Dynamic TMT-Based Quantitative Proteomics Analysis of Critical Initiation Process of Totipotency during Cotton Somatic Embryogenesis Transdifferentiation.

Haixia Guo,Yupeng Fan,Huihui Guo,Zhengmin Tang,Yijie Fan,Fanchang Zeng,Li Zhang

doi:10.3390/ijms20071691

Abstract

The somatic embryogenesis (SE) process of plants, as one of the typical responses to abiotic stresses with hormone, occurs through the dynamic expression of different proteins that constitute a complex regulatory network in biological activities and promotes plant totipotency. Plant SE includes two critical stages: primary embryogenic calli redifferentiation and somatic embryos development initiation, which leads to totipotency. The isobaric labels tandem mass tags (TMT) large-scale and quantitative proteomics technique was used to identify the dynamic protein expression changes in nonembryogenic calli (NEC), primary embryogenic calli (PEC) and globular embryos (GEs) of cotton. A total of 9369 proteins (6730 quantified) were identified; 805, 295 and 1242 differentially accumulated proteins (DAPs) were identified in PEC versus NEC, GEs versus PEC and GEs versus NEC, respectively. Eight hundred and five differentially abundant proteins were identified, 309 of which were upregulated and 496 down regulated in PEC compared with NEC. Of the 295 DAPs identified between GEs and PEC, 174 and 121 proteins were up- and down regulated, respectively. Of 1242 differentially abundant proteins, 584 and 658 proteins were up- and down regulated, respectively, in GEs versus NEC. We have also complemented the authenticity and accuracy of the proteomic analysis. Systematic analysis indicated that peroxidase, photosynthesis, environment stresses response processes, nitrogen metabolism, phytohormone response/signal transduction, transcription/posttranscription and modification were involved in somatic embryogenesis. The results generated in this study demonstrate a proteomic molecular basis and provide a valuable foundation for further investigation of the roles of DAPs in the process of SE transdifferentiation during cotton totipotency.

Highlights

Somatic embryogenesis (SE) is a notable illustration of cell totipotency as one of the typical responses to abiotic stresses with hormone, which processes the developmental reprogramming of somatic cells toward the embryogenesis pathway
We performed quantitative proteomics analysis using the advanced EASY-nLC 1000 UPLC system based on the high-throughput tandem mass tags (TMT)-labeling quantitative detection technique, and we identified 9369 proteins for our samples
Most previous studies have focused on the molecular mechanisms of regulation in the late stage of somatic embryogenesis, and little is known about protein regulation and metabolism in the early stage of embryogenesis

Summary

Introduction

Somatic embryogenesis (SE) is a notable illustration of cell totipotency as one of the typical responses to abiotic stresses with hormone, which processes the developmental reprogramming of somatic cells toward the embryogenesis pathway. Somatic embryogenesis represents a unique phenomenon in the plant kingdom [4]. This developmental pathway is one of the most striking examples of plant cell developmental plasticity [5,6]. It includes a series of characteristic events, including somatic dedifferentiation, cell division activation, metabolism alterations and gene expression pattern reprogramming [4]. During SE, the development of somatic cells is reprogrammed to the embryogenic pathway, and SE forms the basis of cellular totipotency in higher plants [7]. Soybeans and cotton have proven to be the most difficult to regenerate [10]

Methods

Results

Conclusion