Peanut bacterial wilt (BW) caused by Ralstonia solanacearum is one of the most devastating diseases for peanut production in the world. It is believed that breeding and subsequent planting BW-resistant cultivars of peanut (Arachis hypogaea L.) should represent the most effective and economic means of controlling the disease. To illustrate the mo-lecular mechanism of peanut resistant to BW, a BW-resistant cultivar, ‘Yuanza 9102’, and a BW-sensitive one, ‘Zhonghua 12’, were infected with Ralstonia solanacearum and differential expression of the genes related to BW-resistance was analyzed using complementary DNA amplified length polymorphism (cDNA-AFLP) technique. The infected 3-leaflet seedlings were followed for 48 h and root samples were taken at 0, 2, 10, 24 and 48 h after inoculation, respectively. A total of 12596 transcript-derived fragments (TDFs) were amplified with 256 primer combinations, including 709 differential expressed TDFs, which were generated from 119 primer combinations. Ninety-eight TDFs were randomly chosen for DNA sequence analysis. BLASTx analysis of the obtained sequences revealed that 40 TDFs encoded gene products associated with energy, transcription, signal transduction, defense, metabolism, cell growth, cell structure or/and protein synthesis. Analysis of the expression of four genes by qRT-PCR verified the results from cDNA-AFLP. Strikingly, one of the identified TDFs, 32-54-1, occurred for 47 times in a known BW-resistant SSH library. These results suggest that resistance to BW in peanut involves multifaceted biochemical and physiological reactions, including regulation of the genes involved in different pathways, such as defense, singal transduction, metabolism, transcription and abiotic stresses. The TDF 32-54-1 was predicted to be closely related to BW resistance in peanut.
[1] 段乃雄, 谈宇俊, 姜慧芳, 胡端红. 花生种质资源抗花生青枯病鉴定. 中国油料, 1993, 15(1): 22-25, 29.
[2] Gabriëls SHEJ, Takken FLW, Vossen JH, de Jong CF, Liu Q, Turk SCHJ, Wachowski LK, Peters J, Witsenboer HMA, de Wit PJGM, Joosten MHAJ. cDNA-AFLP com-bined with functional analysis reveals novel genes in-volved in the hypersensitive response. Mol Plant-Microbe Interact, 2006, 19(6): 567-576.
[3] Oomen RJFJ, Bergervoet MJEM, Bechem CWB, Visser RGF, Vincken JP. Exploring the use of cDNA-AFLP with leaf protoplasts as a tool to study primary cell wall biosynthesis in potato. Plant Physiol Biochem, 2003, 41(11-12): 965-971.
[4] Mao CZ, Yi KK, Yang L, Zheng BS, Wu YR, Liu FY, Wu P. Identification of aluminium-regulated genes by cDNA- AFLP in rice (Oryza sativa L.): aluminium-regulated genes for the metabolism of cell wall components. J Exp Bot, 2004, 55(394): 137-143.
[5] 王文霞, 褚栋, 高士刚, 闫红飞, 缑丽倩, 杨文香, 刘大群. 小麦抗叶锈近等基因系TcLr38的cDNA-AFLP分析. 中国农业科学, 2010, 43(2): 293-303.
[6] 姜慧芳, 陈本银, 任小平, 廖伯寿, 雷永, 傅廷栋, 马朝芝, Mace E, Crouch JH. 利用重组近交系群体检测花生青枯病抗性SSR标记. 中国油料作物学报, 2007, 29(1): 26-30.
[7] 姜慧芳, 段乃雄. 花生种质资源描述规范和数据标准. 北京: 中国农业出版社, 2006.
[8] 吕建伟. 青枯菌诱导的花生SSH文库的构建及分析
[学位论文]. 北京: 中国农业科学院研究生院, 2010.
[9] 晏立英, 黄家权, 雷永, 王圣玉, 廖伯寿. 花生青枯菌红安分离物的鉴定. 中国油料作物学报, 2010, 32(1): 144-146.
[10] 李乃坚, 黄爱兴, 袁四清, 王得元. 茄科作物抗青枯病水培法鉴定研究 II.液体培养青枯菌的致病力. 广东农业科学, 2000, (3): 38-40.
[11] Bachem CWB, van der Hoeven RS, De Bruijn SM, Vreugdenhil D, Zabeau M, Visser RGF. 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, 9(5): 745-753.
[12] 雷永, 廖伯寿, 王圣玉. 花生AFLP—— 银染体系的建立与优化. 花生学报, 2003, 32 (增刊): 301-305.
[13] Leal-Bertioli SCM, José ACVF, Alves-Freitas DMT, Moretzsohn MC, Guimarães PM, Nielen S, Vidigal BS, Pereira RW, Pike J, Fávero AP, Parniske M, Varshney RK, Bertioli D. Identification of candidate genome regions controlling disease resistance in Arachis. MBC Plant Biol, 2009, (9): 112.
[14] Pfaffl MW. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acid Res, 2001, 29(9): e45.
[15] Luo M, Dang P, Bausher MG, Holbrook CC, Lee RD, Lynch RE, Guo BZ. Identification of transcripts involved in resistance responses to leaf spot disease caused by Cercosporidium personatum in peanut (Arachis hypogaea). Genet Resist, 2005, 95(4): 381-387.
[16] Simões-Araújo JL, Rodrigues RL, de A Gerhardt LB, Mondego JMC, Alves-Ferreira M, Rumjanek NG, Margis-Pinheiro M. Identification of differentially expressed genes by cDNA-AFLP technique during heat stress in cowpea nodules. Federation Eur Biochem Soc Lett, 2002, 515(1-3): 44-50.
[17] 邹艳敏, 于拴仓, 张凤兰, 余阳俊, 赵岫云, 张德双. 白菜抽薹性状相关基因的cDNA-AFLP分析. 遗传, 2009, 31(7): 755-762.
[18] Wang XJ, Tang CL, Zhang G, Li YC, Wang CF, Liu B, Qu ZP, Zhao J, Han QM, Huang LL, Chen XM, Kang ZS. cDNA-AFLP analysis reveals differential gene expression in compatible interaction of wheat challenged with Puccinia striiformis f. sp. tritici. BMC Genomics, 2009, (10): 289.
[19] McClelland M, Mathieu-Daude F, Welsh J. RNA fingerprinting and differential display using arbitrarily primed PCR. Trends Genet, 1995, 11(6): 242-246.
[20] Baldwin D, Crane V, Rice D. A comparison of gel-based, nylon filter and microarray techniques to detect differential RNA expression in plants. Curr Opin Plant Biol, 1999, 2(2): 96-103.
[21] Sztukowska M, Bugno M, Potempa J, Travis J, Kurtz DM. Role of rubrerythrin in the oxidative stress response of Porphyromonas gingivalis. Mol Microb, 2002, 44(2): 479-488.
[22] Lim PO, Lee IC, Kim J, Kim HJ, Ryu JS, Woo HR, Nam HG. Auxin response factor 2 (ARF2) plays a major role in regulating auxin-mediated leaf longevity. J Exp Bot, 2010, 61(5): 1419-14
