|
|
|
Differential Gene and Protein Expression in Soybean at Early Stages of Incompatible Interaction with Phytophthora sojae |
LI Yong-gang1, YANG Ming-xiu1, LI Yan1, LIU Wen-wen1, WEN Jing-zhi1 and LI Yong-hao2 |
1 Department of Plant Protection, Northeast Agricultural University, Harbin 150030, P.R.China
2 Department of Plant Pathology and Ecology, Connecticut Agricultural Experiment Station, 123 Huntington St. New Haven, CT 06511, USA |
|
|
摘要 Soybean root and stem rot caused by Phytophthora sojae is a destructive disease worldwide. Using genetic resistance is an important and major component in the integrated pest management of this disease. To understand molecular mechanisms of root and stem rot resistance in soybeans, the gene and protein expression in hypocotyls and stems of variety Suinong 10 carrying resistance genes Rps1a and Rps2 was investigated by using mRNA differential display reverse transcription PCR and two-dimensional electrophoresis at 0, 0.5, 1, 2, and 4 h after inoculation with P. sojae race 1. The results of the comparison of gene and protein expression showed that at least eight differential fragments at the transcriptional level were related to metabolic pathway, phytoalexin, and signal transduction in defense responses.Sequence analyses indicated that these fragments represented cinnamic acid 4-hydroxylase gene, ATP β gene coding ATP synthase β subunit and ubiquitin-conjugating enzyme gene which upregulated at 0.5 h post inoculation, blue copper protein gene and UDP-N-acetyl-α-D-galactosamine gene which upregulated at 2 h post inoculation, TGA-type basic leucine zipper protein TGA1.1 gene, cyclophilin gene, and 14-3-3 protein gene which upregulated at 4 h post inoculation.Three resistance-related proteins, α-subunit and β-subunit of ATP synthase, and cytochrome P450-like protein, were upregulated at 2 h post inoculation. The results suggested that resistance-related multiple proteins and genes were expressed in the recognition between soybean and P. sojae during zoospore germination, penetration and mycelium growth of P. sojae in soybean.
Abstract Soybean root and stem rot caused by Phytophthora sojae is a destructive disease worldwide. Using genetic resistance is an important and major component in the integrated pest management of this disease. To understand molecular mechanisms of root and stem rot resistance in soybeans, the gene and protein expression in hypocotyls and stems of variety Suinong 10 carrying resistance genes Rps1a and Rps2 was investigated by using mRNA differential display reverse transcription PCR and two-dimensional electrophoresis at 0, 0.5, 1, 2, and 4 h after inoculation with P. sojae race 1. The results of the comparison of gene and protein expression showed that at least eight differential fragments at the transcriptional level were related to metabolic pathway, phytoalexin, and signal transduction in defense responses.Sequence analyses indicated that these fragments represented cinnamic acid 4-hydroxylase gene, ATP β gene coding ATP synthase β subunit and ubiquitin-conjugating enzyme gene which upregulated at 0.5 h post inoculation, blue copper protein gene and UDP-N-acetyl-α-D-galactosamine gene which upregulated at 2 h post inoculation, TGA-type basic leucine zipper protein TGA1.1 gene, cyclophilin gene, and 14-3-3 protein gene which upregulated at 4 h post inoculation.Three resistance-related proteins, α-subunit and β-subunit of ATP synthase, and cytochrome P450-like protein, were upregulated at 2 h post inoculation. The results suggested that resistance-related multiple proteins and genes were expressed in the recognition between soybean and P. sojae during zoospore germination, penetration and mycelium growth of P. sojae in soybean.
|
Received: 10 June 2011
Online: 10 June 2011
Accepted:
|
Corresponding Authors:
WEN Jing-zhi
E-mail: neaulyg@yahoo.cn;jzhwen2000@yahoo.com.cn
|
About author: LI Yong-gang, Ph D, E-mail: neaulyg@yahoo.cn; Correspondence WEN Jing-zhi, Professor, Ph D, Tel: +86-451-55191045, Fax: +86-451-55191045, E-mail:
jzhwen2000@yahoo.com.cn |
Cite this article:
LI Yong-gang, YANG Ming-xiu, LI Yan, LIU Wen-wen, WEN Jing-zhi and LI Yong-hao.
2011.
Differential Gene and Protein Expression in Soybean at Early Stages of Incompatible Interaction with Phytophthora sojae. Journal of Integrative Agriculture, 10(6): 902-910.
|
Bradford M M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein using the principle of protein-dye binding. Analytical Biochemistry, 72, 248-254. Burnham K D, Dorrance A E,van Toai T T, Martin S K S. 2003. Quantitative trait loci for partial resistance to Phytophthora sojae in soybean. Crop Science, 43, 1610-1617. Callow M E, Crawford S, Wetherbee R, Taylor K, Finlay J A, Callow J A. 2001. Brefeldin A affects adhesion of zoospores of the green alga Enteromorpha. Journal of Experimental Botany, 52, 1409-1415. Carlile M J.1986. The zoospore and its problems. In: Ayres P G, Boddy L, eds., Water, Fungi and Plants (11th Symposium of British Mycological Society). Cambridge University Press, Cambridge. pp. 105-118. Connolly M S, Sakihama Y, Phuntumart V, Jiang Y, Warren F, Mourant L, Morris P F. 2005. Heterologous expression of a pleiotropic drug resistance transporter from Phytophthora sojae in yeast transporter mutants. Current Genetics, 48, 356-365. Deacon J W, Saxena G. 1997. Orientated zoospore attachment and cyst germination in Catenaria anguillulae, a facultative endoparasite of nematodes. Mycological Research, 101, 513- 522. Dixon R A, Steele C L. 1999. Flavonoids and isoflavonoids - a gold mine for metabolic engineering. Trends in Plant Science, 4, 394-400. Dixon R A. 2001. Natural products and plant disease resistance. Nature, 411, 843-850. Dorrance A E, McClure S A, de Silva A. 2003. Pathogenic diversity of Phytophthora sojae in Ohio soybean fields. Plant Disease, 87, 139-146. Enkerli K, Haha M Q, Mims C W. 1997. Ultrastructure of compatible and incompatible interaction of soybean roots infected with the plant pathogenic oomycete Phytophthora sojae. Canadian Journal of Botany, 75, 1493-1508. Gibson D M, Stack S, Krell K, House J. 1982. A comparison of soybean agglutinin in cultivars resistant and susceptible to Phytophthora megasperma var. sojae (Race 1). Plant Physiology, 70, 560-566. Giri A V, Anishetty S, Gautam P. 2004. Functionally specified protein signatures distinctive for each of the different blue copper proteins. BMC Bioinformatics, 5, 127-135. Harborne J B. 1994. The Flavonoids - Advances in Research Since 1986. Chapman and Hall, London. Hua C L, Wang Y L, Zheng X B, Dou D L, Zhang Z G, Govers F, Wang Y C. 2008. A Phytophthora sojae G-protein subunit is involved in chemotaxis to soybean isoflavones. Eukaryotic Cell, 7, 2133-2140. Hua C L, Zheng X B, Wang Y C. 2009. G protein α subunit may help zoospore to find the infection site and influence the expression of RGS protein. Communicative and Integrative Biology, 2, 91-93. Ichimura T, Isobe T, Okuyama T, Takahashi N, Araki K, Kuwano R, Takahashi Y. 1988. Molecular cloning of cDNA coding for brain specific 14-3-3 protein, a protein kinase-dependent activator of tyrosine and tryptophan hydroxylases. Proceedings of the National Academy of Sciences of the USA, 85, 7084-7088. Jia H, Kurle J E. 2008. Resistance and partial resistance to Phytophthora sojae in early maturity group soybean plant introductions. Euphytica, 159, 27-34. Kaufmann M J, Gerdemann J W. 1958. Root and stem rot of soybean caused by Phytophthora sojae n. sp. Phytopathology, 48, 201-208. Latunde-Dada A O, Cabello-Hurtado F, Czittrich N, Didierjean L, Schopfer C, Hertkorn N, Werck-Reichhart D, Ebel J. 2001. Flavonoid 6-hydroxylase from soybean (Glycine max L.): a novel plant P-450 monooxygenase. The Journal of Biological Chemistry, 278, 1688-1695. Laxa M, Konig J, Dietz K J, Kandlbinder A. 2007. Role of the cysteine residues in Arabidopsis thaliana cyclophilin CYP20- 3 in peptidyl-prolyl cis-trans isomerase and redox-related functions. Biochemistry Journal, 401, 287-297. Li Y G, Wen J Z, Hao Z N, Sun Y W, Yang M X, Wang L F. 2008. Study on method of identification and screening resistance sources of Phytophthora root rot. Journal of Northeast Agricultural University, 39, 1-5. (in Chinese) Liu W W, Li Y G,Yang M X, Li M, Wen J Z. 2010. Analysis of expressed protein differences in rice leaves induced by probenazole. Acta Phytophylacica Sinica, 37, 109-112. (in Chinese) McDonald K L, David M C. 1999. Influence of abscisic acid and the abscisic acid biosynthesis inhibitor, norflurazon, on interactions between Phytophthora sojae and soybean (Glycine max). European Journal of Plant Pathology, 105, 651-658. Melgar J C, Abney T S, Vierling R A. 2006. Vierling peroxidase activity in soybeans following inoculation with Phytophthora sojae. Mycopathologia, 161, 37-42. Mori H, Inoue M, Yano M, Wakabayashi H, Kido H. 2000. 14- 3-3tau associates with a translational control factor FKBP12- rapamycin-associated protein in T-cells after stimulation by pervanadate. FEBS Letters, 467, 61-64. Morris P F, Bone E, Tyler B M. 1998. Chemotropic and contact responses of Phytophthora sojae hyphae to soybean isoflavonoids and artificial substrates. Plant Pathology, 117, 1171-1178. Moy P, Qutob D, Chapman B P, Atkinson I. 2004. Patterns of gene expression upon infection of soybean plants by Phytophthora sojae. Molecular Plant-Microbe Interactions, 17, 1051-1063. O’Brien T X, Schuyler G T, Rackley M S, Thompson J T. 1999. F1-ATP synthase beta-subunit and cytochrome transcriptional regulation in right ventricular hemodynamic overload and hypertrophically stimulated cardiocytes. Journal of Molecular and Cellular Cardiology, 31, 167-178. Qiu H M, Liu C Y, Zhang D J, Xin X J, Wang J L, Wang J, Shan C Y, Shan D P, Hu G H, Chen Q S. 2009. Proteome analysis of resistance to Phytophora root rot in soybean. Acta Agronomica Sinica, 35, 418-423. Shang F, Gong X, Taylor A. 1997. Activity of ubiquitin-dependent pathway in response to oxidative stress. Ubiquitin-activating enzyme is transiently up-regulated. The Journal of Biological Chemistry, 272, 23086-23093. Shen G F, Zhao Y, Kung S D. 1991. Molecular cloning and sequencing of rice4-coumarate: CoA ligase gene. Scientia Agricultura Sinica, 2, 1-7. (in Chinese) Skalamera D, Wasson A P, Hardham A R. 2004. Genes expressed in zoospores of Phytophthora nicotianae. Molecular Genetics and Genomics, 270, 549-557. Stossel P, Leuba J L. 1984. Effect of chitosan, chitin and some aminosugars on growth of various soilborne phytopathogenic fungi. Journal of Phytopathology, 111, 82-90. Tucker M L, Whitelaw C A, Lyssenko N N, Nath P. 2002. Functional analysis of regulatory elements in the gene promoter for an abscission-specific cellulase from bean and isolation, expression, and binding affinity of three TGA type basic leucine zipper transcription factors. Plant Physiology, 130, 1487-1496. Yano M, Nakamuta S, Wu X, Okumura Y, Kido H. 2006. A novel function of 14-3-3 protein: 14-3-3 zeta is a heat-shock-related molecular chaperone that dissolves thermal-aggregated proteins. Molecular Biology of the Cell, 17, 4769-4779. Zuo Y H, Kang Z S, Huang L L, Han Q M. 2005. Cytology on infection process of soybean hypocotyls by Phytophthora sojae. Acta Phytopathologica Sinica, 35, 235-241. (in Chinese) Ye N, Li Y G, Wen J Z. 2008. Research on molecular mechanisms of soybean resistance to Phytophthora sojae by mRNA differential display PCR. Journal of Northeast Agricultural University, 39, 6-9. (in Chinese) |
No Suggested Reading articles found! |
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
|
Shared |
|
|
|
|
|
Discussed |
|
|
|
|