绿色杜氏藻GGR基因生物信息学分析及表达研究
PDF下载 (297)张 燕,龚一富*,景丹丹,朱帅旗,俞 凯.绿色杜氏藻GGR基因生物信息学分析及表达研究[J].宁波大学学报(理工版),2017,30(1):35-41.DOI:
ZHANG Yan,GONG Yi-fu*,JING Dan-dan,ZHU Shuai-qi,YU Kai.Induced expression and bioinformatic analysis of GGR gene in Dunaliella viridis
[J].Journal of Ningbo University(Natural Science & Engineering Edition),2017,30(1):35-41.DOI:
| Title: | Induced expression and bioinformatic analysis of GGR gene in Dunaliella viridis |
| 作者: | 张 燕, 龚一富*, 景丹丹, 朱帅旗, 俞 凯 |
| Author(s): | ZHANG Yan, GONG Yi-fu*, JING Dan-dan, ZHU Shuai-qi, YU Kai |
| 关键词: | 绿色杜氏藻; 甲基茉莉酸; 叶绿素; 类胡萝卜素 |
| Keywords: | GGR; Dunalilla viridis; MeJA; chlorophyll; carotenoid |
| 分类号: | Q786 |
| 文献标识码: | A |
| 摘要: | 采用Illumina HiSeqTM 2000高通量测序方法获得绿色杜氏藻GGR基因(GenBank序号: KX100480) cDNA全长序列, 研究了甲基茉莉酸(MeJA)对绿色杜氏藻GGR基因表达的影响. 测序结果表明, 绿色杜氏藻GGR基因cDNA全长2356bp, 开放阅读框1539bp, 编码512个氨基酸, 分子量为52.2961kDa, 理论等电点为6.54. 序列分析结果表明, 该蛋白具有保守的ChlP结构域, 为可溶性蛋白, 具有跨膜区域, 无信号肽. TargetP 1.1 Server预测结果表明, 该蛋白可能定位于线粒体. 系统进化树结果表明, 绿色杜氏藻GGR蛋白较其他植物的GGR蛋白亲缘关系较 远. RT-PCR结果表明, 100?mol·L-1 MeJA可显著地提升绿色杜氏藻GGR基因的表达(P<0.01). 同时, 绿色杜氏藻叶绿素和类胡萝卜素含量达到最高, 说明GGR基因与绿色杜氏藻叶绿素和类胡萝卜素的合成存在一定关系 |
| Abstract: | Geranylgeranyl reductase (GGR) is an important enzyme in the upstream of plant tocopherols and chlorophyll biosynthetics pathway. In this study, the full length cDNA of GGR gene (GenBank accession number: KX100480) in Dunaliella viridis is obtained by HiSeqTM Illumina 2000. Transcriptional levels of GGR gene with the treatment of Methyl Jasmonate (MeJA) are examined by Real-Time quantitative PCR (RT-PCR). The results show that the total length of GGR gene is 2356bp, containing an open reading frame of 1539bp encoding 512 amino acids, relative molecular weight is 52.2961kD and isoelectric point is 6.54. Bioinformatics prediction reveals that the protein contains a transmembrane domain without signal peptide. TargetP 1.1 Server prediction results suggest that it is located in mitochondria. Phylogenetic tree results indicate that the protein sequence of GGR in D. viridis is related loosely to other species. RT-PCR result demonstrates that the expression of GGR gene in D. viridis is promoted under the treatment of low concentration of MeJA and limited in high concentration of MeJA. When treated with 100?mol·L-1 MeJA, the expression of GGR gene manifests the significant difference in the control group (P<0.01). And the content of chlorophyll and carotenoids show the same trend. The result implies that the GGR gene is to certain extent correlated with the synthetic of chlorophyll and carotenoids. |
| 参考文献 /References: | [1].OREN A. The alga Dunaliella: Biodiversity, physiology, genomics and biotechnology[J]. Journal of Phycology, 2010, 46(1):216-217. [2].BOLLIVAR D W, WANG S, ALLEN J P, et al. Molecular genetic analysis of terminal steps in bacteriochlorophyll a biosynthesis: Characterization of a Rhodobacter capsulatus strain that synthesizes geranylgeraniol- esterified bacteriochlorophyll a[J]. Biochemistry, 1994, 33(43):12763-12768. [3].SOLL J, SCHULTZ G, R?DIGER W, et al. Hydro- genation of geranylgeraniol: Two pathways exist in spinach chloroplasts[J]. Plant Physiology, 1983, 71(3): 225-230. [4].MARTIN L, CHUNG-SOON I, GROSSMAN A R. Genome-based examination of chlorophyll and carotenoid biosynthesis in Chlamydomonas reinhardtii[J]. Plant Physiology, 2005, 138(1):490-515. [5].MYOUNGRYOUL P, EUNAE C, REHMAN S, et al. Expression of a sesame geranylgeranyl reductase cDNA is induced by light but repressed by abscisic acid and ethylene[J]. Pakistan Journal of Botany, 2010, 42(3): 1815-1826. [6].TANAKA R, KRUSE E, RUDIGER W B, et al. Reduced activity of geranylgeranyl reductase leads to loss of chlorophyll and tocopherol and to partially geranylgeranylated chlorophyll in transgenic tobacco plants expressing antisense RNA for geranylgeranyl reductase[J]. Plant Physiology, 1999, 120(3):695-704. [7].ZHOU Y, GONG Z, YANG Z, et al. Mutation of the light-induced yellow leaf 1 gene, which encodes a geranylgeranyl reductase, affects chlorophyll biosynthesis and light sensitivity in rice[EB/OL]. http://journals.plos.org/ plosone/article/asset?id=10.1371/journal.pone.0075299.PDF. [8].WANG P, LI C, WANG Y, et al. Identification of a geranylgeranyl reductase gene for chlorophyll synthesis in rice[J]. Springerplus, 2014, 3(1):1-9. [9].KELLER Y, BOUVIER F, D’HARLINGUE A, et al. Metabolic compartmentation of plastid prenyllipid biosynthesis: Evidence for the involvement of a multi- functional CHLP[J]. European Journal of Biochemistry, 1998, 251(1/2):413-417. [10].PEUKERT S, GRAY C, TAO J, et al. Design and structure activity relationships of potent and selective inhibitors of undecaprenyl pyrophosphate synthase (GGR): Tetramic, tetronic acids and dihydropyridin-2- ones[J]. Bioorganic & Medicinal Chemistry Letters, 2008, 18(6):1840-1844. [11].BRUNO L, CHIAPPETTA A, MUZZALUPO I, et al. Role of geranylgeranyl reductase gene in organ development and stress response in olive (Olea europaea) plants[J]. Functional Plant Biology, 2009, 36(4):370-381. [12].GIANNINO D, CONDELLO E, BRUNO L G, et al. The gene geranylgeranyl reductase of peach (Prunus persica L Batsch) is regulated during leaf development and responds differentially to distinct stress factors[J]. Journal of Experimental Botany, 2004, 55(405):2063-2073. [13].朱帅旗, 龚一富, 杭雨晴, 等. 绿色杜氏藻转录组分析 [J]. 遗传, 2015, 37(8):828-836. [14].王冬梅, 王玮蔚, 孙雪, 等. 一株杜氏藻的分子鉴定与耐盐特性[J]. 植物生理学报, 2014(3):315- 323. [15].金晓霞, 董延龙, 秦智伟, 等. 黄瓜(Cucumis stavius L.) CSEBF1基因的克隆及其表达特性分析[J]. 中国生物工程杂志, 2014, 34(3):41-47. [16].KANG S M, MIN J Y, KIM Y D, et al. Effects of methyl jasmonate and salicylic acid on the production of bilobalide and ginkgolides in cell cultures of Ginkgo biloba[J]. In Vitro Cellular & Developmental Biology- Plant, 2009, 42(1):44-49. [17].胡英考, 朱敬, 李雅轩, 等. 小麦牻牛儿牻牛儿基脱氢酶基因的克隆及电子定位与表达分析[J]. 麦类作物学报, 2010, 30(5):783-788. [18].潘卫东, 张娟, 张贵星, 等. 藻类及陆生高等植物叶绿体16S rRNA基因进化谱系及同源性分析[J]. 郑州大学学报(医学版), 2004, 39(1):15-20. [19].朱颖, 王丽丽, 柴芸彬. 茉莉酸甲酯对盐生杜氏藻类胡萝卜素含量的影响[J]. 宁波大学学报(理工版), 2010, 23(1):13-16. [20].LASKARIS G, BOUNKHAY M, THEODORIDIS G, et al. Induction of geranylgeranyl diphosphate synthase activity and taxane accumulation in Taxus baccata cell cultures after elicitation by methyl jasmonate[J]. Plant Science, 1999, 147(99):1-8. [21].王鑫威, 王丽丽, 龚一富, 等. 茉莉酸甲酯对雨生红球藻虾青素含量和dxs基因表达的影响[J]. 水产学报, 2011, 35(12):1822-1828. [22].SHPILYOV A V, ZINCHENKO V V, SHESTAKOV S V, et al. Inactivation of the geranylgeranyl reductase (GGR) gene in the Cyanobacterium synechocystis sp. PCC 6803 [J]. Biochimica et Biophysica Acta, 2005, 1706(3):195- 203. [23].CHOI D, HILDEBRAND D F. Lipid-derived signals that discriminate wound-and pathogen-responsive isoprenoid pathways in plants: Methyl jasmonate and the fungal elicitor arachidonic acid induce different 3-hydroxy-3- methylglutaryl-coenzyme A reductase genes and antimicrobial isopren[J]. Proceedings of the National Academy of Sciences, 1994, 91(6):2329-2333. [24].王和勇, 罗恒, 孙敏. 诱导子在药用植物细胞培养中的应用[J]. 中草药, 2004, 35(8):959-963. Induced expression and bioinformatic analysis of GGR gene in Dunaliella viridis [25].ZHANG Yan, GONG Yi-fu*, JING Dan-dan, ZHU Shuai-qi, YU Kai [26].( School of Marine Sciences, Ningbo University, Ningbo 315211, China ) [27].Abstract: Geranylgeranyl reductase (GGR) is an important enzyme in the upstream of plant tocopherols and chlorophyll biosynthetics pathway. In this study, the full length cDNA of GGR gene (GenBank accession number: KX100480) in Dunaliella viridis is obtained by HiSeqTM Illumina 2000. Transcriptional levels of GGR gene with the treatment of Methyl Jasmonate (MeJA) are examined by Real-Time quantitative PCR (RT-PCR). The results show that the total length of GGR gene is 2356bp, containing an open reading frame of 1539bp encoding 512 amino acids, relative molecular weight is 52.2961kD and isoelectric point is 6.54. Bioinformatics prediction reveals that the protein contains a transmembrane domain without signal peptide. TargetP 1.1 Server prediction results suggest that it is located in mitochondria. Phylogenetic tree results indicate that the protein sequence of GGR in D. viridis is related loosely to other species. RT-PCR result demonstrates that the expression of GGR gene in D. viridis is promoted under the treatment of low concentration of MeJA and limited in high concentration of MeJA. When treated with 100?mol·L-1 MeJA, the expression of GGR gene manifests the significant difference in the control group (P<0.01). And the content of chlorophyll and carotenoids show the same trend. The result implies that the GGR gene is to certain extent correlated with the synthetic of chlorophyll and carotenoids. |
| 备注/Memo: | 收稿日期: 2016?04?24. 宁波大学学报（理工版）网址: http://journallg.nbu.edu.cn/ 基金项目: 浙江省科技厅重点科技创新团队项目（2012R10029-07, 2010R50029）; 宁波市科技攻关项目（2013C10018）; 浙江省大学生科技创 新活动计划暨新苗人才计划项目（2014R405050）; 宁波大学优秀学位论文培育基金（py2014020）. 第一作者: 张燕（1991－）, 女, 山东菏泽人, 在读硕士研究生, 主要研究方向: 藻类分子生物学. E-mail: 15005412101@163.com *通信作者: 龚一富（1973－）, 男, 重庆人, 博士/副教授, 主要研究方向: 植物分子生物学. E-mail: gongyifu@163.com 宁波大学学报（理工版）网址：http://journallg.nbu.edu.cn/ |
