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cDNA-AFLP analysis of differentially expressed genes during microspore embryogenesis in non-heading Chinese cabbage

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Abstract

Stress can induce microspores to change their developmental pathway from the gametophytic to the embryogenic pathway. To explore the molecular mechanisms of microspore embryogenesis, complement DNA-amplified fragment length polymorphism was used to isolate the transcript-derived fragments during microspore embryogenesis of the non-heading Brassica campestris L. ssp. chinensis ‘Wuyueman’. With 256 primer combinations screened, a total of 94 transcript-derived fragments were identified, and 15 were successfully sequenced. Based on a BLAST search in the Brassica database, 12 of the 15 sequenced transcript-derived fragments were homologous to genes with annotations; the remaining three transcript-derived fragments did not match any sequences. Transcript-derived fragments with annotations were involved in cell wall formation, hormones, and resistance. Analysis of cis-elements indicated that there were heat shock-related and stress-related cis-elements in the promoter sequences of 12 transcript-derived fragments. TDF1(Bra040720), TDF6(Bra013664), and TDF15(Bra022587) were selected for validation of complement DNA-amplified fragment length polymorphism expression patterns by real-time quantitative PCR. Results confirmed the altered expression patterns of three genes revealed by the complement DNA-amplified fragment length polymorphism. This study provides novel information on the molecular mechanism of microspore embryogenesis in non-heading Chinese cabbage.

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References

  • Abdollahi MR, Moieni A, Mousavi A, Salmanian AH, Jalali Javaran M, Majdi M (2007) Effect of integrated bombardment and agrobacterium transformation system on transient GUS expression in hypocotyls of rapeseed (Brassica napus L. cv. PF704) microspore-derived embryos. Pak J Biol Sci 10:3141–3145

    Article  CAS  PubMed  Google Scholar 

  • Abercrombie JM, Farnham MW, Rushing JW (2005) Genetic combining ability of glucoraphanin level and other horticultural traits of broccoli. Euphytica 143:145–151

    Article  CAS  Google Scholar 

  • Bachem CW, Hoeven RS, Bruijn SM, Vreugdenhil D, Zabeau M, Visser RG (1996) Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP: analysis of gene expression during potato tuber development. Plant J 9:745–753

    Article  CAS  PubMed  Google Scholar 

  • Bélanger S, Marchand S, Jacques PÉ, Meyers B, Belzile F (2018) Differential expression profiling of microspores during the early stages of isolated microspore culture using the responsive barley cultivar gobernadora. G3(Bethesda) 8:1603–1614

    Google Scholar 

  • Belmonte M, Elhiti M, Waldner B, Stasolla C (2010) Depletion of cellular brassinolide decreases embryo production and disrupts the architecture of the apical meristems in Brassica napus microspore-derived embryos. J Exp Bot 61:2779–2794

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Bou Daher F, van Oostende C, Geitmann A (2011) Spatial and temporal expression of actin depolymerizing factors ADF7 and ADF10 during male gametophyte development in Arabidopsis thaliana. Plant Cell Physiol 52:1177–1192

    Article  CAS  PubMed  Google Scholar 

  • Boutilier K, Offringa R, Sharma VK, Kieft H, Ouellet T, Zhang LM, Hattori J, Liu CM, van Lammeren AA, Miki BL, Custers JB, van Lookeren Campagne MM (2002) Ectopic expression of BABY BOOM triggers a conversion from vegetative to embryonic growth. Plant Cell 14:1737–1749

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Cegielska-Taras T, Pniewski T, Szała L (2008) Transformation of microspore-derived embryos of winter oilseed rape (Brassica napus L.) by using agrobacterium tumefaciens. J Appl Genet 49:343–347

    Article  PubMed  Google Scholar 

  • Chan SW (2010) Chromosome engineering: power tools for plant genetics. Trends Biotechnol 28:605–610

    Article  CAS  PubMed  Google Scholar 

  • Chandra S, Kazmi AZ, Ahmed Z, Roychowdhury G, Kumari V, Kumar M, Mukhopadhyay K (2017) Genome-wide identification and characterization of NB-ARC resistant genes in wheat (Triticum aestivum L.) and their expression during leaf rust infection. Plant Cell Rep 36:1097–1112

    Article  CAS  PubMed  Google Scholar 

  • Custers JB, Cordewener JH, Nöllen Y, Dons HJ, Mm VLC (1994) Temperature controls both gametophytic and sporophytic development in microspore cultures of Brassica napus. Plant Cell Rep 13:267–271

    Article  CAS  PubMed  Google Scholar 

  • Dong X, Yi H, Lee J, Nou IS, Han CT, Hur Y (2015) Global gene-expression analysis to identify differentially expressed genes critical for the heat stress response in Brassica rapa. PLoS One 10:e0130451

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Dubas E, Moravčíková J, Libantová J, Matušíková I, Benková E, Żur I, Krzewska M (2014) The influence of heat stress on auxin distribution in transgenic B. napus microspores and microspore-derived embryos. Protoplasma 251:1077–1087

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ehlting J, Sauveplane V, Olry A, Ginglinger JF, Provart NJ, Werck-Reichhart D (2008) An extensive (co-)expression analysis tool for the cytochrome P450 superfamily in Arabidopsis thaliana. BMC Plant Biol 8:47

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • El-Tantawy AA, Solís MT, Da Costa ML, Coimbra S, Risueño MC, Testillano PS (2013) Arabinogalactan protein profiles and distribution patterns during microspore embryogenesis and pollen development in Brassica napus. Plant Reprod 26:231–243

    Article  CAS  PubMed  Google Scholar 

  • Ferrie AMR, Caswell KL (2011) Isolated microspore culture techniques and recent progress for haploid and doubled haploid plant production. Plant Cell Tissue Organ Cult 104:301–309

    Article  Google Scholar 

  • Gong T, Shu D, Zhao M, Zhong J, Deng HY, Tan H (2014) Isolation of genes related to abscisic acid production in Botrytis cinerea, TB-3-H8 by cDNA-AFLP. J Basic Microbiol 54:204–214

    Article  CAS  PubMed  Google Scholar 

  • Guo LH, Chen SN, Liu KH, Liu YF, Ni LH, Zhang KQ, Zhang LM (2008) Isolation of heat shock factor HsfA1a-binding sites in vivo revealed variations of heat shock elements in Arabidopsis thaliana. Plant Cell Physiol 49:1306–1315

    Article  CAS  PubMed  Google Scholar 

  • Haralampidis K, Milioni D, Rigas S, Hatzopoulos P (2002) Combinatorial interaction of cis elements specifies the expression of the Arabidopsis AtHsp90-1 gene. Plant Physiol 129:1138–1149

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Heidari-Zefreh AA, Shariatpanahi ME, Mousavi A, Kalatejari S (2019) Enhancement of microspore embryogenesis induction and plantlet regeneration of sweet pepper (Capsicum annuum L.) using putrescine and ascorbic acid. Protoplasma:13–24

  • Henderson CAP, Pauls KP (1992) The use of haploidy to develop plants that express several recessive traits using light-seeded canola (Brassica napus) as an example. Theor Appl Genet 83:476–479

    Article  CAS  PubMed  Google Scholar 

  • Hoseini M, Ghadimzadeh M, Ahmadi B, Teixeira da Silva JA (2014) Effects of ascorbic acid, alpha-tocopherol, and glutathione on microspore embryogenesis in Brassica napus L. In Vitro Cell Dev Biol-Plant 50:26–35

    Article  CAS  Google Scholar 

  • Jacquard C, Mazeyrat-Gourbeyre F, Devaux P, Boutilier K, Baillieul F, Clémentet C (2009) Microspore embryogenesis in barley: anther pre-treatment stimulates plant defence gene expression. Planta 229:393–402

    Article  CAS  PubMed  Google Scholar 

  • Kitashiba H, Taguchi K, Kaneko I, Inaba K, Yokoi S, Takahata Y, Nishio T (2016) Identification of loci associated with embryo yield in microspore culture of Brassica rapa by segregation distortion analysis. Plant Cell Rep 35:2197–2204

    Article  CAS  PubMed  Google Scholar 

  • Kreps JA, Wu YJ, Chang HS, Zhu T, Wang X, Harper JF (2002) Transcriptome changes for Arabidopsis in response to salt, osmotic, and cold stress. Plant Physiol 130:2129–2141

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kumar A, Ellis BE (2001) The phenylalanine ammonia-lyase gene family in raspberry. Structure, expression, and evolution. Plant Physiol 127:230–239

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li J, Huang Q, Sun MX, Zhang TY, Li H, Chen BY, Xu K, Gao GZ, Li F, Yan GX, Qiao JW, Cai YP, Wu XM (2016) Global DNA methylation variations after short-term heat shock treatment in cultured microspores of Brassica napus cv. Topas Sci Rep 6:38401

    Article  CAS  PubMed  Google Scholar 

  • Li Y, Wang HP, Li XL, Liang G, Yu DQ (2017) Two DELLA-interacting proteins bHLH48 and bHLH60 regulate flowering under long-day conditions in Arabidopsis thaliana. J Ex Bot 68:2757–2767

    Article  CAS  Google Scholar 

  • Liu C, Wang JL, Huang TD, Wang F, Yuan F, Cheng XM, Zhang Y, Shi SW, Wu JS, Liu KD (2010) A missense mutation in the VHYNP motif of a DELLA protein causes a semi-dwarf mutation phenotype in Brassica napus. Theor Appl Genet 121:249–258

    Article  CAS  PubMed  Google Scholar 

  • Liu X, Han FQ, Kong CC, Fang ZY, Yang LM, Zhang YY, Zhuang M, Liu YM, Li ZS, Lv HH (2017) Rapid introgression of the fusarium wilt resistance gene into an elite cabbage line through the combined application of a microspore culture, genome background analysis, and disease resistance-specific marker assisted foreground selection. Front Plant Sci 8:354

    CAS  PubMed  PubMed Central  Google Scholar 

  • 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

    Article  CAS  PubMed  Google Scholar 

  • Lu Y, Dai SY, Gu AX, Liu MY, Wang YH, Luo SX, Zhao YJ, Wang S, Xuan SX, Chen XP, Li XF, Bonnema G, Zhao JJ, Shen SX (2016) Microspore induced doubled haploids production from ethyl Methanesulfonate (EMS) soaked flower buds is an efficient strategy for mutagenesis in Chinese cabbage. Front Plant Sci 7:1780

    PubMed  PubMed Central  Google Scholar 

  • Maheshwari P, Selvaraj G, Kovalchuk I (2011) Optimization of Brassica napus (canola) explant regeneration for genetic transformation. New Biotechnol 29:144–155

    Article  CAS  Google Scholar 

  • Malik MR, Wang F, Dirpaul JM, Zhou N, Hammerlindl J, Keller W, Abrams SR, Ferrie AM, Krochko JE (2008) Isolation of an embryogenic line from non-embryogenic Brassica napus cv. Westar through microspore embryogenesis. J Exp Bot 59:2857–2873

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Malik MR, Wang F, Dirpaul JM, Zhou N, Polowick PL, Ferrie AMR, Krochko JE (2007) Transcript profiling and identification of molecular markers for early microspore embryogenesis in Brassica napus. Plant Physiol 144:134–154

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Maraschin Sde F, Caspers M, Potokina E, Wülfert F, Graner A, Spainke HP, Wang M (2006) cDNA array analysis of stress-induced gene expression in barley androgenesis. Physiol Plant 127:535–550

    Article  CAS  Google Scholar 

  • Maraschin Sde F, Lamers GEM, de Pater BS, Spaink HP, Wang M (2003) 14-3-3 isoforms and pattern formation during barley microspore embryogenesis. J Exp Bot 54:1033–1043

    Article  PubMed  CAS  Google Scholar 

  • Melloul M, Iraqi D, El AM, Erba G, Alaoui S, Ibriz M, Elfahime E (2014) Identification of differentially expressed genes by cDNA-AFLP technique in response to drought stress in Triticum durum. Food Technol Biotechnol 52:479–488

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Mulako I, Farrant JM, Collett H, Illing N (2008) Expression of Xhdsi-1VOC, a novel member of the vicinal oxygen chelate (VOC) metalloenzyme superfamily, is up-regulated in leaves and roots during desiccation in the resurrection plant Xerophyta humilis (Bak) Dur and Schinz. J Exp Bot 59:3885–3901

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Narusaka Y, Narusaka M, Seki M, Umezawa T, Ishida J, Nakajima M, Enju A, Shinozaki K (2004) Crosstalk in the responses to abiotic and biotic stresses in Arabidopsis: analysis of gene expression in cytochrome P450gene superfamily by cDNA microarray. Plant Mol Biol 55:327–342

    Article  CAS  PubMed  Google Scholar 

  • Rieping M, Schöffl F (1992) Synergistic effect of upstream sequences, CCAAT box elements, and HSE sequences for enhanced expression of chimaeric heat shock genes in transgenic tobacco. Mol Gen Genet 231:226–232

    Article  CAS  PubMed  Google Scholar 

  • Rodríguez-Sanz H, Manzanera JA, Solís MT, Gómez-Garay A, Pintos B, Risueño MC, Testillano PS (2014) Early markers are present in both embryogenesis pathways from microspores and immature zygotic embryos in cork oak, Quercus suber L. BMC Plant Biol 14:224

    Article  PubMed  PubMed Central  CAS  Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  • Sato S, Katoh N, Iwai S, Hagimori M (2002) Effect of low temperature pretreatment of buds or inflorescence on isolated microspore culture in Brassica rapa (syn. B. campestris). Breeding Sci 52:23–26

    Article  Google Scholar 

  • Sato T, Nishio T, Hirai M (1989) Plant regeneration from isolated microspore of Chinese cabbage (Brassica campestris ssp. pekinensis). Plant Cell Rep 8:486–488

    Article  CAS  PubMed  Google Scholar 

  • Solís MT, Berenguer E, Risueño MC, Testillano PS (2016) BnPME is progressively induced after microspore reprogramming to embryogenesis, correlating with pectin de-esterification and cell differentiation in Brassica napus. BMC Plant Biol 16:176

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Valdés A, Clemens R, Möllers C (2018) Mapping of quantitative trait loci for microspore embryogenesis-related traits in the oilseed rape doubled haploid population DH4069 × express 617. Mol Breeding 38:65

    Article  CAS  Google Scholar 

  • Vos P, Hogers R, Bleeker M, Reijans M, van de Lee T, Hornes M, Friters A, Pot J, Paleman J, Kuiper M (1995) AFLP: a new technique for DNA fingerprinting. Nucleic Acids Res 23:4407–4414

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang HJ, Yang CG, Zhang C, Wang NY, Lu DH, Wang J, Zhang SS, Wang ZX, Ma H, Wang XL (2011) Dual role of BKI1 and 14-3-3 s in brassinosteroid signaling to link receptor with transcription factors. Dev Cell 21:825–834

    Article  CAS  PubMed  Google Scholar 

  • Wang M, Van Bergen S, Van Duijn B (2000) Insights into a key developmental switch and its importance for efficient plant breeding. Plant Physiol 124:523–530

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Wang XJ, Tang CL, Zhang G, Li YC, Wang CF, Liu B, Qu ZP, Zhao J, Han QM, Huang LL, Chen XM, Kang ZS (2009) CDNA-AFLP analysis reveals differential gene expression in compatible interaction of wheat challenged with Puccinia striiformis f. sp. tritici. BMC Genomics 10:289

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Werck-Reichhart D, Bak S, Paquette S (2002) Cytochromes P450. In: Somerville CR, Meyerowitz EM (eds) The Arabidopsis Book, vol 1. American Society of Plant Biologists, Rockville, p e0028

    Google Scholar 

  • Xiao D, Liu ST, Wei YP, Zhou DY, Hou XL, Li Y, Hu CM (2016) cDNA-AFLP analysis reveals differential gene expression in incompatible interaction between infected non-heading Chinese cabbage and Hyaloperonospora parasitica. Hortic Res 3:16034

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  • Yi SY, Liu J (2009) Combinatorial interactions of two cis- acting elements, AT-rich regions and HSEs, in the expression of tomato Lehsp23.8, upon heat and non-heat stresses. J. Plant Biol 52:560

    Article  CAS  Google Scholar 

  • Yuan SX, Liu YM, Fang ZY, Yang LM, Zhuang M, Zhang YY, Sun PT (2011) Effect of combined cold pretreatment and heat shock on microspore cultures in broccoli. Plant Breed 130:80–85

    Article  Google Scholar 

  • Zeng AS, Yan JY, Song LX, Bing G, Jianqi L (2015) Effects of ascorbic acid and embryogenic microspore selection on embryogenesis in white cabbage (Brassica oleracea L. var. capitata). J Hortic Sci Biotechnol 90:607–612

    Article  CAS  Google Scholar 

  • Zhang XB, Gou MY, Liu CJ (2013) Arabidopsis Kelch repeat F-box proteins regulate phenylpropanoid biosynthesis via controlling the turnover of phenylalanine ammonia-lyase. Plant Cell 25:4994–5010

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Zhao JP, Newcomb W, Simmonds D (2003) Heat-shock proteins 70 kDa and 19 kDa are not required for induction of embryogenesis of Brassica napus L. cv. Topas microspores. Plant Cell Physiol 44:1417–1421

    Article  CAS  PubMed  Google Scholar 

  • Żur I, Dubas E, Golemiec E, Szechyńska-Hebda M, Golebiowska G, Wedzony M (2009) Stress-related variation in antioxidative enzymes activity and cell metabolism efficiency associated with embryogenesis induction in isolated microspore culture of triticale (×Triticosecale Wittm.). Plant Cell Rep 28:1279–1287

    Article  PubMed  CAS  Google Scholar 

  • Żur I, Dubas E, Krzewska M, Janowiak F (2015) Current insights into hormonal regulation of microspore embryogenesis. Front Plant Sci 6:424

    PubMed  PubMed Central  Google Scholar 

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Funding

This work was supported by the National Natural Science Foundation of China (31872106), the National Key Research and Development Program (2018YFD1000800 and 2017YFD0101803), and National vegetable industry technology system (CARS-23-A-06).

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  1. State Key Laboratory of Crop Genetics and Germplasm Enhancement, College of Horticulture, Nanjing Agricultural University, Nanjing, Jiangsu, 210095, People’s Republic of China

    Ya Zhang, Su-yan Gao, Huan-huan Liu, Xi-lin Zhang, Ai-song Zeng, Jian-jun Wang, Xi-lin Hou & Ying Li

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  1. Ya Zhang
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Zhang, Y., Gao, Sy., Liu, Hh. et al. cDNA-AFLP analysis of differentially expressed genes during microspore embryogenesis in non-heading Chinese cabbage. In Vitro Cell.Dev.Biol.-Plant 56, 18–28 (2020). https://doi.org/10.1007/s11627-019-10036-0

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