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Cotton cytosolic pyruvate kinase GhPK6 participates in fast fiber elongation regulation in a ROS-mediated manner

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Abstract

Main conclusion

Cotton cytosolic pyruvate kinase GhPK6 is preferentially expressed in the late stage of fiber elongation process, transgenic experiments indicated that its expression level was negatively correlated to cell expansion rate.

Abstract

Pyruvate kinase (PK) plays vital regulatory roles in rapid cell growth in mammals. However, the function of PK in plant cell growth remains unclear. In allotetraploid upland cotton (Gossypium hirsutum L.), a total of 33 PK genes are encoded by the genome. Analysis of the transcriptome data indicated that only two cytosolic PK genes, GhPK6 and its duplicated gene GhPK26, are preferentially expressed in elongating cotton fiber cells. RT-qPCR and western blot analyses revealed that the expression of GhPK6 was negatively correlated with fiber elongation rate, which well explains the observed sharp increase of cytosolic PK activity at the end of fast fiber elongation process. Furthermore, virus-induced gene silencing of GhPK6 in cotton plants resulted in increased fiber cell elongation and reduced reactive oxygen species (ROS) accumulation. On the contrary, Arabidopsis plants ectopically expressing GhPK6 exhibited ROS-mediated growth inhibition, whereas the addition of ROS scavenging reagents could partly rescue this inhibition. These data collectively suggested that GhPK6 might play an important role in regulating cotton fiber elongation in a ROS-dependent inhibition manner.

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Acknowledgments

We are grateful to Professor Yule Liu for the gift of the VIGS vector pTRV1 and pTRV2. We thank members of our laboratory for their helpful discussions. This study was supported by the State Key Basic Research and Development Plan (2010CB126003), the National Natural Science Foundation of China (90608016), the National Transgenic Animals and Plants Research Project (2009ZX08005-026B) and the China Postdoctoral Science Foundation (2014M550074).

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Authors and Affiliations

  1. Laboratory of Plant Molecular Biology, Center for Plant Biology, School of Life Sciences, Tsinghua University, Beijing, 100084, China

    Bing Zhang & Jin-Yuan Liu

  2. Tsinghua-Peking Center for Life Science, Tsinghua University, Beijing, 100084, China

    Bing Zhang

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Correspondence to Jin-Yuan Liu.

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425_2016_2557_MOESM1_ESM.docx

Figure S1. Phenotype of the elongating cotton fibers. Figure S2. Sequence alignment of GhPK6, GhPK26 and the encoding gene of the differentially expressed pyruvate kinase protein identified by 2-DE. Figure S3. Silencing of GhPK6 expression in cotton plants through VIGS. Figure S4. Phenotypes of transgenic Arabidopsis plants ectopically expressing GhPK6. Figure S5. Dynamic of succinate dehydrogenase and alternative oxidase activities in cotton fiber elongation process. Figure S6. A putative model for the GhPK6-mediated mechanistic links between cellular metabolism and growth control in cotton fibers. (DOCX 1812 kb)

Table S1. Primers used in this study. (DOCX 12 kb)

Table S2. Sequences of peptides for antibody preparation. (DOCX 12 kb)

Table S3. Properties of the pyruvate kinase genes and the deduced proteins in Gossypium hirsutum. (XLSX 12 kb)

425_2016_2557_MOESM5_ESM.xlsx

Table S4. Expression of the pyruvate kinase genes in different cotton tissues based on RNA sequencing data. (XLSX 17 kb)

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Zhang, B., Liu, JY. Cotton cytosolic pyruvate kinase GhPK6 participates in fast fiber elongation regulation in a ROS-mediated manner. Planta 244, 915–926 (2016). https://doi.org/10.1007/s00425-016-2557-8

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