重瓣百合‘Double surprise' AGAMOUS基因的克隆与序列分析
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- 英文论文题名：Gene Cloning and Sequence Analysis of AGAMOUS Genes from Double-flower Lily ‘Double surprise’
- 论文作者：王奇 ; 赵芮 ; 张勤 ; 杨艳余 ; 黄美娟 ; 黄海泉
- 英文论文作者：WANG Qi ; ZHAO Rui ; ZHANG Qin ; YANG Yan-yu ; HUANG Mei-juan ; HUANG Hai-quan ; College of Landscape Architecture ; Southwest Forestry University
- 年：2016
- 作者机构：西南林业大学园林学院;
- 论文关键词：重瓣百合‘Double surprise’ ; AGAMOUS基因 ; 基因克隆 ; 序列分析
- 英文论文关键词：Double flower ‘Double surprise’ ; AGAMOUS gene ; Gene cloning ; Sequence analysis
- 会议召开时间：2016-07-19
- 会议录名称：中国观赏园艺研究进展2016
- 语种：中文
- 分类号：S682.29
- 学会代码：EGYP
- 会议名称：2016年中国观赏园艺学术研讨会
- 会议地点：中国湖南长沙
- 主办单位：中国园艺学会观赏园艺专业委员会、国家花卉工程技术研究中心
- 学会名称：中国园艺学会
- 页数：6
- 文件大小：1785k
- 原文格式：O
- 会议级别：全国
摘要
根据报道的AGAMOUS(AG)基因设计一对简并引物,提取从荷兰引进的重瓣百合‘double surprise'雌蕊总RNA,采用RT-PCR方法扩增到两个特异片段,经克隆测序后发现两个片段均为一个完整的ORF,分别命名为LdsAG1和LdsAG2,其中LdsAG1全长795bp,编码264个氨基酸;LdsAG2全长678bp,编码225个氨基酸;同时发现其核苷酸及氨基酸序列均与其他植物AG基因具有较高的同源性,并对其蛋白质二级结构及等电点等进行了分析。此外,通过软件分析发现两个片段均具有AG基因保守的MADS-box区、K区以及典型的AGⅠ区和AGⅡ区,进一步说明上述两个基因均属于MADS-box基因家族C类基因,为后续探讨该基因在重瓣百合中表达模式及分子机理奠定了一定的基础。
In this study,a pair of degenerate primers was designed according to some reported AGAMOUS(AG) genes in the Genbank,and the total RNA of double-flowered lily ' double surprise' introduced from the Netherlands was isolated from its pistil,two specific fragments were amplified by RT-PCR method and sequenced.The results showed that both of them were a complete open reading frame(ORF),and named LdsAGl and LdsAGl respectively;that the full-length of LdsAGl was 795 bp and encoded 264 amino acids,while the full-length of LdsAGl was 678 bp and encoded 225 amino acids;that both of them in the nucleotide and amino acid sequences had high homology with AG genes from other plants,that the protein secondary structure and isoelectric point of them were analyzed further.Besides,on the basis of comprehensive analysis of two fragments by using the software,it manifested that every fragment contained some conservative regions,e.g.MADS-box region,K region,typical AGI and AGII regions,which further indicated that both of them belonged to the C-class genes of MADS-box gene family.It laid some foundation for the future study of their expression pattern and molecular mechanism in double-flower lily.
In this study,a pair of degenerate primers was designed according to some reported AGAMOUS(AG) genes in the Genbank,and the total RNA of double-flowered lily ' double surprise' introduced from the Netherlands was isolated from its pistil,two specific fragments were amplified by RT-PCR method and sequenced.The results showed that both of them were a complete open reading frame(ORF),and named LdsAGl and LdsAGl respectively;that the full-length of LdsAGl was 795 bp and encoded 264 amino acids,while the full-length of LdsAGl was 678 bp and encoded 225 amino acids;that both of them in the nucleotide and amino acid sequences had high homology with AG genes from other plants,that the protein secondary structure and isoelectric point of them were analyzed further.Besides,on the basis of comprehensive analysis of two fragments by using the software,it manifested that every fragment contained some conservative regions,e.g.MADS-box region,K region,typical AGI and AGII regions,which further indicated that both of them belonged to the C-class genes of MADS-box gene family.It laid some foundation for the future study of their expression pattern and molecular mechanism in double-flower lily.
引文
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11.李娟,薛庆.2005.拟南芥及水稻转录因子MADS密码子的偏好性比较[J].浙江大学学报:农业与生命科学版,31(5):513-517.
12.Pelaz S,Ditta GS,Baumann E,et al.2000.B and C floral organ identity functions require SEPALLATTA MADS-box genes[J].Nature,405:200-203.
13.孙迎坤.2013.山茶花MADS-box家族A类和C类基因克隆及功能分析[D].中国林业科学研究院博士论文.
14.Tan F.C and Swain S.M.2006.Genetics of flower initiation and development in annual and perennial plants[J].Physiologia Plantarum,128:8-17.
15.Yanfsky M.F.,Ma H,Bowman J.H.1990.The protein encoded by the Arabidopsis homeotic gene agamous resembles transcription factors[J].Nature,346:35-39.
16.郑尚永,郭余龙,肖月华,等.2004.棉花MADS框蛋白基因(GhMADS1)的克隆[J].遗传学报,31(10):1136-1141.
2.Annette B,Gunter T.2003.The major clades of MADS-box genes and their role in the development and evolution of flowering plants[J].Molecular Phylogenetics and Evolution,29:464-489.
3.Benedito VA,Visser PB,van Tuyl JM,et al.2004.Ectopic expression of LLAGX,an AGAMOUS homologue from lily(Lilium longiflorum Thunb.)causes floral homeotic modifications in Arabidopsis[J].Journal of Experimental Botany,55:1391-1399.
4.Coen E.S.Meyerowitz E.M.1991.The war of the whorls:genetics interactions controlling flower development[J].Nature,353:31-37.
5.樊金会,李文卿,董秀春,等.2007.风信子AGL6同源基因在拟南芥中异位表达引起提早开花和器官同源转化[J].中国科学,C辑:生命科学,37(04):466-478.
6.龚霞峰,胡江琴,刘姬艳,等.2009.植物AGAMOUS同源基因的表达调控[J].杭州师范大学学报(自然科学版),8(3).
7.Honma T,Goto K.2001.Complexes of MADS-box proteins are sufficient to convert leaves into floral organs[J].Nature,409:525-529.
8.Jack T.2004.Molecular and genetic mechanisms of floral control[J].The Plant Cell Online,16(suppl 1):S1-S17.
9.Kanno A,Nakada M,Akita Y,et al.2007.Class B gene expression and the modified ABC model in nongrass monocots[J].TSWDevelopment&Embryology,2:17-28.
10.Kramer E.M.,Jaramillo M.A.and DiStilio V.S.2004.Patterns of Gene Duplication and Functional Evolution During the Diversification of the AGAMOUS Subfamily of MADS Box Genes in Angiosperms[J].Genetics,166:1011-1023.
11.李娟,薛庆.2005.拟南芥及水稻转录因子MADS密码子的偏好性比较[J].浙江大学学报:农业与生命科学版,31(5):513-517.
12.Pelaz S,Ditta GS,Baumann E,et al.2000.B and C floral organ identity functions require SEPALLATTA MADS-box genes[J].Nature,405:200-203.
13.孙迎坤.2013.山茶花MADS-box家族A类和C类基因克隆及功能分析[D].中国林业科学研究院博士论文.
14.Tan F.C and Swain S.M.2006.Genetics of flower initiation and development in annual and perennial plants[J].Physiologia Plantarum,128:8-17.
15.Yanfsky M.F.,Ma H,Bowman J.H.1990.The protein encoded by the Arabidopsis homeotic gene agamous resembles transcription factors[J].Nature,346:35-39.
16.郑尚永,郭余龙,肖月华,等.2004.棉花MADS框蛋白基因(GhMADS1)的克隆[J].遗传学报,31(10):1136-1141.