菜心BcSVP基因mRNA在异源嫁接体中的运输分析

doi:10.3969/j.issn.1004-1524.2021.07.05

浙江农业学报 ›› 2021, Vol. 33 ›› Issue (7): 1192-1198.DOI: 10.3969/j.issn.1004-1524.2021.07.05

菜心BcSVP基因mRNA在异源嫁接体中的运输分析

陶鹏(), 赵彦婷, 岳智臣, 雷娟利, 李必元^*()

浙江省农业科学院蔬菜研究所,浙江杭州 310021

收稿日期:2020-06-28 出版日期:2021-07-25 发布日期:2021-08-06
通讯作者: 李必元
作者简介:*李必元,E-mail: 16061944@qq.com
陶鹏(1984—),男,江西南昌人,博士,副研究员,主要从事十字花科蔬菜育种与分子生物学研究工作。E-mail: taopeng84@163.com
基金资助:
浙江省自然科学基金(LY21C150006);浙江省农业新品种选育重大科技专项(2016C02051-6-2);国家自然科学基金(31601746)

Analysis of mRNA transport of BcSVP of Chinese flowering cabbage in heterograft

TAO Peng(), ZHAO Yanting, YUE Zhichen, LEI Juanli, LI Biyuan^*()

Institute of Vegetables, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:2020-06-28 Online:2021-07-25 Published:2021-08-06
Contact: LI Biyuan

摘要/Abstract

摘要：

为研究菜心BcSVP基因mRNA的长距离运输,本研究将结球甘蓝茎尖作为接穗嫁接到砧木菜心的花序轴上,构建结球甘蓝/菜心异源嫁接体。基于菜心和结球甘蓝的转录组测序的read文库,拼接获得了菜心的BcSVP和结球甘蓝BoSVP基因编码序列和3’UTR序列。利用菜心BcSVP在3’UTR的种间差异序列(CF和CR),分别在异源嫁接(结球甘蓝/菜心嫁接)的接穗结球甘蓝茎尖的转录组测序read文库T01和T02中筛选到2条和1条来自砧木菜心的BcSVP的read。利用结球甘蓝BoSVP在3’UTR的种间差异序列(GF和GR),计算了结球甘蓝内源BoSVP基因的转录表达量,显示异源嫁接和同源嫁接的接穗结球甘蓝茎尖中BoSVP基因的表达量差异不明显。本研究鉴定了菜心BcSVP基因的mRNA在异源嫁接体中的长距离运输,对进一步研究菜心BcSVP基因的mRNA长距离运输机制及其生物学功能奠定了基础。

关键词: 结球甘蓝, 菜心, 嫁接, BcSVP, mRNA运输

Abstract:

For confirming mRNA long-transport of BcSVP gene, shoot apex of head cabbage was grafted on inflorescence stem of Chinese flowering cabbage to construct head cabbage-Chinese flowering cabbage heterograft. Based on transcriptome sequencing read library of Chinese flowering cabbage and head cabbage, coding sequences and 3’UTR sequences of BcSVP and BoSVP gene were obtained by read joining. According to interspecific differential sequences of BcSVP in 3’UTR (CF and CR), The two and one transported BcSVP reads of Chinese flowering cabbage were identified from transcriptome sequencing read libraries T01 and T02 of shoot apexes of head cabbage scions of head cabbage-Chinese flowering cabbage heterograft, respectively. The endogenous transcriptional expression of BoSVP was analyzed based on interspecific differential sequences of BoSVP(GF and GR), indicating the transcriptional expression of BoSVP did not show obvious difference between homograft and heterograft. The study indicated mRNA long-transport of BcSVP in heterograft. It lays the foundation for further study of molecule mechanism and biological function of BcSVP mRNA transport in head cabbage-Chinese flowering cabbage heterograft.

Key words: head cabbage, Chinese flowering cabbage, grafting, BcSVP, mRNA transport

中图分类号:

S635.1

陶鹏, 赵彦婷, 岳智臣, 雷娟利, 李必元. 菜心BcSVP基因mRNA在异源嫁接体中的运输分析[J]. 浙江农业学报, 2021, 33(7): 1192-1198.

TAO Peng, ZHAO Yanting, YUE Zhichen, LEI Juanli, LI Biyuan. Analysis of mRNA transport of BcSVP of Chinese flowering cabbage in heterograft[J]. Acta Agriculturae Zhejiangensis, 2021, 33(7): 1192-1198.

图/表 7

图1 菜心BcSVP与结球甘蓝BoSVP编码序列及比对起始密码子和终止子分别用下划线和方框表示。

Fig.1 Sequence alignment of BcSVP and BoSVP from Chinese flowering cabbage and head cabbage Start codon and stop codon are marked by underline and box.

表1 菜心与结球甘蓝的SVP基因的同源基因、基因定位及其编码蛋白的等电点和相对分子质量

Table 1 Homologous gene and gene location of the SVP genes from Chinese flowering cabbage and head cabbage and isoelectric points and molecular weight of the corresponding coding proteins

基因名称 Gene name	物种 Species	同源基因定位 Homologous gene location	等电点 Isoelectric point	相对分子质量 Molecular weight/ku
BcSVP	菜心Chinese flowering cabbage	Bra030228(Chr04:10192172-10194736)	5.87	27.24
BoSVP	结球甘蓝head cabbage	Bo4g149800(Chr04:40723422-40726294)	5.87	27.29

图2 菜心BcSVP与结球甘蓝BoSVP基因3’UTR序列及比对终止子用方框表示,黑色背景显示的是检索序列GF的位置,而灰色背景显示的是种间差异序列CF的位置。

Fig.2 Sequence alignment of 3’UTR of BcSVP and BoSVP from Chinese flowering cabbage and head cabbage Stop codon was marked by box. The retrieval sequence GF and CF were marked by black background and gray background respectively.

图3 使用CR序列进行检索在异源嫁接体的接穗结球甘蓝茎尖T01和T02文库中检索到的三个潜在的菜心BcSVP read

Fig.3 The three potential BcSVP reads of Chinese cabbage identified from T01 and T02 library of shoot apexes of head cabbage from heterograft by searching with CR sequence

表2 从异源嫁接的接穗结球甘蓝茎尖的T01和T02文库中识别的来自菜心BcSVP基因read的相关信息

Table 2 Related information of three potential BcSVP reads of Chinese cabbage identified from T01 and T02 library of shoot apexes of head cabbage from heterograft by searching with CR sequence

名称 Name	文库 Library	位置 Location	长度 Length/nt	3’UTR中的核苷酸/nt Nucleotides on 3'UTR/nt	与BcSVP差异核苷酸 Differential nucleotides with BcSVP	与BoSVP的差异核苷酸 Differential nucleotides with BoSVP
Read1	T01	34740498	150	149	0	27
Read2	T01	76178478	150	59	1	5
Read3	T02	59545522	150	71	3	7

图4 长距离运输的read与结球甘蓝BoSVP和菜心BcSVP基因的序列比对检索序列CF和CR用黑色背景标记。

Fig.4 Sequence alignment of long-transport reads, BoSVP and BcSVP from head cabbage and Chinese flowering cabbage The retrieval sequence CF and CR were marked by black background.

表3 不同种间差异序列在文库中检索到的数量

Table 3 The retrieval number in different libraries by searching with interspecific differential sequence

序列名称 Sequence name	序列 Sequence (5’-3’)	T01	T02	T03	T04	T05
CF	AGAAGTTCATGGAGTGAGGAGATGCTCTGTAGTAACAAGTG	0	0	0	0	44
CR	CACTTGTTACTACAGAGCATCTCCTCACTCCATGAACTTCT	2	1	0	0	88
GF	AGAAGTTGTGGAGTGAGGAGATACTCTGTGGTAACAAGTG	103	80	86	46	0
GR	CACTTGTTACCACAGAGTATCTCCTCACTCCACAACTTCT	200	185	103	113	0

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菜心BcSVP基因mRNA在异源嫁接体中的运输分析

Analysis of mRNA transport of BcSVP of Chinese flowering cabbage in heterograft

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