Welcome to Acta Agronomica Sinica,

Acta Agron Sin ›› 2013, Vol. 39 ›› Issue (04): 744-752.doi: 10.3724/SP.J.1006.2013.00744

• RESEARCH NOTES • Previous Articles     Next Articles

Construction and Analysis of SSH Library in Rapeseed (Brassica napus L.) under Drought Stress

XIE Xiao-Yu,ZHANG Bing,ZHANG Xia,MA Zhong-Lian,LI Jia-Na*   

  • Received:2012-08-04 Revised:2012-12-08 Online:2013-04-12 Published:2013-01-28
  • Contact: 李加纳, E-mail: ljn1950@swu.edu.cn E-mail:xiexy8009@163.com

Abstract:

The positive SSH library in leaves of rapeseed (Brassica napus L.) Holiday (with high drought resistance ability) was constructed using the sample under drought stress as Tester and the sample under normal water management as Driver. Twenty-four positive clones selected randomly were verified by using PCR. The result of electrophoresis showed that 23 clones contained inserts whose average length was 750 bp. Ninety-six positive clones were sequenced and analyzed by CAP3 Sequence Assembly Program, showing that four contigs and 82 singletons were contained and the average length of 86 ESTs was 542 bp. Blastx alignment with Nr database showed 11 ESTs without significant similarity and 75 ESTs with similarity in GenBank database. KOBAS suggested that 28 ESTs were mapped to 67 pathways. It could be predicted that the carbon fixation in photosynthetic organism, electron donor III of aerobic respiration, nitrogen metabolism, glyoxylate and dicarboxylate metabolism played a key role in drought tolerance ofrapeseed. The analysis of ESTs showed that a large share of function could be attributed to organelle (58.82%), binding (30.77%), and metabolic process (43.72%), which plays important roles in metabolism of rapeseed under drought stress.

Key words: Rapeseed (Brassica napus L.), Suppression subtractive hybridization (SSH), Drought stress, cDNA library, Express sequence tag

[1]Xiang J(项俊), Chen Z-B(陈兆波), Wang P(王沛), Yu L-J(余龙江), Li M-T(栗茂腾). The effect of CaCl2 treatment on the Chang of drought related physiological and biochemical indices of Brassica napus. J Huazhong Agric Univ (华中农业大学学报), 2007, 26(5): 607–611 (in Chinese with English abstract)



[2]Umezawa T, Yoshida R, Maruyama K, Yamaguchi-Shinozaki K, Shinozaki K. SRK2C, a SNF1-related protein kinase 2, improves drought tolerance by controlling stress-responsive gene expression in Arabidopsis thaliana. Proc Natl Acad Sci USA, 2004, 101: 17306–17311



[3]Selvam J N, Kumaravadivel N, Gopikrishnan A, Kumar B K, Ravikesavan R, Boopathi M N. Identification of a novel drought tolerance gene in Gossypium hirsutum L. cv KC3. Commun Biometry Crop Sci, 2009, 4: 9–13



[4]Yan A H, Zhang L F, Zhang Y W, Wang D M. Early stage SSH library construction of wheat near isogenic line TcLr19 under the stress of Puccinia recondita f. sp. tritici. Front Agric China, 2009, 3: 146–151



[5]Islam M A, Du H, N J, Ye H Y, Xiong L Z. Characterization of Glossyl homologous genes in rice involved in leaf wax accumulation and drought resistance. Plant Mol Biol, 2009, 70: 443–456



[6]Poroyko V, Hejlek L G, Spollen W G, Springer G K, Nguryen H T, Sharp R E, Bohnert H J. The maize root transcriptome by serial analysis of gene expression. Plant Physiol, 2005, 138: 1700–1710



[7]Yue G D, Zhang Y L, Li Z X, Sun L, Zhang J R. Differential gene expression analysis of maize leaf at heading stage in response to water-deficit stress. Bioscie Rep, 2008, 28: 125–134



[8]Zhang G Y, Chen M, Li L C, Xu Z, Chen X P, Guo J M, Ma Y Z. Overexpression of the soybean GmERF3 gene, and AP2/ERF type transcription factor for increased tolerances to salt, drought, and diseases in transgenic tobacco. J Exp Bot, 2009, 60: 3781–3796



[9]Buchanan C D, Lim S, Salzman R A, Kagiampakis I, Morishige D T, Weers B D, Klein R R, Pratt L H, Cordonnier-Pratt M M, Klein P E Mullet, J E. Sorghum bicolor’s transcriptome response to dehydration high salinity and ABA. Plant Mol Biol, 2005, 58: 699–720



[10]Kanneganti V, Gupta A K. Overexpression of OsiSAP8, a member of stress associated protein (SAP) gene family of rice confers tolerance to salt, drought and cold stress in transgenic tobacco and rice. Plant Mol Biol, 2008, 66: 445–462



[11]Yu Q, Hu Y, Li J, Wu Q, Lin Z. Sense and antisense expression of plasma membrane aquaporin BnPl from Brassica napus in tobacco and its effects on plant drought resistance. Plant Sci, 2005, 169: 647–656



[12]Rebrikov D V, Desai S M, Siebert P D, Lukyanov S A. Suppression subtractive hybridization. Methods Mol Biol, 2004, 258: 107–134



[13]Li H-Y(李惠勇), Huang S-H(黄素华), Shi Y-S(石云素), Song Y-C(宋燕春), Zhao J-R(赵久然), Wang F-G(王凤格), Wang T-Y(王天宇), Li Y(黎裕). Isolating soil drought-induced genes from maize seedling leaves through suppression subtractive hybridization. Sci Agric Sin (中国农业科学), 2007, 6(6): 647–651 (in Chinese with English abstract)



[14]Zhang H(张宏), Song G-Q(宋国琦), Ji W-Q(吉万全), Hu Y-G(胡银岗). Gene induction by drought stress in wheat variety Xiaoyan 22 and their expression analysis. J Agric Biotechnol (农业生物技术学报), 2009, 17(4): 670–676 (in Chinese with English abstract)



[15]Clement M, Lambert A, Herouart D, Boncompagni E. Identification of new up-regulated genes under drought stress in soybean nodules. Gene, 2008, 426: 15–22



[16]Wang D-L(王德龙), Ye W-W(叶武威), Wang J-J(王俊娟), Song L-Y(宋丽艳), Fan W-L(樊伟丽), Cui Y-P(崔宇鹏). Construction of SSH library and its analyses of cotton drought associated genes under drought stress. Acta Agron Sin (作物学报), 2010, 36(12): 2035−2044 (in Chinese with English abstract)



[17]Zhang L(张玲), Li F-G(李付广), Liu C-L(刘传亮), Zhang C-J(张朝军), Wu Z-X(武之霞). Isolation and analysis of drought-related gene from cotton (Gossypium arboreum L.) library. Cotton Sci (棉花学报), 2010, 22(2): 110–114 (in Chinese with English abstract)



[18]Liu W-R(刘文荣), Zhang J-S(张积森), Rao J(饶进), Cai Q-H(蔡秋华), Weng X-Y(翁笑艳), Ruan M-H(阮妙鸿), Que Y-X(阙友雄), Chen R-K(陈如凯), Zhang M-Q(张木清). Construction and analysis of suppression subtractive hybridization library for Saccharum arundinaceum Retz. leaves exposed to drought stress. Acta Agron Sin (作物学报), 2007, 33(6): 961–967 (in Chinese with English abstract)



[19]Tao S-H(陶士珩). Bioinformatics (生物信息学). Beijing: Science Press, 2007. pp 98–99 (in Chinese)



[20]Ashburner M, Ball C A, Blake J A, Botstein D, Butler H, Cherry J M, Davis A P, Dolinski K, Dwight S S, Eppig J T, Harris M A, Hill D P, Issel-Tarver L, Kasarskis A, Lewis S, Matese J C, Richardson J E, Ringwald M, Rubin G M, Sherlock G. Gene Ontology: tool for the unification of biology. Nat Genet, 2000, 25: 25–29



[21]Nguyen H T, Leipner J, Stamp P, Guerra-Peraza O. Low temperature stress in maize (Zea mays L.) induces genes involved in photosynthesis and signal transduction as studied by suppression subtractive hybridization. Plant Physiol Biochem, 2009, 47: 116–122



[22]Chaves M M, Flexas J, Pinheiro C. Photosynthesis under drought and salt stress: regulation mechanisms from whole plant to cell. Ann Bot, 2009, 103: 551–560



[23]Turchetto-Zolet A C, Margis-Pinheiro M, Margis R. The evolution of pyrroline-5-carboxylate synthase in plants: a key enzyme in proline synthesis. Mol Genet Genom, 2009, 281: 87–97



[24]Ahmed N U, Park J I, Seo M S, Kumar T S, Lee I H, Park B S, Nou I S. Identification and expression analysis of chitinase genes related to biotic stress resistance in Brassica. Mol Biol Rep, 2012, 39: 3649–3657



[25]Raeini-Sarjanz M, Chalavi V. Effects of water stress and constitutive expression of a drought induced chitinase gene on water-use efficiency and carbon isotope composition of strawberry. J Appl Bot Food Qual, 2011, 84: 90–94

[1] WANG Xia, YIN Xiao-Yu, Yu Xiao-Ming, LIU Xiao-Dan. Effects of drought hardening on contemporary expression of drought stress memory genes and DNA methylation in promoter of B73 inbred progeny [J]. Acta Agronomica Sinica, 2022, 48(5): 1191-1198.
[2] DING Hong, XU Yang, ZHANG Guan-Chu, QIN Fei-Fei, DAI Liang-Xiang, ZHANG Zhi-Meng. Effects of drought at different growth stages and nitrogen application on nitrogen absorption and utilization in peanut [J]. Acta Agronomica Sinica, 2022, 48(3): 695-703.
[3] ZHANG Ming-Cong, HE Song-Yu, QIN Bin, WANG Meng-Xue, JIN Xi-Jun, REN Chun-Yuan, WU Yao-Kun, ZHANG Yu-Xian. Effects of exogenous melatonin on morphology, photosynthetic physiology, and yield of spring soybean variety Suinong 26 under drought stress [J]. Acta Agronomica Sinica, 2021, 47(9): 1791-1805.
[4] LI Jie, FU Hui, YAO Xiao-Hua, WU Kun-Lun. Differentially expressed protein analysis of different drought tolerance hulless barley leaves [J]. Acta Agronomica Sinica, 2021, 47(7): 1248-1258.
[5] LI Peng-Cheng, BI Zhen-Zhen, SUN Chao, QIN Tian-Yuan, LIANG Wen-Jun, WANG Yi-Hao, XU De-Rong, LIU Yu-Hui, ZHANG Jun-Lian, BAI Jiang-Ping. Key genes mining of DNA methylation involved in regulating drought stress response in potato [J]. Acta Agronomica Sinica, 2021, 47(4): 599-612.
[6] QIN Tian-Yuan, LIU Yu-Hui, SUN Chao, BI Zhen-Zhen, LI An-Yi, XU De-Rong, WANG Yi-Hao, ZHANG Jun-Lian, BAI Jiang-Ping. Identification of StIgt gene family and expression profile analysis of response to drought stress in potato [J]. Acta Agronomica Sinica, 2021, 47(4): 780-786.
[7] ZHOU Lian, LIU Chao-Xian, XIONG Yu-Han, ZHOU Jing, CAI Yi-Lin. Functional analysis of plasma membrane intrinsic protein ZmPIP1;1 involved in drought tolerance and photosynthesis in maize [J]. Acta Agronomica Sinica, 2021, 47(3): 472-480.
[8] LIU Ya-Wen, ZHANG Hong-Yan, CAO Dan, LI Lan-Zhi. Prediction of drought and salt stress-related genes in rice based on multi-platform gene expression data [J]. Acta Agronomica Sinica, 2021, 47(12): 2423-2439.
[9] QIN Tian-Yuan, SUN Chao, BI Zhen-Zhen, LIANG Wen-Jun, LI Peng-Cheng, ZHANG Jun-Lian, BAI Jiang-Ping. Identification of drought-related co-expression modules and hub genes in potato roots based on WGCNA [J]. Acta Agronomica Sinica, 2020, 46(7): 1033-1051.
[10] ZHANG Hai-Yan, WANG Bao-Qing, FENG Xiang-Yang, LI Guang-Liang, XIE Bei-Tao, DONG Shun-Xu, DUAN Wen-Xue, ZHANG Li-Ming. Effects of drought treatments at different growth stages on growth and the activity of osmotic adjustment in sweet potato [Ipomoea batatas (L.) Lam.] [J]. Acta Agronomica Sinica, 2020, 46(11): 1760-1770.
[11] LI Xu-Kai,LI Ren-Jian,ZHANG Bao-Jun. Identification of rice stress-related gene co-expression modules by WGCNA [J]. Acta Agronomica Sinica, 2019, 45(9): 1349-1364.
[12] Yi YUAN,Shuang ZHU,Ting-Ting FANG,Jin-Jin JIANG,You-Ping WANG. Analysis of drought resistance and DNA methylation level of resynthesized Brassica napus [J]. Acta Agronomica Sinica, 2019, 45(5): 693-704.
[13] Ping LI,Wan-Wei HOU,Yu-Jiao LIU. Proteomic analysis of drought stress response on drought resistance for Vicia faba L. variety ‘Qinghai 13’ in Qinghai Plateau of China [J]. Acta Agronomica Sinica, 2019, 45(2): 267-275.
[14] LI Peng-Cheng,BI Zhen-Zhen,LIANG Wen-Jun,SUN Chao,ZHANG Jun-Lian,BAI Jiang-Ping. DNA methylation involved in regulating drought stress response of potato [J]. Acta Agronomica Sinica, 2019, 45(10): 1595-1603.
[15] Chao MI,Yan-Ning ZHAO,Zi-Gang LIU,Qi-Xian CHEN,Wan-Cang SUN,Yan FANG,Xue-Cai LI,Jun-Yan WU. Cloning of RuBisCo Subunits Genes rbcL and rbcS from Winter Rapeseed (Brassica rapa) and Their Expression under Drought Stress [J]. Acta Agronomica Sinica, 2018, 44(12): 1882-1890.
Viewed
Full text


Abstract

Cited

  Shared   
  Discussed   
No Suggested Reading articles found!