# Preparation of Chitooligosaccharides with Specific Sequence Arrangement and Their Effect on Inducing Salt Resistance in Wheat Seedlings

## TL;DR

This study shows that specific chitooligosaccharide sequences can help wheat seedlings resist salt stress, with one type showing the best results.

## Contribution

The study is the first to demonstrate the sequence-dependent effect of chitooligosaccharides on plant salt resistance.

## Key findings

- A-COS showed the best activity in inducing salt resistance in wheat seedlings.
- A-COS treatment increased wheat seedling recovery in terms of leaf length, mass, and reduced stress markers.
- A-COS reduced malondialdehyde content and Na+/K+ ratio, indicating reduced salt stress damage.

## Abstract

Chitooligosaccharides (COS) exhibits good activity of inducing plant resistance, but the structure–activity relationship is still unclear. In this study, chitin oligosaccharides (CHOS) with a degree of polymerization (DP) of 2~6 were used as raw materials. Three deacetylases (NodB, VcCOD, and ArCE4A) were employed to prepare three different sequence-arranged COSs, namely N-COS, C-COS, and A-COS, and their structures were characterized by infrared spectroscopy, high-performance liquid chromatography, and mass spectrometry. Further studies were conducted on inducing the plant salt resistance of the three different sequence-arranged COSs on wheat seedlings. The results showed a sequence-dependent effect of COS inducing plant salt resistance. Among them, A-COS exhibited the best activity. When sprayed at a concentration of 10 mg/L on wheat seedlings under salt stress for 3 days, the leaf length of the wheat seedlings sprayed with A-COS was recovered, and the wet mass and dry mass were recovered by 20.40% and 6.64%, respectively. Following the enhancement of proline accumulation, the malondialdehyde content decreased by 34.75%, and the Na+/K+ ratio also exhibited a significant reduction, thereby alleviating salt stress-induced damage. This study was the first to demonstrate the effect of COS with specific sequences on inducing plant salt resistance, providing a theoretical basis for the development of a new generation of efficient COS plant biostimulator.