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GASA5, a regulator of flowering time and stem growth in Arabidopsis thaliana

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Abstract

Flowering is a critical event in the life cycle of plants and is regulated by a combination of endogenous controls and environmental cues. In the present work, we provide clear genetic evidence that GASA5, a GASA family gene in Arabidopsis (Arabidopsis thaliana), is involved in controlling flowering time and stem growth. GASA5 expression was present in all tissues of Arabidopsis plants, as detected by RT-PCR, and robust GUS staining was observed in the shoot apex of 8-day-old seedlings and inflorescence meristems during reproductive development. Phenotypic analysis showed that a GASA5 null mutant (gasa5-1) flowered earlier than wild type with a faster stem growth rate under both long-day (LD) and short-day (SD) photoperiods. In contrast, transgenic plants overexpressing GASA5 demonstrated delayed flowering, with a slower stem growth rate compared to wild-type plants. However, neither the GASA5 null mutants nor the GASA5 overexpressing plants revealed obvious differences in flowering time upon treatment with gibberellic acid (GA3), indicating that GASA5 is involved in gibberellin (GA)-promoted flowering. GAI (GA INSENSITIVE), one of the five DELLAs in Arabidopsis, was more highly expressed in GASA5-overexpressing plants, but it was lower in gasa5-1. Further transcript profiling analysis suggested that GASA5 delayed flowering by enhancing FLOWERING LOCUS C (FLC) expression and repressing the expression of key flowering-time genes, FLOWERING LOCUS T (FT) and LEAFY (LFY). Our results suggest that GASA5 is a negative regulator of GA-induced flowering and stem growth.

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Abbreviations

CO:

CONSTANS

CRPs:

Cysteine-rich peptides

FLC:

FLOWERING LOCUS C

FT:

FLOWERING LOCUS T

GASA:

Gibberellic stimulated in Arabidopsis

GA:

Gibberellin

GA3 :

Gibberellic acid 3

GAI:

GA insensitive

GFP:

Green fluorescent protein

GUS:

β-Glucuronidase

LD:

Long days

LFY:

LEAFY

PAC:

Paclobutrazol

RGA:

Repressor of the ga1-3 mutant

SD:

Short days

SOC1:

SUPPRESSOR OF OVEREXPRESSION OF CO 1

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Acknowledgements

We thank Professor Fu Xiang-dong at the Institute of Genetics and Developmental Biology, Chinese Academy of Sciences for kindly providing the rga-24 and gai-t6 seeds and Professor Huang Tao at Xiamen University in China for the ft-10 seeds. This research was supported by grants from the Nature Science Foundation of China (No. 30570165, U0731006) and the National Key Technology R & D Program in China (No. 2007BAD59B06).

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  1. Guangdong Key Lab of Biotechnology for Plant Development, College of Life Sciences, South China Normal University, Guangzhou, 510631, Guangdong, People’s Republic of China

    Shengchun Zhang, Chengwei Yang, Jianzong Peng, Shulan Sun & Xiaojing Wang

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Fig. S1

GASA5 protein structure and the relationship between GASA5 protein and other GASA homologues. (A) Structure of the GASA5 protein. Gray box represents the GASA domain, and white box represents the cleavable signal peptide (1-27 residues). Prediction of signal peptide in GASA5 protein used the web software PSORT (http://psort.nibb.ac.jp/form.html). (B) Multiple sequence alignment of the GASA5 with several GASA family proteins. Identical residues were labeled with asterisk at the bottom of the sequence. (C) Unrooted phylogenetic tree shows the relationship between the GASA5 protein and other GASA homologues. GASA protein sequences and accession numbers were obtained from public databases (http://www.ncbi.nlm.nih.gov/, http://www.arabidopsis.org). Multiple sequence alignments and phylogenetic tree were constructed by ClustalW (V.1.83) using neighbour-joining (TIFF 433 kb)

Fig. S2

Phenotypes of GASA5-overexpressing plants compared with transgenic plants lacking GASA5 (TIFF 667 kb)

Fig. S3

Phenotypes of gasa5-1 plants after vernalization for three weeks under LD (TIFF 651 kb)

Fig. S4

Phenotypes of GASA5 mutants after being sprayed with 100 µM GA3 under LD (TIFF 757 kb)

Fig. S5

The expression levels of gibberellin 20-oxidase 1 gene (GA20OX1), gibberellin 3-oxidase 1 gene (GA3OX1) and gibberellin 2-oxidase gene 1 (GA2OX1) were detected by RT-PCR. The RNA used was extracted from the inflorescence stem harvested at eight days after bolting under LD (TIFF 241 kb)

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Supplementary material 7 (DOC 26 kb)

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Zhang, S., Yang, C., Peng, J. et al. GASA5, a regulator of flowering time and stem growth in Arabidopsis thaliana . Plant Mol Biol 69, 745–759 (2009). https://doi.org/10.1007/s11103-009-9452-7

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