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. 2019 May 20;8:e42834. doi: 10.7554/eLife.42834

Figure 2. REC8 and TUB4 localization.

(A) Cross section of two neighboring anthers of the KINGBIRD line expressing PROREC8:REC8:GFP and PRORPS5A:TagRFP:TUB4 in wild-type background. A1: premeiosis; A2: leptotene; A3: zygotene; A4: diplotene in the lower anther and metaphase I in the upper anther; A5: telophase I in the lower anther and late prophase II-metaphase II transition in the upper anther; A6: tetrad stage. (B) REC8:GFP localization after metaphase I (B1) in a PROREC8:REC8:GFP plant. The white arrowheads in B2 and B3 indicate centromeric REC8.

Figure 2.

Figure 2—figure supplement 1. Functionality of reporter lines used in this study.

Figure 2—figure supplement 1.

(A) Branches of Col-0 WT, rec8 -/-, REC8:GFP in rec8-/- background, KINGBIRD in Col-0 WT background. While rec8 -/- shows short, thin and sterile siliques (magenta arrow head), REC8:GFP and KINGBIRD present WT-like, elongated and thick siliques (white arrow heads). (B) Open siliques of Col-0 WT, rec8 -/-, REC8:GFP in rec8-/- background, KINGBIRD in Col-0 WT background. Col-0 WT, REC8:GFP and KINGBIRD form round, turgid and fully developed seeds (white arrow heads), whereas rec8 -/- presents dry and aborted seeds (magenta arrow head). (C) Anthers stained with Peterson staining (Peterson et al., 2010). Col-0 WT, REC8:GFP and KINGBIRD only produce viable pollen, while high rates of aborted pollen (blue) produced by rec8 -/- are visible inside the pollen sacs. (D) Rate of seed abortion (%). (E) Rate of aborted pollen in (%). (F) Cell spreads of Col-0 WT, rec8 -/-, REC8:GFP in rec8-/- background, TagRFP-TUB4 in Col-0 WT background and TagRFP:TUA5 in Col-0 WT background. The figure shows a selection of the main meiotic phases. rec8-/- starts to show defects from diplotene onwards, with the presence of univalent at metaphase I, and the consequent miss-segregation of chromosomes, which leads to the formation of micronuclei and unbalanced tetrads. The meiotic progression is restored in rec8-/- expressing REC8:GFP and it is not disrupted in TagRFP:TUB4 and TagRFP:TUA5 in Col-WT background. Both of the constructs generated balanced and viable gametes.