重瓣百合LiSEP3基因克隆与表达分析

doi:10.3969/j.issn.1000-2006.2017.01.007

国家林草科技领军期刊
中国精品科技期刊
中国高校百佳科技期刊
江苏省新闻出版政府奖期刊奖
RCCSE林学权威期刊（A+）
CSCD核心期刊
Scopus数据库收录期刊
中文核心期刊
SCD核心期刊

作者加群：102861116

微信公众号：南京林业大学学报

高级检索

南京林业大学学报（自然科学版） ›› 2017, Vol. 60 ›› Issue (01): 42-48.doi: 10.3969/j.issn.1000-2006.2017.01.007

重瓣百合LiSEP3基因克隆与表达分析

隋娟娟^1,2,李晓昕², 杨秋燕², 吴泽²,曹兴^2,3,何俊娜²,义鸣放²

1.阜阳师范学院生物与食品工程学院,安徽阜阳 236037;
2.中国农业大学观赏园艺与园林系,花卉发育与品质调控北京市重点实验室,北京 100193;
3.聊城大学农学院,山东聊城 252059

出版日期:2017-02-18 发布日期:2017-02-18
基金资助:
基金项目:国家自然科学基金项目(31471904); 安徽省教育厅自然科学研究重点项目(KJ2016A871); 安徽省高校质量工程项目(2015jyxm221)
第一作者:隋娟娟(suijuanjuan@163.com),副教授,博士。

Cloning and expression analysis of gene LiSEP3 in double lily

1. College of Biology and Food Engineering, Fuyang Normal College, Fuyang 236037, China;
2. Beijing Key Laboratory of Development and Quality Control of Ornamental Crops, Department of Ornamental Horticulture and Landscape Architecture, China Agricultural University, Beijing 100193, China;
3. College of Agriculture, Liaocheng University, Liaocheng 252059, China

Online:2017-02-18 Published:2017-02-18

摘要/Abstract

摘要： 【目的】克隆百合LiSEP3基因并确定其在百合不同地上器官及不同花瓣中的表达差异,探讨SEP3基因在重瓣花中的表达模式。【方法】利用RACE方法从重瓣百合品种‘比罗尼卡’中克隆得到LiSEP3基因核苷酸序列,利用SMART软件对其蛋白结构进行分析,以MEGA 5.0软件进行系统进化树分析,用基因枪轰击法进行亚细胞定位分析,采用荧光定量PCR进行不同地上器官及不同花瓣中LiSEP3基因表达差异分析。【结果】LiSEP3基因属于MADS-box家族E类基因,其开放阅读框为729 bp,共编码242个氨基酸; LiSEP3基因编码的蛋白具有典型的MADS结构域、K-box区及2个SEP基序; LiSEP3与同为单子叶植物的六出花的SEP3亲缘关系最近; LiSEP3蛋白定位于细胞核中; LiSEP3基因在花中的相对表达量最高,在茎、叶中几无表达; 在1～7轮花瓣中均能检测到LiSEP3表达,且位于最内侧的第7轮花瓣中相对表达量最高。【结论】LiSEP3基因属于MADS-box家族E类基因,主要在花中表达,其中在最内侧花瓣中表达量最高,其表达模式与双子叶植物花中SEP3基因表达模式存在不同。

Abstract: 【Objetctive】 We cloned LiSEP3 from a double lily and analyzed LiSEP3 expression pattern among different aboveground organs and petals, for probing into SEP3 gene expression pattern in the double flower. 【Method】 LiSEP3 was isolated from double lily cv. ‘Belonica’ by RACE, and protein structure of LiSEP3 was predicted by SMART software. Phylogenetic analysis was performed by MEGA 5.0, subcellular localization of LiSEP3 were detected by particle bombardment. In addition, the expression pattern of LiSEP3 was analyzed among different aboveground organs and petals. 【Result】 LiSEP3 is a member of E class of MADS-box. The open reading frame of LiSEP3 is 729 bp in length, and encodes a protein of 242 amino acids. Protein structure analysis showed that LiSEP3 has a conserved MADS domain, a K-box region and two SEP motifs. Phylogenetic tree analysis suggested LiSEP3 had the highest similarity with monocotyledonous Alstroemeria. Subcellular localization assay revealed LiSEP3 was a nucleus protein. LiSEP3 was abundant in flowers while it couldn’t be detected in stems and leaves. LiSEP3 was expressed in petals from the first whorl to the seventh whorl, and had the highest transcription level in the seventh whorl petals. 【Conclusion】 LiSEP3 belongs to E class of MADS-box family genes and is specifically expressed in flowers and has a higher expression in seventh whorl than that in other whorls. Moreover, the expression pattern of LiSEP3 in flowers is different from its homologous genes in dicotyledons.

中图分类号:

S682
Q785

隋娟娟,李晓昕,杨秋燕,等. 重瓣百合LiSEP3基因克隆与表达分析[J]. 南京林业大学学报（自然科学版）, 2017, 60(01): 42-48.

Cloning and expression analysis of gene LiSEP3 in double lily. Cloning and expression analysis of gene LiSEP3 in double lily[J].Journal of Nanjing Forestry University (Natural Science Edition), 2017, 60(01): 42-48.DOI: 10.3969/j.issn.1000-2006.2017.01.007.

参考文献

[1] COEN E S, MEYEROWITZ E M. The war of the whorls—genetic interactions controlling flower development[J]. Nature, 1991, 353(6339):31-37. DOI: 10.1038/353031a0.
[2] THEISSEN G. Development of floral organ identity: stories from the MADS house[J]. Current Opinion in Plant Biology, 2001, 4(1):75-85. DOI: 10.1016/S1369-5266(00)00139-4.
[3] ROUX F, TOUZET P, CUGUEN J, et al. How to be early flowering: an evolutionary perspective[J]. Trends in Plant Science, 2006, 11(8):375-381. DOI:10.1016/j.tplants.2006.06.006.
[4] MESSENGUY F, DUBOIS E. Role of MADS box proteins and their cofactors in combinatorial control of gene expression and cell development[J]. Gene, 2003, 316(2):1-21. DOI: 10.1016/S0378-1119(03)00747-9.
[5] KIM S, KOH J, YOO M J, et al. Expression of floral MADS-box genes in basal angiosperms: implications for the evolution of floral regulators[J]. Plant Journal, 2005, 43:724-744. DOI:10.1111/j.1365-313X.2005.02487.x.
[6] GONG B H, YI J, WU J, et al. LlHSFA1, a novel heat stress transcription factor in lily(Lilium longiflorum), can interact with LlHSFA2 and enhance the thermotolerance of transgenic Arabidopsis thaliana[J]. Plant Cell Reports, 2014, 33(9):1519-1533. DOI: 10.1007/s00299-014-1635-2.
[7] 席梦利, 刘坡, 吴小萍, 等. 百合花发育相关基因LLGLO1的RNAi载体构建[J]. 南京林业大学学报(自然科学版), 2010, 34(1):1-4. DOI: 10.3969/j.issn.1000-2006.2010.01.001. XI M L, LIU P, WU X P, et al. Construction of RNAi vector of LLGLO1 gene related to flower development of Lilium[J]. Journal of Nanjing Forestry University(Natural Sciences Edition), 2010, 34(1):1-4. DOI: 10.3969/j.issn.1000-2006.2010.01.001.
[8] TONG Z, LI Q H, YANG Y J,et al. Isolation and expression analysis of LoPIP2, a lily (Lilium oriental hybrids)aquaporin gene involved in desiccation-induced anther dehiscence[J]. Scientia Horticulturae, 2013, 164:316-322. DOI: 10.1016/j.scienta.2013.09.022.
[9] SUI J, HE J N, WU J, et al. Characterization and functional analysis of transcription factor LoMYB80 related to anther development in lily(Lilium oriental hybrids)[J]. Journal of Plant Growth Regulation, 2015, 34:1-13. DOI: 10.1007/s00344-015-9489-6.
[10] 吴祝华, 施季森, 池坚, 等. 观赏百合资源与育种研究进展[J]. 南京林业大学学报(自然科学版), 2006, 122: 113-118. DOI:10.3969/j.issn.1000-2006.2006.02.027. WU Z H, SHI J S, CHI J, et al. Research advances on resources and breeding of ornamental lily[J]. Journal of Nanjing Forestry University(Natural Sciences Edition), 2006, 122: 113-118. DOI:10.3969/j.issn.1000-2006.2006.02.027.
[11] HONMA T, GOTO K. Complexes of MADS-box proteins are sufficient to convert leaves into floral organs[J]. Nature, 2001, 409(6819):525-529. DOI:10.1038/35054083.
[12] PELAZ S, GUSTAFSON-BROWN C, KOHALMI S E, et al. APETALA1 and SEPALLATA3 interact to promote flower development[J]. Plant Journal, 2001, 26(4):385-394. DOI: 10.1046/j.1365-313X.2001.2641042.x.
[13] ZAHN L M, KONG H, LEEBENS-MACK J H, et al. The evolution of the SEPALLATA subfamily of MADS-box genes: a preangiosperm origin with multiple duplications throughout angiosperm history[J]. Genetics, 2005, 169(4):2209-2223. DOI:10.1534/genetics.104.037770.
[14] MELZER R, THEISSEN G. Reconstitution of floral quartets in vitro involving class B and class E floral homeotic proteins[J]. Nucleic Acids Research, 2009, 37(8):2723-2736. DOI: 10.1093/Nar/Gkp129.
[15] IMMINK R G H, TONACO I A N, DE FOLYER S, et al. SEPALLATA3: the ‘glue’ for MADS box transcription factor complex formation[J]. Genome Biology, 2009, 10(2):1-16. DOI: 10.1186/Gb-2009-10-2-R24.
[16] 贾春蕾, 向林, 秦德辉,等. 春兰CgSEP3基因的克隆和表达分析[J]. 西北植物学报, 2014, 34(7):1305-1310. DOI:10.7606/j.issn.1000-4025.2014.07.1305. JIA C L, XIANG L, QIN D H, et al. Expression analysis of CgSEP3 gene from Cymbidium goeringii[J]. Acta Botanica Boreali-Occidentalia Sinica, 2014,34(7): 1305-1310.
[17] 徐启江. 草原龙胆花器官MADS-box基因克隆与表达分析[D]. 哈尔滨:东北林业大学, 2007. XU Q J. Molecular colning and expression analysis of floral organ identity MADS-box genes from lisianthus(Eustoma grandiflorum)[D]. Harbin: Northeast Forestry University, 2007.
[18] 黄赫. 洋葱A、B、C、E功能MADS-box基因的克隆与表达分析[D]. 哈尔滨:东北林业大学, 2013. HUANG H. Molecular colning and expression analysis of A、B、C and E-class MADS-box genes from onion(Allium cepa L.)[D]. Harbin: Northeast Forestry University, 2013.
[19] ANGENENT G C, FRANKEN J, BUSSCHER M, et al. Co-suppression of the petunia homeotic gene fbp2 affects the identity of the generative meristem[J]. Plant Journal, 1994, 5(1):33-44. DOI:10.1046/j.1365-313X.1994.5010033.x.
[20] TZENG T Y, HSIAO C C, CHI P J, et al. Two lily SEPALLATA-like genes cause different effects on floral formation and floral transition in Arabidopsis[J], Plant Physiology, 2003, 133:1091-1101. DOI: 10.1104/pp.103.026997.
[21] 宫本贺. 百合热激转录因子L1HSFA1及其下游热激蛋白L1HSP70响应热胁迫的机制解析[D].北京:中国农业大学, 2014. GONG B H. Mechanism analysis of response to heat stress of LlHAFA1 and its downstream LlHSP70 from lily(Lilium longiflorm)[D]. Beijing:China Agricultural University, 2014.
[22] 徐淑平,卫志明. 基因枪的使用方法介绍[J]. 植物生理学通讯, 1998, 34(1): 41-43. DOI: 10.13592/j.cnki.ppj.1998.01.014. XU S P, WEI Z M. Introduction to method of microprojectile bombardment and its application[J]. Plant Physiology Communications, 1998, 34(1): 41-43.
[23] LIVAK K J, SCHMITTGEN T D. Analysis of relative gene expression data using real-time quantitative PCR and the 2(T)(-Delta Delta C)method[J]. Methods, 2001, 25(4):402-408. DOI: 10.1006/meth.2001.1262.
[24] CUI R, HAN J, ZHAO S, et al. Functional conservation and diversification of class E floral homeotic genes in rice(Oryza sativa)[J]. Plant Journal, 2010, 61(5):767-781. DOI:10.1111/j.1365-313X.2009.04101.x.
[25] TZENG T Y, LIU H C, YANG C H. The C-terminal sequence of LMADS1 is essential for the formation of homodimetgrs for B function proteins[J]. Journal of Biological Chemistry, 2004, 279(11):10747-10755. DOI:10.1074/jbc.M311646200.
[26] CHANG Y Y, CHIU Y F, WU J W, et al. Four orchid(Oncidium Gower Ramsey)AP1/AGL9-like MADS box genes show novel expression patterns and cause different effects on floral transition and formation in Arabidopsis thaliana[J]. Plant Cell Physiology, 2009, 50(8):1425-1438. DOI:10.1093/pcp/pcp087.
[27] THEISSEN G, BECKER A, di ROSA A, et al. A short history of MADS-box genes in plants[J]. Plant Molecular Biology, 2000, 42(1):115-149. DOI: 10.1023/A:1006332105728.

重瓣百合LiSEP3基因克隆与表达分析

Cloning and expression analysis of gene LiSEP3 in double lily

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

作者

读者

审稿

[1]	宋雨蒙, 李素艳, 孙向阳. 复合增氧剂对建兰水培苗生长及生理特性的影响[J]. 南京林业大学学报（自然科学版）, 2024, 48(1): 115-123.
[2]	魏绪英, 张瑶, 马美霞, 姜雪茹, 陈慧婷, 吴靖, 杨玉, 蔡军火. 石蒜年生长周期内NSC及其代谢酶活性变化[J]. 南京林业大学学报（自然科学版）, 2024, 48(1): 106-114.
[3]	王纪, 方升佐. 不同抗褐化剂对青钱柳愈伤组织酶活性和生长的影响[J]. 南京林业大学学报（自然科学版）, 2023, 47(6): 167-174.
[4]	肖周宏, 涂蓉慧, 刘昂, 龚佑科, 丁聪, 喻勋林. 湖南省兰科植物新记录(Ⅲ)[J]. 南京林业大学学报（自然科学版）, 2023, 47(5): 255-258.
[5]	张婷, 柏杨, 亓希武, 于盱, 房海灵, 李莉, 刘冬梅, 梁呈元, 李维林. 薄荷MhWRKY57基因克隆及响应茉莉酸信号的表达分析[J]. 南京林业大学学报（自然科学版）, 2022, 46(6): 279-287.
[6]	高燕, 莫建彬, 付艳茹, 奉树成. 铁线莲‘朱卡’组织培养技术及再生体系的建立[J]. 南京林业大学学报（自然科学版）, 2021, 45(3): 109-116.
[7]	闵可怜, 王丹, 湛晓蝶, 陈敏, 岳海燕, 何毅, 刘亮, 黎青, 向毅, 李建伟. 3种辐射保护剂对⁶⁰Co-γ射线辐照小苍兰的保护效应[J]. 南京林业大学学报（自然科学版）, 2020, 44(3): 11-18.
[8]	姜楠南, 张启翔, 王媛, 孙音, 房义福, 徐金光. 赤霉素对‘大富贵’芍药休眠解除及内源激素和糖类代谢的影响[J]. 南京林业大学学报（自然科学版）, 2020, 44(3): 26-32.
[9]	张巧, 李杰, 山萌蒙, 刘鑫, 李媛媛. 春剑种子无菌萌发过程的显微观察[J]. 南京林业大学学报（自然科学版）, 2020, 44(2): 105-110.
[10]	胡晓龙,田绍泽,胡惠蓉. ‘晨光’大花金鸡菊花芽分化及光周期特性研究[J]. 南京林业大学学报（自然科学版）, 2019, 62(04): 43-47.
[11]	李文建,沈永宝,史锋厚,贾文庆. 建兰花色形成的成分检测[J]. 南京林业大学学报（自然科学版）, 2019, 62(04): 57-62.
[12]	韦东山,张秋良,常金宝. 美国薄荷嫩枝扦插繁殖及生根机理研究[J]. 南京林业大学学报（自然科学版）, 2018, 61(05): 39-45.
[13]	蔡军火,李金峰,魏绪英,张露. 石蒜叶期生物量、根系活力的动态变化及更替规律[J]. 南京林业大学学报（自然科学版）, 2018, 61(01): 55-59.
[14]	谢菊英1,张露1*,连芳青1,黄敏仁2,刘青1,陈蕾伊1. 生长调节剂对石蒜无性繁殖的影响[J]. 南京林业大学学报（自然科学版）, 2006, 49(03): 125-127.
[15]	李杰1,2,黄敏仁1,王明庥1,蔡汝2,匡鹏安3. 兰花生物技术的研究及应用[J]. 南京林业大学学报（自然科学版）, 2006, 49(02): 108-112.