Fig. 2: CmoKARI1, but not CsaKARI, exhibits upward movement and is translated by the cucumber scion in response to chilling stress.

a RT-PCR identification of Csa and Cmo mRNA mobility under chilling conditions. Asterisks indicate mobile mRNAs identified in heterologous tissues. Arrows indicate movement direction. Cucumber or pumpkin ACTIN7 was used as an internal reference for mRNA abundance. Lower bands that appeared were primer dimers serving as sample loading controls. See uncropped images in the source file. The samples derive from the same experiment, and the gels were processed in parallel. b Identification of CmoKARI1 mobility from transformed cucumber hairy root to new leaves. The hairy root was transformed with cultures of Agrobacterium rhizogenes (Rhizobium rhizogenes) strain K599 harboring an empty vector (pCambia1300-Super::GFP) or pCambia1300-Super::CmoKARI1-GFP. K599 alone was transformed into a mock control. Fourteen days after transformation, RT-PCR with primers recognizing GFP was used to verify the mobility of GFP or CmoKARI1-GFP. #1, #2, and #3 represent independent transgenic seedlings as biological replicates. Mobility rate was analyzed by mobility seedlings/total independent hairy root transgenic seedlings. CsaACTIN7 was used as an internal reference for mRNA abundance. NPTII was used to exclude the Agrobacterium rhizogenes contamination or mobility. GFP fluorescence was identified using the LUYOR-3415RG fluorescent protein lamp. The samples derive from the same experiment, and the gels were processed in parallel. c Mobile CmoKARI1 from transformed cucumber hairy root was translated into new true leaves, shown by GUS staining. The hairy root was transformed with Agrobacterium rhizogenes (Rhizobium rhizogenes) strain K599 harboring an empty vector (pCambia1305-35S::GUS) or pCambia1305-35S::CmoKARI1-GUS. R: Agrobacterium-transformed cucumber hairy root, L: The emerged new leaves at 14 days after transformation. d RT-PCR with primers recognizing GUS was used to verify the mobility of GUS or CmoKARI1-GUS. #1, #2, and #3 represent independent transgenic seedlings as biological replicates. Mobility rate was analyzed by mobility seedlings/total independent hairy root transgenic seedlings. CsaACTIN7 was used as an internal reference for mRNA abundance. The hygromycin resistance (Hyg R) gene was used to exclude Agrobacterium rhizogenes contamination or mobility. The samples derive from the same experiment, and the gels were processed in parallel. e RT-PCR identification of CsaKARI from cucumber rootstock to pumpkin scion at 6 h, 12 h, and 24 h under chilling conditions. R rootstock, L true leaves. The samples derive from the same experiment, and the gels were processed in parallel. Source data are provided as a Source Data file.