Fig. 1

Schematic illustration of the T-DNA sequence in the constructed VOX vector for the expression of GFP-TaATG8a LB and RB: the left and right border sequences of the T-DNA. P35S: CaMV 35S promoter. Tnos: Nos transcription terminator. The FoMV genome contains five ORFs labeled from 1 to 4 and CP (coat protein)."

Fig. 2

FoMV-mediated expression of GFP and GFP-TaATG8a in N. benthamiana leaves The GFP-expressing and the GFP-TaATG8a-expressing vector plasmids were transformed into Agrobacterium tumefaciens and agroinfiltrated into N. benthamiana leaves, respectively. Control N. benthamiana leaves were infiltrated with the MMA buffer. GFP fluorescence in N. benthamiana leaves was observed at 7 days after agroinfiltration. Bar: 100 μm."

Fig. 3

FoMV-mediated expression of GFP and GFP-TaATG8a in wheat leaf tissues The second leaves of two-leaf-stage wheat seedlings were rub-inoculated with extract prepared from GFP-expressing or GFP-TaATG8a-expressing N. benthamiana leaves containing FoMV virions. For control seedlings, their second leaves were rub-inoculated with extract prepared from N. benthamiana leaves infiltrated with the MMA buffer. GFP fluorescence in the inoculated leaves was observed at 7 days and that in the third uninoculated leaves observed at 20 days after virus inoculation. Bar: 500 μm."

Fig. 4

FoMV-mediated expression of GFP and GFP-TaATG8a in the root tissues of wheat The second leaves of two-leaf-stage wheat seedlings were rub-inoculated with extract prepared from GFP-expressing or GFP-TaATG8a-expressing N. benthamiana leaves containing FoMV virions. For control seedlings, their second leaves were rub-inoculated with extract prepared from N. benthamiana leaves infiltrated with the MMA buffer. GFP fluorescence in the roots was observed at 20 days after virus inoculation. Bar: 500 μm."

Fig. 5

Localization of GFP-TaATG8a proteins in the leaf epidermal cells of N. benthamiana N. benthamiana leaves expressing GFP or GFP-TaATG8a were injected with 100 μmol L-1 E-64D (+E-64D) or an equal volume of 1% solvent DMSO (-E-64D) from the leaf abaxial side. These leaves were then detached and treated with starvation by keeping them in distilled water under dark conditions. GFP fluorescence in leaf epidermal cells was observed under a laser scanning confocal microscope at 24 hours after starvation treatment. Bar: 50 μm."

Fig. 6

Localization of GFP-TaATG8a proteins in the leaf epidermal cells of wheat Wheat leaves expressing GFP or GFP-TaATG8a were injected with 100 μmol L-1 E-64D (+E-64D) or an equal volume of 1% solvent DMSO (-E-64D) from prepared cuts on the main leaf veins. These leaves were then detached and treated with starvation by keeping them in distilled water under dark conditions. GFP fluorescence in the leaf epidermal cells was observed under a laser scanning confocal microscope at 24 hours after starvation treatment. Bar: 20 μm."

Fig. 7

Localization of GFP-TaATG8a proteins in the mesophyll cells of wheat Wheat leaves expressing GFP or GFP-TaATG8a were injected with 100 μmol L-1 E-64D (+E-64D) or an equal volume of 1% solvent DMSO (-E-64D) from prepared cuts on the main leaf veins. These leaves were then detached and treated with starvation by keeping them in distilled water under dark conditions. GFP fluorescence in the mesophyll cells was observed under a laser scanning confocal microscope at 24 hours after starvation treatment. Bar: 20 μm."

Fig. 8

Localization of GFP-TaATG8a proteins in the root cells of wheat Wheat roots expressing GFP or GFP-TaATG8a were detached and treated with starvation (Starvation/-) or starvation plus Concanamycin A (ConA) (Starvation/ConA) by keeping them in distilled water containing 1 μmol L-1 ConA or 1% solvent DMSO under dark conditions. Roots of intact plants that were not treated with starvation and ConA (-/-) were used as controls. GFP fluorescence in root cells was observed under a laser scanning confocal microscope at 24 hours after starvation treatment. Bar: 75 μm."