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作物杂志,2020, 第2期: 20–27 doi: 10.16035/j.issn.1001-7283.2020.02.004

• 遗传育种·种质资源·生物技术 • 上一篇    下一篇

高粱硬脂酰-ACP脱氢酶基因（SbSAD）家族鉴定及不同发育阶段表达分析

赵训超,徐晶宇(),盖胜男,魏玉磊,许晓萱,丁冬,刘梦,张今杰,邵文静   

  1. 黑龙江八一农垦大学农学院/黑龙江省现代农业栽培技术与作物种质改良重点实验室,163319,黑龙江大庆
  • 收稿日期:2019-08-19 修回日期:2019-10-14 出版日期:2020-04-15 发布日期:2020-04-13
  • 通讯作者: 徐晶宇 E-mail:xujingyu2003@hotmail.com
  • 作者简介:赵训超,主要从事作物脂类代谢调控及逆境分子生物学研究,E-mail：zhaoxunchao2017@163.com
  • 基金资助:
    黑龙江八一农垦大学优势特色学科建设项目专项资金(2042070002)

Identification of Stearyl -ACP Desaturase Gene (SbSAD) Family and Their Expression Analysis at Different Developmental Stages in Sorghum

Zhao Xunchao,Xu Jingyu(),Gai Shengnan,Wei Yulei,Xu Xiaoxuan,Ding Dong,Liu Meng,Zhang Jinjie,Shao Wenjing   

  1. College of Agronomy, Heilongjiang Bayi Agricultural University/Key Laboratory of Modern Agricultural Cultivation and Crop Germplasm Improvement of Heilongjiang Province, Daqing 163319, Heilongjiang, China
  • Received:2019-08-19 Revised:2019-10-14 Online:2020-04-15 Published:2020-04-13
  • Contact: Jingyu Xu E-mail:xujingyu2003@hotmail.com

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摘要：

硬脂酰-ACP脱氢酶是形成不饱和脂肪酸的关键酶。利用拟南芥蛋白序列作为比对序列在NCBI和Phytozome数据库进行Blastp同源比对,共鉴定11个SbSADs基因。对SbSAD基因家族进行蛋白特性、进化关系、不同发育阶段表达、基因结构、保守基序、染色体定位、基因二级和三级结构、启动子区的顺式元件及种子发育阶段表达分析,结果显示,SbSADs基因编码区为1 134~1 290bp,其编码蛋白的氨基酸数为377~449,分子量最大为48.0kDa,最小为39.8kDa,等电点为5.26~8.76;根据系统发育树,将SbSAD基因家族分为3个亚族,其中亚族I在不同组织发育阶段表达量较高;11个SbSADs基因不均等地定位在高粱1、2、3、4、6、7和10号染色体上;SbSADs蛋白二级结构均以α-螺旋和无规则卷曲为主,三级结构预测显示,除SbSAD8外,其余SbSADs的蛋白三级结构高度相似;11个SbSADs基因的启动子区与低温胁迫相关的元件数量高度富集。SbSAD5基因在种子发育5和10d均有较高的表达量。

关键词: 高粱, 硬脂酰-ACP脱氢酶, 生物信息学, 基因家族

Abstract:

Stearyl-ACP desaturase is a key enzyme to form unsaturated fatty acids. A total of 11 full-length SbSADs genes were identified by Blastp from the NCBI and Phytozome database using the SbSADs genes as queries. The protein properties, phylogenetics, tissue-specific expression patterns of SbSADs, gene structure, conserved motifs, chromosome mapping, secondary and tertiary structures, cis-elements in promoter regions and their expression level at various seed development stages were analyzed. The results showed that open reading frame of SbSADs genes was among 1 134-1 290bp, the number of amino acids of their encoded proteins ranged from 377 to 449, the maximum molecular weight was 48.0kDa and the minimum was 39.8kDa, and the isoelectric point was 5.26-8.76. According to phylogenetic analysis, SbSADs were divided into 2 sub-groups, and the expression level of sub-group I was higher in different tissues. SbSADs genes were unequally mapped on 1, 2, 3, 4, 6, 7 and 10 chromosomes of sorghum. The prediction of secondary structure showed that SbSADs proteins was mainly composed of α-helix and irregular coil, while the tertiary structures prediction showed that except SbSAD8, the rest of SbSADs proteins were highly similar. In the SbSADs gene family, there were a number of cis-elements related to low temperature stress. The expression of SbSAD5 gene was higher at 5 and 10d of seed development.

Key words: Sorghum, Stearyl-ACP desaturase, Bioinformatics, Gene family

表1

高粱SbSADs蛋白特性分析"

基因
Gene		 登录号
ID		 编码区(bp)
Open reading frame		 氨基酸数
Number of amino acid		 分子量(kDa)
Molecular weight		 等电点
Isoelectric point
SbSAD1 Sobic.001G265300 1 290 429 47.5 8.76
SbSAD2 Sobic.001G280700 1 194 397 44.7 7.15
SbSAD3 Sobic.001G334501 1 350 449 48.0 6.47
SbSAD4 Sobic.002G258500 1 245 414 44.5 6.79
SbSAD5 Sobic.003G377100 1 143 380 42.7 6.62
SbSAD6 Sobic.003G404000 1 194 397 45.1 6.44
SbSAD7 Sobic.004G153300 1 251 416 46.7 6.62
SbSAD8 Sobic.004G157550 1 134 377 39.8 5.26
SbSAD9 Sobic.006G048700 1 179 392 44.7 6.53
SbSAD10 Sobic.007G071600 1 272 423 46.9 8.18
SbSAD11 Sobic.010G235300 1 233 410 45.6 6.71

图1

水稻、玉米、拟南芥和高粱SADs蛋白系统进化关系"

图2

SbSAD基因家族在高粱不同发育阶段不同组织中的表达量"

图3

高粱SbSAD基因家族结构分析"

图4

SbSADs蛋白保守基序分析"

图5

SbSADs基因的染色体分布"

表2

SbSADs蛋白二级结构的氨基酸数目和比例"

蛋白
Protein		 α-螺旋α-helix 延长链Extension chain β-折叠β-sheet 无规则卷曲Random coil
数量
Amount		 比例(%)
Proportion		 数量
Amount		 比例(%)
Proportion		 数量
Amount		 比例(%)
Proportion		 数量
Amount		 比例(%)
Proportion
SbSAD1 221 51.52 46 10.72 45 10.49 117 27.27
SbSAD2 206 51.89 40 10.08 29 7.30 122 30.73
SbSAD3 216 48.11 57 12.69 54 12.03 122 27.17
SbSAD4 212 51.21 49 11.84 38 9.18 115 27.78
SbSAD5 215 56.58 40 10.53 39 10.26 86 22.63
SbSAD6 210 52.90 49 12.34 29 7.30 109 27.46
SbSAD7 219 52.64 32 7.69 30 7.21 135 32.45
SbSAD8 153 40.58 54 14.32 24 6.37 146 38.73
SbSAD9 205 53.66 38 9.95 32 8.38 107 38.73
SbSAD10 225 53.19 51 12.06 39 9.22 108 25.53
SbSAD11 243 59.27 31 7.56 43 10.49 93 22.68

图6

SbSADs蛋白二级结构(A)和三级结构(B)的预测结果"

图7

SbSADs基因启动子逆境相关顺式作用元件数量分布"

图8

SbSADs在种子不同发育时期的表达"

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