摘要
【目的】探究向日葵(Helianthus annuus L.)HaGH3家族成员的理化特征和表达分析,以及其在向日葵花发育中的作用,为向日葵品种选育提供参考。【方法】基于向日葵基因组和转录组数据,利用生物信息学方法筛选鉴定HaGH3家族成员,并对蛋白理化性质、染色体定位、系统进化关系、保守基序、顺式作用元件和组织表达特征等进行分析。利用RT-qPCR分析HaGH3家族成员在野生型向日葵及其花分生组织转变与花器官发生缺陷突变体cb1中St2‒St8时期的表达差异,并对其在St5和St6时期不同花器官中的表达情况进行分析。【结果】向日葵HaGH3基因家族包含19个成员(HaGH3.1‒HaGH3.19),分散分布在12条染色体上,分属于2个亚家族,同一亚家族的成员具有相同的蛋白保守结构域。转录组数据分析显示,HaGH3.1、HaGH3.2、HaGH3.3、HaGH3.4、HaGH3.5、HaGH3.8、HaGH3.11、HaGH3.12、HaGH3.13、HaGH3.14和HaGH3.18等11个基因在向日葵花发育特定时期高水平表达。基于RT-qPCR的表达分析,显示HaGH3.3、HaGH3.4、HaGH3.8、HaGH3.13和HaGH3.14在WT的St2时期表达水平达到峰值,且其表达水平在WT和cb1中存在显著差异;HaGH3.1、HaGH3.2、HaGH3.5、HaGH3.12和HaGH3.18在WT花发育后期(St5‒St6时期)高水平表达,且具有较强的花器官特异性。顺式作用元件分析显示,11个花发育相关HaGH3基因启动子区域内含有大量生长素和赤霉素响应元件和MIKC_MADS识别元件。进一步分析结果显示,HaGH3.8、HaGH3.14和HaGH3.18受到ARF介导的生长素信号调节,HaGH3.1和HaGH3.8受到MYB和bZIP等赤霉素信号相关转录因子的调控,HaGH3.2、HaGH3.8、HaGH3.13、HaGH3.14和HaGH3.18受到花同源异型转录因子MIKC_MADS的调控。【结论】向日葵HaGH3.1和HaGH3.3等11个成员在向日葵花生长发育的调控中发挥重要作用,其中HaGH3.1、HaGH3.2、HaGH3.8、HaGH3.13、HaGH3.14和HaGH3.18在调控花发育过程中,受到生长素或赤霉素信号相关转录因子以及MIKC_MADS成员的调控。
【Objective】Physicochemical property and expression feature of HaGH3 genes were analyzed to explore their functions in the floral development in sunflower(Helianthus annuus L.),and discover some molecular clues for sunflower breeding.【Method】Bioinformatics methods were adapted to identify the HaGH3 candidates based on the genome and transcriptome data of sunflower,and to analyze their physicochemical properties,chromosome localization,phylogenetic relationship,conserved motif,cis-acting elements,and tissue-expressing pattern.Subsequently,RT-qPCR was used to analyze the expression differences of HaGH3 genes between WT sunflower and cb1(crazy broccoli 1)mutant in the phases of reproductive development(from St2 to St8),and to measure the expressions of HaGH3 genes in different flower organs of WT sunflower at St5 and St6 phases.【Result】The 19 members of HaGH3 gene family were in the sunflower genome,and named as HaGH3.1 to HaGH3.19,located on 12 chromosomes and categorized into two distinct subfamilies,and all members of the HaGH3 genes belonging to one subfamily possessed the same conserved domains.Transcription analysis indicated that 11 members presented the high expressions at the specific phase of flower development respectively,including HaGH3.1,HaGH3.2,HaGH3.3,HaGH3.4,HaGH3.5,HaGH3.8,HaGH3.11,HaGH3.12,HaGH3.13,HaGH3.14,and HaGH3.18.RT-qPCR analysis showed that the expressions of HaGH3.3,HaGH3.4,HaGH3.8,HaGH3.13,and HaGH3.14 reached their peak values in the St2 of WT,with significant differences between WT and cb1.HaGH3.1,HaGH3.2,HaGH3.5,HaGH3.12,and HaGH3.18 were highly expressed during St5 and St6 in WT,demonstrating obviously floral organs specificity.Cis-acting element analysis showed that a multitude of hormone response elements and MIKC_MADS recognition elements existed in the promoter regions of these 11 HaGH3 genes associated with floral development.Further analysis showed that HaGH3.8,HaGH3.14 and HaGH3.18 were regulated by ARF-mediated auxin signaling,HaGH3.1 and HaGH3.8 were regulated by gibberellin signal-related transcription factors such as MYB and bZIP,and HaGH3.2,HaGH3.8,HaGH3.13,HaGH3.14 and HaGH3.18 were regulated by the transcription factor MIKC_MADS.【Conclusion】Eleven members including HaGH3.1 and HaGH3.3 play important roles in regulating the growth and development of sunflower inflorescences.Among them,HaGH3.1,HaGH3.2,HaGH3.8,HaGH 3.13,HaGH 3.14 and HaGH 3.18 were regulated by auxin or gibberellin signal-related transcription factors and MADS-box members during flower development.
作者
龙林茜
曾银萍
王茜
邓玉萍
葛敏茜
陈彦灼
李鑫娟
杨军
邹建
LONG Lin-xi;ZENG Yin-ping;WANG Qian;DENG Yu-ping;GE Min-qian;CHEN Yan-zhuo;LI Xin-juan;YANG Jun;ZOU Jian(Key Laboratory of Southwest China Wildlife Resource Conservation(Ministry of Education),College of Life Science,China West Normal University,Nanchong 637002)
出处
《生物技术通报》
北大核心
2026年第1期125-138,共14页
Biotechnology Bulletin
基金
四川省中央引导地方‒自由探索项目(2024ZYD0071)
四川省自然科学基金面上项目(2023NSFSC0226)
西华师范大学科研创新团队项目(KCXTD2023-7)。
关键词
向日葵
花发育
cb1突变体
GH3基因家族
表达水平
顺式作用元件
转录因子
转录调控
sunflower
flower development
cb1 mutant
GH3 gene family
expression level
cis-acting element
transcription factor
transcriptional regulation
