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Microspores of  Brassica napus  can be diverted from normal pollen development into embryogenesis by treating them with a mild heat shock. As microspore embryogenesis closely resembles zygotic embryogenesis, it is used as model for studying the molecular mechanisms controlling embryo formation. A previous study comparing the transcriptomes of three-day-old sorted embryogenic and pollen-like (non-embryogenic) microspores identified a gene homologous to  AT1G74730  of unknown function that was upregulated 8-fold in the embryogenic cells. In the current study, the gene was isolated and sequenced from  B. napus  and named  BnMicEmUP  ( B. napus  microspore embryogenesis upregulated gene). Four forms of  BnMicEmUP  mRNA and three forms of genomic DNA were identified.  BnMicEmUP2,3  was upregulated more than 7-fold by day 3 in embryogenic microspore cultures compared to non-induced cultures.  BnMicEmUP1,4  was highly expressed in leaves. Transient expression studies of  BnMicEmUP3::GFP  fusion protein in  Nicotiana benthamiana  and in stable  Arabidopsis  transgenics showed that it accumulates in chloroplasts. The features of the BnMicEmUP protein, which include a chloroplast targeting region, a basic region, and a large region containing 11 complete leucine-rich repeats, suggest that it is similar to a bZIP PEND (plastid envelope DNA-binding protein) protein, a DNA binding protein found in the inner envelope membrane of developing chloroplasts. Here, we report that the  BnMicEmUP3  overexpression in  Arabidopsis  increases the sensitivity of seedlings to exogenous abscisic acid (ABA). The  BnMicEmUP  proteins appear to be transcription factors that are localized in plastids and are involved in plant responses to biotic and abiotic environmental stresses; as well as the results obtained from this study can be used to improve crop yield.

Identification, Gene Structure, and Expression of BnMicEmUP: A Gene Upregulated in Embryogenic Brassica napus Microspores