|
|
|
|
|
| Modulation of protein expression in alfalfa (Medicago sativa L.) root and leaf tissues by Fusarium proliferatum |
| CONG Li-li1, 3*, SUN Yan2*, LONG Rui-cai1, KANG Jun-mei1, ZHANG Tie-jun1, LI Ming-na2, WANG Zhen1, YANG Qing-chuan1 |
1 Institute of Animal Sciences, Chinese Academy of Agricultural Science, Beijing 100193, P.R.China
2 Institute of Grassland Science, China Agricultural University, Beijing 100193, P.R.China
3 College of Animal Science and Technology, Qingdao Agricultural University, Qingdao 266109, P.R.China |
|
|
|
|
Abstract Alfalfa (Medicago sativa L.) is an important forage crop and is also a target of many fungal diseases including Fusarium spp. As of today, very little information is available about molecular mechanisms that contribute to pathogenesis and defense responses in alfalfa against Fusarium spp. and specifically against Fusarium proliferatum, the causal agent of alfalfa root rot. In this study, we used a proteomic approach to identify inducible proteins in alfalfa during a compatible interaction with F. proliferatum strain YQC-L1. Samples used for the two-dimensional gel electrophoresis (2-DE) and MALDI-TOF/TOF mass spectrometry were from roots and leaves of alfalfa cultivar AmeriGraze 401+Z and WL656HQ. Plants were grown in hydroponic conditions and at 4 days post inoculation with YQC-L1. Our disease symptom assays indicated that AmeriGraze 401+Z was tolerant to YQC-L1 infection while WL656HQ was highly susceptible. Analysis of differentially expressed proteins found in the 2-DE was further characterized using the MASCOT MS/MS ion search software and associated databases to identify multiple proteins that might be involved in F. proliferatum resistance. A total of 66 and 27 differentially expressed proteins were found in the roots and leaves of the plants inoculated with YQC-L1, respectively. These identified proteins were placed in various categories including defense and stress response related metabolism, photosynthesis and protein synthesis. Thirteen identified proteins were validated for their expressions by quantitative reverse transcription (qRT)-PCR. Our results suggested that some of the identified proteins might play important roles in alfalfa resistance against Fusarium spp. These finding could facilitate further dissections of molecular mechanisms controlling root rot disease in alfalfa and potentially other legume crops.
|
|
Received: 22 January 2017
Accepted:
|
| Fund: This work was supported by the earmarked fund for China Agriculture Research System (CARS-35-04), the Chinese Academy of Agricultural Science and Technology Innovation Project (ASTIP-IAS14). |
|
Corresponding Authors:
Correspondence YANG Qing-chuan, Tel/Fax: +86-10-62815996, E-mail: qchyang66@163.com
|
| About author: CONG Li-li, E-mail: congli1985610@126.com
* These authors contributed equally to this study. |
Cite this article:
CONG Li-li, SUN Yan, LONG Rui-cai, KANG Jun-mei, ZHANG Tie-jun, LI Ming-na, WANG Zhen, YANG Qing-chuan.
2017.
Modulation of protein expression in alfalfa (Medicago sativa L.) root and leaf tissues by Fusarium proliferatum. Journal of Integrative Agriculture, 16(11): 2558-2572.
|
| Aranda M A, Escaler M, Wang D, Maule A J. 1996. Induction of HSP70 and polyubiquitin expression associated with plant virus replication. Proceedings of the National Academy of Sciences of the United States of America, 93, 15289–15293.Arias M D, Munkvold G, Leandro L. 2015. First report of Fusarium proliferatum causing root rot on soybean (Glycine max) in the United States. Crop Protection, 67, 52–58.Bhuiyan N H, Liu W, Liu G, Selvaraj G, Wei Y. 2007. Transcriptional regulation of genes involved in the pathways of biosynthesis and supply of methyl units in response to powdery mildew attack and abiotic stresses in wheat. Plant Molecular Biology, 64, 305–318.Brown D E, Rashotte A M, Murphy A S, Tague B W, Peer W A, Taiz L, Muday G K. 2001. Flavonoids act as negative regulators of auxin transport in vivo in Arabidopsis thaliana. Plant Physiology and Biochemistry, 126, 524–535.Cabral C, Melo M P, Fonseca M E N, Boiteux L S, Reis A. 2016. A root rot of chickpea caused by isolates of the Fusarium solani species complex in Brazil. Plant Disease, 100, 2171. Caitriona D, Iain W W, McFadden H. 2004. Gene expression profile changes in cotton root and hypocotyl tissues in response to Infection with Fusarium oxysporum f. sp. vasinfectum. Molecular Plant Microbe Interactions, 17, 654–667.Campo S, Carrascal M, Coca M, Abian J, San Segundo B. 2004. The defense response of germinating maize embryos against fungal infection: A proteomics approach. Proteomics, 4, 383–396.Castillejo M A, Amiour N, Dumas-Gaudot E, Rubiales D, Jorrín J V. 2004. A proteomic approach to studying plant response to crenate broomrape (Orobanche crenata) in pea (Pisum sativum). Phytochemistry, 65, 1817–1828.Chang K, Hwang S, Conner R, Ahmed H, Zhou Q, Turnbull G, Strelkov S, McLaren D, Gossen B. 2015. First report of Fusarium proliferatum causing root rot in soybean (Glycine max L.) in Canada. Crop Protection, 67, 52–58.Chittem K, Mathew F M, Gregoire M, Lamppa R S, Chang Y W, Markell S G, Bradley C A, Barasubiye T, Goswami R S. 2015. Identification and characterization of Fusarium spp. associated with root rots of field pea in North Dakota. European Journal of Plant Pathology, 143, 641–649.Cipriano D J, Wang Y, Bond S, Hinton A, Jefferies K C, Qi J, M F. 2008. Structure and regulation of the vacuolar ATPases. Biochimica et Biophysica Acta (BBA: Bioenergetics), 1777, 599–604.Cong L L, Sun Y, Kang J M, Li M N, Long R C, Zhang T J, Yang Q C. 2016. First report of root rot disease caused by Fusarium proliferatum on alfalfa in China. Plant Disease, 100, 2526. Coumans J V, Poljak A, Raftery M J, Backhouse D, Pereg-Gerk L. 2008. Analysis of cotton (Gossypium hirsutum) root proteomes during a compatible interaction with the black root rot fungus Thielaviopsis basicola. Proteomics, 9, 335–349. Cramer G R, Urano K, Delrot S, Shinozaki K. 2011. Effects of abiotic stress on plants: A systems biology perspective. BMC Plant Biology, 11, 163.Dahal D, Pich A, Braun H P, Wydra K. 2010. Analysis of cell wall proteins regulated in stem of susceptible and resistant tomato species after inoculation with Ralstonia solanacearum: A proteomic approach. Plant Molecular Biology, 73, 643–658.Dao T T, Linthorst H J, Verpoorte R. 2011. Chalcone synthase and its functions in plant resistance. Phytochemistry Reviews, 10, 397–412.Elmer W H. 1990. Fusarium proliferatum as a causal agent in Fusarium crown and root rot of asparagus. Plant Disease, 74, 938.Gal T Z, Glazer I, Koltai H. 2004. An LEA group 3 family member is involved in survival of C. elegans during exposure to stress. Febs Letters, 577, 21–26.Ghabooli M, Khatabi B, Ahmadi F S, Sepehri M, Mirzaei M, Amirkhani A, Jorrín-Novo J V, Salekdeh G H. 2013. Proteomics study reveals the molecular mechanisms underlying water stress tolerance induced by Piriformospora indica in barley. Journal of Proteomics, 94, 289–301.Gloria I C. 2012. Comparing the growth of fungal cultures on groundnut dextrose medium and potatoes dextrose medium. Journal of Science, 1, 46–52.Hoagland D R, Arnon D I. 1950. The Water-Culture Method for Growing Plants Without Soil. California Agricultural Experiment Station, University of California in Berkeley, USA. p. 347.Kolkman J M, Kelly J D. 2000. An indirect test using oxalate to determine physiological resistance to white mold in common bean. Crop Science, 40, 281–285.Kumar Y, Zhang L, Panigrahi P, Dholakia B B, Dewangan V, Chavan S G, Kunjir S M, Wu X, Li N, Rajmohanan P R, Kadoo N Y, Giri A P, Tang H, Gupta V S. 2016. Fusarium oxysporum mediates systems metabolic reprogramming of chickpea roots as revealed by a combination of proteomics and metabolomics. Plant Biotechnology Journal, 14, 1589–1603.Leath K T, Kendall W A. 1977. Fusarium root rot of forage species: Pathogenicity and host range. Phytopathology, 68, 826–831.Li X S, Bai T T, Li Y F, Ruan X L, Li H P. 2013. Proteomic analysis of Fusarium oxysporum f. sp. cubense tropical race 4-inoculated response to Fusarium wilts in the banana root cells. Proteome Science, 11, doi: 10.1186/1477-5956-11-41Liu Y, Liang J, Sun L, Yang X, Li D. 2016. Group 3 LEA protein, ZmLEA3, is involved in protection from low temperature stress. Frontiers in Plant Science, 7, 1–8.Liu Y, Wang L, Xing X, Sun L, Pan J, Kong X. 2013. ZmLEA3, a multifunctional group 3 LEA protein from maize (Zea mays L.), is involved in biotic and abiotic stresses. Plant and Cell Physiology, 54, 944–959.Long R, Li M, Zhang T, Kang J, Sun Y, Cong L, Gao Y, Liu F, Yang Q. 2016. Comparative proteomic analysis reveals differential root proteins in Medicago sativa and Medicago truncatula in response to salt stress. Frontiers in Plant Science, 7, doi: 10.3389/fpls.2016.00424Miller-Garvin J E, Viands D R. 1994. Selection for resistance to Fusarium root rot, and associations among resistances to six diseases in alfalfa. Crop Science, 34, 1461–1465.Moons A, Gerald L. 2005. Regulatory and functional interactions of plant growth regulators and plant glutathione S-transferases (GSTs). Vitamins and Hormones, 72, 155–202.Neuhoff V, Arold N, Taube D, Ehrhardt W. 1988. Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using Coomassie brilliant blue G-250 and R-250. Electrophoresis, 9, 255–262.Pérez B, Berretta M, Carrión E, Wright E. 2016. First report of root rot caused by Fusarium proliferatum on blueberry in Argentina. Journal of Basic Microbiology, 95, 1478.Peter S, Athos B, Katinka B, Regina V L, Roger A A, Jürg L, Andres W, Dongjin K, Douglas R C, Thomas B. 2000. Differential expression of eight chitinase genes in Medicago truncatula roots during Mycorrhiza formation, nodulation, and pathogen infection. Molecular Plant Microbe Interactions, 13, 763–777.Petriccione M, Cecco I D, Arena S, Scaloni A, Scortichini M. 2012. Proteomic changes in Actinidia chinensis shoot during systemic infection with a pandemic Pseudomonas syringae pv. actinidiae strain. Journal of Proteomics, 78, 461–476.Ramírez-Suero M, Khanshour A, Martinez Y, Rickauer M. 2009. A study on the susceptibility of the model legume plant Medicago truncatula to the soil-borne pathogen Fusarium oxysporum. European Journal of Plant Pathology, 126, 517–530.Richard C, Michaud R, Freve A, Gagnon C. 1980. Selection for root and crown rot resistant in alfalfa. Crop Science, 6, 691–695.Román-Avilés B, Kelly J D. 2005. Identification of quantitative trait loci conditioning resistance to Fusarium root rot in common bean. Crop Science, 45, 1881. Salter R, Miller-Garvin J E, Viands D R. 1994. Breeding for resistance to alfalfa root rot caused by Fusarium Species. Crop Science, 34, 1213–1217.Salvalaggio A, Ridao A D C. 2013. First report of Fusarium proliferatum causing rot on garlic and onion in Argentina. Plant Disease, 97, 556.Schneider K A, Grafton K F, Kelly J D. 2001. QTL analysis of resistance to Fusarium root rot in bean. Crop Science, 41, 535–542.Sung D Y, Kaplan F, Guy C L. 2001. Plant Hsp70 molecular chaperones: Protein structure, gene family, expression and function. Physiologia Plantarum, 113, 443–451.Tohge T, Yonekura-Sakakibara K, Niida R, Wantanabe-Takahasi A K S. 2007. Phytochemical genomics in Arabidopsis thaliana: A case study for functional identification of flavonoid biosynthesis genes. Pure and Applied Chemistry, 79, 811–823.Vitale A, Rocco M, Arena S, Giuffrida F, Cassaniti C, Scaloni A, Lomaglio T, Guarnaccia V, Polizzi G, Marra M, Leonardi C. 2014. Tomato susceptibility to Fusarium crown and root rot: Effect of grafting combination and proteomic analysis of tolerance expression in the rootstock. Plant Physiology and Biochemistry, 83, 207–216.Wang Z, Ke Q, Kim M D, Kim S H, Ji C Y, Jeong J C, Lee H S, Park W S, Ahn M J, Li H, Xu B, Deng X, Lee S H, Lim Y P, Kwak S S. 2015. Transgenic alfalfa plants expressing the sweetpotato orange gene exhibit enhanced abiotic stress tolerance. PLoS ONE, 10, e0126050.Watson B S, Lei Z, Dixon R A, Sumner L W. 2004. Proteomics of Medicago sativa cell walls. Phytochemistry, 65, 1709–1720.Xiong J, Yang Q, Kang J, Sun Y, Zhang T, Margaret G, Ding W. 2011. Simultaneous isolation of DNA, RNA, and protein from Medicago truncatula L. Electrophoresis, 32, 321–330.Yang F, Jensen J D, Svensson B, Jorgensen H J, Collinge D B, Finnie C. 2010. Analysis of early events in the interaction between Fusarium graminearum and the susceptible barley (Hordeum vulgare) cultivar Scarlett. Proteomics, 10, 3748–3755.Zhang Y M, Zhao J M, Xiang Y, Bian X C, Zuo Q M, Shen Q, Gai J Y, Xing H. 2011. Proteomics study of changes in soybean lines resistant and sensitive to Phytophthora sojae. Proteome Science, 9, 52–65.Zhou C, Han L, Pislariu C, Nakashima J, Fu C, Jiang Q, Quan L, Blancaflor E B, Tang Y, Bouton J H, Udvardi M, Xia G, Wang Z Y. 2011. From model to crop: Functional analysis of a STAY-GREEN gene in the model legume Medicago truncatula and effective use of the gene for alfalfa improvement. Plant Physiology, 157, 1483–1496. |
| No Suggested Reading articles found! |
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
