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Arabidopsis KHZ1 and KHZ2, two novel non-tandem CCCH zinc-finger and K-homolog domain proteins, have redundant roles in the regulation of flowering and senescence

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The two novel CCCH zinc-finger and K-homolog (KH) proteins, KHZ1 and KHZ2, play important roles in regulating flowering and senescence redundantly in Arabidopsis.

Abstract

The CCCH zinc-finger proteins and K-homolog (KH) proteins play important roles in plant development and stress responses. However, the biological functions of many CCCH zinc-finger proteins and KH proteins remain uncharacterized. In Arabidopsis, KHZ1 and KHZ2 are characterized as two novel CCCH zinc-finger and KH domain proteins which belong to subfamily VII in CCCH family. We obtained khz1, khz2 mutants and khz1 khz2 double mutants, as well as overexpression (OE) lines of KHZ1 and KHZ2. Compared with the wild type (WT), the khz2 mutants displayed no defects in growth and development, and the khz1 mutants were slightly late flowering, whereas the khz1 khz2 double mutants showed a pronounced late flowering phenotype. In contrast, artificially overexpressing KHZ1 and KHZ2 led to the early flowering. Consistent with the late flowering phenotype, the expression of flowering repressor gene FLC was up-regulated, while the expression of flowering integrator and floral meristem identity (FMI) genes were down-regulated significantly in khz1 khz2. In addition, we also observed that the OE plants of KHZ1 and KHZ2 showed early leaf senescence significantly, whereas the khz1 khz2 double mutants showed delayed senescence of leaf and the whole plant. Both KHZ1 and KHZ2 were ubiquitously expressed throughout the tissues of Arabidopsis. KHZ1 and KHZ2 were localized to the nucleus, and possessed both transactivation activities and RNA-binding abilities. Taken together, we conclude that KHZ1 and KHZ2 have redundant roles in the regulation of flowering and senescence in Arabidopsis.

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Abbreviations

TZF:

Tandem CCCH zinc-finger

GA:

Gibberellic acid

ABA:

Abscisic acid

OE:

Overexpression

mRNA:

Messenger RNA

FLK:

Flowering Locus KH Domain

PEP:

PEPPER

JA:

Jasmonate

MS:

Murashige and Skoog

LD:

Long-day

ABRC:

Arabidopsis Biological Resources Center

WT:

Wild type

CDS:

Coding sequences

pSuper 1300:

Super Promoter 1300 vector

GAL4-BD:

GAL4 DNA-binding domain

GST:

Glutathione S-tansferase

GUS:

β-Glucuronidase

X-Glu:

5-Bromo-4-chloro-3-indolyl-β-d-glucuronide

GFP:

Green fluorescent protein

RT-PCR:

Reverse-transcription PCR

qRT-PCR:

Quantitative real-time PCR

COM:

Complementation

FMI:

Floral meristem identity

DAG:

Days after germination

SA:

Salicylic acid

ACC:

1-Aminocyclopropane-1-carboxylic acid

RFP:

Red fluorescent protein

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Acknowledgements

We thank Prof. Junping Gao (China Agricultural University, China) for providing the GAL4-BD, GAL4-BD-VP16 and GAL4-GUS plasmids, Prof. Qijun Chen (China Agricultural University, China) for providing pHEE2A-TRI vector and the assistance of CRISPR/Cas9 technology. The work was supported by a grant from the National Natural Science Foundation of China (No. 21025057).

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  1. State Key Laboratory of Plant Physiology and Biochemistry, College of Biological Sciences, China Agricultural University, Beijing, 100193, China

    Zongyun Yan, Jianheng Jia, Xiaoyuan Yan, Huiying Shi & Yuzhen Han

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  1. Zongyun Yan
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  2. Jianheng Jia
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  3. Xiaoyuan Yan
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YH conceived the research, supervised the experiment. ZY designed and performed experiments, and prepared the figures. JJ contributed materials. XY and HS provided technical assistance. ZY and YH wrote the manuscript.

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Correspondence to Yuzhen Han.

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Yan, Z., Jia, J., Yan, X. et al. Arabidopsis KHZ1 and KHZ2, two novel non-tandem CCCH zinc-finger and K-homolog domain proteins, have redundant roles in the regulation of flowering and senescence. Plant Mol Biol 95, 549–565 (2017). https://doi.org/10.1007/s11103-017-0667-8

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