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作物学报 ›› 2021, Vol. 47 ›› Issue (7): 1297-1308.doi: 10.3724/SP.J.1006.2021.04180

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

甘薯细胞壁蔗糖转化酶基因IbCWIN家族成员鉴定及表达分析

宋天晓1,2, 刘意2,3, 饶莉萍2,3, Soviguidi Deka Reine Judesse2,3, 朱国鹏1,*(), 杨新笋2,*()   

  1. 1海南大学园艺学院海南省热带园艺作物品质调控重点实验室, 海南海口 570228
    2湖北省农业科学院粮食作物研究所, 湖北武汉 430064
    3长江大学农学院, 湖北荆州 434025
  • 收稿日期:2020-08-06 接受日期:2020-11-13 出版日期:2021-07-12 网络出版日期:2020-12-24
  • 通讯作者: 朱国鹏,杨新笋
  • 作者简介:宋天晓, E-mail: 2757318668@qq.com;|刘意, E-mail: 15549421602@163.com
  • 基金资助:
    本研究由国家重点研发计划项目(2018YFD1000700);本研究由国家重点研发计划项目(2018YFD1000705-3);本研究由国家重点研发计划项目(2019YFD1001304);本研究由国家重点研发计划项目(2019YFD1001305);本研究由国家重点研发计划项目(2019YFD1001300);国家现代产业技术体系(甘薯)建设专项(CARS-11-C-15);粮食作物种质创新与遗传改良湖北省重点实验室开放课题(2019lzjj04);湖北省技术创新专项(对外科技合作类)资助(2018AHB012)

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 Published:2021-07-12 Published online:2020-12-24
  • Contact: ZHU Guo-Peng,YANG Xin-Sun
  • 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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摘要：

甘薯是重要的粮食、饲料、工业原料作物和新型的生物能源作物, 细胞壁蔗糖转化酶是植物源、库组织蔗糖代谢的关键酶, 但关于甘薯细胞壁蔗糖转化酶基因IbCWIN家族成员的研究尚未见报道。本研究测定供试品种不同组织部位的蔗糖淀粉含量, 利用生物信息学方法对IbCWIN基因家族的理化性质、保守结构域、系统进化关系、启动子作用元件、组织特异性表达模式进行分析。结果表明, 甘薯茎中蔗糖含量最高, 须根和叶次之, 块根最低; 块根淀粉含量最高, 极显著高于其他部位。甘薯中含有10个IbCWIN基因, 编码氨基酸442~1115个, 蛋白质分子量范围49.56~124.44 kD, 等电点为5.0~9.1。分布在8条染色体上, 都含有Glyco_32保守结构域及相同或相似的保守基序motif, 属于糖基水解酶基因家族GH32。IbCWIN与木薯MeCWINV同源性高, IbCWIN基因家族启动子区域含有多种类型的顺式作用元件。qRT-PCR结果表明, IbCWIN基因家族在甘薯不同组织中均有表达且有多种表达模式, 其中IbCWIN2和IbCWIN9在块根中表达量显著高于其他组织部位。本研究为下一步探索甘薯IbCWIN基因家族的功能及调控甘薯源、库关系机制提供理论指导。

关键词: 甘薯, 细胞壁蔗糖转化酶, 基因家族, 基因表达

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

表1

试验所用引物"

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

图1

甘薯与其他物种CWIN基因家族比较 MeCWINV1~MeCWINV6 GenBank登录号为JQ339929、JX291160、JN801147、JQ792172、JX291159、JQ339930; AtCWINV1~AtCWINV6登录号为At3g13790、At3g52600、At1g55120、At2g36190、At3g13784、At5g11920; StCWIN1~StCWIN4登录号为Z21486、Z22645、AJ133765、AJ133765。"

图2

甘薯IbCWIN基因家族在染色体上的分布"

表2

甘薯IbCWIN基因家族"

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

图3

甘薯IbCWIN基因家族进化树与基因结构"

图4

甘薯IbCWIN基因家族共线性分析"

图 5

甘薯IbCWIN基因家族启动子预测"

图6

鄂11、福18蔗糖和淀粉含量 图中以不同字母表示数据间的显著性差异, 大写字母表示在0.01水平差异显著, 小写字母表示在0.05水平差异显著。"

图7

IbCWIN基因家族的组织特异性表达分析 图中以不同字母表示数据间的显著性差异, 大写字母表示在0.01水平差异显著, 小写字母表示在0.05水平差异显著。"

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