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Downregulation of the mitogen-activated protein kinase SlMAPK7 gene results in pollen abortion in tomato

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Abstract

Mitogen-activated protein kinases (MAPKs) play important roles in various aspects including plant development and stress response. The biological role of MAPKs in tomato development remains unclear. Here, multiple stable transgenetic lines were developed using either gene overexpression or RNA interference (RNAi) technology to investigate the role of SlMAPK7 in tomato reproductive development. Either upregulation or downregulation of SlMAPK7 did not drastically disturb the vegetative growth of tomato. However, in RNAi-based gene silencing plants, approximately 90 % of the pollen grains were identified to be abnormal and lack viability, which were malformed or collapsed. Cytological observations showed that anthers could develop normal microspore mother cells, but the uninucleate microspores failed to form normal mature pollen grains. Transmission electron microscopy examination demonstrated that accelerated degradation of tapetum occurred in the anther and finally the intine layer of pollen appears abnormal. In addition, although inhibition of SlMAPK7 gene did not alter fruit morphology, seed number in RNAi fruits was markedly reduced. Thus, the results demonstrated that SlMAPK7 is involved in tomato pollen development, possibly via regulation the degradation of tapetum.

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Abbreviations

CTAB:

Cetyl trimethyl ammonium bromide

DAPI:

4′,6-Diamidino-2-phenylindole

GFP:

Green fluorescent protein

GUS:

β-Glucuronidase

HYG:

Hygromycin resistant gene

MAPK:

Mitogen-activated protein kinase

MAPKK:

MAPK kinase

MAPKKK:

MAPKK kinase

MMC:

Microspore mother cell

QRT-PCR:

Quantitative reverse transcription polymerase chain reaction

SEM:

Scanning electron microscopy

RNAi:

RNA interference

TEM:

Transmission electron microscopy examination

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Acknowledgments

This work was supported by the Grants from the Nation Natural Science Foundation of China [Grant Numbers 31071337, 31271633], the Natural Science Foundation of Zhejiang Province (R3110209) and Zhejiang Province key science and technology innovation team (2013TD05).

Author contributions

G.L. and L.C. conceived and designed research. X.G., L.C., L.Q., Y.W., and T.Z. conducted experiments. L.C., Y.Z. and C.P. analyzed data. L.C., G.L. wrote the manuscript. All authors read and approved the manuscript.

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    Authors and Affiliations

    1. Key Laboratory of Horticultural Plant Growth, Development and Biotechnology, Agricultural Ministry of China, Department of Horticulture, Zhejiang University, A535 Agriculture building, Zijingang Campus, Hangzhou, 310058, Zhejiang, People’s Republic of China

      Lifei Chen, Xiaoyan Guan, Li Qin, Tao Zou, Youwei Zhang, Jie Wang, Yan Wang, Changtian Pan & Gang Lu

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    1. Lifei Chen
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    Lifei Chen and Xiaoyan Guan have contributed equally to this work.

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    Chen, L., Guan, X., Qin, L. et al. Downregulation of the mitogen-activated protein kinase SlMAPK7 gene results in pollen abortion in tomato. Plant Cell Tiss Organ Cult 126, 79–92 (2016). https://doi.org/10.1007/s11240-016-0979-4

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