cDNA-AFLP analysis reveals differential gene expression in incompatible interaction between infected non-heading Chinese cabbage and Hyaloperonospora parasitica

Dong Xiao , Shi-Tuo Liu , Yan-Ping Wei , Dao-Yun Zhou , Xi-Lin Hou , Ying Li , Chun-Mei Hu

Horticulture Research ›› 2016, Vol. 3 ›› Issue (1) : 16034

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Horticulture Research ›› 2016, Vol. 3 ›› Issue (1) :16034 DOI: 10.1038/hortres.2016.34
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cDNA-AFLP analysis reveals differential gene expression in incompatible interaction between infected non-heading Chinese cabbage and Hyaloperonospora parasitica
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Abstract

Non-heading Chinese cabbage (Brassica rapa ssp. chinensis) is one of the main green leafy vegetables in the world, especially in China, with significant economic value. Hyaloperonospora parasitica is a fungal pathogen responsible for causing downy mildew disease in Chinese cabbage, which greatly affects its production. The objective of this study was to identify transcriptionally regulated genes during incompatible interactions between non-heading Chinese cabbage and H. parasitica using complementary DNA-amplified fragment length polymorphism (cDNA-AFLP). We obtained 129 reliable differential transcript-derived fragments (TDFs) in a resistant line ‘Suzhou Qing’. Among them, 121 upregulated TDFs displayed an expression peak at 24–48 h post inoculation (h.p.i.). Fifteen genes were further selected for validation of cDNA-AFLP expression patterns using quantitative reverse transcription PCR. Results confirmed the altered expression patterns of 13 genes (86.7%) revealed by the cDNA-AFLP. We identified four TDFs related to fungal resistance among the 15 TDFs. Furthermore, comparative analysis of four TDFs between resistant line ‘Suzhou Qing’ and susceptible line ‘Aijiao Huang’ showed that transcript levels of TDF14 (BcLIK1_A01) peaked at 48 h.p.i. and 25.1-fold increased in the resistant line compared with the susceptible line. Similarly, transcript levels of the other three genes, TDF42 (BcCAT3_A07), TDF75 (BcAAE3_A06) and TDF88 (BcAMT2_A05) peaked at 24, 48 and 24 h.p.i. with 25.1-, 100- and 15.8-fold increases, respectively. The results suggested that the resistance genes tended to transcribe at higher levels in the resistance line than in the susceptible line, which may provide resistance against pathogen infections. The present study might facilitate elucidating the molecular basis of the infection process and identifying candidate genes for resistance improvement of susceptible cultivars.

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Dong Xiao, Shi-Tuo Liu, Yan-Ping Wei, Dao-Yun Zhou, Xi-Lin Hou, Ying Li, Chun-Mei Hu. cDNA-AFLP analysis reveals differential gene expression in incompatible interaction between infected non-heading Chinese cabbage and Hyaloperonospora parasitica. Horticulture Research, 2016, 3 (1) : 16034 DOI:10.1038/hortres.2016.34

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References

[1]

Takemoto D, Hardham AR . The cytoskeleton as a regulator and target of biotic interactions in plants. Plant Physiol 2004; 136: 3864-3876.

[2]

Pitino M, Armstrong CM, Duan YP . Rapid screening for citrus canker resistance employing pathogen-associated molecular pattern-triggered immunity responses. Hortic Res 2015; 2: 15042.

[3]

Amrine KCH, Blanco-Ulate B, Riaz S, Pap D, Jones L, Figueroa-Balderas R et al. Comparative transcriptomics of Central Asian Vitis vinifera accessions reveals distinct defense strategies against powdery mildew. Hortic Res 2015; 2: 15037.

[4]

Monot C, Pajot E, Le Corre D, Silué D . Induction of systemic resistance in broccoli (Brassica oleracea var. botrytis) against downy mildew (Peronospora parasitica) by avirulent isolates. Biol Control 2002; 24: 75-81.

[5]

Farinhó M, Coelho P, Carlier J, Svetleva D, Monteiro A, Leitão J . Mapping of a locus for adult plant resistance to downy mildew in broccoli (Brassica oleraceacon var. italica). Theor Appl Genet 2004; 109: 1392-1398.

[6]

Casimiro S, Tenreiro R, Monteiro AA . Identification of pathogenesis-related ESTs in the crucifer downy mildew oomycete Hyaloperonospora parasitica by high-throughput differential display analysis of distinct phenotypic interactions with Brassica oleracea. J Microbiol Methods 2006; 66: 466-478.

[7]

Tang YQ, Yu SC, Zhu YL, Zhang FL, Yu YJ, Zhao XY et al. Construction and analysis of suppression subtractive hybridization cDNA library in Chinese cabbage (Brassica rapa ssp. pekinensis) leaves induced by Peronospora parasitica. Plant Physiol Commun 2010; 46: 453-458.

[8]

De Paepe A, Vuylsteke M, Van Hummelen P, Zabeau M, Van Der Straeten D . Transcriptional profiling by cDNA-AFLP and microarray analysis reveals novel insights into the early response to ethylene in Arabidopsis. Plant J 2004; 39: 537-559.

[9]

Sarosh BR, Meijer J . Transcriptional profiling by cDNA-AFLP reveals novel insights during methyl jasmonate, wounding and insect attack in Brassica napus. Plant Mol Biol 2007; 64: 425-438.

[10]

Chen X, Hou X, Zhang J, Zheng J . Molecular characterization of two important antifungal proteins isolated by downy mildew infection in non-heading Chinese cabbage. Mol Biol Rep 2008; 35: 621-629.

[11]

Livak KJ, Schmittgen TD . Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method . Methods 2001; 25: 402-408.

[12]

Ozturk ZN, Talamé V, Deyholos M, Michalowski CB, Galbraith DW, Gozukirmizi N et al. Monitoring large-scale changes in transcript abundance in drought- and salt-stressed barley. Plant Mol Biol 2002; 48: 551-573.

[13]

Jin S, Cheng Y, Guan Q, Liu D, Takano T, Liu S . A metallothionein-like protein of rice (rgMT) functions in E. coli and its gene expression is induced by abiotic stresses. Biotechnol Lett 2006; 28: 1749-1753.

[14]

Degenhardt J, Al-Masri AN, Kürkcüoglu S, Szankowski I, Gau AE . Characterization by suppression subtractive hybridization of transcripts that are differentially expressed in leaves of apple scab-resistant and susceptible cultivars of Malus domestica. Mol Genet Genomics 2005; 273: 326-335.

[15]

Akashi K, Nishimura N, Ishida Y, Yokota A . Potent hydroxyl radical-scavenging activity of drought-induced type-2 metallothionein in wild watermelon. Biochem Biophys Res Commun 2004; 323: 72-78.

[16]

Das R, Roy A, Dutta N, Majumder H . Reactive oxygen species and imbalance of calcium homeostasis contributes to curcumin induced programmed cell death in Leishmania donovani. Apoptosis 2008; 13: 867-882.

[17]

Rushton PJ, Torres JT, Parniske M, Wernert P, Hahlbrock K, Somssich IE . Interaction of elicitor-induced DNA-binding proteins with elicitor response elements in the promoters of parsley PR1 genes. EMBO J 1996; 15: 5690-5700.

[18]

Cheong YH, Chang HS, Gupta R, Wang X, Zhu T, Luan S et al. Transcriptional profiling reveals novel interactions between wounding, pathogen, abiotic stress, and hormonal responses in Arabidopsis. Plant Physiol 2002; 129: 661-677.

[19]

Zeidler D, Zähringer U, Gerber I, Dubery I, Hartung T, Bors W et al. Innate immunity in Arabidopsis thaliana: Lipopolysaccharides activate nitric oxide synthase (NOS) and induce defense genes. Proc Natl Acad Sci USA 2004; 101: 15811-15816.

[20]

Kim MC, Panstruga R, Elliott C, Muller J, Devoto A, Yoon HW et al. Calmodulin interacts with MLO protein to regulate defence against mildew in barley. Nature 2002; 416: 447-451.

[21]

Hu X, Jiang M, Zhang J, Zhang A, Lin F, Tan M . Calcium-calmodulin is required for abscisic acid-induced antioxidant defense and functions both upstream and downstream of H2O2 production in leaves of maize (Zea mays) plants . New Phytol 2007; 173: 27-38.

[22]

Yang T, Poovaiah BW . Hydrogen peroxide homeostasis: activation of plant catalase by calcium/calmodulin. Proc Natl Acad Sci USA 2002; 99: 4097-4102.

[23]

Turck F, Zhou A, Somssich IE . Stimulus-dependent, promoter-specific binding of transcription factor WRKY1 to its native promoter and the defense-related gene PcPR1-1 in Parsley. Plant Cell 2004; 16: 2573-2585.

[24]

Pandey SP, Somssich IE . The role of WRKY transcription factors in plant immunity. Plant Physiol 2009; 150: 1648-1655.

[25]

Berrocal-Lobo M, Molina A, Solano R . Constitutive expression of ETHYLENE-RESPONSE-FACTOR1 in Arabidopsis confers resistance to several necrotrophic fungi. Plant J 2002; 29: 23-32.

[26]

Singh KB, Foley RC, Oñate-Sánchez L. Transcription factors in plant defense and stress responses. Curr Opin Plant Biol 2002; 5: 430-436.

[27]

Cao Y, Wu Y, Zheng Z, Song F . Overexpression of the rice EREBP-like gene OsBIERF3 enhances disease resistance and salt tolerance in transgenic tobacco. Physiol Mol Plant Pathol 2005; 67: 202-211.

[28]

Lingelbach LB, Kaplan KB . The interaction between Sgt1p and Skp1p is regulated by HSP90 chaperones and is required for proper CBF3 assembly. Mol Cell Biol 2004; 24: 8938-8950.

[29]

Holt BF, Belkhadir Y, Dangl JL . Antagonistic dontrol of disease resistance protein stability in the plant immune system. Science 2005; 309: 929-932.

[30]

Wang K, Uppalapati SR, Zhu X, Dinesh-Kumar SP, Mysore KS . SGT1 positively regulates the process of plant cell death during both compatible and incompatible plant-pathogen interactions. Mol Plant Pathol 2010; 11: 597-611.

[31]

Yokota A, Shigeoka S, Hans J, Bohnert HN, Norman GL . Engineering photosynthetic pathways. Adv Plant Biochem Mol Biol 2008; 1: 81-105.

[32]

Henkes S, Sonnewald U, Badur R, Flachmann R, Stitt M . A small decrease of plastid transketolase activity in antisense tobacco transformants has dramatic effects on photosynthesis and phenylpropanoid metabolism. Plant Cell 2001; 13: 535-551.

[33]

Seki M, Narusaka M, Ishida J, Nanjo T, Fujita M, Oono Y et al. Monitoring the expression profiles of 7000 Arabidopsis genes under drought, cold and high-salinity stresses using a full-length cDNA microarray. Plant J 2002; 31: 279-292.

[34]

Scharte J, SchÖN H, Weis E . Photosynthesis and carbohydrate metabolism in tobacco leaves during an incompatible interaction with Phytophthora nicotianae. Plant Cell Environ 2005; 28: 1421-1435.

[35]

Conklin PL, Last RL . Differential accumulation of antioxidant mRNAs in Arabidopsis thaliana exposed to ozone. Plant Physiol 1995; 109: 203-212.

[36]

Zhao J, Last RL . Coordinate regulation of the tryptophan biosynthetic pathway and indolic phytoalexin accumulation in Arabidopsis. Plant Cell 1996; 8: 2235-2244.

[37]

Mittler R, Vanderauwera S, Gollery M, Van Breusegem F . Reactive oxygen gene network of plants. Trends Plant Sci 2004; 9: 490-498.

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