Abstract
Key message
Mapping and resequencing of two allelic early bolting mutants ebm5-1 and ebm5-2 revealed that the BrSDG8 gene is related to bolting in Chinese cabbage (Brassica rapa ssp. pekinensis).
Abstract
Bolting influences the leafy head formation and seed yield of Chinese cabbage therefore being an important agronomic trait. Herein, two allelic early bolting mutants, ebm5-1 and ebm5-2, stably inherited in Chinese cabbage were obtained from wild-type ‘FT’ seeds by ethyl methane sulfonate mutagenesis. Both mutants flowered significantly earlier than ‘FT,’ and genetic analysis revealed that the early bolting of the two mutants was controlled by one recessive nuclear gene. With BSR-seq, the mutations originating lines ebm5-1 and ebm5-2 were located to the same region in chromosome A07. Using the 1741 F2 individuals with the ebm5-1 phenotype as the mapping population, this region was narrowed to 56.24 kb between markers InDel18 and InDel45. A single-nucleotide polymorphism (SNP) was aligned to the BraA07g040740.3C (BrSDG8) region by whole-genome resequencing of ebm5-1 mutant and ‘FT.’ BrSDG8 is a homolog of Arabidopsis thaliana SDG8 encoding a histone methyltransferase affecting H3K4 trimethylation in FLOWERING LOCUS C chromatin. Comparative sequencing established that the SNP occurred on BrSDG8 17th exon in ebm5-1. Genotype analysis showed full co-segregation of the early bolting phenotype with this SNP. Cloning of allelic mutant ebm5-2 indicated that it harbors a deletion mutation on the 12th exon of BrSDG8. Quantitative real-time PCR analysis indicated that BrSDG8 expression level was observably lower in mutant ebm5-1 than in ‘FT.’ Overall, the present results provide strong evidence that BrSDG8 mutation leads to early bolting in Chinese cabbage, thereby providing a basis to understand the molecular mechanisms underlying this phenotype.
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References
Alexandre CM, Hennig L (2008) FLC or not FLC: the other side of vernalization. J Exp Bot 59(6):1127–1135
Bäurle I, Dean C (2006) The timing of developmental transitions in plants. Cell 125(4):655–664
Behm-Ansmant I, Izaurralde E (2006) Quality control of gene expression: a stepwise assembly pathway for the surveillance complex that triggers nonsense-mediated mRNA decay. Genes Dev 20(4):391–398
Berr A, Shafiq S, Shen WH (2011) Histone modifications in transcriptional activation during plant development. Biochim Biophys Acta 1809(10):567–576
Bolger AM, Lohse M, Usadel B (2014) Trimmomatic: a flexible trimmer for illumina sequence data. Bioinformatics 30(15):2114–2120
Boss PK, Bastow RM, Mylne JS, Dean C (2004) Multiple pathways in the decision to flower: enabling, promoting, and resetting. Plant Cell 16(Suppl):S18–S31
Brogna S, Wen J (2009) Nonsense-mediated mRNA decay (NMD) mechanisms. Nat Struct Mol Biol 16(2):107–113
Dillon SC, Zhang X, Trievel RC, Cheng X (2005) The SET-domain protein superfamily: protein lysine methyltransferases. Genome Biol 6(8):227
He Y, Amasino RM (2005) Role of chromatin modification in flowering-time control. Trends Plant Sci 10(1):30–35
He Y, Doyle MR, Amasino RM (2004) PAF1-complex-mediated histone methylation of FLOWERING LOCUS C chromatin is required for the vernalization-responsive, winter-annual habit in Arabidopsis. Genes Dev 18(22):2774–2784
Huang Y, Liu C, Shen WH, Ying R (2011) Phylogenetic analysis and classification of the Brassica rapa SET-domain protein family. BMC Plant Biol 11:175
Huang SN, Liu ZY, Yao RP, Li DY, Feng H (2015) Comparative transcriptome analysis of the petal degeneration mutant pdm in Chinese cabbage (Brassica campestris ssp. pekinensis) using RNA-seq. Mol Genet Genomics 290(5):1833–1847
Huang SN, Liu ZY, Yao RP et al (2016) Candidate gene prediction for a petal degeneration mutant, pdm, of the Chinese cabbage (Brassica campestris ssp. pekinensis) by using fine mapping and transcriptome analysis. Mol Breed 36(3):26
Huo H, Henry IM, Coppoolse ER, Verhoef-Post M, Schut JW, de Rooij H, Vogelaar A, Joosen RV, Woudenberg L, Comai L, Bradford KJ (2016) Rapid identification of lettuce seed germination mutants by bulked segregant analysis and whole genome sequencing. Plant J 88(3):345–360
Jack T (2004) Molecular and genetic mechanisms of floral control. Plant Cell 16(Suppl):S1–S17
Jiang D, Yang W, He Y, Amasino RM (2007) Arabidopsis relatives of the human lysine-specific demethylase1 repress the expression of FWA and FLOWERING LOCUS C and thus promote the floral transition. Plant Cell 19(10):2975–2987
Jiang D, Wang Y, Wang Y, He Y (2008) Repression of FLOWERING LOCUS C and FLOWERING LOCUS T by the Arabidopsis Polycomb repressive complex 2 components. PLoS ONE 3(10):e3404
Jiang L, Li D, Jin L, Ruan Y, Shen WH, Liu C (2018) Histone lysine methyltransferases BnaSDG8.A and BnaSDG8.C are involved in the floral transition in Brassica napus. Plant J 95:672–685
Jiao Y, Burow G, Gladman N, Acosta-Martinez V, Chen J, Burke J, Ware D, Xin Z (2017) Efficient identification of causal mutations through sequencing of bulked F 2 from two allelic bloomless mutants of sorghum bicolor. Front Plant Sci 8:2267
Johanson U, West J, Lister C, Michaels S, Amasino R, Dean C (2000) Molecular analysis of FRIGIDA, a major determinant of natural variation in Arabidopsis flowering time. Science 290(5490):344–347
Kim SY, Michaels SD (2006) SUPPRESSOR OF FRI 4 encodes a nuclear-localized protein that is required for delayed flowering in winter-annual Arabidopsis. Development 133(23):4699–4707
Kim D, Langmead B, Salzberg SL (2015) HISAT: a fast spliced aligner with low memory requirements. Nat Methods 12(4):357–360
Kosambi DD (1944) The estimation of map distance from recombination values. Ann Eugen 12:172–175
Lee J, Lee I (2010) Regulation and function of SOC1, a flowering pathway integrator. J Exp Bot 61(9):2247–2254
Li H, Durbin R (2009) Fast and accurate short read alignment with Burrows–Wheeler transform. Bioinformatics 25(14):1754–1760
Li H, Handsaker B, Wysoker A, Fennell T, Ruan J, Homer N, Marth G, Abecasis G, 1000 Genome Project Data Processing Subgroup (2009) The sequence alignment/map format and SAMtools. Bioinformatics 25(16):2078–2079
Liu F, Quesada V, Crevillén P, Bäurle I, Swiezewski S, Dean C (2007) The Arabidopsis RNA-binding protein FCA requires a lysine-specific demethylase 1 homolog to downregulate FLC. Mol Cell 28(3):398–407
Liu B, Berr A, Chang C, Liu C, Shen WH, Ruan Y (2016a) Interplay of the histone methyltransferases SDG8 and SDG26 in the regulation of transcription and plant flowering and development. Biochim Biophys Acta 1859(4):581–590
Liu YT, Li CY, Shi XX, Feng H, Wang YG (2016b) Identification of QTLs with additive, epistatic, and QTL x environment interaction effects for the bolting trait in Brassica rapa L. Euphytica 210:427–439
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2- ΔΔCt method. Methods 25:402–408
Maple J, Møller SG (2007) Mutagenesis in Arabidopsis. Methods Mol Biol 362:197–206
McKenna A, Hanna M, Banks E et al (2010) The genome analysis toolkit: a MapReduce framework for analyzing next-generation DNA sequencing data. Genome Res 20:1297–1303
Michaels SD (2004) Flowering time regulation produces much fruit. Curr Opin Plant Biol 12(1):75–80
Michaels SD, Amasino RM (1999) FLOWERING LOCUS C encodes a novel MADS domain protein that acts as a repressor of flowering. Plant Cell 11(5):949–956
Mouradov A, Cremer F, Coupland G (2002) Control of flowering time: interacting pathways as a basis for diversity. Plant Cell 14(Suppl):S111–S130
Murray MG, Thompson WF (1980) Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res 8(19):4321–4325
Niu L, Lu F, Pei Y, Liu C, Cao X (2007) Regulation of flowering time by the protein arginine methyltransferase AtPRMT10. EMBO Rep 8(12):1190–1195
Niu L, Zhang Y, Pei Y, Liu C, Cao X (2008) Redundant requirement for a pair of PROTEIN ARGININE METHYLTRANSFERASE4 homologs for the proper regulation of Arabidopsis flowering time. Plant Physiol 148(1):490–503
Nordström KJ, Albani MC, James GV, Gutjahr C, Hartwig B, Turck F, Paszkowski U, Coupland G, Schneeberger K (2013) Mutation identification by direct comparison of whole-genome sequencing data from mutant and wild-type individuals using k-mers. Nat Biotechnol 31(4):325–330
Pien S, Fleury D, Mylne JS, Crevillen P, Inzé D, Avramova Z, Dean C, Grossniklaus U (2008) ARABIDOPSIS TRITHORAX1 dynamically regulates FLOWERING LOCUS C activation via histone 3 lysine 4 trimethylation. Plant Cell 20(3):580–588
Putterill J, Laurie R, Macknight R (2004) It’s time to flower: the genetic control of flowering time. BioEssays 26(4):363–373
Schmitz RJ, Amasino RM (2007) Vernalization: a model for investigating epigenetics and eukaryotic gene regulation in plants. Biochim Biophys Acta 1769(5–6):269–275
Sheldon CC, Rouse DT, Finnegan EJ, Peacock WJ, Dennis ES (2000) The molecular basis of vernalization: the central role of FLOWERING LOCUS C (FLC). Proc Natl Acad Sci USA 97(7):3753–3758
Simpson GG, Gendall AR, Dean C (1999) When to switch to flowering. Annu Rev Cell Dev Biol 15:519–550
Song YH, Shim JS, Kinmonth-Schultz HA, Imaizumi T (2015) Photoperiodic flowering: time measurement mechanisms in leaves. Annu Rev Plant Biol 66:441–464
Springer NM, Kaeppler SM (2003) Comparative analysis of SET domain proteins in Maize and Arabidopsis reveals multiple duplications preceding the divergence of monocots and dicots. Plant Physiol 132(2):907–925
Strahl BD, Grant PA, Briggs SD, Sun ZW, Bone JR, Caldwell JA, Mollah S, Cook RG, Shabanowitz J, Hunt DF, Allis CD (2002) Set2 is a nucleosomal histone H3-selective methyltransferase that mediates transcriptional repression. Mol Cell Biol 22(5):1298–1306
Su A, Song W, Xing J, Zhao Y, Zhang R, Li C, Duan M, Luo M, Shi Z, Zhao J (2016) Identification of genes potentially associated with the fertility instability of S-type cytoplasmic male sterility in maize via bulked segregant RNA-Seq. PLoS ONE 11(9):e0163489
Tan C, Liu ZY, Huang SN, Feng H (2018) Mapping of the male sterile mutant gene ftms in Brassica rapa L. ssp. pekinensis via BSR-Seq combined with whole–genome resequencing. Theor Appl Genet 132:355–370
Tang X, Wang Y, Zhang Y, Huang S, Liu Z, Fei D, Feng H (2018) A missense mutation of plastid RPS4 is associated with chlorophyll deficiency in Chinese cabbage (Brassica campestris ssp. pekinensis). BMC Plant Biol 18(1):130
Van Ooijen J (2006) Joinmap 4.0 software for the calculation of genetic linkage maps in experimental populations. Kyazma BV, Cambridge
Wang X, Zhang Y, Ma Q, Zhang Z, Xue Y, Bao S, Chong K (2007) SKB1-mediated symmetric dimethylation of histone H4R3 controls flowering time in Arabidopsis. EMBO J 26(7):1934–1941
Wang K, Li M, Hakonarson H (2010) ANNOVAR: functional annotation of genetic variants from high-throughput sequencing data. Nucleic Acids Res 38(16):e164
Wang N, Liu ZY, Zhang Y, Li CY, Feng H (2018) Identification and fine mapping of a stay-green gene (Brnye1) in pakchoi (Brassica campestris L. ssp. chinensis). Theor Appl Genet 131(3):673–684
Xu L, Zhao Z, Dong A, Soubigou-Taconnat L, Renou JP, Steinmetz A, Shen WH (2008) Di- and tri- but not monomethylation on histone H3 lysine 36 marks active transcription of genes involved in flowering time regulation and other processes in Arabidopsis thaliana. Mol Cell Biol 28(4):1348–1360
Yang X, Yu Y, Zhang F, Zou Z, Zhao X, Zhang D, Xu J (2007) Linkage map construction and quantitative trait loci analysis for bolting based on a double haploid population of Brassica rapa. J Integr Plant Biol 49(5):664–671
Zhang X (2012) In: Wendel J, Greilhuber J, Dolezel J, Leitch I (eds) Plant genome diversity, vol 1. Springer, Vienna, pp 237–255
Zhao Z, Yu Y, Meyer D, Wu C, Shen WH (2005) Prevention of early flowering by expression of FLOWERING LOCUS C requires methylation of histone H3K36. Nat Cell Biol 7(12):1256–1260
Acknowledgements
The research was supported by the National Natural Science Foundation of China (Grant No. 31730082). We would like to thank Editage for English language editing.
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WF and SH have equally contributed to this work. HF and ZL designed the experiments. YG, MZ, and GQ helped developing the mutants. WF conducted the experiments and wrote the manuscript. WF and NW performed the data analysis. HF and SH revised the manuscript. All authors reviewed and approved this submission. The authors note that this research was performed and reported in accordance with ethical standards of the scientific conduct.
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Fu, W., Huang, S., Gao, Y. et al. Role of BrSDG8 on bolting in Chinese cabbage (Brassica rapa). Theor Appl Genet 133, 2937–2948 (2020). https://doi.org/10.1007/s00122-020-03647-4
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DOI: https://doi.org/10.1007/s00122-020-03647-4