甘蓝DELLA基因家族鉴定及其mRNA在嫁接体中的运输分析

doi:10.16288/j.yczz.22-330

遗传 ›› 2023, Vol. 45 ›› Issue (2): 156-164.doi: 10.16288/j.yczz.22-330

甘蓝DELLA基因家族鉴定及其mRNA在嫁接体中的运输分析

李必元(), 赵彦婷, 岳智臣, 雷娟利, 胡齐赞, 陶鹏()

浙江省农业科学院蔬菜研究所，杭州 310021

收稿日期:2022-10-18 修回日期:2022-12-31 出版日期:2023-02-20 发布日期:2023-02-06
基金资助:
浙江省自然科学基金项目(LY21C150006);浙江省农业新品种选育重大科技专项(2021C02065-4-4);国家自然科学基金项目(31601746)

Identification of DELLA gene family in head cabbage and analysis of mRNA transport in the heterograft

Biyuan Li(), Yanting Zhao, Zhichen Yue, Juanli Lei, Qizan Hu, Peng Tao()

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

Received:2022-10-18 Revised:2022-12-31 Published:2023-02-20 Online:2023-02-06
Supported by:
the Natural Science Foundation of Zhejiang Province(LY21C150006);the Grand Science and Technology Special Project of Zhejiang Province(2021C02065-4-4);the National Natural Science Foundation of China(31601746)

摘要/Abstract

摘要：

DELLA基因家族参与植物激素信号转导通路的调控，其中GAI (GA insensitive) mRNA还是植物体内长距离运输的信号分子。在全基因组范围内鉴定甘蓝(Brassica oleracea var. capitata) DELLA基因家族成员并分析mRNA运输特性，可为甘蓝DELLA基因家族的开发应用提供基础数据。本研究利用甘蓝基因组数据和转录组数据，在甘蓝中鉴定了5个DELLA基因家族成员(BoRGA1、BoRGA2、BoRGL1、BoRGL2和BoRGL3)，但甘蓝基因组缺失了GAI基因。采用劈接法将甘蓝(自交系G27)接穗和菜心(Brassica campestris L. ssp. chinensis var. utilis Tsen et Lee)(四九菜心)砧木嫁接在一起，构建异源嫁接体。对砧木菜心花序轴和对应位置的实生苗菜心花序轴(对照)取样进行转录组测序。在甘蓝/菜心异源嫁接体的砧木菜心花序轴的转录组测序文库中分别鉴定到8、9、3、5、1个来自甘蓝BoRGA1、BoRGA2、BoRGL1、BoRGL2和BoRGL3的外源read。甘蓝DELLA家族基因mRNA运输并没有提高砧木菜心中DELLA家族基因的转录表达水平。相关性分析显示，甘蓝DELLA家族基因mRNA运输效率与其自身的序列和接穗甘蓝中的DELLA家族基因的转录表达水平相关。本研究为深入探究甘蓝DELLA家族基因mRNA运输的分子机制奠定了基础。

关键词: 甘蓝, 菜心, 嫁接, DELLA基因, mRNA运输

Abstract:

DELLA gene family is involved in the regulation of signal transduction of plant hormones. mRNAs of GA insensitive (GAI), the member of DELLA gene family, are also signaling molecules of long-distance transport in plants. Genome-wide identification and mRNA transport analysis of the members of DELLA gene family in head cabbage (Brassica oleracea var. capitata) can provide basic data for their application in head cabbage. In this study, five members of DELLA gene family (BoRGA1, BoRGA2, BoRGL1, BoRGL2, and BoRGL3) were identified in head cabbage using genome and transcriptome data. However, head cabbage lacked a GAI gene in its genome. The scion (head cabbage, inbred line G27) and the rootstock Chinese flowering cabbage (Brassica campestris L. ssp. chinensis var. utilis Tsen et Lee) (sijiucaixin) were cleft-grafted together to produce the heterograft. Inflorescence stem of the rootstock and the corresponding inflorescence stem in Chinese flowering cabbage seedlings (as controls) were purified and analyzed with transcriptome sequencing. The total of 8, 9, 3, 5, and 1 exogenous read(s), derived respectively from BoRGA1, BoRGA2, BoRGL1, BoRGL2, and BoRGL3, were identified in the transcriptomes of the rootstocks. Nevertheless, mRNA transport of DELLA family genes from scion to rootstock did not increase the transcriptional level of the members of DELLA gene family in the rootstocks. Correlation analysis suggested that mRNA transport efficiency of the DELLA family genes was correlated with the sequence and the transcriptional level of the respective DELLA gene in the scion (head cabbage). This study lays the foundation for further investigation on the molecular mechanism of mRNA transport of the members of DELLA gene family in head cabbage.

Key words: head cabbage, Chinese flowering cabbage, grafting, DELLA genes, mRNA transport

李必元, 赵彦婷, 岳智臣, 雷娟利, 胡齐赞, 陶鹏. 甘蓝DELLA基因家族鉴定及其mRNA在嫁接体中的运输分析[J]. 遗传, 2023, 45(2): 156-164.

Biyuan Li, Yanting Zhao, Zhichen Yue, Juanli Lei, Qizan Hu, Peng Tao. Identification of DELLA gene family in head cabbage and analysis of mRNA transport in the heterograft[J]. Hereditas(Beijing), 2023, 45(2): 156-164.

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名称	检索序列(5′→3′)
BoRGA1a	GGTCAGGTCGTCGGAGATGGCTGAGGTT
BoRGA1b	CGAAACCGAATCTGTGGCGGTTAACTCC
BoRGA1c	CTTGTTTGATTCCTTGGAAGGAGCTCCG
BoRGA2a	CTCCTGATACTAGATTCCCCAACCACGA
BoRGA2b	GTCGAATCATAACGGTCCGGTTTTCCTA
BoRGA2c	ACTGTTTGATTCTTTGGAAGGTGTTCCC
BoRGL1a	AGCTCGCTGACGTGCTCGTTAAGCACCT
BoRGL1b	TCGATAACATTCTGTTAACGATTAAGTC
BoRGL1c	CGAACCACAACGGGGCCGTTTTCCTCGA
BoRGL2a	AACATCAAGGGCAAGATGGCTGATGACA
BoRGL2b	AGCGGCGGAGAGTTTATCGGATCTCGAG
BoRGL2c	ATTAAACCGGTCCGGTTCGGTCGAGAAG
BoRGL3a	GCGGATGTTGCACAGAAGCTTGAGCAGC
BoRGL3b	GAATTCAAAGGTCTTACCGTTGAGAGTT
BoRGL3c	CGACCCGGTTCGATCGAAAAGCTGTTAG

名称	基因号	染色体	拟南芥同源基因	氨基酸数量	分子量(kDa)	等电点
BoRGA1	BolC07g031500.2J	C07	RGA (AT2G01570)	576	62.8	5.52
BoRGA2	BolC09g027430.2J	C09	RGA (AT2G01570)	569	62.4	5.27
BoRGL1	BolC02g022360.2J	C02	RGL1(AT1G66350)	507	55.8	5.37
BoRGL2	BolC05g063590.2J	C05	RGL2 (AT3G03450)	543	59.1	4.73
BoRGL3	BolC09g055750.2J	C09	RGL3 (AT5G17490)	525	57.5	4.84

名称	含外源read的文库数量	可检测到外源read的检索序列数量	外源read的总数	与AtGAI_(998~1771)的序列一致率(%)	在甘蓝/甘蓝嫁接体的茎尖中的表达量(FPKM)
BoRGA1	3	3	8	81.3	133.6
BoRGA2	3	3	9	82.6	147.1
BoRGL1	2	2	3	65.1	20.9
BoRGL2	2	2	5	64.2	23.2
BoRGL3	1	1	1	64.6	12.2

甘蓝DELLA基因家族鉴定及其mRNA在嫁接体中的运输分析

Identification of DELLA gene family in head cabbage and analysis of mRNA transport in the heterograft

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