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Knockdown of NtMed8, a Med8-like gene, causes abnormal development of vegetative and floral organs in tobacco (Nicotiana tabacum L.)

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Abstract

Med8, a subunit of mediator complex, has proved to possess crucial functions in many organisms from yeast to human. In plant, the med8 mutant of Arabidopsis thaliana displayed delayed anthesis and increased number of leaves during the vegetative period. However, the roles of Med8 in other flowering plants are still unknown. To investigate the function of Med8 ortholog in tobacco (Nicotiana tabacum L.; named as NtMed8), we created transgenic tobacco plants with repressed NtMed8 expression mediated by RNA interference (RNAi). Compared with the wild type, the NtMed8-RNAi plants exhibited: more leaves with smaller but thicker blades; larger cells and vascular bundles with lower stomata density in leaves; swelled chloroplasts with thicker and lumen-enlarged thylakoids; weaker root system with fewer lateral roots; larger flowers and floral organs; flowering earlier under long day, but later under short day conditions; and male sterile with larger but less germinable pollens. In addition, quantitative RT-PCR indicated that NtMed8 is expressed in both vegetative and floral tissues. Subcellular localization analysis by transient expression of fusion protein in Nicotiana benthamiana leaves showed that NtMed8 was located in both plasma membrane and nucleus. These results suggest that NtMed8 plays important roles in both vegetative and reproductive development, and the function of Med8 appears to be, at least partially, conserved in flowering plants.

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Abbreviations

CTD:

Carboxy terminal domain

ORF:

Open reading frame

PCR:

Polymerase chain reaction

MED:

Mediator complex subunit

sqRT-PCR:

Semi-quantitative reverse transcription PCR

qRT-PCR:

Quantitative reverse transcription PCR

TBP:

TATA box-binding protein

NLS:

Nuclear localization signal

RNAi:

RNA interference

WT:

Wild type

SD:

Short day

LD:

Long day

eGFP:

Enhanced green fluorescent protein

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Acknowledgments

We thank Ms. Chunmei Meng and Ms. Zhijuan Diao for their help in the experiment. This work was funded by the National Hi-Tech Research and Development Program (863 Program) of China (Grant No.: 2006AA10Z1E2).

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Authors and Affiliations

  1. College of Agriculture and Biotechnology, Zhejiang University, 310029, Hangzhou, Zhejiang, People’s Republic of China

    Fengqing Wang, He Wei, Zhijun Tong, Xiaobo Zhang, Zemao Yang & Weiren Wu

  2. Key Laboratory of Ministry of Education for Genetics, Breeding and Multiple Utilization of Crops, Fujian Agriculture and Forestry University, 350002, Fuzhou, Fujian, People’s Republic of China

    Tao Lan & Yuanlin Duan

  3. School of Life Sciences, Fujian Agriculture and Forestry University, 350002, Fuzhou, Fujian, People’s Republic of China

    Weiren Wu

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  1. Fengqing Wang
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  2. He Wei
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  3. Zhijun Tong
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Correspondence to Weiren Wu.

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Communicated by D. Somers.

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Wang, F., Wei, H., Tong, Z. et al. Knockdown of NtMed8, a Med8-like gene, causes abnormal development of vegetative and floral organs in tobacco (Nicotiana tabacum L.). Plant Cell Rep 30, 2117–2129 (2011). https://doi.org/10.1007/s00299-011-1118-7

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  • DOI: https://doi.org/10.1007/s00299-011-1118-7

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