Subcellular localization and expression analysis of <i>MlSOC1</i> genes during flower bud differentiation period in <i>Magnolia liliflora</i>

doi:10.3785/j.issn.1008-9209.2019.09.291

Journal of Zhejiang University (Agriculture and Life Sciences)

2020, Vol. 46

Issue (4): 407-416 DOI: 10.3785/j.issn.1008-9209.2019.09.291

Horticulture

Subcellular localization and expression analysis of MlSOC1 genes during flower bud differentiation period in Magnolia liliflora

Lingjuan XUAN(

),Shaoyu CHENG,Mengyi DAI,Zhuowei WANG,Yamei SHEN(

)

College of Landscape Architecture, Zhejiang A & F University, Hangzhou 311300, China

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Abstract

In order to clarify the mechanism of SOC1 inthe process of flower formation in Magnolia liliflora, two SOC1 genes were screened out based on the transcriptome datain M. sinostellata, and the complete coding regions of these two SOC1 genes were obtained by homologous gene cloning. The bioinformatics software was used to analyze the information of these two genes, and the expression patterns in different flower bud differentiation periods were verified by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). The results showed that the length of MlSOC1-1 was 666 bp, which encoding 221 amino acids, and the length of MlSOC1-2 was 654 bp, which encoding 217 amino acids. Both of these two MlSOC1 genescontained highly conservative SOC1 motifs, which indicated that the genes belong to the SOC1/TM3 subfamily. Phylogenetic tree analysis showed that MlSOC1-1 was closely related to SOC1 homologous gene of M. praecocissima, while MlSOC1-2 was closely related to SOC1 homologous gene of M. virginiana. Subcellular localization experiments revealed that both genes were located on the nucleus. Besides, the results of expression analysis showed that compared with MlSOC1-2, in addition to participating in the flower transformation of the flower bud in M. liliflora, MlSOC1-1 might also play a role in the flower opening and the synthesis of flower organs. These results show that there are differences in the roles of these two MlSOC1 genes in flower bud differentiation, which provides a theoretical basis for further study of the role of MlSOC1 genes in flower formation of M. liliflora.

Key words： Magnolia liliflora MlSOC1 gene flower bud differentiation subcellular localization expression analysis

Received: 29 September 2019 Published: 11 September 2020

CLC:

Q 785

Corresponding Authors: Yamei SHEN E-mail: 1075855127@qq.com;yameishen@zafu.edu.cn

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Cite this article:

Lingjuan XUAN,Shaoyu CHENG,Mengyi DAI,Zhuowei WANG,Yamei SHEN. Subcellular localization and expression analysis of MlSOC1 genes during flower bud differentiation period in Magnolia liliflora. Journal of Zhejiang University (Agriculture and Life Sciences), 2020, 46(4): 407-416.

URL:

http://www.zjujournals.com/agr/10.3785/j.issn.1008-9209.2019.09.291 OR http://www.zjujournals.com/agr/Y2020/V46/I4/407

紫玉兰MlSOC1基因亚细胞定位及花芽分化时期的表达分析

为了探究SOC1基因在紫玉兰（Magnolia liliflora）成花过程中的作用，通过对景宁木兰转录组数据（NCBI数据库编号：SRP129819）进行筛选，利用同源克隆的方法得到2个SOC1基因的编码区序列；运用在线生物信息学分析工具对这2个基因进行序列分析，并结合实时荧光定量聚合酶链式反应（real-time fluorescence quantitative polymerase chain reaction, qRT-PCR）分析这2个基因在紫玉兰不同花芽分化期的表达模式。将获得的2个SOC1基因分别命名为MlSOC1-1和MlSOC1-2，其中：MlSOC1-1长度为666 bp，编码221个氨基酸；MlSOC1-2长度为654 bp，编码217个氨基酸。2个SOC1基因都具有保守的SOC1基序，属于SOC1/TM3亚家族基因。系统进化树分析结果显示，MlSOC1-1与皱叶木兰（Magnolia praecocissima）中的SOC1同源基因亲缘关系最近，而MlSOC1-2与北美木兰（M. virginiana）中的SOC1同源基因亲缘关系最近，2个基因都与木兰科木兰属植物的遗传距离最近。亚细胞定位试验结果发现，MlSOC1-1和MlSOC1-2都被定位于细胞核上。表达分析结果表明：MlSOC1-1与MlSOC1-2基因相比，前者除了参与紫玉兰芽的成花转变外，还有可能对紫玉兰花器官合成有一定作用。通过对紫玉兰2个MlSOC1基因的研究，发现2个MlSOC1基因在花芽分化过程中的作用存在差异，这为进一步研究MlSOC1基因在紫玉兰成花过程中的作用提供了理论基础。

关键词： 紫玉兰, MlSOC1基因, 花芽分化, 亚细胞定位, 表达分析

Table 1 Primers for cloning, expression analysis and vector construction of MlSOC1 in M. liliflora

Fig. 1 Process of flower bud differentiation in M. lilifloraA-F. External morphological characteristics of bud during flower bud differentiation; G-L. Internal morphological characteristics of bud during flower bud differentiation. Le: Leaf; Fl: Flower primordium; Sc: Scale; Se: Sepal primordium; Pe: Petal primordium; St: Stamen primordium; Te: Tepal; Pi: Pistil primordium.

Fig. 2 Nucleotide sequence alignment of MlSOC1-1 and MlSOC1-2 genes

Fig. 3 Sequence alignment of MlSOC1-1 and MlSOC1-2 with SOC1 amino acid sequences of other speciesACV88635.1: Magnolia virginiana; XP_023900830.1: Quercus suber; NP_001267909.1: Vitis vinifera; AHI_85950.1: Carya cathayensis; NP_001236377.1: Glycine max.

Fig. 4 Phylogenetic tree betweenMlSOC1-1/2proteins and SOC1 proteins of other species

Fig. 5 Schematic diagram of vector construction1: pCMBIA1302-MlSOC1-1/2 vector; 2: pCMBIA1302 vector.

Fig. 6 Subcellular localization of MlSOC1-1/2 proteinsA-C. Subcellular localization of GFP protein; D-F. Subcellular localization of MlSOC1-1 protein; G-I. Subcellular localization of MlSOC1-2 protein.

Fig. 7 Expression analysis of MlSOC1-1 and MlSOC1-2 of leaves and flower buds in different flower bud differentiation periodsA-B. Leaf; C-D. Flower bud. S₁: Undifferentiation period; S₂: Early differentiation period; S₃: Sepal primordium differentiation period; S₄: Petal primordium differentiation period; S₅: Stamen primordium differentiation period; S₆: Pistil primordium differentiation period. Single asterisk (*) indicates significant differences at the 0.05 probability level; double asterisks (**) indicate highly significant differences at the 0.01 probability level; triple asterisks (***) indicate extremely high significant differences at the 0.001 probability level.


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