Identification of Sunflower GH3 Gene Family and Analysis of Their Function in Flower Development

doi:10.13560/j.cnki.biotech.bull.1985.2025-0560

Abstract

Abstract:

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.

Key words: sunflower, flower development, cb1 mutant, GH3 gene family, expression level, cis-acting element, transcription factor, transcriptional regulation

LONG Lin-xi, ZENG Yin-ping, WANG Qian, DENG Yu-ping, GE Min-qian, CHEN Yan-zhuo, LI Xin-juan, YANG Jun, ZOU Jian. Identification of Sunflower GH3 Gene Family and Analysis of Their Function in Flower Development[J]. Biotechnology Bulletin, 2026, 42(1): 125-138.

Figures/Tables 7

Fig. 1 Chromosome localization (A) and phylogenetic analysis (B) of HaGH3 gene family members

Fig. 2 Gene structure and protein conserved motif distribution of HaGH3A: Conserved motif of protein encoded by HaGH3 gene. B: Conserved domain diagram. C: Gene structure map

Fig. 3 Expression analysis of HaGH3 family genes in different tissues of WT sunflowerSt2: Initiation phase of flower primordium. St3: Phase of flower organs starting formation. St4: Meiosis phase. St5: Phase of pollen maturity. St6: Floret opening phase. St7: Fruit filling phase. St8: Seed-full phase. The same below

Fig. 4 Differential expression analysis of HaGH3 gene in WT sunflower and cb1 mutantA: Flower disc of WT sunflower and cb1 mutant. B: Longitudinal section of WT sunflower and cb1 mutant inflorescences. C: Relative expression of HaGH3 gene during inflorescence development of WT and cb1 mutants (St2‒St8). All data are mean ±SD (n=3). * P < 0.05, ** P < 0.01. The same below

Fig. 5 Expressions of the HaGH3 family in flower organs at different phasesA: Anatomy of tubular flowers at St5 phase of sunflower. B: Anatomy of tubular flowers from the St6 phase of sunflower. C Expressions of sunflower GH3 gene family in flower organs at St5 phase. D: Expressions in flower organs at St6 phase. All data are mean ± SD (n=3). Groups marked with different lowercase letters indicate statistically significant differences (P < 0.05)

Fig. 6 Prediction analysis of cis-acting elements and regulatory networks of the promoter region of HaGH3 geneA: Cis-acting element of the HaGH3 gene. B: Prediction of interaction between HaGH3.8 and HaGH3.14 with ARF transcription factors. C: Prediction of interaction between HaGH3.8 and MYB and bZIP transcription factors. D: Prediction of interaction between HaGH3.8, HaGH3.13 and HaGH3.14 genes with MIKC_MADS transcription factors. E: Prediction of interaction between HaGH3.18 and ARF transcription factors. F: Prediction of interaction between HaGH3.1 gene and MYB and bZIP transcription factors. G: Prediction of interaction between HaGH3.18 gene and MIKC_MADS transcription factors

Fig. 7 Relative expressions of HaGH3 gene and its candidate transcription factors involved in the regulation of flower developmentA: Relative expressions of HaGH3.8 and HaGH3.14 and their potential upstream transcriptional regulators ARF. B: Relative expressions of HaGH3.8 and their potential upstream transcriptional regulators MYB and bZIP. C: Relative expressions of HaGH3.8, HaGH3.13 and HaGH3.14 and their potential upstream transcriptional regulators MIKC_MADS. D: Relative expressions of HaGH3.18 and its potential upstream transcriptional regulators ARF. E: Relative expressions of HaGH3.1 and its potential upstream transcriptional regulators MYB and bZIP. F: Relative expressions of HaGH3.18 and its potential upstream transcriptional regulatorsMIKC_MADS

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