Abstract
Spiroplasma eriocheiris, a pathogen that causes mass mortality of Chinese mitten crab Eriocheir sinensis, is a wall less bacteria and belongs to the Mollicutes. This study was designed to investigate the effects of colchicine on S. eriocheiris growth, cell morphology, and proteins expression. We found that in the presence of colchicine, the spiroplasma cells lost their helicity, and the length of the cells in the experimental group was longer than that of the control. With varying concentrations of the colchicine treatment, the total time to achieve a stationary phase of the spiroplasma was increased, and the cell population was decreased. The virulence ability of S. eriocheiris to E. sinensis was effectively reduced in the presence of colchicine. To expound the toxical mechanism of colchicine on S. eriocheiris, 208 differentially expressed proteins of S. eriocheiris were reliably quantified by iTRAQ analysis, including 77 up-regulated proteins and 131 down-regulated proteins. Especially, FtsY, putative Spiralin, and NADH oxidase were down-regulated. F0F1 ATP synthase subunit delta, ParB, DNABs, and NAD(FAD)-dependent dehydrogenase were up-regulated. A qRT-PCR was conducted to detect 7 expressed genes from the iTRAQ results during the incubation. The qRT-PCR results were consistent with the iTRAQ results. All of our results indicate that colchicine have a strong impact on the cell morphology and cellular metabolism of S. eriocheiris.
Highlights
Spiroplasma is a group of bacteria belonging to a class Mollicutes, which includes Mycoplasma, Phytoplasma and so on, and featured by small genome sizes and lack of peptidoglycan layer[1–3]
To study the effect of colchicine on S. eriocheiris cell morphology, 3 g/L colchicine was added into the medium as the final concentrations, and until the cells were in exponential phase
Alterations in cell morphology may be caused by reduction of the amount of membrane proteins, in bacterial cells adapted to high concentrations of colchicine[24]
Summary
Spiroplasma is a group of bacteria belonging to a class Mollicutes, which includes Mycoplasma, Phytoplasma and so on, and featured by small genome sizes and lack of peptidoglycan layer[1–3]. They have a helical cell morphology and swim without flagella when in an appropriate viscous medium[2,4]. Colchicine can be extracted naturally from Gloriosa superba L. and Colchicum autumnale L. plants[12,13]. It is toxic for eukaryotic cells, and is an example of a class of small, tublin-binding molecules[14]. We attempt to explain and establish a significant correlation between the structural and metabolic profile of the S. eriocheiris undergoing challenge by colchicine
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