欧洲千里光花发育相关基因SvGLOBOSA功能研究

doi:10.16288/j.yczz.22-055

遗传 ›› 2022, Vol. 44 ›› Issue (6): 521-530.doi: 10.16288/j.yczz.22-055

欧洲千里光花发育相关基因SvGLOBOSA功能研究

栾思楠¹(), 刘乐乐¹, 周佳圆¹, 努尔阿斯娅·伊马木², 崔敏龙¹, 朴春兰¹()

1. 浙江农林大学园艺科学学院,浙江省山区农业协同创新中心,杭州 311300
2. 新疆乌什县农业农村局,阿克苏 843000

收稿日期:2022-03-22 修回日期:2022-05-05 出版日期:2022-06-20 发布日期:2022-06-06
通讯作者: 朴春兰 E-mail:1842239577@qq.com;2488335116@qq.com
作者简介:栾思楠,在读硕士研究生,专业方向：植物花器官发育。E-mail: 1842239577@qq.com
基金资助:
浙江省农业科学院合作研发项目资助编号(H20210384)

Functional analysis of flower development related gene SvGLOBOSA from Senecio vulgaris

Sinan Luan¹(), Lele Liu¹, Jiayuan Zhou¹, Nurasya Imam², Minlong Cui¹, Chunlan Piao¹()

1. Collaborative Innovation Center for Efficient and Green Production of Agriculture in Mountainous Areas of Zhejiang Province, College of Horticulture Science, Zhejiang A&F University, Hangzhou 311300, China
2. Wushi County Bureau of Agriculture and Rural Affairs, Aksu 843000, China

Received:2022-03-22 Revised:2022-05-05 Online:2022-06-20 Published:2022-06-06
Contact: Piao Chunlan E-mail:1842239577@qq.com;2488335116@qq.com
Supported by:
Supported by Cooperative R & D project of Zhejiang Academy of Agricultural Sciences No(H20210384)

摘要/Abstract

摘要：

菊科(Asteraceae)植物独特的头状花序具有重要的观赏与研究价值,其花发育分子机制复杂,目前相关报道较少。本研究对欧洲千里光(Senecio vulgaris)转录组数据进行筛选,初步得到MADS-box基因SvGLOBOSA (SvGLO),并对其功能进行了初步分析。通过生物信息学分析基因结构并预测其功能;qRT-PCR分析了SvGLO在野生型欧洲千里光不同组织中的相对表达量;在龙葵(Solanum nigrum)中过表达SvGLO并进行形态学观察;通过组织学染色分析转基因龙葵子房的组织细胞形态变化。结果表明,SvGLO的开放阅读框(open reading frame, ORF)长度为591 bp,编码196个氨基酸;具有典型的MADS-box和K-box结构域且在C末端含有PI基序(motif) EPFSFRVQPMQPNLQE,属于B类MADS-box基因的PI/GLO亚家族。qRT-PCR结果表明,SvGLO在花序组织中高表达,在营养器官中不表达;同时在过表达龙葵中观察萼片获得花瓣的部分特征,心皮转化为雄蕊状器官,果实发育异常。组织学染色分析结果表明,转基因龙葵的子房壁细胞形态和野生型雄蕊的花药壁细胞相似。以上结果表明,欧洲千里光SvGLO可能参与调控花瓣与雄蕊发育。

关键词: 欧洲千里光, SvGLO, 花发育, 功能研究

Abstract:

The unique capitulum of Asteraceae has important ornamental and research value. Few studies have described the complex molecular mechanism of flower development. In this study, SvGLOBOSA(SvGLO), the MADS-box gene of Senecio vulgaris, was identified by screening the transcriptome data, and its function was examined. The gene structure was analyzed and its function was predicted through bioinformatics. The relative expression levels in different tissues of wild-type S. vulgaris were analyzed by qRT-PCR. SvGLO was overexpressed in Solanum nigrum and morphological observations were made. Histological staining was used in analyzing the histological changes in the ovary of transgenic S. nigrum. The results showed that the open reading frame of SvGLO was 591 bp long, encoding 196 amino acids. It has typical MADS-box and K-box domains and contains a PI motif at the C-terminal. SvGLO belongs to the PI/GLO subfamily of class B MADS-box genes. qRT-PCR results showed that SvGLO was highly expressed in inflorescence tissues but not in vegetative organs. In SvGLO-overexpressed S. nigrum, the sepals showed some characteristics of petals, carpels transformed into stamen-like organs, and fruit development was abnormal. Histological staining revealed that the morphology of ovary wall cells of transgenic S. nigrum was similar to that of anther wall cells of the stamen of wild-type S. vulgaris. Therefore, SvGLO may be involved in the regulation of petal and stamen development in S. vulgaris.

Key words: S. vulgaris, SvGLO, flower development, functional research

栾思楠, 刘乐乐, 周佳圆, 努尔阿斯娅·伊马木, 崔敏龙, 朴春兰. 欧洲千里光花发育相关基因SvGLOBOSA功能研究[J]. 遗传, 2022, 44(6): 521-530.

Sinan Luan, Lele Liu, Jiayuan Zhou, Nurasya Imam, Minlong Cui, Chunlan Piao. Functional analysis of flower development related gene SvGLOBOSA from Senecio vulgaris[J]. Hereditas(Beijing), 2022, 44(6): 521-530.

图/表 8

表1

图1

图2

图3

图4

图5

图6

图7

参考文献 34

[1]	Chen KL, Piao CL, Hao YM, Feng LJ, Zhou JY, Luan SN, Liu LL, Li FF, Yuan SM, Cui ML. Cloning and functional analysis ofCYCLOIDEA(CYC)-like SvRAY1 gene from Senecio vulgaris. Journal of Zhejiang A&F University, 2021, 38(6):1153-1160.
	陈柯俐, 朴春兰, 郝燕敏, 冯丽君, 周佳圆, 栾思楠, 刘乐乐, 李菲菲, 袁思明, 崔敏龙. 欧洲千里光CYCLOIDEA (CYC)类SvRAY1基因的克隆及功能分析. 浙江农林大学学报, 2021, 38(6):1153-1160.
[2]	Michaels SD, Ditta G, Gustafson-Brown C, Pelaz S, Yanofsky M, Amasino RM. AGL24 acts as a promoter of flowering in Arabidopsis and is positively regulated by vernalization. Plant J, 2003, 33(5):867-874. pmid: 12609028
[3]	Rong DQ, Fan YX, Qiao YM. A newly recorded of semi-escaping introduction in Hebei province, China. Seed, 2016, 35(10):65-66.
	荣冬青, 樊英鑫, 乔永明. 河北省外来逸生种子植物——欧洲千里光. 种子, 2016, 35(10):65-66.
[4]	Yanofsky MF, Ma H, Bowman JL, Drews GN, Feldmann KA, Meyerowitz EM. The protein encoded by the Arabidopsis homeotic gene AGAMOUS resembles transcription factors. Nature, 1990, 346(6279):35-39. doi: 10.1038/346035a0
[5]	Sommer H, Beltrán JP, Huijser P, Pape H, Lönnig WE, Saedler H, Schwarz-Sommer Z. Deficiens, a homeotic gene involved in the control of flower morphogenesis in Antirrhinum majus: the protein shows homology to transcription factors. EMBO J, 1990, 9(3):605-613. doi: 10.1002/j.1460-2075.1990.tb08152.x pmid: 1968830
[6]	Coen ES, Meyerowitz EM. The war of the whorls: genetic interactions controlling flower development. Nature, 1991, 353(6339):31-37. doi: 10.1038/353031a0
[7]	Liu JH, Xu BY, Zhang J, Jin ZQ. The interaction of MADS-box transcription factors and manipulating fruit development and ripening. Hereditas(Beijing), 2010, 32(9):893-902.
	刘菊华, 徐碧玉, 张静, 金志强. MADS-box转录因子的相互作用及对果实发育和成熟的调控. 遗传, 2010, 32(9):893-902.
[8]	Riechmann JL, Krizek BA, Meyerowitz EM. Dimerization specificity of Arabidopsis MADS domain homeotic proteins APETALA1, APETALA3, PISTILLATA, and AGAMOUS. Proc Natl Acad Sci USA, 1996, 93(10):4793-4798. doi: 10.1073/pnas.93.10.4793
[9]	Wang YS, Zhang JL, Hu ZL, Guo XH, Tian SB, Chen GP. Genome-Wide analysis of the MADS-box transcription factor family in Solanum lycopersicum. Int J Mol Sci, 2019, 20(12):2961. doi: 10.3390/ijms20122961
[10]	Zhang M, Huang H, Dai SL. Isolation and expression analysis of proline metabolism-related genes in Chrysanthemum lavandulifolium. Gene, 2014, 537(2):203-213. doi: 10.1016/j.gene.2014.01.002 pmid: 24434369
[11]	Parenicová L, de Folter S, Kieffer M, Horner DS, Favalli C, Busscher J, Cook HE, Ingram RM, Kater MM, Davies B, Angenent GC, Colombo L. Molecular and phylogenetic analyses of the complete MADS-box transcription factor family in Arabidopsis: new openings to the MADS world. Plant Cell, 2003, 15(7):1538-1551. doi: 10.1105/tpc.011544 pmid: 12837945
[12]	Becker A, Theissen G. The major clades of MADS-box genes and their role in the development and evolution of flowering plants. Mol Phylogenet Evol, 2003, 29(3):464-489. doi: 10.1016/S1055-7903(03)00207-0
[13]	Xia Y, Shi M, Chen WW, Hu RQ, Jing DL, Wu D, Wang SM, Li QF, Deng HH, Guo QG, Liang GL. Expression pattern and functional characterization of PISTILLATA ortholog associated with the formation of petaloid sepals in double-flower Eriobotrya japonica (Rosaceae). Front Plant Sci, 2020, 10:1685. doi: 10.3389/fpls.2019.01685
[14]	Davies B, Di Rosa A, Eneva T, Saedler H, Sommer H. Alteration of tobacco floral organ identity by expression of combinations ofAntirrhinum MADS-box genes. Plant J, 1996, 10(4):663-677. pmid: 8893543
[15]	Vandenbussche M, Zethof J, Royaert S, Weterings K, Gerats T. The duplicated B-class heterodimer model: whorl-specific effects and complex genetic interactions in Petunia hybrida flower development. Plant Cell, 2004, 16(3):741-754. pmid: 14973163
[16]	Sasaki K, Aida R, Yamaguchi H, Shikata M, Niki T, Nishijima T, Ohtsubo N. Functional divergence within class B MADS-box genes TfGLO and TfDEF in Torenia fournieri Lind. Mol Genet Genomics, 2010, 284(5):399-414. doi: 10.1007/s00438-010-0574-z
[17]	Sasaki K, Yamaguchi H, Nakayama M, Aida R, Ohtsubo N. Co-modification of class B genes TfDEF and TfGLO in Torenia fournieri Lind. alters both flower morphology and inflorescence architecture. Plant Mol Biol, 2014, 86(3):319-334. doi: 10.1007/s11103-014-0231-8
[18]	Park S, Lee E, Heo J, Kim DH, Chun HJ, Kim MC, Bang WY, Lee YK, Park SJ. Rapid generation of transgenic and gene-edited Solanum nigrum plants using Agrobacterium- mediated transformation. Plant Biotechnol Rep, 2020, 14:497-504. doi: 10.1007/s11816-020-00616-7
[19]	Yu D, Kotilainen M, Pöllänen E, Mehto M, Elomaa P, Helariutta Y, Albert VA, Teeri TH. Organ identity genes and modified patterns of flower development in Gerbera hybrida(Asteraceae). Plant J, 1999, 17(1):51-62. pmid: 10069067
[20]	Ai Y, Zhang CL, Sun YL, Wang WN, He YH, Bao MZ. Characterization and functional analysis of five MADS- Box B Class genes related to floral organ identification in Tagetes erecta. PLoS One, 2017, 12(1):e0169777. doi: 10.1371/journal.pone.0169777
[21]	Goto K, Meyerowitz EM. Function and regulation of the Arabidopsis floral homeotic gene PISTILLATA. Genes Dev, 1994, 8(13):1548-1560. doi: 10.1101/gad.8.13.1548
[22]	Nakatsuka T, Saito M, Nishihara M. Functional characterization of duplicated B-class MADS-box genes in Japanese gentian. Plant Cell Rep, 2016, 35(4):895-904. doi: 10.1007/s00299-015-1930-6 pmid: 26769577
[23]	Zeng ZH, Chen S, Xu MR, Wang M, Chen ZH, Wang LL, Pang JL. Cloning, expression, and tobacco overexpression analyses of a PISTILLATA/GLOBOSA-like(OfGLO1) gene from Osmanthus fragrans. Genes (Basel), 2021, 12(11):1748. doi: 10.3390/genes12111748
[24]	Tröbner W, Ramirez L, Motte P, Hue I, Huijser P, Lönnig WE, Saedler H, Sommer H, Schwarz-Sommer Z. GLOBOSA: a homeotic gene which interacts with DEFICIENS in the control of Antirrhinum floral organogenesis. EMBO J, 1992, 11(13):4693-4704. doi: 10.1002/j.1460-2075.1992.tb05574.x pmid: 1361166
[25]	Wang BQ, Liu J, Chu L, Jing X, Wang HD, Guo J, Yi B. Exogenous promoter triggers APETALA3 silencing through RNA-directed DNA methylation pathway in Arabidopsis. Int J Mol Sci, 2019, 20(18):4478. doi: 10.3390/ijms20184478
[26]	Matsunaga S, Isono E, Kejnovsky E, Vyskot B, Dolezel J, Kawano S, Charlesworth D. Duplicative transfer of a MADS box gene to a plant Y chromosome. Mol Biol Evol, 2003, 20(7):1062-1069. doi: 10.1093/molbev/msg114
[27]	Shchennikova AV, Shulga OA, Skryabin KG. Ectopic expression of the homeotic MADS-Box gene HAM31 (Helianthus annuus L.) in transgenic plants Nicotiana tabacum L. affects the gynoecium identity. Dokl Biochem Biophys, 2018, 483(1):363-368. doi: 10.1134/S1607672918060182 pmid: 30607740
[28]	Chen KL. Study on the function of CYCLOIDEA (CYC) SvCYC12 and AGAMUS-LIKE 43 (SvAGL43) genes in Asteraceae [Dissertation]. Zhejiang A&F University, 2021.
	陈柯俐. 菊科欧洲千里光CYCLOIDEA(CYC)类 SvCYC12及AGAMOUS-LIKE 43(SvAGL43)基因的功能研究[学位论文]. 浙江农林大学, 2021.
[29]	Hao YM, Chen KL, Feng LJ, Li FF, Cui ML, Piao CL. Cloning and functional analysis of SvAPETALA1 in Senecio vulgaris. Journal of Zhejiang A&F University, 2022, doi: 10.11833/j.issn.2095-0756.20210651. doi: 10.11833/j.issn.2095-0756.20210651
	郝燕敏, 陈柯俐, 冯丽君, 李菲菲, 崔敏龙, 朴春兰. 欧洲千里光 SvAPETALA1 基因的克隆及功能分析. 浙江农林大学学报, 2022, doi: 10.11833/j.issn.2095-0756.20210651. doi: 10.11833/j.issn.2095-0756.20210651
[30]	Kumar S, Stecher G, Li M, Knyaz C, Tamura K. MEGA X: molecular evolutionary genetics analysis across computing platforms. Mol Biol Evol, 2018, 35(6):1547-1549. doi: 10.1093/molbev/msy096
[31]	Letunic I, Bork P. Interactive Tree Of Life (iTOL) v5: an online tool for phylogenetic tree display and annotation. Nucleic Acids Res, 2021, 49(W1):W293-W296. doi: 10.1093/nar/gkab301
[32]	Wu JG. Cloning and functional study of an R2R3-MYB SnMYB11 involved in anthocyanin synthesis in Solanum nigrum L.[Dissertation]. Zhejiang A&F University, 2021.
	吴金国. 龙葵中花青素合成相关R2R3-MYB基因SnMYB11克隆与功能研究[学位论文]. 浙江农林大学, 2021.
[33]	Poupin MJ, Federici F, Medina C, Matus JT, Timmermann T, Arce-Johnson P. Isolation of the three grape sub-lineages of B-class MADS-box TM6, PISTILLATA and APETALA3 genes which are differentially expressed during flower and fruit development. Gene, 2007, 404(1-2):10-24. doi: 10.1016/j.gene.2007.08.005
[34]	Lou H. Functional analysis of MADS-box gene related to flower organ development in Allium sativum L. [Dissertation]. Northeast Forestry University, 2019.
	娄虎. 大蒜花器官发育相关MADS-box基因的功能分析[学位论文]. 东北林业大学, 2019.

编辑推荐

Metrics

www.chinagene.cn
备案号：京ICP备09063187号-4
总访问:,今日访问:,当前在线:

引物名称	引物序列(5′→3′)	用途
SvGLO	F: ATGGGTAGAGGAAAGATAGA	DNA鉴定
SvGLO	R: TCACATCCTCTCTTGCAAGTTA	DNA鉴定
NPT Ⅱ	F: AGATGGATTGCACGCAGGTTC	DNA鉴定
NPT Ⅱ	R: GAGGTCGAATGGGCAGGTAG	DNA鉴定
qSvGLO	F: CTTGAAAATGGGCTCACCAACA	qRT-PCR
qSvGLO	R: TCGTGACCTTGATAATCTCCCAAT	qRT-PCR
qSn18s	F: TTCTGTCGGCGATGCGTCC	qRT-PCR
qSn18s	R: GCCTCAAACTTCCGCGGCCT	qRT-PCR
qSv18s	F: ATAGCAGAACGACCTGTGAA	qRT-PCR
qSv18s	R: GAAGCAAGATCCAACGCAAT	qRT-PCR

欧洲千里光花发育相关基因SvGLOBOSA功能研究

Functional analysis of flower development related gene SvGLOBOSA from Senecio vulgaris

RichHTML

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

图/表 8

参考文献 34

相关文章 6

编辑推荐

Metrics

[1]	李菲菲, 郝燕敏, 崔敏龙, 朴春兰. 金鱼草RADIALIS-like 1基因克隆与功能研究[J]. 遗传, 2023, 45(6): 526-535.
[2]	郭文雅,崔艳梅,王婷婷,喻德跃,黄方. 野生大豆花发育相关基因GsLFY的功能研究[J]. 遗传, 2017, 39(1): 56-65.
[3]	周坤, 张今今. 植物中的NO及其对花发育的调节[J]. 遗传, 2014, 36(7): 661-668.
[4]	张向前，邹金松，朱海涛，李晓燕，曾瑞珍. 水稻早熟多子房突变体fon5的遗传分析和基因定位[J]. 遗传, 2008, 30(10): 1349-1355.
[5]	袁晓萌，周云涛，张红岩，薛华，周琳，赵云. 甘蓝型油菜小核糖核蛋白BnSmD1编码区全长cDNA 的克隆与表达分析[J]. 遗传, 2007, 29(12): 1525-1528.
[6]	王利琳，梁海曼，庞基良，朱睦元. 拟南芥LEAFY基因在花发育中的网络调控及其生物学功能[J]. 遗传, 2004, 26(1): 137-142.