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Int J Mol Sci
2022 Jul 14;2314:. doi: 10.3390/ijms23147756.
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Camellia sinensis Chloroplast Fluoride Efflux Gene CsABCB9 Is Involved in the Fluoride Tolerance Mechanism.
Luo B
,
Guang M
,
Yun W
,
Ding S
,
Ren S
,
Gao H
.
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Soil is a main source of fluoride for plants. The tea plants (Camellia sinensis) accumulate excessive amounts of fluoride in their leaves compared to other plants, but their fluoride tolerance mechanism is poorly understood. A chloroplast fluoride efflux gene (CsABCB9) was newly discovered by using transcriptome analysis, cloned from Camellia sinensis, and its function was demonstrated in the fluoride detoxication mechanism in Escherichia coli/Xenopus laevis oocytes and Arabidopsis thaliana. CsABCB9 is expressed in tea leaves upon F− treatment. The growth of tea, E. coli, and Arabidopsis were inhibited by F− treatment. However, growth of CsABCB9-overexpression in E. coli was shown to increase with lower fluoride content under F− treatment compared to the control. Furthermore, chlorophyll, xanthophyll and soluble sugar contents of CsABCB9-overexpression in Arabidopsis were improved under F− treatment compared to the wild type. CsABCB9 functions in fluoride transport, and the mechanism by which CsABCB9 improves fluoride resistance in tea is mainly chloroplast protection through fluoride efflux.
Figure 4. Functional analysis of CsABCB9 in Escherichia coli and Xenopus oocytes. (A) Phenotypes of Escherichia coli colonies harboring empty vector (pET28a), CsFEX or CsABCB9. Washed bacteria solution of 10-fold serial gradient dilutions were spotted on culture plates containing different F− concentrations (0, 5, 50, 100 mM). (B) Diagram of the growth curves of Escherichia coli cells transformed with pET28a, CsFEX or CsABCB9 in different concentrations of F− liquid medium. (C) Intracellular F− content in Escherichia coli harboring pET28a, CsFEX or CsABCB9 growing in different F− concentration media. Data are shown as means ± SD (n = 4). n.s. indicates no significant difference compared with pET28a in 5 mM F group. Asterisks indicate significant differences (***, p < 0.001, one-way ANOVA) compared with the pET28a in 50 mM or 100 mM F− concentration group. (D) F− efflux activity of CsABCB9 in Xenopus oocytes. Oocytes were injected with water, CsFEX or CsABCB9 mRNA and cultured in cultural solution containing 1mM F− for 8 h. Water-injected or CsFEX-expressing oocytes were used as a negative or positive controls, respectively. Data are shown as means ± SD (n = 8). Asterisks indicate significant differences compared with water-injected oocytes (**, p < 0.01, one-way ANOVA).
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