Cytoplasmic Fusion Detection at the Grafting Junction Based on Split sfGFP System

doi:10.7668/hbnxb.20192980

Abstract

Abstract:

In order to explore how cells from different plant individuals reestablish the connection between scion and stock to form a functional graft,self-assembling split super-folder greenfluorescent protein(sfGFP)system was used to detect the cytoplasm fusion at the graft junction.Transgenic Arabidopsis thaliana and Nicotiana benthamiana plants expressing sfGFP1-10 and mCherry-11 in cytoplasm were obtained by floral dip transformation and leaf disc transformation respectively,and 8-day old T1 transgenic plant were used for grafting.Interfamilial grafts(At-1-10/Nb-11),autografts(At-1-10/At-11,Nb-1-10/Nb-11)and control grafts(At-11/Nb-11,At-11/At-11,Nb-11/Nb-11)were constructed.On the fourth day after grafting,the sfGFP signal at the junction was observed under confocal microscope,and the fluorescence intensity of the grafts was calculated.The results showed that the interfamilial grafts and antografts displayed GFP signal in junction.GFP fluorescence intensity of interfamilial At/Nb grafts was significantly higher than that of autografts.GFP signal was not observed in cells of transgenic Arabidopsis thaliana and Nicotiana benthamiana expressing sfGFP1-10 or mCherry-11 alone and control grafts.These results indicated that at the fourth day,the fusion of cytoplasm at junction readily occurred.The sfGFP fragments were reassembled at the junction through the contact between the stock and the scion targeting in cytoplasm.Moreover,the reassembled fluorescence intensity in the interfamilial At/Nb heterografts was stronger than autografts,suggesting an extensive cellular fusion for At/Nb distant graft.This study provides a new perspective for the cellular events occurring at the graft junction.

Key words: Arabidopsis thaliana, Nicotiana benthamiana, Interfamilial grafting, Healing mechanism, sfGFP system

LI Dongyi, DENG Zhuying, LIANG Dacheng. Cytoplasmic Fusion Detection at the Grafting Junction Based on Split sfGFP System[J]. Acta Agriculturae Boreali-Sinica, 2022, 37(4): 151-157. doi: 10.7668/hbnxb.20192980.

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Figures/Tables 5

Fig.1 Transgenic Arabidopsis thaliana and Nicotiana benthamiana plants and constructed grafts

Fig.2 GFP signal observation of transgenic Arabidopsis thaliana plants and At/At grafts

Fig.3 GFP signal observation of transgenic Nicotiana benthamiana plants and Nb/Nb grafts

Fig.4 GFP signal observation of At/Nb grafts

Fig.5 GFP fluorescence intensity of heterograft and autografts Different lowercase letters indicated significant differences between the data(P<0.05).

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