Welcome to Acta Agronomica Sinica，

Acta Agron Sin ›› 2009, Vol. 35 ›› Issue (1): 71-78.doi: 10.3724/SP.J.1006.2009.00071

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Cloning of Cu/Zn-Superoxide Dismutase of Brassica napus and Its Induced Expression by Sclerotinia slerotiorum

YANY Yang-Yang,LI Yun,DING Yong,XI Chun-Lei,ZHANG Cheng-Gui,LIU Ying,GAN Li   

  1. College of Plant Science and Technology,Huazhong Agricultural University, Wuhan 430070,China
  • Received:2008-04-20 Revised:2008-06-15 Online:2009-01-12 Published:2008-11-17
  • Contact: YANY Yang-Yang E-mail:yangyang401@126.com

PDF

1593

Cited

4

Abstract:

Bythe primers designed from conserved homologous sequences of the genes for superoxide dismutase (SOD) among Arabidopsis thaliana, Brassica juncea and B. rapa ssp. pekinensis, Cu/ZnSOD and FeSOD genes in B. napus were cloned using the methods of homology-based candidate gene and RACE-PCR. After cloning andsequencing, the full lengths of cDNA of Cu/ZnSOD and FeSOD genes (accession no. AY970822 and EF634058 in GenBank) were 756 and 1 037 bp, respectively. Their corresponding genomic sequences were 1 322 bp for Cu/ZnSOD gene and 1 659 bp for FeSOD gene with accession nos. DQ431853 and EF634057 in GenBank, respectively. Bioinformatic analyses showed that the ORF of Cu/ZnSOD gene with 459 bp contained six introns with seven extrons and a polyA tail and could express a protein of 152 amino acids; and the one of FeSOD gene with 792 bp contained seven introns with eight extrons and could determine a protein of 263 amino acids. With cDNA of Cu/ZnSOD gene as probe, Northern blotting analysis showed that its mRNA expression in B. napus cultivars increased after the infection by Sclerotinia sclerotiorum, the expression amount in resistant or tolerant cultivars was higher than that in sensitive ones. The same trend was observed for the SOD activity in leaves. These results suggested that SOD genes were related with the resistance to the disease.

Key words: Cu/ZnSOD, FeSOD, Brassica napus, Sclerotinia sclerotionrum, Gene expression analysis

[1]Fink R C, Scandalios J G. Molecular evolution and structure-function relationships of the superoxide dismutase gene families in angiosperms and their relationship to other eukaryotic and prokaryotic superoxide dismutases. Arch Biochem Biophys, 2002, 399: 19–36
[2]McCord J M, Fridovich I. Superoxide dismutase: An enzymatic function for erythrocuprein (hemocuprein). J Biol Chem, 1969, 244: 6049–6055
[3]Perl A, Perl-Treves R, Galili S. Enhanced oxidative- stress defense in transgenic potato expressing tomato Cu/Zn superoxide dismutase. Theor Appl Genet, 1993, 85: 568–576
[4]McKersie B D, Murnaghan J, Jones K S, Bowley S R. Iron superoxide dismutase expression in transgenic alfalfa increases winter survival without a detectable increase in photosynthetic oxidative stress tolerance. Plant Physiol, 2000, 122: 1427–1437
[5]Kliebenstein D J, Gershenzon J, Mitchellolds T. Comparative quantitative trait loci mapping of aliphatic, indolic and benzylic glucosinolate production in Arabidopsis thaliana leaves and seeds. Genetics, 2001, 159: 359–370
[6]Baek K H, Skinner D Z. Differential expression of manganese superoxide dismutase sequence variants in near isogenic lines of wheat during cold acclimation. Plant Cell Rep, 2006, 25: 223–30
[7]Liu J J, Goh C J, Loh C S, Tay E B H, Pua E C. Cloning of two cDNAs encoding Cu/Zn-superoxide dismutase (accession No. X95726, X95728) of mustard (Brassica juncea L. Czern & Coss). Plant Physiol, 1998, 116: 867–867
[8]Chen J-Y(陈锦云), Lin X-Y(林祥永), Lan Z-B(兰志斌). The effect of superoxide dismutase on weath fleck prevention. Fujian Agric Sci Technol (福建农业科技), 1996, (5): 13(in Chinese)
[9]Wen C-Y(文才艺), Hao Y-H(昊元华), Li H-Y(李浩戈). Changes of SOD activity and MDA in tobacco leaves infected by potato virus Y vein necrosis. Chin Tob Sci (中国烟草科学), 1999, (1): 12–14(in Chinese with English abstract)
[10]Wong Y P, Liu Y Q, Shi L. SOD activity of wheat varieties with different resistance to scab. Acta Phytophysiol Sin, 1993, 19: 358–358
[11]Yun X F, LI R X. Studies of induced resistance to downy mildew of cucumber with SOD isozyme protein in cotyledons. Acta Phytopathol Sin, 1997, 27: 221–224
[12]Niu L-Y(牛立元), Wang H-S(王鸿升), Shi M-W(石明旺). Changes of SOD and POD activities in wheat leaves infected by wheat powdery mildew and their relations to resistance. J Henan Voction-Technical Teachers Coll (河南职业技术师范学院学报), 2004, 32(4): 5–8(in Chinese with English abstract)
[13]Li R-H(李荣花), Chen J(陈捷), Gao Z-G(高增贵), Tang B-H(唐保宏), Liu J-H(刘军华). Relations between resistance to maize sheath blight and resistant enzymes. J Tianjin Norm Univ (Nat Sci Edn)(天津师范大学学报·自然科学版), 2005, 25(4): 32–36(in Chinese with English abstract)
[14]Wu J-J(吴俊江). Studies on the changes of SOD, POD activity and soluble sugar content in leaves of soybean varieties inoculated with Pseudomonas syringae pv. glycinea. Heilongjiang Agric Sci (黑龙江农业科学), 2006, (2): 32–34(in Chinese with English abstract)
[15]Purdy L H. Sclerotinia sclerotiorum: History, disease and symptomatology, host range, geographic distribution and impact. Phytopathology, 1979, 69: 875–890
[16]Gan L(甘莉), Wu X-L(伍新玲), Jin L(金良), Feng S-Q(丰胜求), Chen C-L(陈翠莲), Tang H(汤华). The establishment of sclerotinia sclerotiorum resistant near isogenic lines. J Wuhan Univ (Nat Sci Edn) (武汉大学学报·理学版), 2002, 48(6): 761–764(in Chinese with English abstract)
[17]Hu B-C(胡宝成), Rimmer S R. Preliminary study of artificial inoculation for resistance (tolerance) to sclerotinia sclerotiorum in rapeseed using detached leaves. J Anhui Agric Sci (安徽农业科学), 1989, (3): 56–58(in Chinese with English abstract)
[18]Yoon K, Kim H, Lee W. Cloning and DNA sequencing of cytosolic Cu/Zn superoxide dismutase gene from Chinese cabbage. J Plant Biol, 1998, 41: 68–71
[19]Li H-S(李合生), Sui Q(孙群), Zhao S-J(赵世杰), Zhang W-H(章文华). Principles and Techniques for Plant Physiological and Biochemical Experiment(植物生理生化实验原理和技术). Beijing: Higher Education Press, 2000. pp 167–169(in Chinese)
[20]Patel R N, Nripendra S, Shukla K K, Gundla V L, Chauhan U K. Synthesis, structure and biomimetic properties of Cu(II)-Cu(II) and Cu(II)-Zn(II) binuclear complexes: Possible models for the chemistry of Cu/Zn superoxide dismutase. J Inorganic Biochem, 2005, 99: 651–663
[1] Huan TAN,Yu-Hui LIU,Li-Xia LI,Li WANG,Yuan-Ming LI,Jun-Lian ZHANG. Cloning and Functional Analysis of R2R3 MYB Genes Involved in Anthocyanin Biosynthesis in Potato Tuber [J]. Acta Agronomica Sinica, 2018, 44(7): 1021-1031.
[2] CHEN Xue-Ping**,JING Ling-Yun**,WANG Jia,JIAN Hong-Ju,MEI Jia-Qin,XU Xin-Fu,LI Jia-Na,LIU Lie-Zhao*. Correlation Analysis of Sclerotinia Resistance with Lignin Content and Monomer G/S and its QTL Mapping in Brassica napus L. [J]. Acta Agron Sin, 2017, 43(09): 1280-1289.
[3] HOU Lin-Tao,WANG Teng-Yue,JIAN Hong-Ju,WANG Jia,LI Jia-Na,LIU Lie-Zhao. QTL Mapping for Seedling Dry Weight and Fresh Weight under Salt Stress and Candidate Genes Analysis in Brassica napus L. [J]. Acta Agron Sin, 2017, 43(02): 179-189.
[4] LU Kun,Shen Ge-Zi,LIANG Ying,FU Ming-Lian,HE Bin,TIE Lin-Mei,ZHANG Ye, PENG Liu,LI Jia-Na. Analysis of Yield Components with High Harvest Index in Brassica napusunder Environments Fitting Different Yield Levels [J]. Acta Agron Sin, 2017, 43(01): 82-96.
[5] WANG Wen-Xiang,HU Qiong,MEI De-Sheng,LI Yun-Chang,ZHOU Ri-Jin,WANG Hui,CHENG Hong-Tao,FU Li,LIU Jia*. Genetic Effects of Branch Angle Using Mixture Model of Major Gene Plus Polygene in Brassica napus L. [J]. Acta Agron Sin, 2016, 42(08): 1103-1111.
[6] TAN Tai-Long,FENG Tao,LUO Hai-Yan,PENG Ye,LIU Rui-Yang,GUAN Chun-Yun. Cloning and Characterization of Phospholipids:Diacylglycerol Acyltransferase (BnPDAT1) cDNA from Brassica napus L. [J]. Acta Agron Sin, 2016, 42(05): 658-666.
[7] ZHOU Tian-Shan,WANG Xin-Chao,YU You-Ben,XIAO Yao,QIAN Wen-Jun,XIAO Bin,YANG Ya-Jun. Correlation Analysis between Total Catechins (or Anthocyanins) and Expression Levels of Genes Involved in Flavonoids Biosynthesis in Tea Plant with Purple Leaf [J]. Acta Agron Sin, 2016, 42(04): 525-531 .
[8] KUAI Jie,DU Xue-Zhu,HU Man,ZENG Jiang-Xue,ZUO Qing-Song,WU Jiang-Sheng,ZHOU Guang-Sheng. Effect of Symbiosis Periods and Plant Densities on Growth and Yield of Rapeseed Intercropping Cotton [J]. Acta Agron Sin, 2016, 42(04): 591-599.
[9] LU Kun,QU Cun-Min,LI Sha,ZHAO Hui-Yan,WANG Rui,XU Xin-Fu,LIANG Ying,LI Jia-Na. Expression Analysis and eQTL Mapping of BnTT3 Gene in Brassica napus L. [J]. Acta Agron Sin, 2015, 41(11): 1758-1766.
[10] JIAO Cong-Cong,HUANG Ji-Xiang,WANG Yi-Long,ZHANG Xiao-Yu,XIONG Hua-Xin,NI Xi-Yuan,ZHAO Jian-Yi. Genetic Analysis of Yield-Associated Traits by Unconditional and Conditional QTL in Brassica napus [J]. Acta Agron Sin, 2015, 41(10): 1481-1489.
[11] TANG Min-Qiang,CHENG Xiao-Hui,TONG Chao-Bo,LIU Yue-Ying,ZHAO Chuan-Ji,DONG Cai-Hua,YU Jing-Yin,MA Xiao-Gen,HUANG Jun-Yan,LIU Sheng-Yi. Genome-wide Association Analysis of Plant Height in Rapeseed (Brassica napus) [J]. Acta Agron Sin, 2015, 41(07): 1121-1126.
[12] WANG Jia,JING Ling-Yun,JIAN Hong-Ju,QU Cun-Min,CHEN Li,LI Jia-Na,LIU Lie-Zhao. Quantitative Trait Loci Mapping for Plant Height, the First Branch Height, and Branch Number and Possible Candidate Genes Screening in Brassica napus L. [J]. Acta Agron Sin, 2015, 41(07): 1027-1038.
[13] ZHENG Xiao-Min,GUO Nan,GAO Tian-Shu,GONG Hui-Ming,ZHANG Tao. Cloning and Expression Analysis of Defensin Genes from Brassica napus [J]. Acta Agron Sin, 2015, 41(05): 725-732.
[14] ZHANG Ya-Jie,LI Jing,PENG Hong-Kun,CHEN Xiu-Bin,ZHENG Hong-Yu,CHEN Sheng-Bei,LIU An-Guo,HU Li-Yong. Dynamic Simulation Model for Growth Duration of Rapeseed (Brassica napus) [J]. Acta Agron Sin, 2015, 41(05): 766-777.
[15] Lü Yan-Yan,FU San-Xiong,CHEN Song,ZHANG Wei,QI Cun-Kou*. Cloning of BnADH3 Gene from Brassica napus L. and Submergence Tolerance of BnADH3 Transgenic Arabidopsis [J]. Acta Agron Sin, 2015, 41(04): 565-573.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
Add: No. 80 Xueyuan South Rd, Beijing 100081, P. R. China E-mail: zwxb301@caas.cn
Supported by Beijing Magtech Co.Ltd