Phylogenetic and Transcription Analysis of Chrysanthemum WRKY Transcription Factors

Song, Aiping; Li, Peiling; Jiang, Jiafu; Chen, Sumei; Li, Huiyun; Zeng, Jun; Shao, Yafeng; Zhu, Li; Zhang, Zhaohe; Chen, Fadi

doi:10.3390/ijms150814442

Cited by 40 publications

(46 citation statements)

References 48 publications

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“…The conditions for the PCR amplification were as follows: polymerase activation at 95°C for 3 min, followed by 40 cycles of 95°C for 20 s, 60°C for 20 s and 72°C for 45 s. All reactions were performed in triplicate on a Mastercycler ep realplex device (Eppendorf, Hamburg, Germany) according the kit’s protocol. The EF1α gene was used as the reference gene [ 39 ], and relative expression levels were calculated by the 2 -△△CT method [ 40 ].…”

Section: Methodsmentioning

confidence: 99%

Identification of MicroRNAs and their Targets Associated with Embryo Abortion during Chrysanthemum Cross Breeding via High-Throughput Sequencing

et al. 2015

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BackgroundMicroRNAs (miRNAs) are important regulators in plant development. They post-transcriptionally regulate gene expression during various biological and metabolic processes by binding to the 3’-untranslated region of target mRNAs to facilitate mRNA degradation or inhibit translation. Chrysanthemum (Chrysanthemum morifolium) is one of the most important ornamental flowers with increasing demand each year. However, embryo abortion is the main reason for chrysanthemum cross breeding failure. To date, there have been no experiments examining the expression of miRNAs associated with chrysanthemum embryo development. Therefore, we sequenced three small RNA libraries to identify miRNAs and their functions. Our results will provide molecular insights into chrysanthemum embryo abortion.ResultsThree small RNA libraries were built from normal chrysanthemum ovules at 12 days after pollination (DAP), and normal and abnormal chrysanthemum ovules at 18 DAP. We validated 228 miRNAs with significant changes in expression frequency during embryonic development. Comparative profiling revealed that 69 miRNAs exhibited significant differential expression between normal and abnormal embryos at 18 DAP. In addition, a total of 1037 miRNA target genes were predicted, and their annotations were defined by transcriptome data. Target genes associated with metabolic pathways were most highly represented according to the annotation. Moreover, 52 predicted target genes were identified to be associated with embryonic development, including 31 transcription factors and 21 additional genes. Gene ontology (GO) annotation also revealed that high-ranking miRNA target genes related to cellular processes and metabolic processes were involved in transcription regulation and the embryo developmental process.ConclusionsThe present study generated three miRNA libraries and gained information on miRNAs and their targets in the chrysanthemum embryo. These results enrich the growing database of new miRNAs and lay the foundation for the further understanding of miRNA biological function in the regulation of chrysanthemum embryo abortion.

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Section: Methodsmentioning

confidence: 99%

Identification of MicroRNAs and their Targets Associated with Embryo Abortion during Chrysanthemum Cross Breeding via High-Throughput Sequencing

et al. 2015

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“…To our knowledge, little information on the isolation and functional identification of WRKY transcription factors in chrysanthemum is available. We have previously isolated 15 WRKY genes, which are induced by various stresses (Song et al 2014a). Overexpression of DgWRKY3, which was isolated from chrysanthemum, increased salt stress tolerance in tobacco ).…”

Section: Introductionmentioning

confidence: 99%

Chrysanthemum WRKY gene CmWRKY17 negatively regulates salt stress tolerance in transgenic chrysanthemum and Arabidopsis plants

Song

Gao

et al. 2015

Plant Cell Rep

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CmWRKY17 was induced by salinity in chrysanthemum, and it might negatively regulate salt stress in transgenic plants as a transcriptional repressor. WRKY transcription factors play roles as positive or negative regulators in response to various stresses in plants. In this study, CmWRKY17 was isolated from chrysanthemum (Chrysanthemum morifolium). The gene encodes a 227-amino acid protein and belongs to the group II WRKY family, but has an atypical WRKY domain with the sequence WKKYGEK. Our data indicated that CmWRKY17 was localized to the nucleus in onion epidermal cells. CmWRKY17 showed no transcriptional activation in yeast; furthermore, luminescence assay clearly suggested that CmWRKY17 functions as a transcriptional repressor. DNA-binding assay showed that CmWRKY17 can bind to W-box. The expression of CmWRKY17 was induced by salinity in chrysanthemum, and a higher expression level was observed in the stem and leaf compared with that in the root, disk florets, and ray florets. Overexpression of CmWRKY17 in chrysanthemum and Arabidopsis increased the sensitivity to salinity stress. The activities of superoxide dismutase and peroxidase and proline content in the leaf were significantly lower in transgenic chrysanthemum than those in the wild type under salinity stress, whereas electrical conductivity was increased in transgenic plants. Expression of the stress-related genes AtRD29, AtDREB2B, AtSOS1, AtSOS2, AtSOS3, and AtNHX1 was reduced in the CmWRKY17 transgenic Arabidopsis compared with that in the wild-type Col-0. Collectively, these data suggest that CmWRKY17 may increase the salinity sensitivity in plants as a transcriptional repressor.

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“…The heat stress experiment (40 °C) was conducted as previously described [39]. The second leaf (counted from the shoot apex) at 0 h, 4 h, 8 h, 12 h and 24 h after heat stress, respectively, was sampled for RNA isolation.…”

Section: Methodsmentioning

confidence: 99%

Over-expression of chrysanthemum CmDREB6 enhanced tolerance of chrysanthemum to heat stress

Sheng

et al. 2018

BMC Plant Biol

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BackgroundChrysanthemum is among the top ten traditional flowers in China, and one of the four major cut flowers in the world, but the growth of chrysanthemum is severely restricted by high temperatures which retard growth and cause defects in flowers. DREB (dehydration-responsive element-binding) transcription factors play important roles in the response to abiotic and biotic stresses. However, whether the DREB A-6 subgroup is involved in heat tolerance has not been reported conclusively.ResultIn the present study, CmDREB6 was cloned from chrysanthemum (Chrysanthemum morifolium) ‘Jinba’. CmDREB6, containing a typical AP2/ERF domain, was classed into the DREB A-6 subgroup and shared highest homology with Cichorium intybus L. CiDREB6 (73%). CmDREB6 was expressed at its highest levels in the leaf. The CmDREB6 protein localized to the nucleus. Based on the yeast one hybrid assay, CmDREB6 showed transcription activation activity in yeast, and the transcriptional activation domain was located in the 3 ‘end ranging from 230 to 289 amino acids residues. CmDREB6 overexpression enhanced the tolerance of chrysanthemum to heat. The survival rate of two transgenic lines was as high as 85%, 50%, respectively, in contrast to 3.8% of wild-type (WT). Over-expression of CmDREB6 promoted the expression of CmHsfA4, CmHSP90, and the active oxygen scavenging genes CmSOD and CmCAT.ConclusionIn this study, DREB A-6 subgroup gene CmDREB6 was cloned from chrysanthemum ‘Jinba’. Overexpression of CmDREB6 enhanced heat tolerance of chrysanthemum by regulating genes involved in the heat shock response and ROS homeogenesis.Electronic supplementary materialThe online version of this article (10.1186/s12870-018-1400-8) contains supplementary material, which is available to authorized users.

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Phylogenetic and Transcription Analysis of Chrysanthemum WRKY Transcription Factors

Cited by 40 publications

References 48 publications

Identification of MicroRNAs and their Targets Associated with Embryo Abortion during Chrysanthemum Cross Breeding via High-Throughput Sequencing

Identification of MicroRNAs and their Targets Associated with Embryo Abortion during Chrysanthemum Cross Breeding via High-Throughput Sequencing

Chrysanthemum WRKY gene CmWRKY17 negatively regulates salt stress tolerance in transgenic chrysanthemum and Arabidopsis plants

Over-expression of chrysanthemum CmDREB6 enhanced tolerance of chrysanthemum to heat stress

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