Identification and characterization of the karrikins signaling gene SsSMAX1 in Sapium sebiferum

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Plant Biology

Main article text

 

Introduction

Materials and Methods

Bioinformatic analysis

Cloning of Sapium sebiferum SMAX1 gene

Expression vector design

Plant materials and growth conditions

Seed germination, hypocotyl length, and growth parameters analysis

Primer design, RNA extraction, cDNA synthesis, and RT–qPCR conditions

Statistical analysis

Results

Bio-informatics analysis of SsSMAX1

SsSMAX1 distribution in Sapium sebiferum

SsSMAX1 involvement in seed germination and seedling development

SsSMAX1 expression in Arabidopsis and its role in plant growth and development

Discussion

Conclusion

Supplemental Information

SsSMAX1 gene’s full-length CDS and protein sequences.

SsSMAX1 gene’s full-length CDS and protein sequences

DOI: 10.7717/peerj.16610/supp-1

List of Primers.

Primers used for qPCR were designed by using primer Premier 6. The Tm of the primers was between 59.0 and 61.0° C . Primers used for qPCR were designed by using primer Premier 5.

DOI: 10.7717/peerj.16610/supp-2

Supplementary Figure 1: Diagram of over-expression vector.

SsSMAX1 was cloned by using primers given in supplementary table 1. Ligated to blunt end vector, sequenced, and then digested at Sal1 and Sma1 positions. Overexpression vector pOCA30 was also double-digested at Sal1 and Sma1 positions. The Full-length SsSMAX1 gene was ligated to pOCA30.

DOI: 10.7717/peerj.16610/supp-3

Figure 2 data.

Gene expression determination by qPCR. Data represent the 2−∆∆Ct value of each gene. SsACTIN2 was used as a reference gene.

DOI: 10.7717/peerj.16610/supp-4

Figure 3 data.

Hypocotyl length, Root length, Seed germination of SsSMAX1 transgenic lines; overexpression 1 (OE1), 2 (OE2), Atsmxl1, and wild-type (WT)

DOI: 10.7717/peerj.16610/supp-5

Figure 4 data.

Expression levels in wild-type and SsSMAX1 transgenic line 1 and line 2 (OE1 and OE2 respectively). Leaves of 15-day-old seedlings of wild-type (WT), AtSMAX1, and SsSMAX1 transgenic Arabidopsis line 1(OE1) and 2 (OE2) were collected from gene expression analysis.

DOI: 10.7717/peerj.16610/supp-6

Figure 5 data.

Leaf length, Leaf width, and Petiol length of SsSMAX1 transgenic lines; overexpression 1 (OE1), 2 (OE2), Atsmxl1, and wild-type (WT)

DOI: 10.7717/peerj.16610/supp-7

Additional Information and Declarations

Competing Interests

The authors declare that they have no competing interests.

Author Contributions

Fang Ni conceived and designed the experiments, performed the experiments, analyzed the data, prepared figures and/or tables, and approved the final draft.

Faheem Afzal Shah performed the experiments, analyzed the data, prepared figures and/or tables, authored or reviewed drafts of the article, and approved the final draft.

Jie Ren conceived and designed the experiments, analyzed the data, authored or reviewed drafts of the article, and approved the final draft.

DNA Deposition

The following information was supplied regarding the deposition of DNA sequences:

The sequences are available at GenBank: SRX656554.

Data Availability

The following information was supplied regarding data availability:

The raw measurements are available in the Supplemental Files.

Funding

This work was supported by Research on the Landscape Application and Rapid propagation of Strong Seedings of Hydrangea in Anhui province (Project No: KJ2020A0815; XZR2020A03). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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