Abstract
Hazel (Corylus spp.) has unique ovary and ovule development characteristics, as the ovary, ovule, and mature embryo sac only begin to develop after pollination. Frequent development failure of ovary and ovule causes yield losses in hazelnut. Despite the economic importance, proteins involved in the regulation of ovary and ovule differentiation remain largely unknown. To address these gaps, total protein was extracted from the pistillate inflorescences or young ovaries at three stages: stage F (unpollinated inflorescences), stage S (beginning of a rudimentary ovary formation), and stage T (ovule differentiation). Protein profiling analysis was carried out using isobaric tags for relative and absolute quantitation (iTRAQ), leading to the identification of 2586 proteins. In total, 815 differentially expressed proteins (DEPs) were identified, including 386, 173, and 697 DEPs in S-vs-F, T-vs-S, and T-vs-F paired comparisons, respectively. The proteins and DEPs using iTRAQ technology showed substantially higher transcript abundance than differentially expressed genes (DEGs) and unigenes in previous comparative transcriptome analysis at mRNA level. Pathway mapping results revealed that nine pathways were significantly enriched, sharing two common KEGG pathways, ko00940 (phenylpropanoid biosynthesis) and ko00941 (flavonoid biosynthesis), which were reported previously in a comparative transcriptome analysis. Of 815 DEPs, 236 were simultaneously differentially expressed at mRNA level, and a set of DEPs including chalcone synthase was predicted to participate in ovary and ovule differentiation based on the results at expression and mRNA level. Our findings provide new insight into the molecular mechanisms of ovary differentiation and development after pollination at the expression level.
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Data archiving statement
The data used for analysis in this study was provided as supplementary files of Figs. S1–S3 and Tables S1–S11.
Funding
This work was financially supported by the National Natural Science Foundation of China (No. 31670681; 31770723) and Jilin Provincial Department of Education (No. JJKH20191012KJ; JJKH20190996KJ).
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Supplementary Figure 1
Protein identification overview (PNG 58 kb)
Supplementary Figure 2
Quantitation of chalcone synthase (CHS) and apyrase 1 (APY1) by western blot at three developmental stages. A: Quantitation of CHS in Fig. 9; B: Quantitation of APY1 in Fig. 9. The relative ratios of the signal intensity between target proteins (CHS and APY1) and internal reference β-actin were quantified. β-actin was used for normalization. Error bar represents the SE of three independent measurements. Different lowercase letters above each column denote significant differences determined by Student’s t test at the 5% level of significance. (JPG 29 kb)
Supplementary Figure 3
Original pictures of western blot at three developmental stages. Note: Set 1 to set 3 are three replicates of the western blot. (JPG 60 kb)
Supplementary Table 1
Detailed information of protein identification (XLSX 204 kb)
Supplementary Table 2
DEPs in S-vs-F paired comparison (XLSX 70 kb)
Supplementary Table 3
DEPs in T-vs-S paired comparison (XLSX 18 kb)
Supplementary Table 4
DEPs in T-vs-F paired comparison (XLS 3578 kb)
Supplementary Table 5
FPKM for 15 randomly chosen DEPs (XLSX 304 kb)
Supplementary Table 6
236 DEGs which were simultaneously differentially expressed at mRNA level (XLSX 142 kb)
Supplementary Table 7
FPKM for all identified unigenes (XLSX 565 kb)
Supplementary Table 8
FPKM for all identified DEGs (XLSX 12 kb)
Supplementary Table 9
FPKM for all identified proteins (XLSX 27 kb)
Supplementary Table 10
FPKM for all identified DEPs (XLSX 6264 kb)
Supplementary Table 11
FPKM for all interest proteins (XLSX 1073 kb)
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Cheng, Y., Mou, Y., Zhang, X. et al. iTRAQ protein profiling reveals candidate proteins regulating ovary and ovule differentiation in pistillate inflorescences after pollination in hazel. Tree Genetics & Genomes 15, 21 (2019). https://doi.org/10.1007/s11295-019-1328-7
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DOI: https://doi.org/10.1007/s11295-019-1328-7