Capillary electrophoresis study of systemin peptides spreading in tomato plant

Abstract

Systemin (Sys) is an 18‐aa plant peptide hormone involved in the regulation of plant's defensive response. Sys is considered as a fast‐spreading systemic wound signal. We developed a simple and rapid CE method to monitor the spreading of Sys peptides through tomato plant. A 1,2,3‐triazole‐linked AZT‐systemin conjugate was designed as a model to study the possibility of translocating small cargo molecules 3'‐Azido‐2',3'‐dideoxythymidine by systemin. The Sys peptides (Sys, N‐propiolyl Sys, and AZT‐systemin conjugate) were injected into the stem and leaves of mature tomato plant. Its transportation throughout the plant tissue was traced by CE. The peptides were clearly visible in the crude tomato exudates and an optimum separation was achieved in 25 mM phosphate “buffer” at pH 2.5 and a voltage of 20 kV using uncoated fused silica capillary. CE analysis showed that Sys peptides are well separated from tomato plant exudates ingredients and are stable in tomato stem and leaf exudates for up to 24 h. CE study revealed that the Sys peptides are effectively spreading throughout tomato stem and leaves and the peptides could be directly detected in the crude plant matrixes. The translocation was strongly inhibited by sodium azide. The results showed that the established CE method can be used to characterize plant peptides spreading under plant physiological conditions.

Arabidopsis Plants Sense Non-self Peptides to Promote Resistance Against Plectosphaerella cucumerina

“…Few methods for small peptides determination in solanaceous are found along the literature (Mucha et al, 2019). To further confirm the uptake and the presence of the non-self peptides that were able to induce resistance in Arabidopsis we developed a multi-residue analytical method based on the one described in Pastor et al (2018).…”

Recent developments in capillary and microchip electroseparations of peptides (2019–mid 2021)

“…CE represents a suitable method for analysis of plant bioactive peptides [232]. Particular example is a CE study of spreading of systemin (18‐AA plant peptide hormone involved in the regulation of plant's defensive response) and its derivatives in tomato plant [233]. CE analyses were performed in 75 μm id, 60/50 cm total/effective length capillary, at 20 kV separation voltage, 25°C, 214 nm detection wavelength, and using 25 mM phosphate buffer as BGE.…”

Systemin‐mediated long‐distance systemic defense responses

“…Thus, multiple mobile signals are integrated to modulate the SYS‐mediated systemic defense responses in plants. Note that the mobility of SYS peptide and Pro‐SYS mRNA in tomato was demonstrated only for SYS peptide applied at high concentrations into a wound or injected into the stem (Narváez‐Vásquez et al , 1995; Dobkowski et al , 2014; Mucha et al , 2019), or Pro‐SYS mRNA agroinfiltrated into the leaf epidermis (Zhang & Hu, 2017), and therefore, whether endogenous SYS peptide and Pro‐SYS mRNA are mobile in tomato remains to be determined.…”

“…The results of these grafting experiments suggested that SYS is not the mobile defense signal but is required for local biosynthesis of the signal (JA) in the wounded tomato leaf (Li et al , 2002). Very recent work, however, found that SYS peptide injected into the stem and leaves of mature tomato plants was transported throughout the plant tissue, as detected using capillary electrophoresis (Mucha et al , 2019). This is consistent with the observation that SYS peptide was easily translocated through the tomato stem (Dobkowski et al , 2014).…”