水稻蛋白激酶基因启动子OsSRLp的分离及特性分析

doi:10.16819/j.1001-7216.2018.7127

摘要/Abstract

摘要：

【目的】为研究水稻中I型酪蛋白激酶基因OsSRL启动子的结构及功能,【方法】以水稻中花11基因组DNA为模板,经PCR扩增获得基因上游约1600 bp序列,命名为启动子OsSRLp。应用PLACE在线软件,分析序列中的顺式作用元件,同时构建含有GUS报告基因的植物表达载体,转化水稻。【结果】OsSRL为组成型表达,OsSRLp含有调控生殖发育、激素应答及逆境响应等多种顺式作用元件。OsSRLp驱动的GUS报告基因在根、茎中均有所表达,在小穗中表达丰度较高,而在其他组织中表达丰度较低。【结论】OsSRLp为组成型启动子,其下游调控基因OsSRL可能参与水稻发育及生殖过程。

关键词: 水稻, 启动子OsSRLp, 表达特性, GUS活性, 发育及生殖

Abstract:

【Objective】To study the construction and function of OsSRL, a typeⅠcasein kinase, 【Method】OsSRL and about 1600 bp up-stream of the gene named as OsSRLp were isolated using genomic DNA of rice Zhonghua 11. The regulatory elements of OsSRLp were analyzed using online software of PLACE, and the results showed that several key cis-elements which associated with reproductive process, signal and stress response were included. 【Results】The expression pattern analysis showed that OsSRL was constitutively expressed in different tissues of rice. The fusion expression vector of OsSRLp and GUS was constructed, and the vector was transformed into rice callus. After positive transgenic lines were obtained, the GUS activities were further detected, and the results showed that relative higher level expression was detected in spikelet, root and stem, as well as low expression levels in other rice tissues. The results indicated that OsSRLp possess constitutive activities with slightly variations in different rice tissues. 【Conclusion】 Combined with the characteristic elements and their potential functions of OsSRLp, the results implied that OsSRL and its promoter OsSRLp could participate in and regulate the development and reproduction progresses of plants.

Key words: rice, promoter OsSRLp, expression pattern, GUS activity, development and reproduction

中图分类号:

Q755
S511.01

王庆国, 王莹莹, 李臻, 潘教文, 王一帆, 李颖秀, 刘炜. 水稻蛋白激酶基因启动子OsSRLp的分离及特性分析[J]. 中国水稻科学, 2018, 32(4): 335-341.

Qingguo WANG, Yingying WANG, Zhen LI, Jiaowen PAN, Yifan WANG, Yingxiu LI, Wei LIU. Isolation and Characteristic Analysis of Protein Kinase Promoter OsSRLp from Rice[J]. Chinese Journal OF Rice Science, 2018, 32(4): 335-341.

图/表 5

参考文献 20

[1]	Liu W, Xu Z H, Luo D, Xue H W.Roles of OsCKI1, a rice casein kinase I, in root development and plant hormone sensitivity.Plant J, 2003, 36(2): 189-202.
[2]	Liu W, Ji S X, Fang X L, Wang Q G, Li Z, Yao F Y, Hou L, Dai S J.Protein Kinase LTRPK 1 influences cold adaptation and microtubule stability in rice. J Plant Growth Regul, 2013, 32(3): 483-490.
[3]	Borgal L, Rinschen M M, Dafinger C, Hoff S, Reinert M J, Lamkemever T, Lienkamp S S, Benzing T, Schermer B.Casein Kinase 1α phosphorylates the Wnt regulator Jade-1 and modulates its activity. J Biol Chem, 2012, 289(38): 26344-26356.
[4]	Kwon C T, Kim S H, Kim D, Paek N C.The rice floral repressor Early flowering1 affects spikelet fertility by modulating gibberellin signaling. Rice, 2015, 8(1): 58.
[5]	Hori K, Oqiso-Tanaka E, Matsubara K, Yamanouchi U, Ebana K, Yano M.Hd16 , a gene for casein kinase I, is involved in the control of rice flowering time by modulating the day-length response. Plant J, 2013, 76(1): 36-46.
[6]	张中保, 卢敏, 李会勇, 张登峰, 刘颖慧, 石云素, 宋燕春, 王天宇, 黎裕. 玉米干旱诱导表达基因ZmCKS2的克隆与表达分析. 作物学报, 2010, 36(6): 945-952.
	Zhang Z B, Lu M, Li H Y, Zhang D F, Liu Y X, Chi Y S, Song Y C, Wang T Y, Li S.Isolation and expression analysis of a drought-induced gene ZmCKS2 in maize(Zea mays L.). Acta Agron Sin, 2010, 36(6): 945-952. (in Chinese with English abstract).
[7]	Wingender E, Chen X, Hehl R, Karas H, Liebich I, Matys V, Meinhardt T, Pruss M, Reuter I, Schacherer F: TRANSFAC: An integrated system for gene expression regulation.Nucleic Acids Res, 2000, 28(1): 316-319.
[8]	Tao Y B, He L L, Niu L J, Xu Z F.Isolation and characterization of the Jatropha curcas PETALA1 (JcAP1) promoter conferring preferential expression in inflorescence buds. Planta, 2016, 244(2): 467-478.
[9]	Srinath T, Reddy V D, Rao KV.Isolation and functional characterization of a novel stress inducible promoter from pigeonpea ( Cajanus cajan L). Plant Cell Tiss Organ Cult, 2017, 128(2): 457-468.
[10]	Hofgen R, Willmitzer L.Storage of competent cells for Agrobacterium transformation. Nucleic Acids Res, 1988, 16(20): 9877.
[11]	Murray M G, Thompson W F.Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res, 1980, 8(19): 4321.
[12]	Jefferson R A.Assaying chimeric genes in plants: The GUS gene fusion system. Plant Mol Biol Rep, 1987, 5(4): 387-405.
[13]	房孝良, 刘炜, 安静, 王庆国. 水稻种胚特异性启动子OsESP1的克隆及其表达特性. 作物学报, 2011, 37(10): 1904-1909.
	Fang X L, Liu W, An J, Wang Q G.Isolation and characterization of an embryo-specific promoter OsESP1 from rice. Acta Agron Sin, 2011, 37(10): 1904-1909. (in Chinese with English abstract)
[14]	Qi X, Zhang Y, Chai T.Characterization of a novel plant promoter specifically induced by heavy metal and identification of the promoter regions conferring heavy responsiveness. Plant Physiol, 2007, 143(1): 50-59.
[15]	Yanagisawa S.Dof1 and Dof2 transcription factors are associated with expression of multiple genes involved in carbon metabolism in maize. Plant J, 2000, 21(3): 281-288.
[16]	蔡晓锋, 张余洋, 张俊红, 李汉霞, 叶志彪. 植物Dof基因家族功能研究进展. 植物生理学报, 2013, 49(1): 1-12.
	Cai X F, Zhang Y Y, Zhang J H, Li H X, Ye Z B.Advances in research on function of the Dof gene family in plant. Plant Physiol J, 2013, 49(1): 1-12. (in Chinese with English abstract)
[17]	Green P J, Yong M H, Cuozzo M, Kano-Murakami Y, Silverstein P, Chua N H.Binding site requirements for pea nuclear protein factor GT-1 correlate with sequences required for light-dependent transcriptional activation of the rbcS-3A gene. EMBO J, 1988, 7(13): 4035-4044.
[18]	Lam E, Chua N H.GT-1 binding site confers light responsive expression in transgenic tobacco.Science, 1990, 248(4954): 471-474.
[19]	Gourrierec J L, Delaporte V, Ayadi M, Li Y F, Zhou D X.Functional analysis of Arabidopsis transcription factor GT-1 in the expression of light-regulated genes. Genom Lett, 2002, 1(2): 77-82.
[20]	Filichkin S A, Leonard J M, Monteros A, Liu P P, Nonogaki H.A novel endo-β-mannanase gene in tomato LeMAN5 is associated with anther and pollen development. Plant Physiol, 2004, 134(3): 1080-1087.

顺式作用元件 cis-element	位置及序列 Sites(bp) and sequence	生物学功能 Biological function
生长素响应 Auxin response
CATATGGMSAUR	133, 326 CATATG	大豆SAUR (Small Auxin-Up RNA) 15A基因启动子NDE元件中序列; 参与生长素响应Sequence in NDE element in soybean SAUR (Small Auxin-Up RNA) 15A gene promoter; Involved in auxin responsiveness
NTBBF1ARROLB	277 ACTTTA	发根农杆菌rolB基因中NtBBF1(烟草中Dof蛋白)结合位点,参与组织表达特异性及生长素诱导表达调控Tissue-specific and auxin regulated expression of the rolB oncogene in plants
胚及贮藏蛋白相关 Embryo and storage protein related
CANBNNAPA	13 CNAACAC	油菜中贮藏蛋白基因核心(CA)元件; 调控基因表达的胚及胚乳特异性 Core of (CA)_n element in storage protein genes in Brasica napus; embryo- and endosperm-specific transcription of napin gene
EBOXBNNAPA	133, 326, 632, 1484 CANNTG	油菜中napA贮藏蛋白基因的E盒 E-box of napA storage-protein gene of Brassica napus
DOFCOREZM	78, 82, 88, 120, 172, 842, 923, 1514 AAAG	Dof1和Dof2转录因子,参与玉米碳代谢相关基因的表达调控 Dof1 and Dof2 transcription factors are associated with expression of multiple genes involved in carbon metabolism in maize
生物节律相关 Biorhythm related
CIACADIANLELHC	311 CAANNNNATC	番茄LHC启动子区元件,参与昼夜节律调控 Identification of tomato LHC promoter regions necessary for circadian expression
光响应 Light response
GT1CONSENSUS	35, 75, 85, 91, 180, 181, 331, 377, 907 GRWAAW	光调节基因保守GT-1结合位点 Consensus GT-1 binding site in many light-regulated genes
HBOXCONSENSUSPV	599 CCTACCNNNNNNNCT	菜豆chs15启动子区中H盒保守序列; 在光调节及激发子诱导中起作用 Consensus sequence of H-boxes found in bean chs15 gene promoter; Essential for both light regulation and elicitor induction
IBOX	700, 856 GATAAG	光调节基因上游保守序列Conserved sequence upstream of light-regulated genes
IBOXCORE	35, 85, 700, 856 GATAA	单子叶及双子叶植物中高度保守的光调节基因上游序列 Conserved upstream-sequence of light-regulated genes of monocots and dicots
开花调控 Flowering process control
MYBPLANT	103 MACCWAMC	激活类苯基丙烷生物合成基因转录的花特异的MYB蛋白 A flower-specific MYB protein activates transcription of RT phenylpropanoid biosynthetic genes
MYBPZM	54, 599, 1033 CCWACC	通过激活类黄酮从而调控玉米花器官鞣红色素沉积的MYB同源P基因 The MYB-homologous P gene controls phlobaphene pigmentation in maize floral organs by directly activating a flavonoid
MYBST1	34, 699 GGATA	MybSt1(土豆中MYB同源基因)结合位点核心元件 A novel DNA binding protein with homology to MYB oncoproteins containing only one repeat can function as a transcriptional activator
花粉发育 Pollen development
POLLEN1LELAT52	80, 90, 170, 376, 586, 840 AGAAA	受各阶段特异性及依赖性激活元件调控,负责调控花粉特异表达的调控元件 Pollen-specific transcription is developmentally regulated by multiple stage- specific and co-dependent activator elements
其他 Others
TATA BOX2	340 TATAAAT	豌豆legA 5′上游TATA盒;与甘薯贮藏蛋白A相关,调控基因转录起始TATA box in the 5' upstream region of pea legA gene; Related with Sporamin A protein of sweet potato
CAAT BOX1	299, 322, 354, 1484 CAAT	烟草中决定豆球蛋白启动子的组织特异性Sequences responsible for the tissue specific promoter activity of a pea legumin gene in tobacco

顺式作用元件 cis-element	位置及序列 Sites(bp) and sequence	生物学功能 Biological function
生长素响应 Auxin response
CATATGGMSAUR	133, 326 CATATG	大豆SAUR (Small Auxin-Up RNA) 15A基因启动子NDE元件中序列; 参与生长素响应Sequence in NDE element in soybean SAUR (Small Auxin-Up RNA) 15A gene promoter; Involved in auxin responsiveness
NTBBF1ARROLB	277 ACTTTA	发根农杆菌rolB基因中NtBBF1(烟草中Dof蛋白)结合位点,参与组织表达特异性及生长素诱导表达调控Tissue-specific and auxin regulated expression of the rolB oncogene in plants
胚及贮藏蛋白相关 Embryo and storage protein related
CANBNNAPA	13 CNAACAC	油菜中贮藏蛋白基因核心(CA)元件; 调控基因表达的胚及胚乳特异性 Core of (CA)_n element in storage protein genes in Brasica napus; embryo- and endosperm-specific transcription of napin gene
EBOXBNNAPA	133, 326, 632, 1484 CANNTG	油菜中napA贮藏蛋白基因的E盒 E-box of napA storage-protein gene of Brassica napus
DOFCOREZM	78, 82, 88, 120, 172, 842, 923, 1514 AAAG	Dof1和Dof2转录因子,参与玉米碳代谢相关基因的表达调控 Dof1 and Dof2 transcription factors are associated with expression of multiple genes involved in carbon metabolism in maize
生物节律相关 Biorhythm related
CIACADIANLELHC	311 CAANNNNATC	番茄LHC启动子区元件,参与昼夜节律调控 Identification of tomato LHC promoter regions necessary for circadian expression
光响应 Light response
GT1CONSENSUS	35, 75, 85, 91, 180, 181, 331, 377, 907 GRWAAW	光调节基因保守GT-1结合位点 Consensus GT-1 binding site in many light-regulated genes
HBOXCONSENSUSPV	599 CCTACCNNNNNNNCT	菜豆chs15启动子区中H盒保守序列; 在光调节及激发子诱导中起作用 Consensus sequence of H-boxes found in bean chs15 gene promoter; Essential for both light regulation and elicitor induction
IBOX	700, 856 GATAAG	光调节基因上游保守序列Conserved sequence upstream of light-regulated genes
IBOXCORE	35, 85, 700, 856 GATAA	单子叶及双子叶植物中高度保守的光调节基因上游序列 Conserved upstream-sequence of light-regulated genes of monocots and dicots
开花调控 Flowering process control
MYBPLANT	103 MACCWAMC	激活类苯基丙烷生物合成基因转录的花特异的MYB蛋白 A flower-specific MYB protein activates transcription of RT phenylpropanoid biosynthetic genes
MYBPZM	54, 599, 1033 CCWACC	通过激活类黄酮从而调控玉米花器官鞣红色素沉积的MYB同源P基因 The MYB-homologous P gene controls phlobaphene pigmentation in maize floral organs by directly activating a flavonoid
MYBST1	34, 699 GGATA	MybSt1(土豆中MYB同源基因)结合位点核心元件 A novel DNA binding protein with homology to MYB oncoproteins containing only one repeat can function as a transcriptional activator
花粉发育 Pollen development
POLLEN1LELAT52	80, 90, 170, 376, 586, 840 AGAAA	受各阶段特异性及依赖性激活元件调控,负责调控花粉特异表达的调控元件 Pollen-specific transcription is developmentally regulated by multiple stage- specific and co-dependent activator elements
其他 Others
TATA BOX2	340 TATAAAT	豌豆legA 5′上游TATA盒;与甘薯贮藏蛋白A相关,调控基因转录起始TATA box in the 5' upstream region of pea legA gene; Related with Sporamin A protein of sweet potato
CAAT BOX1	299, 322, 354, 1484 CAAT	烟草中决定豆球蛋白启动子的组织特异性Sequences responsible for the tissue specific promoter activity of a pea legumin gene in tobacco