Fig. 1: ATG5- and ATG7-mediated autophagy is involved in pollen tube growth in Arabidopsis thaliana.

a Schematic illustration of the T-DNA insertion sites of atg5-1 and atg7-2 of Arabidopsis thaliana. Red arrows mark the locations of primers used for RT-qPCR. b RT-qPCR analysis was performed to quantify ATG5 expression in flowers at developmental stages 11–14 of WT, atg5-1, atg5-1/atg7-2 and pATG5::GFP-ATG5/atg5-1. Statistical analysis was performed using two-tailed paired Student’s t-test, with n = 3 independent experimental replicates (error bars ± SD, ∗∗∗p < 0.001, ns, no significant). Source data are provided in Source data file. c Similarly, ATG7 expression was analyzed in flowers of WT, atg7-2, atg5-1/atg7-2 and pATG7::GFP-ATG7/atg7-2. Actin2 was used as an internal control. Statistical analysis was performed using two-tailed paired Student’s t-test, with n = 3 independent experimental replicates (error bars ± SD, ∗∗∗p < 0.001, ns, no significant). Source data are provided in Source data file. d Western blot detection of ATG8 and ATG8-PE in germinated pollen of WT, atg5-1 and atg7-2 using anti-ATG8 antibody. Three independent experimental replicates were conducted, yielding consistent results. Source data of blotting are provided in Source data file. e Co-expression of GFP-ATG5 with mCherry-ATG7, ATG6-RFP, YFP-ATG8e, GFP-ATG9 and SH3P2-RFP in growing tobacco pollen tubes respectively. The punctate dots of ATG5 and ATG7 colocalize with one another and they mainly localize in the pollen tube shank. Results of colocalization ratios are calculated by either Pearson correlation coefficients or as Spearman’s rank correlation coefficients with ImageJ. The generated r values in the range −1 to 1, where 0 indicates no discernable correlation and +1 or −1 indicate strong positive or negative correlations, respectively. Consistent results were obtained across three independent experimental replicates. Scale bar  = 25 μm.