基于转录组分析内源激素在调控枇杷花发育进程中的作用

doi:10.3969/j.issn.1004-1524.20230143

浙江农业学报 ›› 2023, Vol. 35 ›› Issue (7): 1648-1661.DOI: 10.3969/j.issn.1004-1524.20230143

基于转录组分析内源激素在调控枇杷花发育进程中的作用

徐红霞¹(), 李晓颖¹, 葛航¹, 朱启轩¹^,², 陈俊伟¹^,^*()

1.浙江省农业科学院园艺研究所,浙江杭州 310021
2.河北科技师范学院园艺科技学院,河北秦皇岛 066600

收稿日期:2023-02-14 出版日期:2023-07-25 发布日期:2023-08-17
作者简介:徐红霞(1980—),女,浙江台州人,博士,副研究员,研究方向为枇杷发育与抗逆机理。E-mail:xuhongxia@zaas.ac.cn
通讯作者: *陈俊伟,E-mail:chenjunwei@zaas.ac.cn
基金资助:
浙江省农科院地方科技合作项目(CA202009);浙江省重大协同推广计划项目(2022XTTGGP02);浙江省农业新品种选育重大科技专项(2021C02066-3)

Transcriptome-based analysis of the role of endogenous hormones in regulating flower development in loquat (Eriobotrya japonica Lindl.)

XU Hongxia¹(), LI Xiaoying¹, GE Hang¹, ZHU Qixuan¹^,², CHEN Junwei¹^,^*()

1. Institute of Horticulture, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China
2. College of Horticultural Science & Technology, Hebei Normal University of Science & Technology, Qinhuangdao 066600, Hebei, China

Received:2023-02-14 Online:2023-07-25 Published:2023-08-17
Contact: CHEN Junwei

摘要/Abstract

摘要：

为研究枇杷(Eriobotrya japonica Lindl.‘Ninghaibai’)花发育机制,以花芽生理分化期(EjS1)、花芽形态分化期(EjS2)、花穗发育期(EjS3)、小花发育期(EjS4)和开花期(EjS5)这5个时期的当年生春梢上顶芽、花芽或花穗为试验材料进行转录组测序分析,筛选出相关性最高的代谢通路。测定花发育过程中生长素(IAA)、脱落酸(ABA)、赤霉素(GA)和玉米素核苷(ZR)等激素水平的变化,并对激素代谢与信号转导相关基因,以及花发育相关MADS-box家族基因的花发育时期特异性表达模式进行分析;同时,通过实时荧光定量PCR(qRT-PCR)对部分花发育相关基因进行验证。结果表明:不同发育时期差异表达基因在植物激素代谢和信号转导途径中显著富集;低含量IAA、GA与高含量ABA有利于枇杷花芽的形成,较高水平GA和ABA能促进枇杷开花;高水平ZR能促进枇杷花芽分化和花穗形成,并可能与花器官分化有关。植物激素代谢和信号转导途径相关基因相互作用调节激素水平变化,并与MADS-box家族基因共同调控枇杷花发育,研究结果有助于阐明内源激素在枇杷花发育进程中的作用机制。

关键词: 枇杷, 花发育, 转录组, 内源激素

Abstract:

To study the flower development mechanism in Eriobotrya japonica Lindl. ‘Ninghaibai’, transcriptome analysis was performed on the apical buds, floral buds, or inflorescences at 5 different stages, including physiological differentiation stage (EjS1), morphological differentiation stage (EjS2), inflorescence emergence and expansion stage (EjS3), single flower development stage (EjS4) and blossoming stage (EjS5). The aim was to identify the highest correlated metabolic pathways associated with the flower developoment. The changes of hormone levels of auxin (IAA), abscisic acid (ABA), gibberellin (GA), and zeatin riboside (ZR) during flower development were determined, and the specific expression patterns of hormone metabolism and signal transduction genes as well as MADS-box family genes were analyzed. Some flower development-related genes were verified by real-time quantitative PCR (qRT-PCR) analysis. The results showed that differentially expressed genes at different developmental stages were significantly enriched in plant hormone metabolism and signal transduction pathways. Low content of IAA and GA and high content of ABA were beneficial to the formation of loquat flower buds, while high level of GA and ABA promoted the flowering of loquat. High levels of ZR promoted flower bud differentiation and panicle formation in loquat, and may be related to flower organ formation. The genes related to plant hormone metabolism and signal transduction pathway interact with each other to regulate hormone levels, and together with MADS-box family genes to regulate the development process of loquat flowers. This study may contribute to clarifying the mechanism of endogenous hormones in the development process of loquat flowers.

Key words: loquat, flower development, transcriptome, endogenous hormone

中图分类号:

S667.3

徐红霞, 李晓颖, 葛航, 朱启轩, 陈俊伟. 基于转录组分析内源激素在调控枇杷花发育进程中的作用[J]. 浙江农业学报, 2023, 35(7): 1648-1661.

XU Hongxia, LI Xiaoying, GE Hang, ZHU Qixuan, CHEN Junwei. Transcriptome-based analysis of the role of endogenous hormones in regulating flower development in loquat (Eriobotrya japonica Lindl.)[J]. Acta Agriculturae Zhejiangensis, 2023, 35(7): 1648-1661.

图/表 9

表1 荧光定量PCR中所用差异表达基因及其引物序列

Table 1 Differentially expressed genes and their primer sequences in qRT-PCR

基因名称 Gene name	基因ID Gene ID	正向引物 Forward primer (5’→3’)	反向引物 Reverse primer(5’→3’)
Ejactin	c255911_g1	GGATTTGCTGGTGATGATGC	CCGTGCTCAATGGGATACTT
SOC1	c131331_g2	TTGGGAAATGGTTTGGAT	CTGCTTTGCACGACACTC
SOC1	c123324_g2	GGTTCATGCCATTGTCTG	TTCTCCAACTGGTGCTCT
LEAFY	c125200_g1	AAGGGAGCACCCGTTCAT	CGCATCTTTGGCTTGTTGA
FT	c132831_g2	GGACGAGTGGTAGGTGAT	TTGGTGGACAACTTGAGAAG
AGL24	c118626_g2	ACCAGCTAAAGCAGAGGA	GGAGCAGGCGGTGACATT
AP1	c125934_g2	GCAAATCCTTGAACGCTAT	GAATCCAGGTCTTCTCCC
AP2	c129345_g1	GAGGACGGATCAGACGAAG	TCACGGAGAACCCGAATAT
SEP	c126115_g3	TTTCTCCTCCTTTCTTCTTTAGC	AAGTTCCTTTGCAGGTATGTTG
TFL	c113629_g1	ATTGGCATCCACAGGTTT	AAGTAGACGGCAGCGACA
CAL	c128672_g1	GAGCATGATCAGGTGCAGGT	GTGTGTGTTTGTAGCAGCGG
FLC	c128866_g4	CGGGCTGATGAAGAAGGC	GCGGTGGAAGGAAGAAGAAA
SVP	c132152_g5	GAGCAAAGGCAGAGGATA	ACAAGATGTCGGAGCAGT

图1 不同发育时期枇杷花的石蜡切片图和表型EjS1,花芽生理分化期;EjS2,花芽形态分化期;EjS3,花穗发育期;EjS4,小花发育期;EjS5,开花期。下同。标尺为200 μm。

Fig.1 Paraffin section and phenotype of loquat flower at different development stages EjS1, Physiological differentiation stage; EjS2, Morphological differentiation stage; EjS3, Inflorescence emergence and expansion stage; EjS4, Single flower development stage; EjS5, Blossoming stage. The same as below. Bar=200 μm.

图2 枇杷花发育进程中激素含量变化柱上无相同字母表示差异显著(P<0.05)。

Fig.2 Changes of phytohormone content in loquat flower development process Data on the bars marked without the same letter indicated significant differences at P<0.05.

表2 Unigene在7个公共数据库的注释情况统计

Table 2 Annotation of assembled unigenes in 7 public databases

注释数据库 Annotation database	Unigene注释数量 Number of annotated unigenes	占比 Percentage/%
NR	115 604	35.80
NT	92 121	28.53
KEGG	52 186	16.16
Swissprot	118 766	36.78
Pfam	118 886	36.82
GO	120 666	37.37
KOG	79 750	24.70

图3 枇杷花发育进程中差异表达基因分析 A,不同发育进程差异表达基因聚类情况;B,EjS2 vs EjS1、EjS3 vs EjS1、EjS4 vs EjS1和EjS5 vs EjS1的差异基因上调和下调表达情况;C,EjS2 vs EjS1、EjS3 vs EjS1、EjS4 vs EjS1和EjS5 vs EjS1比较组之间差异表达基因的韦恩图。

Fig.3 Analysis of the differentially expressed genes during flower development process in loquat A, Hierarchical clustering of the differentially expressed genes in different flower development process; B, Up-and down-regulated DEGs at stages EjS2, EjS3, EjS4, and EjS5 compared with stage EjS1; C, Venn diagram showed the number of DEGs bewteen EjS2 vs EjS1, EjS3 vs EjS1, EjS4 vs EjS1, and EjS5 vs EjS1.

图4 枇杷花发育进程中差异表达基因KEGG代谢通路富集分析

Fig.4 KEGG pathway enrichment of the annotated differentially expressed genes between different developmental stage comparisons

图5 枇杷花发育进程中激素代谢和信号转导相关基因热图分析

Fig.5 Heat map analysis of genes related to hormone metabolism and signal transduction in loquat flower development process

图6 枇杷花发育进程中MADS基因家族成员基因表达热图

Fig.6 Gene expression heat map of MADS gene family members in loquat flower development process

图7 十二个花发育相关基因在不同发育时期的表达丰度

Fig.7 Expression abundance of 12 flower development related genes at different flower development stages

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基于转录组分析内源激素在调控枇杷花发育进程中的作用

Transcriptome-based analysis of the role of endogenous hormones in regulating flower development in loquat (Eriobotrya japonica Lindl.)

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