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Ectopic expression of GhCOBL9A, a cotton glycosyl-phosphatidyl inositol-anchored protein encoding gene, promotes cell elongation, thickening and increased plant biomass in transgenic Arabidopsis

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Abstract

Cellulose is a major component of plant cell walls and is necessary for plant morphogenesis and biomass. COBL (COBRA-Like) proteins have been shown to be key regulators in the orientation of cell expansion and cellulose crystallinity status. To clarify the role of a cotton COBL gene, GhCOBL9A, we conducted the ectopic expression and functional analysis in Arabidopsis. Previous study showed that GhCOBL9A was preferentially expressed during secondary cell wall biosynthesis in cotton fibers, and showed a significant co-expression pattern with cellulose synthase genes. Here, we detected that overexpression of GhCOBL9A induced the up-regulation of genes related to cellulose synthesis and enhanced the cellulose deposition. As a result, GhCOBL9A transgenic plants displayed increased hypocotyl and root lengths in early development, and cell wall thickening at the SCW stage. Notably, overexpression of GhCOBL9A led to an erect, robust-stature phenotype and brought higher biomass in mature plants. In addition, overexpression of GhCOBL9A in Arabidopsis AtCOBL4 mutants, a paralogous gene of GhCOBL9A, also led to a stronger growth potential, but the Atcobl4 mutant phenotype could not be rescued, implying the functional divergence of GhCOBL9A and AtCOBL4 paralogs. Taken together, these results suggest that overexpression of GhCOBL9A contributes to plant cell elongation and thickening, and increased biomass, which provides references for further utilizing GhCOBL9A to improve yield and quality traits in cotton and other species.

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Abbreviations

PCW:

Primary cell wall

SCW:

Secondary cell wall

Susy:

Sucrose synthase

KOR:

Korrigan

CTL:

Chitinase-like

CesAs:

Cellulose synthases

COBL :

COBRA-Like

GPI:

Glycosyl-phosphatidyl inositol

CBM:

Carbohydrate-binding module

CSCs:

Cellulose synthesizing complexes

CaMV:

Cauliflower mosaic virus

DPA:

Days post anthesis

qRT-PCR:

Quantitative real-time PCR

PIECE:

Plant intron exon comparison and evolution database

CDD:

Conserved domain database

ORF:

Open reading frame

MS:

Murashige and Skoog

BLAST:

Basic local alignment search tool

ph:

Phloem

xy:

Xylem

if:

Interfascicular fiber

pi:

Pith cell

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Acknowledgements

This program was financially supported in part by National Natural Science Foundation of China (31701472), Natural Science Foundation in Jiangsu Province (BK20160712), and Jiangsu Collaborative Innovation Center for Modern Crop Production (No. 10).

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Authors and Affiliations

  1. State Key Laboratory of Crop Genetics and Germplasm Enhancement, Hybrid Cotton R & D Engineering Research Center, Ministry of Education, Nanjing Agricultural University, Nanjing, 210095, Jiangsu, China

    Erli Niu, Shuai Fang, Xiaoguang Shang & Wangzhen Guo

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  1. Erli Niu
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  2. Shuai Fang
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  3. Xiaoguang Shang
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  4. Wangzhen Guo
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Correspondence to Wangzhen Guo.

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The experiments in this manuscript complied with the current laws of the country in which they were performed.

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Communicated by S. Hohmann.

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Fig. S1

Identification of GhCOBL9A transgenic lines in Arabidopsis. Detection on GhCOBL9A overexpression transgenic lines in DNA (a) and transcription (b) levels. WT, Arabidopsis Columbia-0 (Col-0); OE1 to OE10, overexpression GhCOBL9A transgenic lines in WT plants. The expression level of AtUbq5 (NM_116090.3) was used as the internal control and the relative expression level was calculated using the 2-△CT method (Livak and Schmittgen 2001) (TIFF 144 KB)

Fig. S2

Number of different types of branches in GhCOBL9A transgenic plants. Different Arabidopsis branches were referred to the descriptions as Sugimoto et al. (2014) (TIFF 95 KB)

Fig. S3

Identification of GhCOBL9A transgenic lines in Atcobl4 mutant. Detection on GhCOBL9A overexpression transgenic lines in DNA (a) and transcription (b) levels. Atcobl4, T-DNA insertion mutant of AtCOBL4 (IRX6; N431557); RM1 to RM12, overexpression GhCOBL9A transgenic lines in Atcobl4 mutant. The expression level of AtUbq5 (NM_116090.3) was used as the internal control and the relative expression level was calculated using the 2-△CT method (Livak and Schmittgen 2001) (TIFF 139 KB)

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Niu, E., Fang, S., Shang, X. et al. Ectopic expression of GhCOBL9A, a cotton glycosyl-phosphatidyl inositol-anchored protein encoding gene, promotes cell elongation, thickening and increased plant biomass in transgenic Arabidopsis. Mol Genet Genomics 293, 1191–1204 (2018). https://doi.org/10.1007/s00438-018-1452-3

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  • DOI: https://doi.org/10.1007/s00438-018-1452-3

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