Welcome to Acta Agronomica Sinica，

Acta Agron Sin ›› 2012, Vol. 38 ›› Issue (01): 62-70.doi: 10.3724/SP.J.1006.2012.00062

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

cDNA-AFLP Analysis of Differentially Expressed Genes in Tobacco Infected by TMV

CHEN Rong-Ping1,2,LIU Lie3,WAN Xiu-Qing2,QIU En-Jian2,WANG Chun-Jun2,SONG Bao-Gang2,YAN Pei-Qiang2,YANG Tie-Zhao1,*   

  1. 1 College of Tobacco Science, Henan Agricultural University, Zhengzhou 450002, China; 2 Mudanjiang Tobacco Science and Research Institute of Heilongjiang Province Tobacco Company, Mudanjiang 157011, China; 3 Centers for Disease Control and Prevention in Mudanjiang, Mudanjiang 157022, China
  • Received:2011-05-10 Revised:2011-09-12 Online:2012-01-12 Published:2011-11-07
  • Contact: 杨铁钊, E-mail: yangtiezhao@126.com, Tel: 0371-63358030

PDF

1527

Cited

8

Abstract: Flue-cured tobacco variety Longjiang 925 with high resistance to tobacco mosaic virus (TMV) was used as the tested material, and we selected 240 pairs of primers and amplified cDNA from the tobacco leaf inoculated by TMV. The results showed that approximately 9500 gene transcript fragments were obtained, and twelve inducible expressed gene fragments were selected out by cloning and sequencing. The inducible fragments functions, involve in the nucleic acid metabolism, protein synthesis and modulation, energy metabolism, stress responding, intracellular transport, metabolism of carbohydrates. To validate the functions of these differentially expressed gene sequences obtained, we analyzed TIF2 fragment by real-time PCR with the samples collected at 0, 12, 24, 48, and 72 hours after inoculation, and the mRNAs samples were isolated. The result of real-time PCR indicated that the genes isolated were related to TMV-resistance. Then, both 5′- and 3′-rapid amplification of cDNA ends (RACE) were preformed. A full length cDNA sequence of TIF2 contained 875 bp, with a coding zone of 101 bp to 613 bp conjecturably, encoding 170 amino acids. Analyses of Blastn and Blastp showed that the gene was not homologous to tobacco, and probably was the TMV-resistance related novel gene. The results will be helpful for the future research on tobacco resistant-breeding to TMV in molecule level and so on.

Key words: Tobacco, TMV, Differentially Expressed Genes, cDNA-AFLP

[1]Fang Y-C(方宇澄). Pictures Book of Tobacco Diseases in China (中国烟草病虫害彩色图志). Hefei: Anhui Science and Technology Publishing House, 1992. pp 24–15 (in Chinese)
[2]Li Y-P(李永平), Wang Y-K(王颖宽), Ma W-G(马文广), Wang S-K(王绍坤), Duan Y-Q(段玉琪). Preliminary research report on TMV resistant breeding of flue-cured Tobacco. J Yunnan Agric Univ (云南农业大学学报), 2003, (3): 94–96 (in Chinese with English abstract)
[3]Su D-C(苏德成) ed. Tobacco Breeding (烟草育种). Beijing: China Financial & Economic Publishing House, 2000. pp 117–119 (in Chinese)
[4]Stavely J R. Disease resistance. In: Durbin R D ed. Nicotiana. Procedures for Experimental Use, Technical Bulletin 1586, US Department of Agriculture, 1979. pp 87–110
[5]Zhang H-M(张恒木), Chen J-P(陈剑平), Cheng Y(程晔). Study on the resistant gene of tobacco to tobacco mosaic virus. Acta Agric Zhejiangensis (浙江农业学报), 2001, 13(2): 55–60 (in Chinese with English abstract)
[6]Buchanan B, Gruissem W, Jones R, eds. Qu L-J(瞿礼嘉), Gu H-Y(顾红雅), Bai S-N(白书农), Zhao J-D(赵进东), Chen Z-L(陈章良), Deng X-W(邓兴旺) trans. Biochemistry & Molecular Biology of Plants (植物生物化学与分子生物学). Beijing: Science Press, 2004. pp 119–121 (in Chinese)
[7]Canto T, Palukaitis P. Novel N gene-Associated-temperature-independent resistance to the movement of tobacco mosaic virus vectors neutralized by a cucumber mosaic virus RNA1 transgene. J Virol, 2002, 12: 12908–12916
[8]Stange C, Matus J T, Elorza A, Arce-Johnson P. Identification and characterization of a novel tobacco mosaic virus resistance N gene homologue in Nicotiana tabacum plants. Funct Plant Biol, 2004, 31: 149–158
[9]Whitham S, McCormick S, Backer B. The N gene of tobacco confers resistance to tobacco mosaic virus in transgenic tomato. Plant Biol, 1996, 93: 8776–8781
[10]Zhang G(张岗), Dong Y-L(董艳玲), Xia N(夏宁), Zhang Y(张毅), Wang X-J(王晓杰), Qu Z-P(屈志鹏), Li Y-M(李依民), Huang L-L(黄丽丽), Kang Z-S(康振生). cDNA-AFLP analysis reveals differential gene expression in wheat adult-plant resistance to stripe rust. Acta Agron Sin (作物学报), 2010, 36(3): 401–409 (in Chinese with English abstract)
[11]Zheng X W, Chen X W, Zhang X H, Lin Z Z, Shang J J, Xu J C, Zhai W X, Zhu L H. Isolation and identification of a gene in response to rice blast disease in rice. Plant Mol Biol, 2004, 54: 99–109
[12]Xie R(谢荣), He Z-S(何志水), Liu F-H(刘芳华), Zhou H-S(皱华松), Zhu J-B(朱家壁), Yu G-Q(俞冠翘). Using cDNA-AFLP identify the genes relation to root nodules development in Medicago sativa. Chin Sci Bull (科学通报), 2006, 51(15): 1794–1801 (in Chinese with English abstract)
[13]Chen G-P(陈桂平), Huang Z-J(黄占景), Ma W-S(马闻师), Shen Y-Z(沈银柱). Isolation and characterization of the cDNA fragments of wheat involved in saIt stress. Sci Agric Sin (中国农业科学), 2003, 36(9): 996–999 (in Chinese with English abstract)
[14]Chen G P, Ma W S, Huang Z J, Xu T, Xue Y B, Shen Y Z. Isolation and characterization of TaGSKl involved in wheat salt tolerance. Plant Sci, 2003, 165: 1369–1375
[15]Mi C-L(秘彩莉), Zhang X-Y(张学勇), Wen X-J(温小杰), Liu X(刘旭). Isolation of TaVHA-C, a gene in wheat related to salt-tolerance via cDNA-AFLP. Sci Agric Sin (中国农业科学), 2006, 39(9): 1736–1742 (in Chinese with English abstract)
[16]Liu L(刘丽), Ma Y-Y(马永毅), Zhang Z-M(张志明), Pan G-T(潘光堂), Zhao M-J(赵茂俊). cDNA-AFLP analysis of Rhizoctonia solani AG1-lA-induced maize differential gene expression. Acta Phytopathol Sin (植物病理学报), 2009, 39(4): 385–391 (in Chinese with English abstract)
[17]Zhao J-R(赵继荣), Li N(李宁), Liu H-X(刘红霞), Bi Y(毕阳), Zhang Z-Y(张增艳). Confirmation of cDNA-AFLP differential expressed fragments of related defense genes against BYDV in wheat. J Plant Genet Resour (植物遗传资源学报), 2009, 10(1): 16–20 (in Chinese with English abstract)
[18]Bachem C W, van derHoeven R S, de Bruijn S M, Vreugdenhil D, Zabeau M, Visser R G. Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP: Analysis of gene expression during potato tuber development. Plant J, 1996, 19: 745–753
[19]Guo Z-K(郭兆奎), Yan P-Q(颜培强), Wan X-Q(万秀清), Li L-J(李丽杰), Yang Q(杨谦). Study on the real-time PCR quantity detecting methods of genetic modified tobacco. China Biotechnol (中国生物工程杂志), 2005, 25(1): 76–80 (in Chinese with English abstract)
[20]Zhou J(周佳), Li F-X(李凤霞), Chen S(陈帅), Luo C-G(罗成刚), Liu G-S(刘贯山), Jiang C-H(蒋彩虹), Yang A-G(杨爱国), Su Z-G(苏振刚), Wang Y-Y(王元英). Cloning and expression analysis of the PVY resistance-related gene NtPsaN in tobacco (Nicotiana tabacum). Sci Agric Sin (中国农业科学), 2010, 43(16): 3323–3330 (in Chinese with English abstract)
[21]Chen S(陈帅), Liu G-S(刘贯山), Yang A-G(杨爱国), Wang Y-Y(王元英), Sun Y-H(孙玉合). Gene isolation and expression analysis of NtERD1 induced by PVY in tobacco (Nicotiana tabacum L.). Chin Tob Sci (中国烟草科学), 2010, (5): 62–67 (in Chinese with English abstract)
[1] LI A-Li, FENG Ya-Nan, LI Ping, ZHANG Dong-Sheng, ZONG Yu-Zheng, LIN Wen, HAO Xing-Yu. Transcriptome analysis of leaves responses to elevated CO2 concentration, drought and interaction conditions in soybean [Glycine max (Linn.) Merr.] [J]. Acta Agronomica Sinica, 2022, 48(5): 1103-1118.
[2] HUANG Wen-Gong, JIANG Wei-Dong, YAO Yu-Bo, SONG Xi-Xia, LIU Yan, CHEN Si, ZHAO Dong-Sheng, WU Guang-Wen, YUAN Hong-Mei, REN Chuan-Ying, SUN Zhong-Yi, WU Jian-Zhong, KANG Qing-Hua. Transcriptome profiling of flax (Linum usttatissimum L.) response to low potassium stress [J]. Acta Agronomica Sinica, 2021, 47(6): 1070-1081.
[3] XU Jing, PAN Li-Juan, LI Hao-Yuan, WANG Tong, CHEN Na, CHEN Ming-Na, WANG Mian, YU Shan-Lin, HOU Yan-Hua, CHI Xiao-Yuan. Expression pattern analysis of genes related to lipid synthesis in peanut [J]. Acta Agronomica Sinica, 2021, 47(6): 1124-1137.
[4] WANG Rui-Li, WANG Liu-Yan, LEI Wei, WU Jia-Yi, SHI Hong-Song, LI Chen-Yang, TANG Zhang-Lin, LI Jia-Na, ZHOU Qing-Yuan, CUI Cui. Screening candidate genes related to aluminum toxicity stress at germination stage via RNA-seq and QTL mapping in Brassica napus L. [J]. Acta Agronomica Sinica, 2021, 47(12): 2407-2422.
[5] LI Peng, LIU Che, SONG Hao, YAO Pan-Pan, SU Pei-Lin, WEI Yao-Wei, YANG Yong-Xia, LI Qing-Chang. Identification and analysis of non-specific lipid transfer protein family in tobacco [J]. Acta Agronomica Sinica, 2021, 47(11): 2184-2198.
[6] LIU Qing-Li,JIANG Yu-Zhou,ZOU Yan,ZHANG Yun-Gui,ZHANG Heng,SHI Jun-Xiong,LI Zhi-Hong. The study of carbon budget on field-tobacco ecosystem [J]. Acta Agronomica Sinica, 2020, 46(8): 1258-1265.
[7] REN Meng-Meng, ZHANG Hong-Wei, WANG Jian-Hua, WANG Guo-Ying, ZHENG Jun. Fine mapping of a major QTL qMES20-10 associated with deep-seeding tolerance in maize and analysis of differentially expressed genes [J]. Acta Agronomica Sinica, 2020, 46(7): 1016-1024.
[8] DONG Qing-Yuan,MA De-Qing,YANG Xue,LIU Yong,HUANG Chang-Jun,YUAN Cheng,FANG Dun-Huang,YU Hai-Qin,TONG Zhi-Jun,SHEN Jun-Ru,XU Yin-Lian,LUO Mei-Zhong,LI Yong-Ping,ZENG Jian-Min. Construction and characterization of a BAC library for flue-cured tobacco line with high resistance to blank shank [J]. Acta Agronomica Sinica, 2020, 46(6): 869-877.
[9] HENG You-Qiang,YOU Xi-Long,WANG Yan. Pathogenesis-related protein gene SfPR1a from Salsola ferganica enhances the resistances to drought, salt and leaf spot disease in transgenic tobacco [J]. Acta Agronomica Sinica, 2020, 46(4): 503-512.
[10] Shan-Bin CHEN, Si-Fan SUN, Nan NIE, Bing DU, Shao-Zhen HE, Qing-Chang LIU, Hong ZHAI. Cloning of IbCAF1 and identification on tolerance to salt and drought stress in sweetpotato [J]. Acta Agronomica Sinica, 2020, 46(12): 1862-1869.
[11] JIA Xiao-Xia,QI En-Fang,LIU Shi,WEN Guo-Hong,MA Sheng,LI Jian-Wu,HUANG Wei. Effects of over-expression of AtDREB1A gene on potato growth and abiotic stress resistance gene expression [J]. Acta Agronomica Sinica, 2019, 45(8): 1166-1175.
[12] Xiao-Han MA,Jie ZHANG,Huan-Wei ZHANG,Biao CHEN,Xin-Yi WEN,Zi-Cheng XU. Exogenous MeJA improves cold tolerance of tobacco by inhibiting H2O2 accumulation [J]. Acta Agronomica Sinica, 2019, 45(3): 411-418.
[13] Zhi-Jun TONG,Yi-Han ZHANG,Xue-Jun CHEN,Jian-Min ZENG,Dun-Huang FANG,Bing-Guang XIAO. Mapping of quantitative trait loci conferring resistance to brown spot in cigar tobacco cultivar Beinhart1000-1 [J]. Acta Agronomica Sinica, 2019, 45(3): 477-482.
[14] Cheng-Jiang LI,Da-Fei LI,Gui-Su ZHOU,Long XU,Tian-Yang XU,Zheng-Xiong ZHAO. Effects of different types of biochar on soil microorganism and rhizome diseases occurrence of flue-cured tobacco [J]. Acta Agronomica Sinica, 2019, 45(2): 289-296.
[15] Jian-Fei ZHOU,Yun-Jie WU,Gang XUE,An-Qian ZHANG,Pei TIAN,Yu-Fu PENG,Tie-Zhao YANG. Relationship between GS isoenzyme activity and nitrogen transportation in flue-cured tobacco leaves [J]. Acta Agronomica Sinica, 2019, 45(1): 111-117.
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