Abstract
As one of the most important resistance (R) gene families in plants, the NBS–LRR genes, encoding proteins with nucleotide-binding site (NBS) and leucine-rich repeat (LRR) domains, play significant roles in resisting pathogens. The published genomic data for cabbage (Brassica oleracea L.) provide valuable data to identify and characterize the genomic organization of cabbage NBS–LRR genes. Ultimately, we identified 105 TIR (N-terminal Toll/interleukin-1 receptor)-NBS–LRR (TNL) genes and 33 CC (coiled-coil)-NBS–LRR (CNL) genes. Further research indicated that 50.7% of the 138 NBS–LRR genes exist in 27 clusters and there are large differences among the gene structures and protein characteristics. Conserved motif and phylogenetic analysis showed that the structures of TNLs and CNLs were similar, with some differences. These NBS–LRRs are evolved under negative selection and mostly arose from whole-genome duplication events during evolution. Tissue-expression profiling of NBS–LRR genes revealed that 37.1% of the TNL genes are highly or specifically expressed in roots, especially the genes on chromosome 7 (76.5%). Digital gene expression and reverse transcription PCR analyses revealed the expression patterns of the NBS–LRR genes upon challenge by Fusarium oxysporum f.sp. conglutinans: nine genes were upregulated, and five were downregulated. The major resistance gene Foc1 probably works together with the other four genes in the same cluster to resist F. oxysporum infection.
Similar content being viewed by others
Abbreviations
- DGE:
-
Digital gene expression
- R:
-
Resistance
- NBS:
-
Nucleotide-binding site
- LRR:
-
Leucine-rich repeat
- TIR:
-
N-terminal Toll/interleukin-1 receptor
- CC:
-
Coiled-coil
References
Bai J, Pennill L, Ning J, Weon L, Jegadeesan R, Webb C, Zhao B, Sun Q, Nelson J, Leach J, Hulbert S (2003) Diversity in nucleotide binding site-leucine-rich repeat genes in cereals. Genome Res 12:1871–1884. https://doi.org/10.1101/gr.454902
Bailey T, Bodén M, Buske F, Frith M, Grant C, Clementi L, Ren J, Li WS, Noble W (2009) Meme suite: tools for motif discovery and searching. Nucleic Acids Res 37:W202–W208. https://doi.org/10.1093/nar/gkp335
Chen CJ, Xia R, Chen H, He YH (2018) TBtools, a Toolkit for Biologists integrating various biological data handling tools with a user-friendly interface. bioRxiv. https://doi.org/10.1101/289660
Conesa A, Madrigal P, Tarazona S, Gomez-Cabrero D, Cervera A, McPherson A, Szcześniak MJ, Gaffney D, Elo L, Zhang X, Mortazavi A (2016) A survey of best practices for RNA-seq data analysis. Genome Biol 17:181. https://doi.org/10.1186/s13059-016-0881-8
Dangl JL, Jones JDG (2001) Plant pathogens and integrated defence responses to infection. Nature 411:826–833. https://doi.org/10.1038/35081161
Dennis C, Surridge C (2000) Arabidopsis thaliana genome. Introduction. Nature 408(6814):791. https://doi.org/10.1038/35048677
Deyoung BJ, Innes RW (2006) Plant NBS–LRR proteins in pathogen sensing and host defense. Nat Immunol 7:1243–1249. https://doi.org/10.1038/ni1410
Die JV, Román B, Qi X, Rowland LJ (2018) Genome-scale examination of NBS-encoding genes in Blueberry. Sci Rep 8:3429. https://doi.org/10.1038/s41598-018-21738-7
Finn RD, Penelope C, Eberhardt RY (2016) The Pfam protein families database: towards a more sustainable future. Nucleic Acids Res 44:D279–D285. https://doi.org/10.1093/nar/gkv1344
Friedman AR, Baker BJ (2007) The evolution of resistance genes in multi-protein plant resistance systems. Curr Opin Genet Dev 17:493–499. https://doi.org/10.1016/j.gde.2007.08.014
Fujita M, Fujita Y, Noutoshi Y, Takahashi F, Narusaka Y, Yamaguchi-Shinozaki K, Shinozaki K (2006) Crosstalk between abiotic and biotic stress responses: a current view from the points of convergence in the stress signaling networks. Curr Opin Plant Biol 9:436–442. https://doi.org/10.1016/j.pbi.2006.05.014
Gaut BS, Morton BR, Mccaig BC, Clegg MT (1996) Substitution rate comparisons between grasses and palms: synonymous rate differences at the nuclear gene Adh parallel rate differences at the plastid gene rbcL. PNAS 93:10274–10279. https://doi.org/10.1073/pnas.93.19.10274
Głowacki S, Macioszek VK, Kononowicz AK (2011) R proteins as fundamentals of plant innate immunity. Cell Mol Biol Lett 16:1–24. https://doi.org/10.2478/s11658-010-0024-2
Guo AY, Zhu QH, Chen X, Luo JC (2007) GSDS: a gene structure display server. Hereditas 29:1023–1026. https://doi.org/10.1360/yc-007-1023
Guo CJ, Sun XG, Chen X, Yang SH, Li J, Wang L, Zhang XH (2016) Cloning of novel rice blast resistance genes from two rapidly evolving NBS–LRR gene families in rice. Plant Mol Biol 90:95–105. https://doi.org/10.1007/s11103-015-0398-7
Haron S, Gong W, He S, Sun G, Sun J, Du X (2016) Genome-wide characterization and expression analysis of MYB transcription factors in Gossypium hirsutum. BMC Genet 17:129. https://doi.org/10.1186/s12863-016-0436-8
Iyer LM, Aravind L (2012) ALOG domains: provenance of plant homeotic and developmental regulators from the DNA-binding domain of a novel class of DIRS1-type retroposons. Biol Direct 7:39. https://doi.org/10.1186/1745-6150-7-39
Jia Y, Mcadams SA, Bryan GT, Hershey HP, Valent B (2014) Direct interaction of resistance gene and avirulence gene products confers rice blast resistance. EMBO J 19:4004–4014. https://doi.org/10.1093/emboj/19.15.4004
Kim YW, Jung HJ, Park JI, Hur Y, Nou IS (2015) Response of NBS encoding resistance genes linked to both heat and Fungal stress in Brassica oleracea. Plant Physiol Biochem 86:130–136. https://doi.org/10.1016/j.plaphy.2014.11.009
Kohler A, Guinet C, Duplessis S, Baucher M, Geelen D, Duchaussoy F, Meyers B, Boerjan W, Martin F (2008) Genome-wide identification of NBS resistance genes in Populus trichocarpa. Plant Mol Biol 66:619–636. https://doi.org/10.1007/s11103-008-9293-9
Kozák L, Szilágyi Z, Vágó B, Kakuk A, Tóth L, Molnár I, Pócsi I (2018) Inactivation of the indole-diterpene biosynthetic gene cluster of Claviceps paspali, by Agrobacterium-mediated gene replacement. Appl Microbiol Bio 102:3255–3266. https://doi.org/10.1007/s00253-018-8807-x
Krzywinski M, Schein J, Birol I, Connors J, Gascoyne R, Horsman DJ, Jones S, Marra M (2009) Circos: an information aesthetic for comparative genomics. Genome Res 19:1639–1645. https://doi.org/10.1101/gr.092759.109
Landolfo S, Ianiri G, Camiolo S, Porceddu A, Mulas G, Chessa R, Zara G, Mannazzu I (2018) CAR gene cluster and transcript levels of carotenogenic genes in Rhodotorula mucilaginosa. Microbiology 164:78–87. https://doi.org/10.1099/mic.0.000588
Leister D (2004) Tandem and segmental gene duplication and recombination in the evolution of plant disease resistance gene. Trends Genet Tig 20:116–122. https://doi.org/10.1016/j.tig.2004.01.007
Letunic I, Doerks T, Bork P (2012) SMART 7: recent updates to the protein domain annotation resource. Nucleic Acids Res 40:D302–D305. https://doi.org/10.1093/nar/gkr931
Librado P, Rozas J (2009) DnaSP v5: a software for comprehensive analysis of DNA polymorphism data. Bioinformatics 25:1451–1452. https://doi.org/10.1093/bioinformatics/btp187
Liu S, Liu Y, Yang X, Tong C, Edwards D, Parkin I, Zhao M, Ma J, Yu J, Shunmou H, Wang X, Wang J, Lu K, Fang Z, Bancroft I, Yang TJ, Hu Q, Wang X, Yue ZH, Paterson A (2014) The Brassica oleracea genome reveals the asymmetrical evolution of polyploid genomes. Nat Commun. 5:3930. https://doi.org/10.1038/ncomms4930
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(-delta delta c(t)) method. Methods 25:402–408. https://doi.org/10.1006/meth.2001.1262
Lozano R, Hamblin MT, Prochnik S, Jannink JL (2015) Identification and distribution of the NBS–LRR gene family in the Cassava genome. BMC Genom 16:360. https://doi.org/10.1186/s12864-015-1554-9
Luck JE, Lawrence GJ, Dodds PN, Shepherd KW, Ellis JG (2000) Regions outside of the leucine-rich repeats of flax rust resistance proteins play a role in specificity determination. Plant Cell 12:1367–1377. https://doi.org/10.1105/tpc.12.8.1367
Lv HH, Yang LM, Kang JG, Wang QB, Wang XW, Fang ZY, Liu YM, Zhuang M, Zhang YY, Lin Y, Yang YH, Xie BY, Liu B, Liu JS (2013) Development of Indel markers linked to Fusarium wilt resistance in cabbage. Mol Breed 32:961–967. https://doi.org/10.1007/s11032-013-9925-x
Lv S, Zhang CW, Tang J, Li Y, Wang Z, Jiang D, Hou XL (2015) Genome-wide analysis and identification of TIR-NBS–LRR genes in Chinese cabbage (Brassica rapa, ssp. pekinensis) reveal expression patterns to TuMV infection. Physiol Mol Plant P 90:89–97. https://doi.org/10.1016/j.pmpp.2015.04.001
Lyons E, Freeling M (2008) How to usefully compare homologous plant genes and chromosomes as DNA sequences. Plant J 53:661–673. https://doi.org/10.1111/j.1365-313x.2007.03326.x
Martin GB, Bogdanove AJ, Sessa G (2003) Understanding the functions of plant disease resistance proteins. Annu Rev Plant Biol 54:23. https://doi.org/10.1146/annurev.arplant.54.031902.135035
Meyers BC, Kozik A, Griego A, Kuang H, Michelmore RW (2003) Genome-wide analysis of NBS–LRR-encoding genes in Arabidopsis. Plant Cell 15:809–834. https://doi.org/10.1105/tpc.009308
Meyers BC, Morgante M, Michelmore RW (2010) TIR-X and TIR-NBS proteins: two new families related to disease resistance TIR-NBS–LRR proteins encoded in Arabidopsis and other plant genomes. Plant J 32:77–92. https://doi.org/10.1046/j.1365-313x.2002.01404.x
Miller R, Bertioli D, Baurens FC, Santos C, Alves PC, Martins N, Togawa R, Souza JM, Pappas G (2008) Analysis of non-TIR-NBS–LRR resistance gene analogs in Musa acuminata Colla: isolation, RFLP marker development, and physical mapping. BMC Plant Biol 8:15. https://doi.org/10.1186/1471-2229-8-15
Peele HM, Guan N, Fogelqvist J, Dixelius C (2014) Loss and retention of resistance genes in five species of the Brassicaceae family. BMC Plant Bio 14:1–11. https://doi.org/10.1186/s12870-014-0298-z
Pennisi E (2009) Stressed Out Over a Stress Hormone. Science 324:1012–1013. https://doi.org/10.1126/science.324-1012
Richly E, Kurth J, Leister D (2002) Mode of amplification and reorganization of resistance genes during recent Arabidopsis thaliana evolution. Mol Biol Evol 19:76–84. https://doi.org/10.1093/oxfordjournals.molbev.a003984
Saraste M (1990) The P-loop-a common motif in ATP- and GTP-binding proteins. Trends Biochem Sci 15:430–434. https://doi.org/10.1016/0968-0004(90)90281-F
Shao ZQ, Zhang YM, Hang YY, Xue JY, Zhou GC, Wu P, Wu X, Wu XZ, Wang Q, Wang B, Chen J (2014) Long-term evolution of nucleotide-binding site-leucine-rich repeat genes: understanding gained from and beyond the legume family. Plant Physiol 166:217–234. https://doi.org/10.1104/pp.114.243626
Sharma R, Rawat V, Suresh CG (2017) Genome-wide identification and tissue-specific expression analysis of nucleotide binding site-leucine rich repeat gene family in Cicer arietinum (kabuli chickpea). Genom Data 14:24–31. https://doi.org/10.1016/j.gdata.2017.08.004
Shazia AK, Abdur RM, Jong-In P et al (2018) Identification of NBS-encoding genes linked to black rot resistance in cabbage (Brassica oleracea var. capitata). Mol Biol Rep 45:773–778. https://doi.org/10.1007/s11033-018-4217-5
Shimizu M, Pu ZJ, Kawanabe T, Kitashiba H, Matsumoto S, Ebe Y, Sano M, Funaki T, Fukai E, Fujimoto R, Okazaki K (2014) Map-based cloning of a candidate gene conferring Fusarium yellows resistance in Brassica oleracea. Theor Appl Genet 128:119–130. https://doi.org/10.1007/s00122-014-2416-6
Tamura K, Dudley J, Nei M, Kumar S (2011) Mega 5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731. https://doi.org/10.1093/molbev/msr121
Tan X, Meyers B, Kozik AAL, West M, Morgante MA, St CD, Bent A, Michelmore R (2007) Global expression analysis of nucleotide binding site-leucine rich repeat-encoding and related genes in Arabidopsis. BMC Plant Biol 7:56. https://doi.org/10.1186/1471-2229-7-56
Tarr DEK, Alexander HM (2009) TIR-NBS–LRR genes are rare in monocots: evidence from diverse monocot orders. BMC Res Notes 2:197. https://doi.org/10.1186/1756-0500-2-197
Tian RP, Kang JG, Geng LH, Xie JM, Jian YC, Ding YH (2009) Study on the method of Fusarium wilts resistance in cabbage. CASB 5:39–42
Van RDH, Kamoun S (2008) From guard to decoy: a new model for perception of plant pathogen effectors. Plant Cell 20:2009–2017. https://doi.org/10.1105/tpc.108.060194
Van OG, Ha VDB, Cornelissen BJ, Takken F (2007) Structure and function of resistance proteins in Solanaceous plants. Annu Rev Phytopathol 45:43–72. https://doi.org/10.1146/annurev.phyto.45.062806.094430
Wan H, Yuan W, Bo K, Shen J, Pang X, Chen J (2013) Genome-wide analysis of NBS-encoding disease resistance genes in Cucumis sativus and phylogenetic study of NBS-encoding genes in Cucurbitaceae crops. BMC Genom 14:109. https://doi.org/10.1186/1471-2164-14-109
Wang L, Feng Z, Wang X, Wang X, Zhang X (2010) DEGseq: an R package for identifying differentially expressed genes from RNA-seq data. Bioinformatics 26:136–138. https://doi.org/10.1093/bioinformatics/btp612
Wang X, Wang H, Wang J et al (2014) The genome of the mesopolyploid crop species Brassica rapa. Nat Genet 43:1035–1039. https://doi.org/10.1038/ng.919
Wu J, Zhu J, Wang L, Wang S (2017) Genome-wide association study identifies NBS–LRR-encoding genes related with anthracnose and common bacterial blight in the common bean. Front Plant Sci 8:1398. https://doi.org/10.3389/fpls.2017.01398
Xing MM, Lv HH, Ma J, Xu DH, Li HL, Yang LM, Kang JG, Wang X, Fang ZY (2016) Transcriptome Profiling of Resistance to Fusarium oxysporum f.sp. conglutinans in Cabbage (Brassica oleracea) Roots. Plos One 11:e0148048. https://doi.org/10.1371/journal.pone.0148048
Yang S, Xi Zhang, Yue JX, Tian D, Chen JQ (2008) Recent duplications dominate NBS-encoding gene expansion in two woody species. Mol Genet Genom 280:187–198. https://doi.org/10.1007/s00438-008-0355-0
Yu J, Tehrim S, Zhang F, Tong C, Huang J, Cheng X, Dong C, Zhou Y, Qin R, Hua W, Liu S (2014) Genome-wide comparative analysis of NBS-encoding genes between Brassica species and Arabidopsis thaliana. BMC Genom 15:3. https://doi.org/10.1186/1471-2164-15-3
Yue JX, Meyers B, Chen JQ, Tian DC, Yang SH (2012) Tracing the origin and evolutionary history of plant nucleotide-binding site-leucine-rich repeat (NBS–LRR) genes. New Phytol 193:1049–1063. https://doi.org/10.1111/j.1469-8137.2011.04006.x
Zhang X, Liang P, Ming R (2016a) Genome-wide identification and characterization of nucleotide-binding site (NBS) resistance genes in pineapple. Trop Plant Biol 9:187–199. https://doi.org/10.1007/s12042-016-9178-z
Zhang YM, Sha ZQ, Wang Q, Hang YY, Xue JY, Wang B, Chen JQ (2016b) Uncovering the dynamic evolution of nucleotide-binding site-leucine-rich repeat (NBS–LRR) genes in Brassicaceae. J Integr Plant Biol 58:165–177. https://doi.org/10.1111/jipb.12365
Acknowledgements
This work was supported financially by grants from the National key research and development program (2016YFD0101804, 2016YFD0101702), the National Natural Science Foundation of China (31171958), and the Science & Technology Innovation Program of BAAFS (KJCX20180427, KJCX20170102, KJCX20170710).
Author information
Authors and Affiliations
Contributions
ZCL, JHY, JGK, JMX, and HPW designed the research. ZCL and HPW completed the experiments. JMX and HLL performed the data analysis and prepared the manuscript. ZCL, XHZ, JGK, and JX revised the manuscript. All authors had read and approved the final version of the manuscript.
Corresponding authors
Ethics declarations
Conflict of interest
The authors have no conflict of interest to disclose.
Electronic supplementary material
Below is the link to the electronic supplementary material.
13205_2019_1714_MOESM1_ESM.tif
Exon-intron structures of CNLs. The green bars indicate the exons, and the black lines indicate the introns (TIFF 6142 kb)
13205_2019_1714_MOESM2_ESM.tif
Exon-intron structures of TNLs. The green bars indicate the exons, and the black lines indicate the introns (TIFF 7235 kb)
Rights and permissions
About this article
Cite this article
Liu, Z., Xie, J., Wang, H. et al. Identification and expression profiling analysis of NBS–LRR genes involved in Fusarium oxysporum f.sp. conglutinans resistance in cabbage. 3 Biotech 9, 202 (2019). https://doi.org/10.1007/s13205-019-1714-8
Received:
Accepted:
Published:
DOI: https://doi.org/10.1007/s13205-019-1714-8