Abstract
BackgroundGrafting with rootstocks is essential for the culture of many perennial fruit crops and is increasing being used in the production of annual fruits and vegetables. Our previous work based on microarrays showed that transcripts encoding enzymes of both primary and secondary metabolism were differentially expressed during graft union formation in both homo-grafts (a genotype grafted with itself) and hetero-grafts (two different genotypes grafted together). The aim of this study was to profile primary and secondary metabolites, and quantify the activity of phenylalanine ammonia lyase (PAL) and neutral invertase (NI) in the scion and rootstock tissues and the graft interface of homo and hetero-grafts of grapevine 1 month after grafting. Table-top grafting was done on over-wintering stems (canes) of grapevine and the graft interface tissues (containing some woody stem tissues and callus) were compared to the surrounding rootstock and scion tissues. The objective was to identify compounds involved in graft union formation and hetero-grafting responses.ResultsA total of 54 compounds from primary and secondary metabolism (19 amino acids, five primary and 30 secondary compounds metabolites) and the activity of two enzymes were measured. The graft interface was associated with an increase in the accumulation of the branched-chain amino acids, basic amino acids, certain stilbene compounds and higher PAL and NI activity in comparison to the surrounding woody stem tissues. Some amino acids and stilbenes were identified as being accumulated differently between the graft interfaces of the scion/rootstock combinations in a manner which was unrelated to their concentrations in the surrounding woody stem tissues.ConclusionsThis study revealed the modification of primary metabolism to support callus cell formation and the stimulation of stilbene synthesis at the graft interface, and how these processes are modified by hetero-grafting. Knowledge of the metabolites and/or enzymes required for successful graft union formation offer us the potential to identify markers that could be used by nurseries and researchers for selection and breeding purposes.
Highlights
Grafting with rootstocks is essential for the culture of many perennial fruit crops and is increasing being used in the production of annual fruits and vegetables
Graft union formation begins with the formation of a necrotic layer and adhesion between the two grafted partners; this is followed by the proliferation of callus tissue and the formation of functional xylem and phloem connections between the scion and rootstock [2]
Cabernet Sauvignon (CS/CS) 28 d after grafting showed a distinct primary metabolite profile in comparison with the scion and rootstock tissues. This graft interface tissue had higher water, protein and Glc contents, and a lower concentration of starch compared to the surrounding tissues (Fig. 1., Additional file 1: Table S1)
Summary
Grafting with rootstocks is essential for the culture of many perennial fruit crops and is increasing being used in the production of annual fruits and vegetables. The aim of this study was to profile primary and secondary metabolites, and quantify the activity of phenylalanine ammonia lyase (PAL) and neutral invertase (NI) in the scion and rootstock tissues and the graft interface of homo and hetero-grafts of grapevine 1 month after grafting. Grafting is a considerable stress to plants and triggers wounding responses such as the production of reactive oxygen species [3], the expression of defense related genes [4,5,6], the accumulation of defense related and antioxidant enzymes [3, 7, 8], phenolic compounds [9] and the production of wound-induced callus [10]. Increased PAL activity at the graft interface has the potential to increase the production of stress-related secondary metabolites with antioxidant functions well as providing precursors for the synthesis of lignin (necessary for xylem formation)
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