Kawakatsu, Y. | bioRxiv Science

bioRxiv2020

Autophagy is induced during plant grafting for wound healing

O_LIGrafting is an important technique in agriculture to obtain several good traits such as high disease tolerance and high yield by exchanging root system. However, the underlined cellular processes to compensate the wound damage and repair tissues were largely unknown. C_LIO_LIWe analyzed two graft combinations: Nicotiana benthamiana (Nb) homograft as a compatible, wound repairing model and Nb heterograft with Arabidopsis thaliana (At) as an incompatible and more stressful model, which we recently identified as an exceptional maintainable interfamily grafting. C_LIO_LIIn both graft combinations, nutrient loss was observed in gene expression after grafting, where the level of nutrient loss was more sever in heterografts. Transmission electron micrographs of Nb/At heterografts suggested that microautophagy was induced in cells near the graft boundary. In At seedling micrografting, the fluorescence of autophagy protein marker GFP-ATG8 was highly observed at graft junction especially in cambial region. In At atg2 mutant homografts, growth after grafting decreased compared with wild-type homografts. Moreover, when NbATG5 knocked-down Nb scion was grafted to At stock, the successful rate of grafting was significantly decreased. C_LIO_LIAltogether, these results suggest that component of autophagy is induced during grafting and has a role in wound healing. C_LI

plant biology↗

bioRxiv2019

Micrografting device for testing environmental conditions for grafting and systemic signaling in Arabidopsis

SummaryGrafting techniques have been applied in studies of systemic, long-distance signaling in several model plants. Seedling grafting in Arabidopsis, known as micrografting, enables investigation of the molecular mechanisms of systemic signaling between shoots and roots. However, conventional micrografting requires a high level of skill, limiting its use. Thus, an easier user-friendly method is needed. Here, we developed a silicone microscaled device, the micrografting chip, to obviate the need for training and to generate less stressed and more uniformly grafted seedlings. The chip has tandemly arrayed units, each of which consists of a seed pocket for seed germination and a micro-path with pairs of pillars for hypocotyl holding. Grafting, including seed germination, micrografting manipulation, and establishment of tissue reunion, is performed on the chip. Using the micrografting chip, we evaluated the effect of temperature and the carbon source on grafting and showed that a temperature of 27{degrees}C and a sucrose concentration of 0.5% were optimal. We also used the chip to investigate the mechanism of systemic signaling of iron status using a quadruple nicotianamine synthase (nas) mutant. The constitutive iron-deficiency response in the nas mutant because of aberrant partitioning was significantly rescued by grafting of wild-type shoots or roots, suggesting that shoot-and root-ward translocation of nicotianamine-iron complexes is essential for iron mobilization. Thus, our micrografting chip will promote studies of long-distance signaling in plants. Significance StatementA number of micrografting studies on systemic, long-distance signaling have been performed, but the technique is not yet used widely. Here, we developed a silicone-based micrografting chip to improve the ease-of-use, efficiency, and success rate of micrografting, even for untrained users.

plant biology↗