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

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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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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Correspondence to Ying Li.

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The authors declare that they have no conflicts of interest.

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Editor: Bin Tian

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Zhang, Y., Gao, S., Liu, H. 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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Keywords

  • cDNA-AFLP
  • Microspore embryogenesis
  • Transcript-derived fragments (TDFs)
  • Cis-elements
  • Non-heading Chinese cabbage