两种水杨酸代谢相关酶在逆境龙须菜中的活性研究

doi:10.13560/j.cnki.biotech.bull.1985.2016.05.026

摘要/Abstract

摘要： 旨在探索水杨酸（Salicylic acid，SA）信号通路相关的两种酶——苯丙氨酸解氨酶（PAL）和β-1，3-葡聚糖酶（β-1，3-GA）在病烂、盐度、温度逆境和添加SA条件下在龙须菜中的活性变化。结果表明，3组病烂龙须菜中PAL和β-1，3-GA活性分别增加为对照组的1.27-1.42倍和1.26-1.35倍；高盐条件下PAL活性与对照组无显著差别，而β-1，3-GA活性在24 h和48 h分别增加为对照组的1.90倍和1.42倍；高温组PAL和β-1，3-GA活性在24 h均显著升高，分别是对照组的1.25倍和1.27倍；100 μmol/L SA添加后PAL和β-1，3-GA的活性升高，但高浓度SA却抑制了两种酶的活性。结果表明，在逆境胁迫下龙须菜中PAL和β-1，3-GA的活性增加与水杨酸的抗逆作用有关，而100 μmol/L SA诱导两种酶的效果最显著。

关键词: 龙须菜, 水杨酸, 苯丙氨酸解氨酶, β-1, 3-葡聚糖酶

Abstract: The article mainly studied two enzymes related to Salicylic acid（SA） signaling pathway：phenylalanine ammonia-lyase（PAL）and β-1，3-glucanase（β-1，3-GA）in marine algae Gracilariopsis lemaneiformis，and measured their activity change under sick，high salt，high temperature stresses and addition of SA. Results showed that the activities of PAL and β-1，3-GA in G. lemaneiformis of 3 sick groups increased 1.27-1.42 times and 1.26-1.35 times compared to healthy groups，respectively. For high salt stress，the activities of PAL had no significant difference from the control，whereas β-1，3-GA increased 1.90 times and 1.42 times at 24 h and 48 h compared to the control，respectively. The activities of PAL and β-1，3-GA under high temperature stress increased 1.25 times and 1.27 times compared to the control groups. The activities of PAL and β-1，3-GA increased after adding 100 μmol/L SA，but high concentration of SA inhibited the activities of 2 enzymes. This study indicated that the increases of activities of PAL and β-1，3-GA in G. lemaneiformis might be associated with the SA resistance to environment. In addition，100 μmol/L SA had the most significant effect on the induction of PAL and β-1，3-GA activities.

Key words: Gracilariopsis lemaneiformis, salicylic acid, phenylalanine ammonia-lyase, β-1, 3-glucanase

邹同雷, 汪芳俊, 侯赛男, 孙雪, 徐年军. 两种水杨酸代谢相关酶在逆境龙须菜中的活性研究[J]. 生物技术通报, 2016, 32(5): 194-199.

ZOU Tong-lei,WANG Fang-jun,HOU Sai-nan,SUN Xue,XU Nian-jun. On the Activities of Two Enzymes Related to Salicylic Acid Metabolism in Gracilariopsis lemaneiformis Under Adverse Environment[J]. Biotechnology Bulletin, 2016, 32(5): 194-199.

参考文献

[1]Raskin I. Role of salicylic acid in plant[J]. Annu Rev Plants Physiol Plant Mol Biol, 1992, 43（1）：439-463.
[2]Rivas-San Vicente M, Plasencia J. Salicylic acid beyond defence：its role in plant growth and development[J]. Journal of Experimental Botany, 2011, 62（10）：3321-3338.
[3]Hayat Q, Hayat S, Irfan M, et al. Effect of exogenous salicylic acid under changing environment：a review[J]. Environmental and Experimental Botany, 2010, 68（1）：14-25.
[4]Kumar D. Salicylic acid signaling in disease resistance[J]. Plant Science, 2014, 228：127-134.
[5]Chen Z, Zheng Z, Huang J, et al. Biosynthesis of salicylic acid in plants[J]. Plant Signaling & Behavior, 2009, 4（6）：493-496.
[6]Métraux JP. Recent breakthroughs in the study of salicylic acid biosynthesis[J]. Trends in Plant Science, 2002, 7（8）：332-334.
[7]Zhang X, Liu CJ. Multifaceted regulations of gateway enzyme phenylalanine ammonia-lyase in the biosynthesis of phenylpropanoids[J]. Molecular Plant, 2015, 8（1）：17-27.
[8]Dehghan S, Sadeghi M, Poppel A, et al. Differential inductions of phenylalanine ammonia-lyase and chalcone synthase during wounding, salicylic acid treatment, and salinity stress in safflower, Carthamus tinctorius[J]. Bioscience Reports, 2014, 34（3）：273-282.
[9]Moffitt MC, Louie GV, Bowman ME, et al. Discovery of two cyanobacterial phenylalanine ammonia lyases：kinetic and structural characterization[J]. Biochemistry, 2007, 46（4）：1004-1012.
[10]Zamora-Carreras H, Torres M, Bustamante N, et al. The C-terminal domains of two homologous Oleaceae β-1, 3-glucanases recognise carbohydrates differently：Laminarin binding by NMR[J]. Archives of Biochemistry and Biophysics, 2015, 580：93-101.
[11]Dogra V, Sreenivasulu Y. Cloning and functional characterization of β-1, 3-glucanase gene from Podophyllum hexandrum-A high altitude Himalayan plant[J]. Gene, 2015, 554（1）：25-31.
[12]Van Loon LC, Rep M, Pieterse CMJ. Significance of inducible defense-related proteins in infected plants[J]. Annu Rev Phytopathol, 2006, 44：135-162.
[13]Kim J S, Lee J, Lee C, et al. Activation of pathogenesis-related genes by the Rhizobacterium, Bacillus sp. JS, which induces systemic resistance in tobacco plants[J]. The Plant Pathology Journal, 2015, 31（2）：195-201.
[14]Xie YR, Raruang Y, Chen ZY, et al. ZmGns, a maize class I β-1, 3-glucanase, is induced by biotic stresses and possesses strong antimicrobial activity[J]. Journal of Integrative Plant Biology, 2015, 57（3）：271-283.
[15]Provasoli L. Media and prospects for the cultivation of marine algae[M]//Watarnable A, Hattori R. Culture and collection of algae. Proceedings of the US-Japan Conference, Hakone：Japan Soc Plant Physiol, 1968：63-75.
[16]高峰, 李国英, 王钦英. 水杨酸诱导棉花耐黄萎病的效应[J]. 新疆农业科学, 2004, 41（5）：333-336.
[17]Bariya HS, Thakkar VR, Thakkar AN, et al. Induction of systemic resistance in different varieties of Solanum tuberosum by pure and crude elicitor treatment[J]. Index Medicus for South-East Asia Region, 2011, 49（2）：151-162.
[18]Dong CJ, Li L, Shang QM, et al. Endogenous salicylic acid accumulation is required for chilling tolerance in cucumber（Cucumis sativus L. ）seedlings[J]. Planta, 2014, 240（4）：687-700.
[19]Siboza XI, Bertling I, Odindo AO. Salicylic acid and methyl jasmonate improve chilling tolerance in cold-stored lemon fruit（Citrus limon）[J]. Journal of Plant Physiology, 2014, 171（18）：1722-1731.
[20]Misra N, Misra R, Mariam A, et al. Salicylic acid alters antioxidant and phenolics metabolism in catharanthus roseus grown under salinity stress[J]. African Journal of Traditional, Complementary and Alternative Medicines, 2014, 11（5）：118-125.
[21]Wada KC, Mizuuchi K, Koshio A, et al. Stress enhances the gene expression and enzyme activity of phenylalanine ammonia-lyase and the endogenous content of salicylic acid to induce flowering in pharbitis[J]. Journal of Plant Physiology, 2014, 171（11）：895-902.
[22]Hwang H, Kim ST, Kim SG, et al. Comprehensive analysis of the expression of twenty-seven beta-1, 3-glucanase genes in rice（Oryza sativa L. ）[J]. Molecules and Cells, 2007, 23（2）：207-214.
[23]Tian S, Wan Y, Qin G, et al. Induction of defense responses against Alternaria rot by different elicitors in harvested pear fruit[J]. Applied Microbiology and Biotechnology, 2006, 70（6）：729-734.
[24]Zhang Y, Fu X, Hao X, et al. Molecular cloning and promoter analysis of the specific salicylic acid biosynthetic pathway gene phenylalanine ammonia-lyase（AaPAL1）from Artemisia annua[J]. Biotechnology and Applied Biochemistry, 2015, doi：10. 1002/bab. 1403.
[25]Gunter EA, Kapustina OM, Popeyko OV, et al. Induction of β-1, 3-glucanase in callus cultures in vitro[J]. Biochemistry（Moscow）, 2008, 73（7）：826-832.
[26]焦蒙丽, 曹蓉蓉, 陈红艳, 等. 水杨酸对丹参培养细胞中迷迭香酸生物合成及其相关酶的影响[J]. 生物工程学报, 2012, 28（3）：320-328.
[27]Hao W, Guo H, Zhang J, et al. Hydrogen peroxide is involved in salicylic acid-elicited rosmarinic acid production in Salvia miltiorrhiza cell cultures[J]. The Scientific World Journal, 2014, doi. org/10. 1155/2014/843764.
[28]李颖章, 郑晓华, 唐海林, 等. 水杨酸和黄萎病菌毒素处理棉花愈伤组织使β-1, 3-葡聚糖酶和几丁质酶的活性增加[J]. 植物学报, 2003, 45（7）：802-808.