Welcome to Acta Agronomica Sinica,

Acta Agron Sin ›› 2012, Vol. 38 ›› Issue (04): 606-613.doi: 10.3724/SP.J.1006.2012.00606

Previous Articles     Next Articles

Cloning and Expression Analysis of Fusarium Wilt Resistance-related Gene PvCaM1 in Common Bean (Phaseolus vulgaris L.)

XUE Ren-Feng,ZHU Zhen-Dong,WANG Xiao-Ming,WANG Lan-Fen,WU Xiao-Fei,WANG Shu-Min*   

  1. Institute of Crop Sciences, Chinese Academy of Agricultural Sciences / National Key Facility for Crop Gene Resources and Genetic Improvement, Beijing 100081 China
  • Received:2011-09-22 Revised:2011-12-19 Online:2012-04-12 Published:2012-02-13
  • Contact: 王述民, E-mail: smwang@mail.caas.net.cn, Tel: 010-82108567

Abstract: Calmodulin (CaM) is a multifunctional Ca2+-binding protein in plant cells. It plays important roles in regulating the growth, development and disease resistance in plants. A full-length cDNA sequence coding for CaM in common bean was cloned based on expressed sequence tags from common bean. Sequence analysis showed that the isolated fragment was 713 bp, it contained an open reading frame (ORF) of 453 bp encoding 150 amino acids with a theoretical molecular weight of 17.16 kD, designated PvCaM1 (GenBank accession number JN418801),. Online ScanProsite tool analysis showed that PvCaM1 had four Ca2+-binding domains with the function of combining with free Ca2+. Homology analysis indicated that PvCaM1 gene was similar to CaM genes in other plant species including Lotus japonicus (CAB63264.3), watermelon (BAI52955.1), Populus (ADC80735.1) and castor bean (XP_002533357.1).Phylogenetic analysis based on the amino acids sequence of PvCaM1 with other nine species showed that the protein encoded by this gene had the closest relationship with the CaM in Lotus japonicus and watermelon, the homology were 77% and 76%, respectively. Real time-PCR analysis indicated that the expression level of PvCaM1 in the interactions between the resistant cultivars and Fusarium wilt pathogen FOP-DM01 (Fusarium oxysporum f. sp. phaseoli) increased significantly and reached the peak at 96 h, however, the susceptible one touched the bottom under the same conditions.The expression level of PvCaM1 in 260205 leaves was higher than that in BRB-130 at all different time points. PvCaM1 expressed differently in the leaves, stems and roots, the expression level in leaves was higher than that in roots and stems. Transcriptional level of PvCaM1 was up-regulated by exogenous abscisic acid, methyl jasmonate and ethephon, but not increased significantly by salicylic acid and 3-indoleacetic acid. These results suggested that PvCaM1 is probably involved in the abscisic acid, jasmonic acid and ethylene-regulated resistant response pathways, but not closely related to the salicylic acid and 3-indoleacetic acid pathway. The study indicated that the function of PvCaM1 should be closely related to the resistant response pathways against Fusarium wilt pathogen FOP-DM01 with involvement of abscisic acid, jasmonic acid and ethylene in common bean.

Key words: Common bean, Fusarium wilt, PvCaM1 gene, Expression analysis

[1]Buruchara R A, Camacho L. Common bean reaction to Fusarium oxysporum f. sp. phaseoli, the cause of severe vascular wilt in Central Africa. J Phytopathol, 2000, 148(1): 39–45

[2]Pastor C, Abawi G S. Reactions of selected bean germplasm to infection by Fusarium oxysporum f. sp. phaseoli. Plant Dis, 1987, 71: 990–993

[3]Salgado M O, Schwartz H F, Brick M A. Inheritance of resistance to a Colorado race of Fusarium oxysporum f. sp. phaseoli in common beans. Plant Dis, 1995, 79: 279–281

[4]Miklas P N, Kelly J D, Beebe S E, Blair M W. Common bean breeding for resistance against biotic and abiotic stresses: From classical to MAS breeding. Euphytica, 2006, 147: 105–131

[5]Zielinski R E. Calmodulin and calmodulin-binding proteins in plants. Annu Rev Plant Biol, 1998, 49: 697–725

[6]Yang T, Segal G, Abbo S, Feldman M, Fromm H. Characterization of the calmodulin gene family in wheat: structure, chromosomal location, and evolutionary aspects. Mol Gen Genet, 1996, 252: 684–694

[7]Snedden W A, Fromm H. Calmodulin, calmodulin-related proteins and plant responses to the environment. Trends Plant Sci, 1998, 3: 299–304

[8]Barnett M J, Long S R. Nucleotide sequence of an alfalfa calmodulin cDNA. Nucl Acids Res, 1990, 18: 3395

[9]Chandra A, Thungapathra M, Upadhyaya K C. Molecular cloning and characterization of a calmodulin gene from Arabidopsis thaliana. J Plant Biochem Biotachnol, 1994, 3: 31–35

[10]Chye M L, Liu C M, Tan C T. A cDNA clone encoding Brassica calmodulin. Plant Mol Biol. 1995, 27: 419–423

[11]Duval F D, Renard M, Jaquinod M, Biou V, Montrichard F, Macherel D. Differential expression and functional analysis of three calmodulin isoforms in germinating pea (Pisum sativum L.) seeds. Plant J, 2002, 32: 481–493

[12]Lee S H, Kim J C, Lee M S, Heo, W D, Seo H Y, Yoon H W, Hong J C, Lee S Y, Bahk J D, Hwang I. Identification of a novel divergent calmodulin isoform from soybean which has differential ability to activate calmodulin-dependent enzymes. J Biol Chem, 1995, 270: 21806–21812

[13]Ling V, Assmann S M. Cellular distribution of calmodulin and calmodulin-binding proteins in Vicia faba L. Plant Physiol. 1992, 100: 970–978

[14]Nath M, Goel A, Taj G, Kumar A. Molecular cloning and comparative in silico analysis of calmodulin genes from cereals and millets for understanding the mechanism of differential calcium accumulation. J Prot Bioinform, 2010, 3: 294–301

[15]Takezawa D, Liu Z H, An G, Poovaiah B W. Calmodulin gene family in potato: developmental and touch-induced expression of the mRNA encoding a novel isoform. Plant Mol Biol, 1995, 27: 693–703

[16]Watillon B, Kettmann R, Boxus P, Burny A. Cloning and characterization of an apple (Malus domestica L. Borkh) calmodulin gene. Plant Sci, 1992, 82: 201–212

[17]Wang Y-H(王艳辉), Jia H(贾慧), Si H-L(司贺龙), Ma J-F(马继芳), Hao H-F(郝会芳), Dong J-G(董金皋). Change of calmodulin in corn leaf cell with different resistant genes under stress of HT-toxin from Exserohilum turcicum. J Hebei Agric Univ (河北农业大学学报), 2007, 30: 4–7 (in Chinese with English abstract)

[18]Hong Bo S, Li Ye C, Ming A S. Calcium as a versatile plant signal transducer under soil water stress. Bioessays, 2008, 30: 634–641

[19]Zhu J K. Salt and drought stress signal transduction in plants. Annu Rev Plant Biol, 2002, 53: 247–273

[20]Kidd B N, Kadoo N Y, Dombrecht B, Tekeo Lu M, Gardiner D M, Thatcher L F, Aitken E A B, Schenk P, Manners J, Kazan K. Auxin signaling and transport promote susceptibility to the root infecting fungal pathogen Fusarium oxysporum in Arabidopsis. Mol Plant Microbe Interact, 2011, 24: 733–748

[21]Leon-Reyes A, Du Y, Koornneef A, Proietti S, Körbes A P, Memelink J, Pieterse C M J, Ritsema T. Ethylene signaling renders the jasmonate response of Arabidopsis insensitive to future suppression by salicylic acid. Mol Plant Microbe Interact, 2010, 23(2): 187–197

[22]Kim M C, Chung W S, Yun D J, Cho M J. Calcium and calmodulin-mediated regulation of gene expression in plants. Mol Plant, 2009, 2(1): 13–21

[23]Wang Q, Chen B, Liu P, Zheng M, Wang Y, Cui S, Sun D, Fang X, Liu C M, Lucas W J. Calmodulin binds to extracellular sites on the plasma membrane of plant cells and elicits a rise in intracellular calcium concentration. J Biol Chem, 2009, 284: 12000–12007

[24]Heo W D, Lee S H, Kim M C, Kim J C, Chung W S, Chun H J, Lee K J, Park C Y, Park H C, Choi J Y. Involvement of specific calmodulin isoforms in salicylic acid-independent activation of plant disease resistance responses. Proc Natl Acad Sci USA, 1999, 96: 766–771

[25]Yamakawa H, Mitsuhara I, Ito N, Seo S, Kamada H, Ohashi Y. Transcriptionally and post-transcriptionally regulated response of 13 calmodulin genes to tobacco mosaic virus-induced cell death and wounding in tobacco plant. Eur J Biochem, 2001, 268: 3916–3929

[26]Huo J-F(霍建飞), Song S-S(宋水山), Li X(李星), Yang W-X(杨文香), Liu D-Q(刘大群). Research on CaM and its isform genes involved in the resistance response of wheat to Puccinia triticina. Acta Agron Boreali-Sin (华北农学报), 2010, 25(4): 175–179 (in Chinese with English abstract)

[27]Liu X-Y(刘新颖), Wang X-J(王晓杰), Xue J(薛杰), Xia N(夏宁), Deng L(邓麟), Cai G-L(蔡高磊), Tang C-L(汤春蕾), Wei G-R(魏国荣), Huang L-L(黄丽丽), Kang Z-S(康振生). Cloning and expression analysis of a novel calmodulin isoform TaCaM5 from wheat. Acta Agron Sin (作物学报), 2010, 36(6): 953–960 (in Chinese with English abstract)

[28]Herman R, Zvirin Z, Kovalski I, Freeman S, Denisov Y, Zuri G, Katzir N, Perl-Treves R, Pitrat M. Characterization of Fusarium race 1, 2 resistance in melon and mapping of a major QTL for this trait near a fruit netting locus. In: Pitrat M ed. The IXth EUCARPIA Meeting on Genetics and Breeding of Cucurbitaceae. Avignon (France): INRA. Centre de Recherche d'Avignon. Unité Génétique et Amélioration des Fruits et Légumes, Montfavet, 2008. pp 149–156

[29]Berrocal-Lobo M, Molina A. Arabidopsis defense response against Fusarium oxysporum. Trends Plant Sci, 2008, 13: 145–150

[30]Kazan K, Manners J M. Jasmonate signaling: toward an integrated view. Plant Physiol, 2008, 146: 1459–1468

[31]Koornneef A, Pieterse C M J. Cross talk in defense signaling. Plant Physiol, 2008, 146: 839–844

[32]Lorenzo O, Solano R. Molecular players regulating the jasmonate signalling network. Curr Opin Plant Biol, 2005, 8: 532–540

[33]Yang T, Lev-Yadun S, Feldman M, Fromm H. Developmentally regulated organ-, tissue-, and cell-specific expression of calmodulin genes in common wheat. Plant Mol Biol, 1998, 37(1): 109–120

[34]Yue H-L(岳海林), Deng X-X(邓秀新), Peng S-A(彭抒昂). Expression of calmodulin mRNAs in ovaries and fruitlets of pear. Sci Agric Sin (中国农业科学), 2008, 41(1): 176–181 (in Chinese with English abstract)
[1] CHEN Song-Yu, DING Yi-Juan, SUN Jun-Ming, HUANG Deng-Wen, YANG Nan, DAI Yu-Han, WAN Hua-Fang, QIAN Wei. Genome-wide identification of BnCNGC and the gene expression analysis in Brassica napus challenged with Sclerotinia sclerotiorum and PEG-simulated drought [J]. Acta Agronomica Sinica, 2022, 48(6): 1357-1371.
[2] JIN Min-Shan, QU Rui-Fang, LI Hong-Ying, HAN Yan-Qing, MA Fang-Fang, HAN Yuan-Huai, XING Guo-Fang. Identification of sugar transporter gene family SiSTPs in foxtail millet and its participation in stress response [J]. Acta Agronomica Sinica, 2022, 48(4): 825-839.
[3] YIN Ming, YANG Da-Wei, TANG Hui-Juan, PAN Gen, LI De-Fang, ZHAO Li-Ning, HUANG Si-Qi. Genome-wide identification of GRAS transcription factor and expression analysis in response to cadmium stresses in hemp (Cannabis sativa L.) [J]. Acta Agronomica Sinica, 2021, 47(6): 1054-1069.
[4] JIA Xiao-Ping, LI Jian-Feng, ZHANG Bo, QUAN Jian-Zhang, WANG Yong-Fang, ZHAO Yuan, ZHANG Xiao-Mei, WANG Zhen-Shan, SANG Lu-Man, DONG Zhi-Ping. Responsive features of SiPRR37 to photoperiod and temperature, abiotic stress and identification of its favourable allelic variations in foxtail millet (Setaria italica L.) [J]. Acta Agronomica Sinica, 2021, 47(4): 638-649.
[5] YUE Jie-Ru, BAI Jian-Fang, ZHANG Feng-Ting, GUO Li-Ping, YUAN Shao-Hua, LI Yan-Mei, ZHANG Sheng-Quan, ZHAO Chang-Ping, ZHANG Li-Ping. Cloning and potential function analysis of ascorbic peroxidase gene of hybrid wheat in seed aging [J]. Acta Agronomica Sinica, 2021, 47(3): 405-415.
[6] HE Xiao, LIU Xing, XIN Zheng-Qi, XIE Hai-Yan, XIN Yu-Feng, WU Neng-Biao. Molecular cloning, expression, and enzyme kinetic analysis of a phenylalanine ammonia-lyase gene in Pinellia ternate [J]. Acta Agronomica Sinica, 2021, 47(10): 1941-1952.
[7] LI Guo-Ji, ZHU Lin, CAO Jin-Shan, WANG You-Ning. Cloning and functional analysis of GmNRT1.2a and GmNRT1.2b in soybean [J]. Acta Agronomica Sinica, 2020, 46(7): 1025-1032.
[8] Jin-Feng ZHAO,Yan-Wei DU,Gao-Hong WANG,Yan-Fang LI,Gen-You ZHAO,Zhen-Hua WANG,Yu-Wen WANG,Ai-Li YU. Identification of PEPC genes from foxtail millet and its response to abiotic stress [J]. Acta Agronomica Sinica, 2020, 46(5): 700-711.
[9] LIANG Si-Wei,JIANG Hao-Liang,ZHAI Li-Hong,WAN Xiao-Rong,LI Xiao-Qin,JIANG Feng,SUN Wei. Genome-wide identification and expression analysis of HD-ZIP I subfamily genes in maize [J]. Acta Agronomica Sinica, 2020, 46(4): 532-543.
[10] Tong-Hong ZUO, He-Cui ZHANG, Qian-Ying LIU, Xiao-Ping LIAN, Qin-Qin XIE, Deng-Ke HU, Yi-Zhong ZHANG, Yu-Kui WANG, Xiao-Jing BAI, Li-Quan ZHU. Molecular cloning and expression analysis of BoGSTL21 in self-incompatibility Brasscia oleracea [J]. Acta Agronomica Sinica, 2020, 46(12): 1850-1861.
[11] WANG Yan-Hua,XIE Ling,YANG Bo,CAO Yan-Ru,LI Jia-Na. Flowering genes in oilseed rape: identification, characterization, evolutionary and expression analysis [J]. Acta Agronomica Sinica, 2019, 45(8): 1137-1145.
[12] Hong-Ju JIAN,Bo YANG,Yang-Yang LI,Hong YANG,Lie-Zhao LIU,Xin-Fu XU,Jia-Na LI. Identification and expression analysis of PEBP gene family in oilseed rape [J]. Acta Agronomica Sinica, 2019, 45(3): 354-364.
[13] Xiao-Hong ZHANG,Gen-Hai HU,Han-Tao WANG,Cong-Cong WANG,Heng-Ling WEI,Yuan-Zhi FU,Shu-Xun YU. Expression and promoter activity of GhTFL1a and GhTFL1c in Upland cotton [J]. Acta Agronomica Sinica, 2019, 45(3): 469-476.
[14] Pi-Biao SHI,Bing HE,Yue-Yue FEI,Jun WANG,Wei-Yi WANG,Fu-You WEI,Yuan-Da LYU,Min-Feng GU. Identification and expression analysis of GRF transcription factor family of Chenopodium quinoa [J]. Acta Agronomica Sinica, 2019, 45(12): 1841-1850.
[15] 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.
Full text



No Suggested Reading articles found!