甘蓝型油菜蔗糖磷酸合酶(SPS)基因家族成员鉴定及表达分析

doi:10.3724/SP.J.1006.2018.00197

作物学报 ›› 2018, Vol. 44 ›› Issue (02): 197-207.doi: 10.3724/SP.J.1006.2018.00197

甘蓝型油菜蔗糖磷酸合酶(SPS)基因家族成员鉴定及表达分析

张莉^**, 荐红举^**, 杨博, 张翱翔, 张超, 杨鸿, 张立源, 刘列钊, 徐新福, 卢坤, 李加纳^*()

西南大学农学与生物科技学院, 重庆 400715;

收稿日期:2017-05-03 接受日期:2017-09-10 出版日期:2018-02-12 网络出版日期:2017-10-27
通讯作者: 张莉,荐红举,李加纳
作者简介:
第一作者联系方式: 张莉, E-mail: windylili@foxmail.com; 荐红举, E-mail: jianhongju1989@126.com;^**同等贡献(Contributed equally to this work)
基金资助:
本研究由国家自然科学基金项目(U1302266), 国家科技支撑计划项目(2013BAD01B03-12), 国家现代农业产业技术体系建设专项(CARS-13), 重庆市社会事业与民生保障科技创新专项(cstc2016shms-ztzx80020)和重庆市研究生科研创新项目(CYS16082)资助

Genome-wide Analysis and Expression Profiling of SPS Gene Family in Brassica nupus L.

Li ZHANG^**, Hong-Ju JIAN^**, Bo YANG, Ao-Xiang ZHANG, Chao ZHANG, Hong YANG, Li-Yuan ZHANG, Lie-Zhao LIU, Xin-Fu XU, Kun LU, Jia-Na LI^*

College of Agronomy and Biotechnology, Southwest University, Chongqing 400715, China;

Received:2017-05-03 Accepted:2017-09-10 Published:2018-02-12 Published online:2017-10-27
Contact: Li ZHANG,Hong-Ju JIAN,Jia-Na LI
Supported by:
This study was supported by the Natural Science Foundation of China (U1302266), the National Key Technologies R&D Program of China (2013BAD01B03-12), the China Agriculture Research System (CARS-13), the Science and Technology Innovation Project of Social Undertakings and Livelihood Protection of Chongqing (cstc2016shms-ztzx80020), and the Scientific Research Innovation Projects for Graduate Student of Chongqing (CYS16082).

摘要/Abstract

摘要：

蔗糖磷酸合酶(sucrose phosphate synthase, SPS)是调控植物蔗糖合成的限速酶, 对光合产物的转运和积累有重要影响。利用拟南芥SPS蛋白保守结构域在甘蓝型油菜基因组数据库鉴定出11个甘蓝型油菜SPS基因家族成员, 根据系统进化分析将其分成A、B和C共3个亚家族。基因结构预测表明, SPSC-1有5个外显子, 其他SPS基因均有11~15个外显子。顺式作用元件分析表明, 油菜SPS基因除含基本的启动子保守元件外, 还含有许多与逆境和激素响应相关的顺式作用元件。实时荧光定量PCR结果表明, BnSPSA1在花中表达量最高, BnSPSA2在各组织中均有不同程度的表达, BnSPSB只在叶、蕾和花中表达, BnSPSC在叶中表达量最高, 在蕾和花中有少量表达而在其他组织中基本不表达, 说明SPS基因在甘蓝型油菜中的表达具有明显的组织特异性; BnSPSA1和BnSPSC在高生物产量油菜叶片中的表达高于低生物产量油菜, BnSPSB则在低生物产量油菜中的表达量更高, 说明SPS基因与油菜生物产量密切相关。本研究为油菜SPS基因的功能研究和利用奠定了基础。

关键词: 甘蓝型油菜, 蔗糖磷酸合酶, 全基因组鉴定, 表达分析

Abstract:

Sucrose phosphate synthase (SPS) is the rate-limiting enzyme that controls the sucrose biosynthesis in plants and has great influence on transportation and accumulation of photosynthate. In this study, we identified 11 members of the SPS gene family in the oilseed rape (Brassica nupus L.) genome. These SPS genes were classed into clusters A, B, and C according to gene structure and phylogenetic relationship. Gene structure prediction indicated that BnSPS genes were highly conserved, in which BnSPSC-1 consisted five exons and others consisted 11-15 exons. Promoter cis-element analysis indicated that BnSPS genes had not only essential responsive elements but also some types of elements potentially responsive to stresses or hormone responses. The qRT-PCR assay showed tissue-specific expressions of BnSPS genes with rich expressions of BnSPSA1 in flower, BnSPSB in leaf, bud and flower, BnSPSC in leaf, and BnSPSA2 in various tissues. The relative expression levels of BnSPSA1 and BnSPSC were higher in high-biomass rapeseed varieties than in low-biomass rapeseed varieties, whereas, that of BnSPSB was higher in low-biomass rapeseed varieties, suggesting that SPS genes are closely ralated to biological yield of rapeseed. This study provides basic information for functional study and utilization of BnSPS genes.

Key words: Brassica napus, sucrose phosphate synthase, genome-wide analysis, expression analysis

张莉, 荐红举, 杨博, 张翱翔, 张超, 杨鸿, 张立源, 刘列钊, 徐新福, 卢坤, 李加纳. 甘蓝型油菜蔗糖磷酸合酶(SPS)基因家族成员鉴定及表达分析[J]. 作物学报, 2018, 44(02): 197-207.

Li ZHANG, Hong-Ju JIAN, Bo YANG, Ao-Xiang ZHANG, Chao ZHANG, Hong YANG, Li-Yuan ZHANG, Lie-Zhao LIU, Xin-Fu XU, Kun LU, Jia-Na LI. Genome-wide Analysis and Expression Profiling of SPS Gene Family in Brassica nupus L.[J]. Acta Agronomica Sinica, 2018, 44(02): 197-207.

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自交系编号 Code of inbred line	品种名称 Variety name	生物产量 Biological yield (g plant^-1)	自交系编号 Code of inbred line	品种名称 Variety name	生物产量 Biological yield (g plant^-1)
L1	云花油9号Yunhuayou 9	198	L18	SWU63	121
L2	中双11 Zhongshuang 11	180	L19	SWU82	119
L3	SWU47	172	L20	秦油5号 Qinyou 5	118
L4	WH-30	171	L21	WH-30	115
L5	SWU106	164	L22	SWU71	113
L6	盐油2号 Yanyou 2	160	L23	SWU80	112
L7	油研10选 Youyan 10 xuan	157	L24	SWU97	111
L8	中双4号 Zhongshuang 4	155	L25	2012-8380	110
L9	沪油14 Huyou 14	154	L26	WH-62	105
L10	中双12 Zhongshuang 12	152	L27	华油2号 Huayou 2	99
L11	宁油12 Ningyou 12	148	L28	丰油9号 Fengyou 9	96
L12	Wx10315	146	L29	中油821 Zhongyou 821	95
L13	SWU94	140	L30	SWU108	93
L14	云油双1号 Yunyoushuang 1	139	L31	SWU101	89
L15	2012-9323	134	L32	10-1047	87
L16	华双128 Huashuang 128	131	L33	花油8号 Huayou 8	59
L17	SWU68	128

基因 Gene	引物序列 Primer sequence (5′-3′)
Actin7	F: TGGGTTTGCTGGTGACGAT; R: TGCCTAGGACGACCAACAATACT
BnSPSA1-1	F: AGGATAAGTATATCCCGAAGGAACTC; R: CGTCACCAGCATCAGCGTAGT
BnSPSA1-2	F: CACATGATGGTGATATGGAAGACG; R: GATATTCTTTTTTGGGTCGGGC
BnSPSA1-3	F: GAGACTGATCTTCACAAGTCATGGA; R: TCCTTTTCCTCATGCTGCTTCT
BnSPSA2-1	F: AAATGGCAGAGAGTCGAGTTCG; R: AGCTTAGCTTTTCTTTCAGCGGT
BnSPSA2-2	F: TGCTAGTGCGATGGGGGAT; R: GGTAGCTTCTGTTGCCGTGC
BnSPSA2-3	F: TGACTGGAGCTATGCCGAACCT; R: GGACATCTGCATCACATGGCTAAG
BnSPSA2-4	F: CGGTTTGATACGAGGTGAGAACAT; R: GCATCTCAGACGGTTCAGCG
BnSPSB-1	F: ATGTCGTATGAGACATCCCCAGT; R: TATTCGGTTCAAGAGACGCATTC
BnSPSB-2	F: CGAGGAACTTATCTCTGGCACAC; R: CCAATAAAAGGACTCTCTGACGG
BnSPSC-1	F: ACGTGAAAAAGGACGCAATGAT; R: GCAGATAGAGGCAGGCACAATG
BnSPSC-2	F: GTGATTACTGCTGATTCCTACGATG; R: TATTTTTCCCTTGCCCGATGT

基因名称 Gene name	基因序列号 Gene ID	氨基酸数量 Amino acid number	分子量 Molecular weight (kD)	等电点 Isoelectric point	染色体 Chromosome
BnSPSA1-1	BnaC09g37470D	966	109.2	6.35	C09
BnSPSA1-2	BnaA10g15120D	966	109.0	6.29	A10
BnSPSA1-3	BnaA02g04800D	972	109.3	5.52	A02
BnSPSA2-1	BnaA03g03230D	1039	116.3	6.13	A03
BnSPSA2-2	BnaC03g04660D	962	107.4	6.66	C03
BnSPSA2-3	BnaC07g29460D	967	107.9	6.78	C07
BnSPSA2-4	BnaA06g27590D	960	107.2	6.66	A06
BnSPSB-1	BnaA10g03060D	1064	119.5	5.74	A10
BnSPSB-2	BnaC05g02930D	1065	119.6	5.74	C05
BnSPSC-1	BnaC02g29300D	473	53.8	8.06	C02
BnSPSC-2	BnaA02g23460D	1047	118.5	6.24	A02

基因 Gene	ABRE	ARE	CGTCA-motif	GARE-motif	MBS	TCA-element	TGACG-motif
SPSA1-1	0	4	3	3	0	2	3
SPSA1-2	0	1	3	2	1	2	3
SPSA1-3	0	1	0	0	0	1	0
SPSA2-1	1	3	1	2	1	0	1
SPSA2-2	1	2	1	3	1	0	1
SPSA2-3	0	1	0	1	2	1	0
SPSA2-4	6	1	3	1	2	2	3
SPSB-1	5	6	1	1	1	2	1
SPSB-2	4	4	2	1	1	1	2
SPSC-1	0	0	2	1	1	0	2
SPSC-2	0	0	2	2	1	0	2
总计Total	17	23	18	17	11	11	18

甘蓝型油菜蔗糖磷酸合酶(SPS)基因家族成员鉴定及表达分析

Genome-wide Analysis and Expression Profiling of SPS Gene Family in Brassica nupus L.

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