PDF(1102 KB)
Excavation and expression analysis of miRNA related to oil synthesis during seed development in sunflower
Fei ZHOU, Wen-jun WANG, Xu-tang HUANG, Jun MA, Li-ren WU, Jing WANG, Peng-yuan XIE, Yan LIU
CHINESE JOURNAL OF OIL CROP SCIENCES ›› 2023, Vol. 45 ›› Issue (5) : 965-971.
PDF(1102 KB)
PDF(1102 KB)
Excavation and expression analysis of miRNA related to oil synthesis during seed development in sunflower
In order to explore the role of miRNA （microRNA） in seed oil synthesis of sunflower, miRNA sequencing in 3 development stages （7 d, 22 d, 37 d after flowering） of high oleic acid sunflower seeds were mined. Results showed that 22 differentially expressed miRNAs were involved in oil metabolism pathway based on comparisons of L7d vs L22d, L7d vs L37d and L22d vs L37d. Clustering heat map showed that most miRNAs in the 22d vs 37d comparison group were down-regulated, indicating that the expression of most miRNAs related to oil metabolism decreased at 37 d after flowering compared with 22 d after flowering. The 22 differentially expressed miRNAs annotated with target gene into fatty acid and triacylglycerol synthesis pathway were analyzed for the correlation between expression level and fatty acid components at different stages of seed development. Results also showed that caproic acid and miR172a3, tridecylic acid and miR171b2 were both extremely significantly negatively correlated, and oil content was significantly positively correlated with miR167d5p, indicating that these miRNAs might be involved in oil synthesis. Through co-expression analysis, it was found that 6 miRNAs related to oil metabolism （miR166u, miR171b_2, miR397a_3, miR157d, miR167a-5p, miR169v_1） were negatively correlated with their target genes, indicating that the differential expression of these genes might be the result of miRNA regulation.
sunflower / high oleic acid / seed development / fatty acid / microRNA sequencing {{custom_keyword}} /
Table 1 Differentially expressed miRNA and their target genes in lipid metabolism pathways表1 油脂代谢通路中差异表达miRNA及其靶基因 |
| miRNA ID | 靶基因ID Target gene ID | 基因名 Gene name | KEGG注释 KEGG annotation |
|---|---|---|---|
| miR157a-5p（22d vs 37d） miR157d（7d vs 22d，7d vs 37d） | XM_022145135.1 XM_022145134.1 XM_022145138.1 XM_022145137.1 | ACOX1， ACOX3 | K00232 acyl-CoA oxidase |
| miR6300（7d vs 22d） | XM_022144203.1 | ACOX1， ACOX3 | K00232 acyl-CoA oxidase |
| miR171a_3（7d vs 37d） miR171b_2（7d vs 22d，7d vs 37d） | XM_022180675.1 | FAD2 | K10256 Delta-12 fatty acid desaturase |
| miR167a（7d vs 22d，7d vs 37d，22d vs 37d） miR167a-5p（7d vs 37d，22d vs 37d） miR167a_2（7d vs 22d，7d vs 37d） miR167d-5p（7d vs 22d，7d vs 37d，22d vs 37d） miR167d_1（7d vs 22d，7d vs 37d） | XM_022138859.1 XM_022138860.1 XM_022138861.1 XM_022138855.1 XM_022138863.1 | KAS-II | K09458 3-oxoacyl-［acyl-carrier-protein］ synthase II |
| miR6111-5p（7d vs 22d） | XM_022173634.1 XM_022173633.1 | ECI1_2 | K07517 Delta3-Delta2-enoyl-CoA isomerase |
| miR6111-5p（7d vs 22d） | XM_022170833.1 XM_022147236.1 | PLD1_2 | K01115 phospholipase D1/2 |
| miR166u（7d vs 22d，7d vs 37d） | XM_022179947.1 | KCS | K15397 3-ketoacyl-CoA synthase |
| miR164c_1（7d vs 37d，22d vs 37d） | XM_022172057.1 | HAD | K02372 3-hydroxyacyl-［acyl-carrier-protein］ dehydratase |
| miR169v_1（7d vs 37d，22d vs 37d） | XM_022154766.1 XM_022154765.1 | LACS | K01897 long-chain acyl-CoA synthetase |
| miR397a_3（7d vs 22d，7d vs 37d） | XM_022173175.1 | DPP1， DPPL， PLPP4_5 | K18693 diacylglycerol diphosphate phosphatase |
| miR172a_2（7d vs 22d，7d vs 37d，22d vs 37d） miR172a_3（7d vs 37d，22d vs 37d） miR172c_2（7d vs 22d，7d vs 37d） | XM_022122517.1 | glpQ， ugpQ | K01126 glycerophosphoryl diester phosphodiesterase |
| miR396b（7d vs 22d，7d vs 37d，22d vs 37d） miR396b-5p（7d vs 22d，7d vs 37d，22d vs 37d） miR396e-5p（7d vs 22d，7d vs 37d，22d vs 37d） | XM_022153737.1 XM_022153734.1 XM_022153735.1 | MGD | K03715 1，2-diacylglycerol 3-beta-galactosyltransferase |
| miR171i_1（7d vs 22d，7d vs 37d，22d vs 37d） | XM_022121938.1 | LCAT3 | K22389 phospholipase A1 |
Fig. 1 Clustering heat map of expression of differentially expressed miRNA related to lipid metabolism图1 油脂代谢相关差异表达miRNA的表达聚类热图 |
Table 2 Fatty acid composition and oil content analysis of sunflower seeds with high oleic acid at three development stages表2 高油酸向日葵三个发育时期种子脂肪酸组成和油分含量分析 |
| 脂肪酸种类 /% Fatty acid | 开花后7 d 7 DAF | 开花后22 d 22 DAF | 开花后37 d 37 DAF |
|---|---|---|---|
| 己酸 C6：0 | 0 | 0 | 0.37±0.03 |
| 癸酸 C10：0 | 6.61±0.09 | 1.31±0.19 | 0 |
| 月桂酸 C12：0 | 1.27±0.21 | 0.08±0.01 | 0 |
| 十三烷酸 C13：0 | 6.92±1.56 | 0.75±0.10 | 0 |
| 十五烷酸 C15：0 | 0 | 0.29±0.04 | 0 |
| 棕榈酸 C16：0 | 17.19±1.89 | 10.24±2.03 | 6.83±0.76 |
| 硬脂酸 C18：0 | 2.01±0.32 | 3.12±0.46 | 2.06±0.31 |
| 油酸 C18：1 | 11.45±2.1 | 50.84±4.68 | 70.19±5.89 |
| 亚油酸 C18：2 | 47.44±5.98 | 28.28±3.68 | 19.69±2.68 |
| 亚麻酸 C18：3 | 6.29±0.78 | 2.44±0.36 | 0.55±0.07 |
| 花生酸 C20：0 | 0.81±0.10 | 1.18±0.21 | 0 |
| 山嵛酸 C22：0 | 0 | 0.70±0.09 | 0.36±0.06 |
| 木蜡酸 C24：0 | 0 | 0.84±0.10 | 0 |
| 含油量 Oil content | 3.51±0.4 | 15.34±1.80 | 31.95±2.23 |
Table 3 Correlation analysis between expression level of miRNAs in lipid metabolism pathway表3 油脂代谢通路中miRNA表达量与脂肪酸含量的相关性分析 (and fatty acid content) |
| miRNA | 己酸 C6：0 | 癸酸 C10：0 | 月桂酸 C12：0 | 十三烷酸 C13：0 | 十五烷酸 C15：0 | 棕榈酸 C16：0 | 硬脂酸 C18：0 | 油酸 C18：1 | 亚油酸 C18：2 | 亚麻酸 C18：3 | 花生酸 C20：0 | 山嵛酸 C22：0 | 木蜡酸 C24：0 | 油分 Oil |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| miR157a5p | -0.574 | -0.245 | -0.371 | -0.331 | 0.996 | -0.107 | 0.992 | 0.107 | -0.129 | -0.107 | 0.797 | 0.810 | 0.996 | -0.183 |
| miR157d | 0.160 | -0.852 | -0.914 | -0.895 | 0.775 | -0.770 | 0.799 | 0.770 | -0.784 | -0.770 | 0.150 | 0.990 | 0.775 | 0.555 |
| miR6300 | 1.000* | -0.638 | -0.531 | -0.567 | -0.517 | -0.740 | -0.483 | 0.740 | -0.725 | -0.740 | -0.958 | -0.003 | -0.517 | 0.902 |
| miR171a3 | 0.465 | -0.974 | -0.995 | -0.990 | 0.534 | -0.933 | 0.567 | 0.933 | -0.941 | -0.933 | -0.172 | 0.893 | 0.534 | 0.790 |
| miR171b2 | 0.580 | -0.996 | -0.999* | -1.000** | 0.416 | -0.973 | 0.452 | 0.973 | -0.978 | -0.973 | -0.302 | 0.824 | 0.416 | 0.865 |
| miR167a | -0.052 | 0.790 | 0.864 | 0.842 | -0.839 | 0.696 | -0.860 | -0.696 | 0.712 | 0.697 | -0.256 | -0.999* | -0.839 | -0.461 |
| miR167a5p | 0.822 | -0.968 | -0.927 | -0.942 | 0.083 | -0.994 | 0.123 | 0.994 | -0.991 | -0.994 | -0.607 | 0.584 | 0.083 | 0.984 |
| miR167a2 | 0.317 | -0.925 | -0.967 | -0.955 | 0.663 | -0.862 | 0.692 | 0.863 | -0.874 | -0.863 | -0.011 | 0.954 | 0.663 | 0.681 |
| miR167d5p | 0.879 | -0.936 | -0.881 | -0.900 | -0.026 | -0.976 | 0.014 | 0.976 | -0.971 | -0.976 | -0.690 | 0.492 | -0.026 | 0.997* |
| miR167d1 | 0.563 | -0.994 | -1.000* | -1.000* | 0.434 | -0.968 | 0.469 | 0.968 | -0.973 | -0.968 | -0.283 | 0.835 | 0.434 | 0.855 |
| miR61115p | -0.444 | 0.969 | 0.993 | 0.987 | -0.554 | 0.924 | -0.586 | -0.924 | 0.932 | 0.924 | 0.149 | -0.903 | -0.554 | -0.775 |
| miR166u | -0.381 | -0.451 | -0.565 | -0.529 | 0.991 | -0.321 | 0.996 | 0.321 | -0.342 | -0.322 | 0.646 | 0.918 | 0.991 | 0.036 |
| miR164c1 | -0.836 | 0.131 | -0.001 | 0.042 | 0.893 | 0.269 | 0.874 | -0.269 | 0.248 | 0.269 | 0.964 | 0.535 | 0.893 | -0.534 |
| miR169v1 | 0.631 | -1.000* | -0.995 | -0.998* | 0.357 | -0.985 | 0.394 | 0.985 | -0.989 | -0.986 | -0.362 | 0.786 | 0.357 | 0.895 |
| miR397a3 | 0.313 | -0.924 | -0.966 | -0.954 | 0.666 | -0.860 | 0.695 | 0.861 | -0.872 | -0.861 | -0.007 | 0.955 | 0.666 | 0.678 |
| miR172a2 | -0.530 | -0.296 | -0.419 | -0.380 | 0.999* | -0.159 | 0.997* | 0.159 | -0.181 | -0.159 | 0.764 | 0.839 | 0.999* | -0.131 |
| miR172a3 | -1.000** | 0.653 | 0.548 | 0.583 | 0.500 | 0.753 | 0.465 | -0.753 | 0.738 | 0.753 | 0.952 | -0.016 | 0.500 | -0.910 |
| miR172c2 | 0.999* | -0.628 | -0.520 | -0.556 | -0.528 | -0.731 | -0.494 | 0.731 | -0.716 | -0.731 | -0.961 | -0.016 | -0.528 | 0.896 |
| miR396b | -0.855 | 0.951 | 0.902 | 0.920 | -0.021 | 0.985 | -0.061 | -0.985 | 0.981 | 0.985 | 0.655 | -0.532 | -0.021 | -0.993 |
| miR396b5p | -0.965 | 0.430 | 0.308 | 0.348 | 0.711 | 0.553 | 0.682 | -0.553 | 0.534 | 0.553 | 0.999* | 0.248 | 0.711 | -0.769 |
| miR396e5p | -0.987 | 0.523 | 0.406 | 0.444 | 0.633 | 0.637 | 0.602 | -0.637 | 0.620 | 0.637 | 0.989 | 0.145 | 0.633 | -0.832 |
| miR171i1 | -0.864 | 0.946 | 0.895 | 0.913 | -0.04 | 0.982 | -0.044 | -0.982 | 0.978 | 0.982 | 0.668 | -0.518 | -0.004 | -0.995 |
Table 4 Differential co-expression statistics of negatively correlated miRNA and target genes表4 miRNA与靶基因呈负相关的差异共表达统计 |
| 比较组 Comparison group | 共表达数量 Coexpression number | miRNA数量 Number of miRNAs | 靶基因数量 Number of target genes |
|---|---|---|---|
| L7d vs L22d | 407 | 56 | 239 |
| L7d vs L37d | 549 | 79 | 251 |
| L22d vs L37d | 121 | 31 | 80 |
Table 5 Negatively correlated co-expression of miRNA and target genes in lipid metabolism pathway表5 油脂代谢途径中呈负相关的共表达miRNA和靶基因 |
| mRNA ID | miRNA ID | KEGG注释 KEGG annotation | 比较组 Comparison group |
|---|---|---|---|
| XM_022179947.1 | miR166u | 3-ketoacyl-CoA synthase | L7d vs L22d |
| XM_022180675.1 | miR171b_2 | delta（12） fatty acid desaturase | L7d vs L22d |
| XM_022173175.1 | miR397a_3 | diacylglycerol diphosphate phosphatase | L7d vs L22d L7d vs L37d |
| XM_022145134.1 | miR157d | acyl-CoA oxidase | L7d vs L37d |
| XM_022138855.1 | miR167a-5p | 3-oxoacyl-［acyl-carrier-protein］ synthase II | L7d vs L37d L22d vs L37d |
| XM_022154765.1 | miR169v_1 | long-chain acyl-CoA synthetase | L22d vs L37d |
| 1 |
周菲， 王文军， 刘岩， 等. 向日葵籽仁脂肪和脂肪酸含量近红外光谱模型的建立 ［J］. 作物杂志， 2021， 2： 200-206. DOI： 10.16035/j.issn.1001-7283.2021.02.029 .
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| 2 |
马方， 杨宜婷， 陈则华. 不同类型n-3多不饱和脂肪酸对心血管疾病的防治作用及其机制研究进展 ［J］. 中国油脂， 2018， 43（2）： 65-69. DOI： 10.3969/j.issn.1003-7969.2018.02.015 .
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| 3 |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| 4 |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| 5 |
黄俊骏， 刘文文， 郭亚如， 等， microRNA在植物生长发育中的研究进展 ［J］. 生物技术通报， 2019， 35（11）： 141-149. DOI： 10.13560/j.cnki.biotech.bull.1985.2019-0526 .
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| 6 |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| 7 |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| 8 |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| 9 |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| 10 |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| 11 |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| 12 |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| 13 |
崔良基， 王德兴， 宋殿秀. 国内外向日葵遗传改良成就与发展趋势［J］. 杂粮作物， 2006， 26（6）： 402-406. DOI： 10.3969/j.issn.2095-0896.2006.06.009 .
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| 14 |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| 15 |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| 16 |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| 17 |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| 18 |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| 19 |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| 20 |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| 21 |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| 22 |
张天豹. 胡麻种子发育和脂肪酸代谢相关microRNAs的鉴定和功能分析 ［D］. 北京： 中国农业科学院， 2020.
{{custom_citation.content}}
{{custom_citation.annotation}}
|
| {{custom_ref.label}} |
{{custom_citation.content}}
{{custom_citation.annotation}}
|
PDF(1102 KB)
Table 1 Differentially expressed miRNA and their target genes in lipid metabolism pathways
Fig. 1 Clustering heat map of expression of differentially expressed miRNA related to lipid metabolismTable 2 Fatty acid composition and oil content analysis of sunflower seeds with high oleic acid at three development stagesTable 3 Correlation analysis between expression level of miRNAs in lipid metabolism pathwayTable 4 Differential co-expression statistics of negatively correlated miRNA and target genesTable 5 Negatively correlated co-expression of miRNA and target genes in lipid metabolism pathway/
| 〈 |
|
〉 |
