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Acta Agronomica Sinica ›› 2021, Vol. 47 ›› Issue (7): 1297-1308.doi: 10.3724/SP.J.1006.2021.04180

• CROP GENETICS & BREEDING · GERMPLASM RESOURCES · MOLECULAR GENETICS • Previous Articles     Next Articles

Identification and expression analysis of cell wall invertase IbCWIN gene family members in sweet potato

SONG Tian-Xiao1,2, LIU Yi2,3, RAO Li-Ping2,3, Soviguidi Deka Reine Judesse2,3, ZHU Guo-Peng1,*(), YANG Xin-Sun2,*()   

  1. 1Key Laboratory for Quality Regulation of Tropical Horticultural Plants of Hainan Province, Horticulture College, Hainan University, Haikou 570228, Hainan, China
    2Food Crops Institute, Hubei Academy of Agricultural Sciences, Wuhan 430064, Hubei, China
    3College of Agriculture, Yangtze University, Jingzhou 434025, Hubei, China
  • Received:2020-08-06 Accepted:2020-11-13 Online:2021-07-12 Published:2020-12-24
  • Contact: ZHU Guo-Peng,YANG Xin-Sun E-mail:guopengzhu@163.com;yangxins013@163.com
  • Supported by:
    This study was supported by the National Key Research and Development Program of China(2018YFD1000700);This study was supported by the National Key Research and Development Program of China(2018YFD1000705-3);This study was supported by the National Key Research and Development Program of China(2019YFD1001304);This study was supported by the National Key Research and Development Program of China(2019YFD1001305);This study was supported by the National Key Research and Development Program of China(2019YFD1001300);the National Modern Sweet Potato Industrial Technology System Construction Project(CARS-11-C-15);the Open Project of Hubei Provincial Key Laboratory of Food Crop Germplasm Innovation and Genetic Improvement(2019lzjj04);the Hubei Province Technological Innovation Project (Foreign Science and Technology Cooperation)(2018AHB012)

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Abstract:

Sweet potato is an important food, feed, industrial raw material crop and a new type of bioenergy crop. Cell wall sucrose invertase is a key enzyme for sucrose metabolism in plant sources and sink. However, the research of sweet potato cell wall sucrose gene invertase (IbCWIN) family members has not been reported. In this study, the content of sucrose starch in different tissues of the tested varieties was determined, and the physicochemical properties, conserved domains, phylogenetic relationships, promoter acting elements and tissue-specific expression patterns of IbCWIN gene family were analyzed by bioinformatics methods. The results showed that the sucrose content of sweet potato stems was the highest, followed by fibrous roots and leaves, and the lowest in tuber root; the starch content of tubers was the highest, which was significantly higher than other parts. There were 10 IbCWIN genes in sweet potato, encoding 442-1115 amino acids, protein molecular weight range 49.56-124.44 kD with isoelectric point of 5.0-9.1. Distributed on 8 chromosomes, they all contained the Glyco_32 conserved domain and the same or similar conserved motif, belonging to the glycosyl hydrolase gene family GH32. IbCWIN has a high homology relationship with cassava MeCWINV. The promoter region of the IbCWIN gene family contained many types of cis-acting elements. The results of qRT-PCR showed that the IbCWIN gene family was expressed in different tissues of sweet potato and had multiple expression patterns. Among them, the expression levels of IbCWIN2 and IbCWIN9 in tubers were significantly higher than those in other tissues. This study provides a theoretical guidance for the next step in exploring the functions of the sweet potato IbCWIN gene family and regulating the relationship between the source and sink of sweet potato.

Key words: sweet potato, cell wall sucrose invertase, gene family, gene expression

Table 1

Primers used in this study"

基因名称
Gene name		 正向引物序列
Forward primer sequence (5'-3')		 反向引物序列
Reverse primer sequence (5'-3')
IbCWIN1 GGGTGTTGCATGCTGTTCC ACCCAACTGCACCGATTGTC
IbCWIN2 ACTTGGATGGGTGTGG TTGATACCCGGCCCGTTTG
IbCWIN3 GTATGTGGGAGTGCGTGGAT CGTCGTCGTAACTTCCCAGG
IbCWIN4 ACCCTCATTTGCTGGATTTG TGCCTCTGTGCCATTGTTG
IbCWIN5 GGTTAGTTACAATAGGGTCCAAGG TCCACGCATTCCCACATA
IbCWIN6 GTTCTTGCATTCGGTTCCGC ATCGAGTCTCAACCCGTGTC
IbCWIN7 TAGGGAGCAAAGTTGAGCGG AGAGGGTGTTCGGCTTTGAC
IbCWIN8 GGATCACAACACTGGTCGGT ATTAAGTTCGTGCATGCGGC
IbCWIN9 GACACCGAGGACTTCAAGCA GGTGGTCGAAACCGGGTAAA
IbCWIN10 TGGGGAACGAGAGAAAGCAC AAAAGTCAGGGCACTCCCAC
β-Actin AGCAGCATGAAGATTAAGGTTGTAGCAC TGGAAAATTAGAAGCACTTCCTGTGAAC

Fig. 1

Comparison of CWIN gene family between sweet potato and other species Accession numbers of MeCWINV1-MeCWINV6 in GenBank are JQ339929, JX291160, JN801147, JQ792172, JX291159, JQ339930; Accession numbers of AtCWINV1-AtCWINV6 are At3g13790, At3g52600, At1g55120, At2g36190, At3g13784, At5g11920; and accession numbers of StCWIN1-StCWIN4 are Z21486, Z22645, AJ133765, and AJ133765, respectively."

Fig. 2

Distribution of IbCWIN gene family on chromosomes in sweet potato"

Table 2

IbCWIN gene family in sweet potato"

基因名称
Gene name		 基因序列号
Gene ID		 氨基酸
Amino acid		 分子量
Molecular weight (kD)		 等电点
Isoelectric point		 预测亚细胞定位
Predicting subcellular localization
IbCWIN1 g1732 657 72.01 5.19 液泡Vacuole
IbCWIN2 g1764 680 74.64 5.00 液泡Vacuole
IbCWIN3 g15600 1115 124.44 5.96 液泡Vacuole
IbCWIN4 g25102 467 52.68 6.47 细胞壁Cell wall
IbCWIN5 g25369 792 86.78 5.58 液泡Vacuole
IbCWIN6 g49879 552 61.99 8.30 细胞壁Cell wall
IbCWIN7 g55209 442 49.56 9.01 细胞壁Cell wall
IbCWIN8 g55220 913 100.07 8.61 细胞壁Cell wall
IbCWIN9 g55625 591 66.10 5.53 细胞壁Cell wall
IbCWIN10 g60985 577 64.50 8.78 细胞壁Cell wall

Fig. 3

Phylogenetic tree and gene structure of IbCWIN gene family in sweet potato"

Fig. 4

Collinearity analysis of IbCWIN gene family in sweet potato"

Fig. 5

Promoter prediction of IbCWIN gene family in sweet potato"

Fig. 6

Contents of sucrose and starch in E11 and Fu18 Different letters in the figure indicate significant differences among the treatments. Uppercase letter indicates significant differences at the 0.01 probability level, and lowercase letter indicates significant differences at the 0.05 probability level."

Fig. 7

Tissue specific expression analysis of IbCWIN gene family Different letters in the figure indicate significant differences among the treatments. Uppercase letter indicates significant differences at the 0.01 probability level, and lowercase letter indicates significant differences at the 0.05 probability level."

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