低温短日照诱导五叶草莓成花诱导的机理研究

doi:10.3969/j.issn.1004-1524.2022.08.10

浙江农业学报 ›› 2022, Vol. 34 ›› Issue (8): 1661-1668.DOI: 10.3969/j.issn.1004-1524.2022.08.10

低温短日照诱导五叶草莓成花诱导的机理研究

向淅(), 王思悦, 蒲俊宏, 唐雯璐, 陈清^*()

四川农业大学园艺学院,四川成都 611130

收稿日期:2021-02-24 出版日期:2022-08-25 发布日期:2022-08-26
作者简介:*陈清,E-mail: supnovel@sicau.edu.cn
向淅(1999—),女,四川仁寿人,主要从事果树种质资源与遗传育种研究。E-mail: xiangxi0727@foxmail.com
通讯作者: 陈清
基金资助:
国家级大学生创新训练计划(201910626030);国家自然科学基金(31972387)

Flowering transition of Fragaria pentaphylla under low-temperature and short-day conditions

XIANG Xi(), WANG Siyue, PU Junhong, TANG Wenlu, CHEN Qing^*()

College of Horticulture, Sichuan Agricultural University, Chengdu 611130, China

Received:2021-02-24 Online:2022-08-25 Published:2022-08-26
Contact: CHEN Qing

摘要/Abstract

摘要：

光周期和温度是影响植物成花诱导的两个主要因素。为研究短日照和低温共同处理对五叶草莓(Fragaria pentaphylla)成花诱导的影响,我们分析了CONSTANS(FpCO)、FLOWERING LOCUS T(FpFT)、LEAFY(FpLFY)、APETALA1(FpAP1)4种重要成花基因在叶片、茎尖分生组织、果实不同发育阶段以及在低温(15 ℃)短日照(8 h·d^-1)下诱导不同时间点的表达情况,同时,剥离处理不同时间点的茎尖进行花芽分化进程的石蜡切片观测。结果表明:FpFT1、FpLFY3、FpCO和FpAP1是草莓中的关键成花因子。FpCO基因于处理28 d后进入表达高峰,随后表达量大幅下降。FpFT1基因自42 d起整体表达量较高,之后虽呈现下降趋势,但维持较高表达水平;FpAP1与FpLFY3基因表达趋势相似,在第6周进入表达高峰,达到高峰后基因表达水平下降;五叶草莓在此条件下处理28 d后启动成花诱导,42 d已完成了由营养生长到生殖生长的转变过程。该结果与切片观察的形态分化过程一致。

关键词: 五叶草莓, 成花基因, 生物信息学分析, 表达定量, 石蜡组织切片

Abstract:

Photoperiod and temperature are the two main factors that affect the flowering induction in plants. In order to study the effect of the combination of short-day and low temperature treatments on the flowering induction of Fragaria pentaphylla, four important determinants CONSTANS (FpCO), FLOWERING LOCUS T (FpFT), LEAFY (FpLFY), and APETALA1 (FpAP1) were analyzed. Gene expressions in different developmental stages of leaves, stem apex meristems, fruits, and after induction at different time points under low temperature (15 ℃) and short day (8 h·d^-1) were analyzed. At the same time, the apex meristem at different time points was stripped, fixed and paraffin embedded for microscopic observation of fluorescence differentiation process. The results showed that: FpFT1, FpLFY3, FpCO and FpAP1 were the key flower-forming factors in wild strawberries. The FpCO gene reached the peak of expression after 28 d of treatment, and then the expression level dropped sharply. Since 42 d, the overall expression level of FpFT1 gene was relatively high, after which it showed a downward trend, but maintained a high expression level. The FpAP1 and FpLFY3 gene expression trends were similar, both peaking at 42 d, and decreasing after the peak. These results implied that Fragaria pentaphylla started the flower induction process after four weeks of treatment under the tested condition and finished the transition process after 42 d. These results were consistent with the morphological differentiation process observed by slices.

Key words: Fragaria pentaphylla, floral gene, bioinformatic analysis, gene quantification, paraffin tissue section

中图分类号:

S668.4

向淅, 王思悦, 蒲俊宏, 唐雯璐, 陈清. 低温短日照诱导五叶草莓成花诱导的机理研究[J]. 浙江农业学报, 2022, 34(8): 1661-1668.

XIANG Xi, WANG Siyue, PU Junhong, TANG Wenlu, CHEN Qing. Flowering transition of Fragaria pentaphylla under low-temperature and short-day conditions[J]. Acta Agriculturae Zhejiangensis, 2022, 34(8): 1661-1668.

图/表 4

表1 用于实时荧光定量的引物

Table 1 Primers for real-time quantitative PCR

引物名称 Primer name	序列 Sequences(5'-3')
FpActin 2-F	GCTAATCGTGAGAAGATGAC
FpActin 2-R	AGCACAATACCAGTAGTACG
FpAP1-F	GCACGAGAAGAATGTAGC
FpAP1-R	GTTCCTCCTCACCTGAAG
FpLFY3-F	TGATGATGATGAGGATGAAGA
FpLFY3-R	CGTAGAGATGGAAGAGGTAAT
FpCO-F	AACAACAACGGATTCTTCTT
FpCO-R	TGGTCATTAGTAGCAGTAGT
FpFT1-F	TCTCAGGGTGACTTACACTTCT
FpFT1-R	GTTGGCCGTGGACTTTCATA

图1 四种成花关键基因生物信息学分析 A,成花基因结构分析;B,成花基因在茎部分生组织中的表达量;C,五叶草莓果实发育3个阶段中成花基因表达量热图。

Fig.1 Bioinformatics analysis of the four key genes for flowering initiation A, Analysis of the structure of flowering genes; B, The expression level of floral genes in the Fragaria vesca shoot meristematic tissues; C, Heat map of floral gene expression in three stages of Fragaria pentaphylla fruit development.

图2 短日照低温共同处理不同时间点四种成花基因在叶片中的相对表达量

Fig.2 The relative expression levels of the four flowering genes in Fragaria pentaphylla after different time duration under the short-day and low-temperature treatment

图3 五叶草莓的花芽分化进程 A,未分化时期;B,花序分化期;C,顶花芽花萼分化期;D,雌蕊分化期。标尺:500 μm。

Fig.3 Flower bud differentiation process of Fragaria pentaphylla A, Undifferentiated period; B, Inflorescence differentiation period; C, Apical flower bud calyx differentiation period; D, Pistil differentiation period. Scale bars: 500 μm.

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低温短日照诱导五叶草莓成花诱导的机理研究

Flowering transition of Fragaria pentaphylla under low-temperature and short-day conditions

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