首页 > 分享 > 向日葵YABBY基因家族鉴定及表达分析

向日葵YABBY基因家族鉴定及表达分析

花匠小妙招
2024-11-25 20:35

摘要：

【目的】鉴定向日葵中花发育相关的HaYABBY基因，有助于探究其在向日葵花发育过程中的生物学功能和调控机制，为向日葵品种选育提供重要的分子线索。【方法】基于向日葵基因组和转录组数据，使用生物信息学方法，分析向日葵YABBY基因家族成员的染色体定位、系统进化关系、基因结构、保守基序、蛋白结构及理化性质、启动子顺式作用元件和组织表达模式等，并通过RT-qPCR分析HaYABBY基因在WT向日葵和花分生组织决定和花器官分化缺陷突变体cb1中st2-st8时期的表达差异。【结果】在向日葵基因组中鉴定到12个YABBY基因家族成员，分属于4个亚家族，分布在8条染色体上，并且同一亚家族中成员在基因结构和保守基序组成上高度相似。其中8个基因HaYABBY1a、HaYABBY1c、HaYABBY3、HaYABBY4b、HaYABBY5a、HaYABBY5b、HaCRABS CLAWa、HaCRABS CLAWb在向日葵花发育时期特异性高表达。在这8个基因中，HaYABBY3、HaYABBY5a和HaYABBY5d在突变体cb1花发育后期表达上调，且与WT花发育早期水平相当；而HaYABBY4b和HaCRABS CLAWa在cb1中的表达被强烈抑制。顺式作用元件分析显示，HaYABBY均含有多个启动子顺式作用元件，部分元件仅存在于特定基因中。其中，胚乳表达元件存在于HaCRABS CLAWa和HaYABBY4a中；生长素响应元件存在于HaCRABS CLAWa、HaCRABS CLAWb、HaYABBY4a、HaYABBY4b和HaYABBY5c中；分生组织表达元件存在于HaYABBY1a、HaYABBY1c、HaYABBY4b、HaYABBY5a和HaYABBY5d中；赤霉素响应元件存在于HaCRABS CLAWa、HaYABBY5b、HaYABBY5c和HaYABBY5d中；种子特异性调控元件存在于HaYABBY5a、HaYABBY5c和HaYABBY5d中。【结论】HaYABBY3、HaYABBY4b、HaYABBY5a、HaYABBY5d和HaCRABS CLAWa等5个HaYABBY基因在花分生组织的转变和花器官发生过程中可能发挥重要作用，它们在花分生组织转变和花器官发生调控中可能受到生长素响应因子ARF、赤霉素响应因子MYB以及MADS-box等关键因子调节。

关键词: 向日葵, YABBY基因家族, 花发育, 花分生组织转变, 花器官发生, 基因表达

Abstract:

【Objective】The identification of HaYABBY gene related to flower development in sunflower（Helianthus annuus L.）is conductive to exploring its biological function and regulatory mechanism in the floral development of sunflower, and to provide important molecular clues for sunflower variety breeding.【Method】Based on sunflower genome database and our transcriptome data, bioinformatics analysis was conducted to analyze the physicochemical properties, chromosome localization, phylogenetic relationship, gene structure, conserved motif, protein structure, promoter cis-action regulatory element, and tissue expression pattern, etc. RT-qPCR was used to analyze the differences of HaYABBY gene expression between WT sunflower and cb1（crazy broccoli 1）mutant with the defection of meristem determinacy and floral organ differentiation at st2-st8.【Result】Twelve members of YABBY family were identified, belonging to 4 subfamilies, and located in 8 chromosomes； meanwhile, the members in the same subfamily shared similar gene structure and conserved motifs. Results of tissue expression pattern showed that 8 members presented high expressions during flower development in sunflower, including HaYABBY1a, HaYABBY1c, HaYABBY3, HaYABBY4b, HaYABBY5a, HaYABBY5b, HaCRABS CLAWa, and HaCRABS CLAWb, among which HaYABBY3, HaYABBYY5a, and HaYABBYY5d expressed with up-regulation at the late stages of flower development in cb1 mutant; while the expression of HaYABBYY4b and HaCRABS CLAWa was strongly inhibited in cb1 mutants, corresponding to their expressions at early floral developmental in WT plants. The analysis of cis-regulatory elements showed that all the HaYABBY members contained multiple promoter cis-acting elements, some acting elements only existing in specific genes. Among them, endosperm expression elements were presented in HaCRABS CLAWa and HaYABBY4a. Auxin responders were in HaCRABS CLAWa, HaCRABS CLAWb, HaYABBY4a, HaYABBY4b and HaYABBY5c. Meristem expression elements existed in HaYABBY1a, HaYABBY1c, HaYABBY4b, HaYABBY5a, and HaYABBY5d. Gibberellin response elements were included in HaCRABS CLAWa, HaYABBY5b, HaYABBY5c, and HaYABBY5d. Seed-specific regulatory elements presented in HaYABBY5a, HaYABBY5c, and HaYABBY5d.【Conclusion】Five HaYABBY genes of HaYABBY3, HaYABBY4b, HaYABBY5a, HaYABBY5d and HaCRABS CLAW play important roles in the process of floral meristem transformation and floral organ formation in sunflower, and they might depend on the regulation of auxin, gibberellin and meristem expression related regulators during meristem transformation and floral organogenesis, and need the involvement of the auxin response factor ARFs, gibberellin response factor MYBs and flower organ development regulator MADS-boxes.

Key words: sunflower, YABBY gene family, flower development, floral meristem transformation, floral organogenesis, gene expression