Identification and Expression Analysis of the Members of the Asparagus officinalis DUF247 Gene Family

doi:10.13560/j.cnki.biotech.bull.1985.2024-0596

Abstract

Abstract:

【Objective】To establish a foundation for the molecular functional study of this gene family, particularly AoSOFF's role in sex determination, a comprehensive identification and analysis of the A. officinalis DUF247 family was conducted.【Method】The bioinformatics methods were used to identify the members of the DUF247 gene family in A. officinalis, and conducted analyses on their encoded protein physicochemical properties, clustering, protein conserved domains, cis-acting elements in the promoter, and expression patterns. Real-time quantitative PCR（RT-qPCR）and tissue in situ hybridization were used to validate the expression pattern of the A. officinalis sex determination gene AoSOFF.【Result】The 24 members of the DUF247 family were unevenly distributed on 7 chromosomes of A. officinalis, and showed differential expressions in various organs. The encoded proteins of these family members had differences in their physicochemical properties, including amino acid composition, molecular weight, theoretical isoelectric point, instability index, and aliphatic index. RT-qPCR and tissue in situ hybridization results demonstrated that AoSOFF had high expressions in the pistil, stamen, and corolla of male flowers during the early stage of sex differentiation but decreased its expression as sex differentiation progressed. Its expression in hermaphrodite flowers was low, and extremely low or undetectable in the female flowers, indicating that AoSOFF had male-specific expression in male asparagus, which was closely related to its female inhibition function in A. officinalis sex determination.【Conclusion】The members of the A. officinalis DUF247 family have obvious conservation and exhibit significantly different expression patterns in different tissues of A. officinalis of different sexes. In particular, AoSOFF is a male-specific gene, and its homologs are involved in regulating plant self-incompatibility, suggesting that its similar reaction to self-incompatibility may lead to the inhibition of female development.

Key words: Asparagus officinalis, DUF247, AoSOFF, sex differentiation, male specificity, female inhibition, in situ hybridization

LI Yun-bin, LI Yu-ping, CHENG Qin, LIN Chun, MAO Zi-chao, LIU Zheng-jie. Identification and Expression Analysis of the Members of the Asparagus officinalis DUF247 Gene Family[J]. Biotechnology Bulletin, 2025, 41(1): 198-209.

Figures/Tables 8

Fig. 1 Phylogenetic tree of DUF247 protein from A. officinalis, A. setaceus, A. taliensis and A. thaliana V1: A. officinalis; OF: A. setaceus; Ata: A. taliensis; AT: A. thaliana

Fig. 2 Cis-acting elements (A) and conserved motifs map (B) of encoded proteins of DUF247 family genes in A. officinalis

Fig. 3 Chromosome distribution（A）and collinearity analysis（B）of DUF247 family genes in A. officinalis

Fig. 4 Heatmaps of expressions of gene members of A. officialis DUF247 family in different genders and tissues or organs MR: Male root; MS: male stem; MF: male flower; FR: female root; FS: female stem; FF: female flower; HR: hermaphrodite root; HS: hermaphrodite stem; HF: hermaphrodite flower; R1, R2, and R3 indicate the three replicates of the sample. The same below

Fig. 5 Different expressions of V1_01.231, V1_05.1843, and V1_05.2202 in early development buds of different genders ffs: Female flower'stamen; mfs: male flower'stamen; fft: female flower'tapal; mft: male flower'tapal; ffp: female flower'pistil; mfp: male flower'pistil. Different lowercase letters indicatet significant differences（P<0.05）, the same below

Fig. 6 Primary structure comparison and 3D structure prediction of protein encoded by AoSOFF gene in A. officinalis A: 3D structure of AoSOFF-M protein; B: 3D structure of AoSOFF-F protein; C: 3D structure of AoSOFF-H protein; D: comparison of three proteins in primary structure, M for male, F for female, and H for hermaphroditic

Fig. 7 RT-qPCR analysis of A. officinalis AoSOFF in different samples

Fig. 8 Detection of A. officinalis AoSOFF in male, female and hermaphroditic flowers by in situ hybridization A-C: Male flower; D-F: male flower control; G-I: hermaphroditic flower; J-L: hermaphroditic flower control; M-O: female flower in situ hybridization results; P-R: female flower control. I, II, and III: Three stages of flower development. White arrow: Ovule; blue arrow: ovary; red arrows: stigma; yellow arrow: outer wall of pollen sac. Bar=500 μm

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