Abstract
Here is reported a novel histological marker line that strongly and specifically labels the male gametophyte in Arabidopsis flowers, from the uninucleate microspore to the pollination stage. In this marker line, the expression of the UidA and GFP marker genes is driven by the promoter of the At5g17340 Arabidopsis gene, a gene highly expressed in the male gametophyte. Strong GUS histochemical staining is detected in the uninucleate microspores. Strong GUS, and GFP, histological labelling is also detected in the subsequent stages of male gametophyte development. The potential applications of this marker line are discussed.Electronic supplementary materialThe online version of this article (doi:10.1186/2193-1801-2-237) contains supplementary material, which is available to authorized users.
Highlights
The Arabidopsis male gametophyte development is well characterised at the cytological level, and is a good system to study mitotic cell divisions
The At5g17340 promoter sequence promotes strong glucuronidase induced (GUS) staining in anthers One of the selected genes, the At5g17340, is a gene predicted to code for a plant specific 160 aminoacids transmembrane peptide
PAt5g17340:UidA:green fluorescence protein (GFP) is a good marker line to induce strong male gametophyte specific GUS staining, in uninucleate microspores and pollen grains In this report, it is shown that the flowers from the new pAt5g17340:UidA:GFP marker line plants exhibit strong and anther specific GUS staining (Additional file 1: Figure S1b-e), in uninucleate microspores and bicellular and tricellular pollen grains, after incubation with X-Gluc (Figure 1; Additional file 2: Figure S2)
Summary
The Arabidopsis male gametophyte development is well characterised at the cytological level, and is a good system to study mitotic cell divisions. It is a good system to indirectly characterise meiosis, since all four products of a single male meiotic cell division are viable, giving rise to microspores and pollen grains. The first mitotic cell division gives rise to bicellular pollen grains. As a result of the second mitotic cell division, tricellular pollen grains containing two gamete cells (the sperm cells) are formed. The bicellular pollen grain contains two nuclei, a condensed germinative nucleus and a large vegetative nucleus. The tricellular pollen contains a large vegetative nucleus and two condensed sperm nuclei (da Costa-Nunes and Grossniklaus 2003; Johnson-Brousseau and McCormick 2004; Borg et al 2009).
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