Identification of CslD Gene Family in Pineapple and Functional Analysis of AcoCslD2a

doi:10.16420/j.issn.0513-353x.2021-0518

Acta Horticulturae Sinica ›› 2022, Vol. 49 ›› Issue (8): 1650-1662.doi: 10.16420/j.issn.0513-353x.2021-0518

• Research Papers • Previous Articles Next Articles

Identification of CslD Gene Family in Pineapple and Functional Analysis of AcoCslD2a

LIU Chaoyang(), LIAO Zhichan, LU Xinxin, HE Yehua

Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in South China,Ministry of Agriculture and Rural Affairs,College of Horticulture,South China Agricultural University,Guangzhou 510642,China

Received:2022-02-22 Revised:2022-05-09 Online:2022-08-25 Published:2022-09-05

Abstract

Abstract:

In this study,eight CslD genes were characterized in pineapple genome. Chromosomal mapping showed that they were distributed on seven chromosomes and one scaffold. Three to five exons were identified in these genes. Three subgroups were divided according to the phylogenetic analysis,and the number of Group Ⅲ was significantly increased in pineapple. The potential biological functions of the pineapple CslD gene family members were further predicted,basing on the collinearity and gene expression analysis. The full length cDNA sequence of AcoCslD2a was cloned and the subcellular localization analysis indicated that its encoding protein was localized in Golgi apparatus. Two expression vector for AcoCslD2a gene,which were driven by the promoters including ProAtUbq10 and ProAtCslD3,respectively,were constructed and transformed into the Arabidopsis atcsld3 root hair deficient mutant,mediated by Agrobacterium tumefaciens GV3101. The phenotype analysis for the transgenic lines of different expression vectors both indicated that the defect root hair number phenotype of mutant could be recovered to the wild-type phenotype by AcoCslD2a,while the defect root hair length phenotype could be partially recovered.

Key words: pineapple, cellulose synthase-like, CslD, cell wall, root hair

CLC Number:

S668.3

LIU Chaoyang, LIAO Zhichan, LU Xinxin, HE Yehua. Identification of CslD Gene Family in Pineapple and Functional Analysis of AcoCslD2a[J]. Acta Horticulturae Sinica, 2022, 49(8): 1650-1662.

Figures/Tables 10

Table 1 The primer sequences used in this study

引物用途 Primer use	引物名称 Primer name	引物序列（5′-3′） Primer sequence
基因ORF克隆 ORF Cloning	AcoCSLD2a-F	ATGGCATCGAACAGCGCGCTT
基因ORF克隆 ORF Cloning	AcoCSLD2a-R	TTAAGGGAATGTGAAGGAACC
植物表达载体构建 Plant expression vector construction	EcoRⅠ-ProAtUBQ10-F	GAATTCCGACGAGTCAGTAATAAAC
	BamHⅠ-ProAtUBQ10-R	GGATTCTGTTAATCAGAAAAACTCAG
	EcoRⅠ-ProAtCSLD3-F	GAATTCCACTTGTGTCCTGATACTCTC
	BamHⅠ-ProAtCSLD3-R	GGATTCTGTCTAATAATAACACTAT
	PstⅠ-AcoCSLD2a-F	CTGCAGATGGCATCGAACAGCGCGCTT
	SpeⅠ-AcoCSLD2a-R	ACTAGTTTAAGGGAATGTGAAGGAACC
亚细胞定位载体构建 Subcellular localization vector construction	XbaⅠ-GFP-AcoCSLD2a-F	TCTAGAATGGCATCGAACAGCGCGCTT
亚细胞定位载体构建 Subcellular localization vector construction	SacⅠ-GFP-AcoCSLD2a-R	GAGCTCAGGGAATGTGAAGGAACC

Fig. 1 The gene structure of the pineapple CslD genes and conserved domains of the corresponding protein

Fig. 2 Phylogenetic tree of CslD proteins in pineapple and other ten species At：Arabidopsis thaliana;Os：Oryza sativa;Zm：Zea mays;Sb：Sorghum bicolor;Bd：Brachypodium distachyon;Ma：Musa acuminate;Gr：Gossypium raimondii;Prupe：Prunus persica;Solyc：Solanum lycopersicum;Ptr：Populus trichocarpa.

Fig. 3 The chromosomal localization and synteny analysis of pineapple CslD genes The red lines indicate the segmental duplicated CslD gene pairs.

Fig. 4 Synteny analysis of CslD genes from pineapple,Brachypodium distachyon and rice（A）and synteny analysis of CslD genes from pineapple,banana,maize and Arabidopsis（B） Lines with different colors indicated the syntenic CslD gene pairs between pineapple and other species. Ac：Ananas comosus;Bd：Brachypodium distachyon;Os：Oryza sativa;Ma：Musa acuminata;Zm：Zea mays;At：Arabidopsis thaliana.

Fig. 5 Heat map of pineapple CslD gene expression in various pineapple tissues during different developmental stages M：Stamen;T：Style;P：Petal;L：Leaf;S：Stem;R：Root;S1-S5 indicate different development stages for flower and fruit of pineapple,respectively.

Fig. 6 The cDNA full length cloning（A）,protein structure prediction（B）and expression features in different pineapple tissues（C）of AcoCslD2a gene

Fig. 7 The subcellular localization of AcoCslD2a

Fig. 8 The phenotype for the transgenic lines（culture for 7 days）of AcoCslD2a gene

Fig. 9 The average root hair length and root hair number for the transgenic lines（culture for 7 days）of AcoCslD2a gene

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Identification of CslD Gene Family in Pineapple and Functional Analysis of AcoCslD2a

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