418 and 2 689 bp, respectively. The amino acid sequences contain two functional domains of PAL-HAL and PAL, and the enzyme active site (GTITASGDLVPLSYIA). The expression analysis showed that the three ClPALs differentially expressed in different organs. The expression level of ClPAL1 in lignified stem was higher than that in needle, male cone, root and non-lignified stem, and ClPAL3 was mainly expressed in lignified stem. Both genes were predominantly expressed in xylem, and their expression levels in intermediate wood were over 20 times higher than that in sapwood. ClPAL2 exhibited the high transcript abundance in phloem-enriched needles and bark. IAA, GA₃ and BR can promote the differentiation of xylem cells, and thus they should directly or indirectly associate with lignin biosynthesis. After application of the three hormones, ClPAL1 and ClPAL3 were induced to different extents, and changes in the expressions were relatively more significant with IAA and BR, indicating that ClPAL1 and ClPAL3 were likely to play important roles in lignin biosynthesis of Chinese Fir wood tissue. During the compression wood formation, the expression level of ClPAL1 in the compression area of xylem showed decreased then increased, and reached to 1.4 times of the control level after 10 days bending treatment. Its expression levels in the opposite area of the compressed zone were lower than that of the control. The ClPAL2 expression was up-regulated in the compression wood, and its value in the opposite wood area was obviously higher than that of the control after 10 days bending. The expression changes of ClPAL3 were not significant compared with the control. In addition, the expression level of the ClPAL2 in intermediate wood was higher than that in sapwood. It deserves further study whether this gene would participate in the formation of the heartwood color.

Key words: Cunninghamia lanceolata, PAL gene, gene cloning, gene expression, wood formation

CLC Number:

Xu Lili, Tong Zaikang, Lin Erpei, Huang Huahong. Isolation and Expression of ClPAL Genes in Chinese Fir (Cunninghamia lanceolata)[J]. Scientia Silvae Sinicae, 2013, 49(12): 64-72.

References

宋坤霖. 2012. 杉木边心材转变过程中细胞壁结构与性能的变化. 北京: 中国林业科学研究院硕士学位论文.
王桂凤, 高燕, 杨立伟, 等. 2007. 杉木木材形成过程特异表达基因的鉴定与分析. 遗传, 29(4): 483-489.
Bagal U R, Leebens-Mack J H, Lorenz W W, et al. 2012. The phenylalanine ammonia lyase (PAL) gene family shows a gymnosperm-specific lineage. BMC Genomics, 13(Suppl 3): S1.
Beritognolo I, Magel E, Abdel-Latif A, et al. 2002. Expression of genes encoding chalcone synthase, flavanone 3-hydroxylase and dihydroflavonol 4-reductase correlates with flavanol accumulation during heartwood formation in Juglans nigra. Tree Physiology, 22(5): 291-300.
Boerjan W, Ralph J, Baucher M. 2003. Lignin biosynthesis. Annual Review of Plant Physiology, 54(10): 519-546.
Butland S L, Chow M L, Ellis B E. 1998. A diverse family of phenylalanine ammonia-lyase genes expressed in pine trees and cell cultures. Plant Molecular Biology, 37(1):15-24.
Cochrane F C, Davin L B, Lewis N G. 2004. The Arabidopsis phenylalanine ammonia lyase gene family: kinetic characterization of the four PAL isoforms. Phytochemistry, 65(11):1557-1564.
Demura T, Fukuda H. 1993. Molecular cloning and characterization of cDNAs associated with tracheary element differentiation in cultured zinnia cells. Plant Physiology, 103(2): 815-821.
Hamberger B, Ellis M, Friedmann M, et al. 2007. Genome-wide analyses of phenylpropanoid-related genes in Populus trichocarpa, Arabidopsis thaliana, and Oryza sativa: the Populus lignin toolbox and conservation and diversification of angiosperm gene families. Canadian Journal of Botany, 85(12):1182-1201.
Hillis W E. 1996. Formation of robinetin crystals in vessels of Intsia species. IAWA Journal, 17(4): 405-419.
Hossain Z, McGarvey B, Amyot L, et al. 2012. DIMINUTO1 affects the lignin profile and secondary cell wall formation in Arabidopsis. Planta, 235(3): 485-498.
Huang Huahong, Xu Lili, Tong Zaikang, et al. 2012. De novo characterization of the Chinese fir (Cunninghamia lanceolata) transcriptome and analysis of candidate genes involved in cellulose and lignin biosynthesis. BMC Genomics, 13(1): 648.
Huang Junli, Gu Min, Lai Zhibing, et al. 2010. Functional analysis of the Arabidopsis PAL gene family in plant growth, development, and response to environmental stress. Plant Physiology, 153(4): 1526-1538.
IAWA Committee. 1989. IAWA list of microscopic features for hardwood identification. IAWA Bulletin, 10(3): 219-332.
Israelsson M, Eriksson M E, Hertzberg M, et al. 2003. Changes in gene expression in the wood-forming tissue of transgenic hybrid aspen with increased secondary growth. Plant Molecular Biology, 52(4): 893-903.
Kao Y Y, Harding S A, Tsai C J. 2002. Differential expression of two distinct phenylalanine ammonia-lyase genes in condensed tannin-accumulating and lignifying cells of quaking aspen. Plant Physiology, 130(2): 796-807.
Karpinska B, Karlsson M, Srivastava M, et al. 2004. MYB transcription factors are differentially expressed and regulated during secondary vascular tissue development in hybrid aspen. Plant Molecular Biology, 56(2):255-70.
Li Laigeng, Lu Shanfa, Chiang V. 2006. A genomic and molecular view of wood formation. Critical Reviews in Plant Sciences, 25(3): 215-233.
Minami E, Ozeki Y, Matsuoka M, et al. 1989. Structure and some characterization of the gene for phenylalanine ammonia-lyase from rice plants. European Journal of Biochemistry, 185(1):19-25.
Nagai N, Kitauchi F, Shimosaka M, et al. 1994. Cloning and sequencing of a full-length cDNA coding for phenylalanine ammonia-lyase from tobacco cell culture. Plant Physiology, 104(3): 1091-1092.
Onaka F. 1949. Studies on compression and tension wood. Wood Research, 24(1):1-88.
Pallardy S G. 2008. Physiology of woody plants. London: Academic Press, 454.
Ritter H, Schulz G E. 2004. Structural basis for the entrance into the phenylpropanoid metabolism catalyzed by phenylalanine ammonia-lyase. Plant Cell, 16(12): 3426 -3436.
Rohde A, Morreel K, Ralph J, et al. 2004. Molecular phenotyping of the pal1 and pal2 mutants of Arabidopsis thaliana reveals far-reaching consequences on phenylpropanoid, amino acid, and carbohydrate metabolism. Plant Cell, 16(10): 2749-2771.
Shang Qingmao, Li Liang, Dong Chunjuan. 2012. Multiple tandem duplication of the phenylalanine ammonia-lyase genes in Cucumis sativus L. Planta, 236(4):1093-1105.
Sitbon F, Hennion S, Little C H A, et al. 1999. Enhanced ethylene production and peroxidase activity in IAA-overproducing transgenic tobacco plants is associated with increased lignin content and altered lignin composition. Plant Science, 141(2): 165-173.
Tamura K, Dudley J, Nei M, et al. 2007. MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4. 0. Molecular Biology and Evolution, 24(8): 1596 -1599.
Timell T. 1986. Compression wood in gymnosperms: Vol. 1-3. Berlin: Springer-Verlag, 1339-2150.
Ursache R, Nieminen K, Helariutta Y. 2012. Genetic and hormonal regulation of cambial development. Physiologia Plantarum, 147(1): 36-45.
Wang Guifeng, Gao Yan, Wang Jinjun, et al. 2011. Overexpression of two cambium-abundant Chinese fir (Cunninghamia lanceolata) α-expansin genes ClEXPA 1 and ClEXPA 2 affect growth and development in transgenic tobacco and increase the amount of cellulose in stem cell walls. Plant Biotechnology Journal, 9(4): 486-502.
Wang Guifeng, Gao Yan, Yang Liwei, et al. 2007. Identification and analysis of differentially expressed genes in differentiating xylem of Chinese fir (Cunninghamia lanceolata) by suppression subtractive hybridization. Genome, 50(12): 1141-1155.
Wardrop A B, Dadswell H E. 1950. The nature of reaction woodⅡ: the cell wall organization wood tracheid. Australian Journal of Scientific Research, 3(1): 1-13.
Whetten R, Sederoff R. 1992. Phenylalanine ammonia-lyase from loblolly pine: Purification of the enzyme and isolation of complementary DNA clones. Plant Physiology, 98(1): 380-386.
Wilson B F, Archer R R. 1977. Reaction wood: induction and mechanical action. Annual Review of Plant Physiology, 28(6): 23-43.
Zhong Ruiqin, Morrison W H, Himmelsbach D S, et al. 2000. Essential role of caffeoyl coenzyme A O-methyltransferase in lignin biosynthesis in woody poplar plants. Plant Physiology, 124(2): 563-577.

[1]	Liu Daofeng, Wang Xia, Dai Yin, Yang Jianfeng, Ma Jing, Li Mingyang, Sui Shunzhao. Cloning and Function Analysis of CpTAF10 from Wintersweet (Chimonanthus praecox) [J]. Scientia Silvae Sinicae, 2019, 55(6): 176-183.
[2]	Lu Huijun, Li Ziyi, Liang Hanyu, Yue Yuanzhi, Zhou Tianchang, Yang Yuzhang, Wang Yucheng, Ji Xiaoyu. Expression and Stress Tolerance Analysis of NAC24 from Tamarix hispida [J]. Scientia Silvae Sinicae, 2019, 55(3): 54-63.
[3]	Fan Weijian, Xiang Weifang, Wang Jing, Bai Penghua, Pan Lina, Yang Yixin, Zhu Gengping, Li Min. Effects of Insecticides and Ultraviolet on the Expression of sHSP12.2 Gene in Chouioia cunea [J]. Scientia Silvae Sinicae, 2019, 55(2): 128-136.
[4]	Zhang Yang, Du Lin, Tang Xianfeng, Liu Huanhuan, Zhou Gongke, Chai Guohua. Identification and Expression Pattern Analyses of Populus BAG Genes [J]. Scientia Silvae Sinicae, 2019, 55(1): 138-145.
[5]	Wei Mingke, Yu Jinjian, Huang Xiaolong, Liu Qiongyao, Huang Huahong, Lin Erpei, Tong Zaikang. Cloning, Expression and Single Nucleotide Polymorphisms Analysis of NAC Transcription Factor Gene ClNAC1 in Cunninghamia lanceolata [J]. Scientia Silvae Sinicae, 2018, 54(9): 49-59.
[6]	Ren Jian, Yin Yuqing, Zhang Huihui, Chen Dian, Wang Kexin, Wang Yong. Cloning and Analysis of AmCesA1 Gene in Acacia mangium [J]. Scientia Silvae Sinicae, 2018, 54(8): 79-87.
[7]	Wang Juan, Chen Hui. Identification and Expression of Cold Shock Domain-containing Protein Gene in Dendroctonus armandi [J]. Scientia Silvae Sinicae, 2018, 54(4): 58-66.
[8]	Zhang Hehua, Liu Jiaxin, Luo Chaobing, Zhang Lingyun. Cloning and Analysis of a Transcription Factor PwERF8 and the Promoter Sequences in Picea wilsonii [J]. Scientia Silvae Sinicae, 2018, 54(3): 48-60.
[9]	Li Jianbo, Jia Huixia, Zhang Jin, Liu Bobin, Hu Jianjun, Wang Lijuan, Lu Mengzhu. Effect of Overexpression of Populus tomentosa WUSCHEL-related homeobox 4 (PtoWOX4a) on the Secondary Growth of Poplar [J]. Scientia Silvae Sinicae, 2018, 54(2): 52-59.
[10]	Zhang Qin, Xu Zongda, Zhao Kai, Li Xiaowei, Zhang Luosha, Zhang Qixiang. Isolation and Biological Function Analysis of Anthocyanin Regulatory Gene PmMYB1 from Prunus mume [J]. Scientia Silvae Sinicae, 2018, 54(10): 64-72.
[11]	Sun Huayu, Li Lichao, Zhao Hansheng, Yang Yihong, Wang Sining, Gao Zhimin. Molecular Characteristics and SSR Marker Development of Two NAC Genes from Daemonorops jenkinsiana [J]. Scientia Silvae Sinicae, 2017, 53(8): 132-140.
[12]	Tang Feifei, Zhao Yulin, Wang Peilong, Feng Deming, Song Yi, Gao Caiqiu. Cloning and Stress Tolerance Analysis of ThP5CR from Tamarix hispida [J]. Scientia Silvae Sinicae, 2017, 53(7): 1-9.
[13]	Li Nan, Zheng Yongqi, Ding Hongmei, Liu Xinhong, Sheng Weitong, Jiang Bo, Li Haibo. Analysis on Differential Expression of Cold Resistance Related Genes of Casuarina equisetifolia under Low Temperature Stress [J]. Scientia Silvae Sinicae, 2017, 53(7): 62-71.
[14]	You Hanli, Yuan Yihang, Li Changjiang, Zhang Lingyun. Cloning and Expression Analysis of MYB Homologous Gene PwMYB20 from Picea wilsonii [J]. Scientia Silvae Sinicae, 2017, 53(5): 23-32.
[15]	Yu Jian, Zhao Aichun, Liu Changying, Liang Yanmei, Zhu Panpan, Cai Yuxiang, Wang Xiling, Li Zhengang, Yu Maode. Effects of Exogenous Ethylene and 1-MCP Treatments on the Expression of Genes Involved in Ethylene and Anthocyanin in Mulberry Fruit [J]. Scientia Silvae Sinicae, 2017, 53(2): 138-148.

Isolation and Expression of ClPAL Genes in Chinese Fir (Cunninghamia lanceolata)

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments