青杄干旱诱导基因PwWDS1的cDNA分离与表达分析

doi:10.11707/j.1001-7488.20140419

摘要/Abstract

Abstract:

Drought-induced proteins, which are induced or up-regulated by water deficient stress, play an important role in drought resistance process of plants. In this research, full length cDNA of PwWDS1 (GenBank accession number KJ526353), a drought induced gene, was obtained by RACE PCR method based on the full length cDNA library of Picea wilsonii. Several software and tools such as Expasy, DNAMAN, ClustalX and MEGA were used to analyze the physical and chemical properties of PwWDS1. Then expression pattern of PwWDS1 was identified by RT-qPCR and semi-quantitative RT-PCR under drought, salt stress, abscisic acid (ABA) and ethrel treatments. We found that the full length cDNA of PwWDS1 was 1 077 bp encoding a protein which consists of 207 amino acids residues. The theoretical molecular weight of PwWDS1 was 22.76 kDa with isoeletric point of 5.02. Sequence analysis indicated that C terminal of PwWDS1 had a conserved ABA/WDS domain and N terminal had a strong hydrophilic property. RT-qPCR and semi-quantitative RT-PCR showed that PwWDS1 was mainly expressed in needles of P. wilsonii and the expression level of PwWDS1 increased with the growth of cotyledon during seed germination. The PwWDS1 expression was up-regulated by drought, salt stress, ethrel and ABA treatments. PwWDS1 was expressed highly in 6 h under drought and ABA treatments. Under salt and ethrel treatments, PwWDS1 was expressed most in 12 h. These results indicated that PwWDS1 would play a role in response to signals of abiotic stresses and plant hormones in P. wilsonii.

Key words: Picea wilsonii, water deficiency stress(WDS), bioinformatic analysis, tissue expression, stress response

中图分类号:

S718.46

李长江, 崔晓燕, 孙帆, 张凌云. 青杄干旱诱导基因PwWDS1的cDNA分离与表达分析[J]. 林业科学, 2014, 50(4): 129-136.

Li Changjiang, Cui Xiaoyan, Sun Fan, Zhang Lingyun. Isolation and Expression Analysis of PwWDS1 in Picea wilsonii[J]. Scientia Silvae Sinicae, 2014, 50(4): 129-136.

参考文献

李长江, 曹一博, 张凌云. 2012. 青杄PSAK的克隆及生物信息学分析. 生物技术, 22(3): 4-9.
(Li C J, Cao Y B, Zhang L Y. 2012. Cloning and bioinformatic analysis of PSAK from Picea wilsonii. Biotechnology, 22(3): 4-9. [in Chinese] )
李长江, 孙帆, 张通, 等. 2013. 青杄PwPSAF基因的克隆与组织表达分析. 林业科学, 49(10): 40-47.
(Li C J, Sun F, Zhang T, et al. 2013. Cloning and tissue expression analysis of PwPSAF in Picea wilsonii. Scientia Silvae Sinicae, 49(10): 40-47. [in Chinese] )
张盾, 刘亚静, 李长江, 等. 2012. 青杄均一化cDNA文库构建及EST序列分析. 生物技术通报, (6): 71-76.
(Zhang D, Liu Y J, Li C J, et al. 2012. Construction of normalized cDNA library and analysis of corresponding EST sequences in Picea wilsonii. Biotechnology Bulletin, (6): 71-76. [in Chinese] )
Arc E, Sechet J, Corbineau F, et al. 2013. ABA crosstalk with ethylene and nitric oxide in seed dormancy and germination. Frontiers in Plant Science, 4: 63.
Cao Y R, Chen S Y, Zhang J S. 2008. Ethylene signaling regulates salt stress response: An overview. Plant Signaling & Behavior, 3 (10): 761-763.
Dalal M, Tayal D, Chinnusamy V, et al. 2009. Abiotic stress and ABA-inducible group 4 LEA from Brassica napus plays a key role in salt and drought tolerance. Journal of Biotechnology, 139(2): 137-145.
Desclos M, Dubousset L, Etienne P, et al. 2008. A proteomic profiling approach to reveal a novel role of Brassica napus drought 22 kD/water-soluble chlorophyll-binding protein in young leaves during nitrogen remobilization induced by stressful conditions. Plant Physiology, 147(4): 1830-1844.
Du Q S, Fan X W, Wang C H, et al. 2011. A possible CO₂ conducting and concentrating mechanism in plant stomata SLAC1 channel. PloS One, 6(9): e24264.
George S, Usha B, Parida A. 2009. Isolation and characterization of an atypical LEA protein coding cDNA and its promoter from drought-tolerant plant Prosopis juliflora. Applied Biochemistry and Biotechnology, 157(2): 244-253.
Goel D, Singh A K, Yadav V, et al. 2010. Overexpression of osmotin gene confers tolerance to salt and drought stresses in transgenic tomato (Solanum lycopersicum L.). Protoplasma, 245(1-4): 133-141.
Guo L, Yang H, Zhang X, et al. 2013. Lipid transfer protein 3 as a target of MYB96 mediates freezing and drought stress in Arabidopsis. Journal of Experimental Botany, 64(6): 1755-1767.
Ho S L, Huang L F, Lu C A, et al. 2013. Sugar starvation-and GA-inducible calcium-dependent protein kinase 1 feedback regulates GA biosynthesis and activates a 14-3-3 protein to confer drought tolerance in rice seedlings. Plant Molecular Biology, 81(4/5): 347-361.
Hong Y, Zhang W, Wang X. 2010. Phospholipase D and phosphatidic acid signalling in plant response to drought and salinity. Plant, Cell & Environment, 33(4): 627-635.
Hundertmark M, Hincha D K. 2008. LEA (late embryogenesis abundant) proteins and their encoding genes in Arabidopsis thaliana. BMC Genomics, 9: 118.
Jiang Y, Liang G, Yu D. 2012. Activated expression of WRKY57 confers drought tolerance in Arabidopsis. Molecular Plant, 5(6): 1375-1388.
Jin Z, Shen J, Qiao Z, et al. 2011. Hydrogen sulfide improves drought resistance in Arabidopsis thaliana. Biochemical and Biophysical Research Communications, 414(3): 481-486.
Karaba A, Dixit S, Greco R, et al. 2007. Improvement of water use efficiency in rice by expression of HARDY, an Arabidopsis drought and salt tolerance gene. Proceedings of the National Academy of Sciences of the United States of America, 104(39): 15270-15275.
Kizis D, Pages M. 2002. Maize DRE-binding proteins DBF1 and DBF2 are involved in rab17 regulation through the drought-responsive element in an ABA-dependent pathway. The Plant Journal, 30(6): 679-689.
Liu A L, Zou J, Liu C F, et al. 2013.Over-expression of OsHsfA 7 enhanced salt and drought tolerance in transgenic rice. BMB Reports, 46(1): 31-36.
Lorenz W W, Alba R, Yu Y S, et al. 2011. Microarray analysis and scale-free gene networks identify candidate regulators in drought-stressed roots of loblolly pine (P. taeda L.). BMC Genomics, 12: 264.
Lu S, Bahn S C, Qu G, et al. 2013. Increased expression of phospholipase Dalpha1 in guard cells decreases water loss with improved seed production under drought in Brassica napus. Plant Biotechnology Journal, 11(3): 380-389.
Osakabe Y, Arinaga N, Umezawa T, et al. 2013. Osmotic stress responses and plant growth controlled by potassium transporters in Arabidopsis. The Plant Cell, 25(2): 609-624.
Padmanabhan V, Dias D M, Newton R J. 1997. Expression analysis of a gene family in loblolly pine (Pinus taeda L.) induced by water deficit stress. Plant Molecular Biology, 35(6): 801-807.
Poor P, Kovacs J, Szopko D, et al. 2013. Ethylene signaling in salt stress-and salicylic acid-induced programmed cell death in tomato suspension cells. Protoplasma, 250(1): 273-284.
Soderman E, Hjellstrom M, Fahleson J, et al. 1999. The HD-Zip gene ATHB 6 in Arabidopsis is expressed in developing leaves, roots and carpels and up-regulated by water deficit conditions. Plant Molecular Biology, 40(6): 1073-1083.
Sun L, Wang Y P, Chen P, et al. 2011. Transcriptional regulation of SlPYL, SlPP 2 C, and SlSnRK 2 gene families encoding ABA signal core components during tomato fruit development and drought stress. Journal of Experimental Botany, 62(15): 5659-5669.
Urao T, Katagiri T, Mizoguchi T, et al. 1994. Two genes that encode Ca²⁺-dependent protein kinases are induced by drought and high-salt stresses in Arabidopsis thaliana. Molecular & General Genetics: MGG, 244(4): 331-340.
Wang T Z, Zhang J L, Tian Q Y, et al. 2013. A Medicago truncatula EF-Hand family gene, MtCaMP 1 , is involved in drought and salt stress tolerance. PloS One, 8(4): e58952.
Wilmowicz E, Kesy J, Kopcewicz J. 2008. Ethylene and ABA interactions in the regulation of flower induction in Pharbitis nil. Journal of Plant Physiology, 165(18): 1917-1928.
Xiao B, Huang Y, Tang N, et al. 2007. Over-expression of a LEA gene in rice improves drought resistance under the field conditions. Theor Appl Genet, 115(1): 35-46.
Yoshida T, Fujita Y, Sayama H, et al. 2010. AREB1, AREB2, and ABF3 are master transcription factors that cooperatively regulate ABRE-dependent ABA signaling involved in drought stress tolerance and require ABA for full activation. The Plant Journal, 61(4): 672-685.
Yamaguchi-Shinozaki K, Shinozaki K. 1994. A novel cis-acting element in an Arabidopsis gene is involved in responsiveness to drought, low-temperature, or high-salt stress. The Plant Cell, 6(2): 251-264.
Yu Y, Li Y, Huang G, et al. 2011. PwHAP5, a CCAAT-binding transcription factor, interacts with PwFKBP12 and plays a role in pollen tube growth orientation in Picea wilsonii. Journal of Experimental Botany, 62(14): 4805-4817.
Yu Y, Zhang H, Li W, et al. 2012. Genome-wide analysis and environmental response profiling of the FK506-binding protein gene family in maize (Zea mays L.). Gene, 498(2): 212-222.
Zegzouti H, Jones B, Marty C, et al. 1997. ER 5 , a tomato cDNA encoding an ethylene-responsive LEA-like protein: characterization and expression in response to drought, ABA and wounding. Plant Molecular Biology, 35(6): 847-854.
Zhang M, Yuan B, Leng P. 2009. The role of ABA in triggering ethylene biosynthesis and ripening of tomato fruit. Journal of Experimental Botany, 60(6): 1579-1588.
Zhang Y, Li Y, Lai J, et al. 2012. Ectopic expression of a LEA protein gene TsLEA 1 from Thellungiella salsuginea confers salt-tolerance in yeast and Arabidopsis. Molecular Biology Reports, 39(4): 4627-4633.
Zhao L, Hu Y, Chong K, et al. 2010. ARAG 1 , an ABA-responsive DREB gene, plays a role in seed germination and drought tolerance of rice. Annals of Botany, 105(3): 401-409.
Zhou M L, Ma J T, Zhao Y M, et al. 2012. Improvement of drought and salt tolerance in Arabidopsis and Lotus corniculatus by overexpression of a novel DREB transcription factor from Populus euphratica.Gene, 506(1): 10-17."

[1]	张鹤华, 刘嘉欣, 罗朝兵, 张凌云. 青杄转录因子PwERF8及其启动子序列的克隆与分析[J]. 林业科学, 2018, 54(3): 48-60.
[2]	刘长英, 张梦, 魏从进, 许亚珍, 曹博宁, 郑莎, 范伟, 向仲怀, 赵爱春. 转桑树MAPK5基因拟南芥在不同逆境胁迫下的抗性分析[J]. 林业科学, 2017, 53(9): 35-44.
[3]	徐嘉娟, 李火根. 鹅掌楸LcPAT8基因的克隆及功能初步分析[J]. 林业科学, 2017, 53(9): 45-54.
[4]	游韩莉, 袁义杭, 李长江, 张凌云. 青杄MYB转录因子基因PwMYB20的克隆及表达分析[J]. 林业科学, 2017, 53(5): 23-32.
[5]	罗群凤, 胥猛, 冯源恒, 徐嘉娟, 李火根. 北美鹅掌楸LtCHS基因的克隆及生物信息学与组织表达特征分析[J]. 林业科学, 2015, 51(5): 37-45.
[6]	李艳艳, 杨艳芳, 王俊青, 王帅, 刘洪伟, 邱德有. 红豆杉TcLBDs基因的克隆与表达分析[J]. 林业科学, 2015, 51(10): 126-133.
[7]	李林峰, 张金屯, 周兰, 邵丹. 自组织特征映射网络在芦芽山自然保护区青杄林分类和排序中的应用[J]. 林业科学, 2014, 50(5): 1-7.
[8]	张通, 李巧玲, 张凌云. PwEXP1在青杄种子萌发及逆境响应中的表达特征[J]. 林业科学, 2014, 50(12): 56-62.
[9]	刘长英;赵爱春;李军;吕蕊花;余亚圣;王茜龄;鲁成;余茂德. 桑树查尔酮异构酶基因的克隆与原核表达分析[J]. , 2013, 49(2): 39-45.
[10]	李长江, 孙帆, 张通, 张凌云. 青杄PwPSAF基因的克隆与组织表达分析[J]. 林业科学, 2013, 49(10): 40-47.
[11]	彭梅芳阳义健杨春贤陈敏廖志华. 银杏IspF基因的克隆与功能分析*[J]. 林业科学, 2008, 44(10): 49-54.