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


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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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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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).

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