生物技术通报 ›› 2026, Vol. 42 ›› Issue (3): 362-373.doi: 10.13560/j.cnki.biotech.bull.1985.2025-0563
李天源(
), 亓新亮, 刘珊, 张建成, 王鹏飞, 张帅, 贾璐婷, 穆霄鹏(
)
收稿日期:2025-05-31
出版日期:2026-03-26
发布日期:2026-04-23
通讯作者:
穆霄鹏,男,博士,副教授,研究方向 :园艺植物种质利用与创新;E-mail: 15110671026@163.com作者简介:李天源,女,硕士研究生,研究方向 :园艺植物种质利用与创新;E-mail: 17735426816@163.com
基金资助:
LI Tian-yuan(
), QI Xin-liang, LIU Shan, ZHANG Jian-cheng, WANG Peng-fei, ZHANG Shuai, JIA Lu-ting, MU Xiao-peng(
)
Received:2025-05-31
Published:2026-03-26
Online:2026-04-23
摘要:
目的 研究欧李SPL基因家族成员的结构功能和表达分析,为后续深入研究欧李SPL基因家族的功能奠定基础。 方法 基于欧李全基因组和转录组数据,系统鉴定欧李SPL基因家族成员,并对ChSPL基因家族进行生物信息学分析以及表达模式分析。 结果 共鉴定出17个欧李SPL基因家族成员(ChSPL01‒ChSPL17),ChSPLs不均等地分布在7条染色体上,各亚族内的基因结构及保守结构域均表现出高度相似性,ChSPL14‒ChSPL16定位在细胞质,其余的家族成员则定位在细胞核。顺式作用元件分析表明,ChSPLs的光响应元件数量最多,其次为厌氧诱导响应元件(ARE)、脱落酸响应元件(ABRE)和茉莉酸甲酯响应元件(CGTCA-motif)。基于转录组和RT-qPCR分析表明,ChSPL01、ChSPL02、ChSPL06、ChSPL09和ChSPL11等多数ChSPLs在果实发育早期呈现较高的表达量,且随着果实成熟表达量逐渐下降,而ChSPL12、ChSPL14和ChSPL16随果实发育表达量逐渐上调,其中ChSPL12在果实成熟期表达量最高。ChSPLs还存在明显的组织特异性表达,ChSPL02、ChSPL05、ChSPL06、ChSPL10和ChSPL17等在茎和叶中表达量相对较高;ChSPL01、ChSPL04、ChSPL09在叶中表达量相对较高;而ChSPL14、ChSPL15和ChSPL16在花和果实中表达量相对较高。 结论 共鉴定出17个欧李ChSPL基因,推测ChSPL基因家族在果实发育中发挥重要作用,其中ChSPL12、ChSPL14和ChSPL16可能为促进果实成熟的关键候选基因,为未来研究ChSPL基因功能提供参考。
李天源, 亓新亮, 刘珊, 张建成, 王鹏飞, 张帅, 贾璐婷, 穆霄鹏. 欧李SPL基因家族的鉴定及在果实发育过程中的表达分析[J]. 生物技术通报, 2026, 42(3): 362-373.
LI Tian-yuan, QI Xin-liang, LIU Shan, ZHANG Jian-cheng, WANG Peng-fei, ZHANG Shuai, JIA Lu-ting, MU Xiao-peng. Identification of Cerasus humilisSPL Gene Family and Expression Analysis during Fruit Development[J]. Biotechnology Bulletin, 2026, 42(3): 362-373.
基因名称 Gene name | 上游引物序列 Forward primer sequence (5′‒3′) | 下游引物序列 Reverse primer sequence (5′‒3′) |
|---|---|---|
| ChSPL01 | AAGAGGAGGAGGAGGAGGAG | TGCTTGACAAGATGGCCTTG |
| ChSPL02 | GCTGTGCTCTCTCTCTTCTGTCAA | TTGAGTCAGTGACTCCTGTGACG |
| ChSPL03 | CCAAGGAAAACGTAGTTGTCGTAGA | TCAAATTCAGGAAGCTGATGAAATC |
| ChSPL04 | TGGTGCTGATTTTGTTGAGGC | CACTTCTCTGCTTGACAACATGGT |
| ChSPL05 | GCCACTGCTCAGTCTAAAGC | TTCAACTTGGCAGTGTGGTG |
| ChSPL06 | CAAGAGAGGCAGAGGAGGAG | ACTCCTTTGCGTTCATCAGC |
| ChSPL07 | ATTCTGGAGCTGGGACTCAC | CGTTGGAGCTGTGAGGAAAG |
| ChSPL08 | GCAAACAGGCAAATGCTTCC | CTGAACTCCTGCTGAATGGC |
| ChSPL09 | GGGTGATAATAGTGGAAGAATTCGG | TGGCCACATACATCCTTCAACTG |
| ChSPL10 | CTTCTGCCTGCCAACAACTT | CTGAGCTGGCCAGATTGAAC |
| ChSPL11 | CATAACCAGCATCTCCAACTGC | AAGACCGGTATCAGCCTCAGC |
| ChSPL12 | CCAAGCAGATGCAGAGGTTC | GAGAGTGAGCCGTGAGGTAA |
| ChSPL13 | GGCCTGTGGAAGCCAAAAAG | TCTGCTTTCTGCAGGTGAGG |
| ChSPL14 | ATGCCAAGGATTACCACCGA | TATTATGGCCAGCCAAACGC |
| ChSPL15 | ATTGTGATGGTGGAGCAGGA | CAGGCACTGAACTGAGGGTA |
| ChSPL16 | GCGTTTGGCTGGCCATAATA | TGGCTAGGCTCCTCAGTAGA |
| ChSPL17 | CCTTCAACAACCTCGTCCCT | TAGATGGAGTGGAGCTGCTG |
| ChActin | TTCAAAGACCAGCTCATCTGTGG | CAATGCCAGGGAACATAGTGGA |
表1 RT-qPCR引物序列
Table 1 Sequences of primers used for RT-qPCR
基因名称 Gene name | 上游引物序列 Forward primer sequence (5′‒3′) | 下游引物序列 Reverse primer sequence (5′‒3′) |
|---|---|---|
| ChSPL01 | AAGAGGAGGAGGAGGAGGAG | TGCTTGACAAGATGGCCTTG |
| ChSPL02 | GCTGTGCTCTCTCTCTTCTGTCAA | TTGAGTCAGTGACTCCTGTGACG |
| ChSPL03 | CCAAGGAAAACGTAGTTGTCGTAGA | TCAAATTCAGGAAGCTGATGAAATC |
| ChSPL04 | TGGTGCTGATTTTGTTGAGGC | CACTTCTCTGCTTGACAACATGGT |
| ChSPL05 | GCCACTGCTCAGTCTAAAGC | TTCAACTTGGCAGTGTGGTG |
| ChSPL06 | CAAGAGAGGCAGAGGAGGAG | ACTCCTTTGCGTTCATCAGC |
| ChSPL07 | ATTCTGGAGCTGGGACTCAC | CGTTGGAGCTGTGAGGAAAG |
| ChSPL08 | GCAAACAGGCAAATGCTTCC | CTGAACTCCTGCTGAATGGC |
| ChSPL09 | GGGTGATAATAGTGGAAGAATTCGG | TGGCCACATACATCCTTCAACTG |
| ChSPL10 | CTTCTGCCTGCCAACAACTT | CTGAGCTGGCCAGATTGAAC |
| ChSPL11 | CATAACCAGCATCTCCAACTGC | AAGACCGGTATCAGCCTCAGC |
| ChSPL12 | CCAAGCAGATGCAGAGGTTC | GAGAGTGAGCCGTGAGGTAA |
| ChSPL13 | GGCCTGTGGAAGCCAAAAAG | TCTGCTTTCTGCAGGTGAGG |
| ChSPL14 | ATGCCAAGGATTACCACCGA | TATTATGGCCAGCCAAACGC |
| ChSPL15 | ATTGTGATGGTGGAGCAGGA | CAGGCACTGAACTGAGGGTA |
| ChSPL16 | GCGTTTGGCTGGCCATAATA | TGGCTAGGCTCCTCAGTAGA |
| ChSPL17 | CCTTCAACAACCTCGTCCCT | TAGATGGAGTGGAGCTGCTG |
| ChActin | TTCAAAGACCAGCTCATCTGTGG | CAATGCCAGGGAACATAGTGGA |
基因 ID Gene ID | 基因 Gene | 染色体位置 Chr. No. | 编码氨基酸长度 Length (aa) | 分子量Molecular weight (kD) | 等电点 pI | 不稳定系数 Instability index | 脂肪族氨基酸指数 Aliphatic index | 亲水性 Grand average of hydropathicity | 亚细胞定位 Subcellular localization |
|---|---|---|---|---|---|---|---|---|---|
| ouLi_011577-RA | SPL01 | Chr3 | 489 | 38.03 | 5.26 | 28.89 | 30.88 | 0.681 | Nucleus |
| ouLi_006828-RA | SPL02 | Chr2 | 1 152 | 95.94 | 5.01 | 58.50 | 23.96 | 0.800 | Nucleus |
| ouLi_019229-RA | SPL03 | Chr6 | 1 143 | 95.19 | 5.01 | 58.64 | 23.80 | 0.797 | Nucleus |
| ouLi_010488-RA | SPL04 | Chr3 | 489 | 38.23 | 5.24 | 19.82 | 26.99 | 0.648 | Nucleus |
| ouLi_002187-RA | SPL05 | Chr1 | 1 446 | 119.72 | 4.99 | 44.37 | 28.56 | 0.805 | Nucleus |
| ouLi_017116-RA | SPL06 | Chr5 | 1 230 | 100.61 | 5.01 | 45.43 | 24.07 | 0.786 | Nucleus |
| ouLi_024093-RA | SPL07 | Chr8 | 1 656 | 137.64 | 4.99 | 43.99 | 28.02 | 0.724 | Nucleus |
| ouLi_002823-RA | SPL08 | Chr1 | 1 497 | 124.27 | 5.01 | 42.47 | 29.33 | 0.743 | Nucleus |
| ouLi_013789-RA | SPL09 | Chr4 | 570 | 44.99 | 5.22 | 23.54 | 30.88 | 0.724 | Nucleus |
| ouLi_002175-RB | SPL10 | Chr1 | 951 | 78.33 | 5.08 | 38.32 | 28.08 | 0.802 | Nucleus |
| ouLi_024712-RA | SPL11 | Chr8 | 1 227 | 102.04 | 5.02 | 52.67 | 29.01 | 0.858 | Nucleus |
| ouLi_026147-RA | SPL12 | Chr8 | 3 213 | 265.03 | 4.84 | 42.23 | 27.23 | 0.725 | Cytoplasm/Nucleus |
| ouLi_012144-RA | SPL13 | Chr4 | 1 176 | 97.79 | 5.02 | 47.01 | 26.53 | 0.799 | Nucleus |
| ouLi_000956-RA | SPL14 | Chr1 | 3 105 | 255.36 | 4.86 | 41.47 | 28.28 | 0.716 | Cytoplasm |
| ouLi_001341-RA/RB | SPL15 | Chr1 | 2 451 | 202.05 | 4.91 | 39.97 | 27.46 | 0.696 | Cytoplasm |
| ouLi_000956-RD | SPL16 | Chr1 | 3 033 | 248.56 | 4.87 | 40.26 | 28.39 | 0.699 | Cytoplasm |
| ouLi_017119-RA | SPL17 | Chr5 | 1 137 | 97.28 | 4.94 | 62.14 | 21.37 | 1.115 | Nucleus |
表2 ChSPL家族成员的基本信息
Table 2 Basic information of ChSPL family members
基因 ID Gene ID | 基因 Gene | 染色体位置 Chr. No. | 编码氨基酸长度 Length (aa) | 分子量Molecular weight (kD) | 等电点 pI | 不稳定系数 Instability index | 脂肪族氨基酸指数 Aliphatic index | 亲水性 Grand average of hydropathicity | 亚细胞定位 Subcellular localization |
|---|---|---|---|---|---|---|---|---|---|
| ouLi_011577-RA | SPL01 | Chr3 | 489 | 38.03 | 5.26 | 28.89 | 30.88 | 0.681 | Nucleus |
| ouLi_006828-RA | SPL02 | Chr2 | 1 152 | 95.94 | 5.01 | 58.50 | 23.96 | 0.800 | Nucleus |
| ouLi_019229-RA | SPL03 | Chr6 | 1 143 | 95.19 | 5.01 | 58.64 | 23.80 | 0.797 | Nucleus |
| ouLi_010488-RA | SPL04 | Chr3 | 489 | 38.23 | 5.24 | 19.82 | 26.99 | 0.648 | Nucleus |
| ouLi_002187-RA | SPL05 | Chr1 | 1 446 | 119.72 | 4.99 | 44.37 | 28.56 | 0.805 | Nucleus |
| ouLi_017116-RA | SPL06 | Chr5 | 1 230 | 100.61 | 5.01 | 45.43 | 24.07 | 0.786 | Nucleus |
| ouLi_024093-RA | SPL07 | Chr8 | 1 656 | 137.64 | 4.99 | 43.99 | 28.02 | 0.724 | Nucleus |
| ouLi_002823-RA | SPL08 | Chr1 | 1 497 | 124.27 | 5.01 | 42.47 | 29.33 | 0.743 | Nucleus |
| ouLi_013789-RA | SPL09 | Chr4 | 570 | 44.99 | 5.22 | 23.54 | 30.88 | 0.724 | Nucleus |
| ouLi_002175-RB | SPL10 | Chr1 | 951 | 78.33 | 5.08 | 38.32 | 28.08 | 0.802 | Nucleus |
| ouLi_024712-RA | SPL11 | Chr8 | 1 227 | 102.04 | 5.02 | 52.67 | 29.01 | 0.858 | Nucleus |
| ouLi_026147-RA | SPL12 | Chr8 | 3 213 | 265.03 | 4.84 | 42.23 | 27.23 | 0.725 | Cytoplasm/Nucleus |
| ouLi_012144-RA | SPL13 | Chr4 | 1 176 | 97.79 | 5.02 | 47.01 | 26.53 | 0.799 | Nucleus |
| ouLi_000956-RA | SPL14 | Chr1 | 3 105 | 255.36 | 4.86 | 41.47 | 28.28 | 0.716 | Cytoplasm |
| ouLi_001341-RA/RB | SPL15 | Chr1 | 2 451 | 202.05 | 4.91 | 39.97 | 27.46 | 0.696 | Cytoplasm |
| ouLi_000956-RD | SPL16 | Chr1 | 3 033 | 248.56 | 4.87 | 40.26 | 28.39 | 0.699 | Cytoplasm |
| ouLi_017119-RA | SPL17 | Chr5 | 1 137 | 97.28 | 4.94 | 62.14 | 21.37 | 1.115 | Nucleus |
蛋白 Protein | α-螺旋 Alpha helix (%) | 延伸链 Extended strand (%) | β-折叠 Beta turn (%) | 无规则卷曲 Random coil (%) |
|---|---|---|---|---|
| SPL01 | 31.48 | 5.56 | 0 | 62.96 |
| SPL02 | 9.40 | 4.96 | 0 | 85.64 |
| SPL03 | 11.05 | 6.05 | 0 | 82.89 |
| SPL04 | 31.48 | 5.56 | 0 | 62.96 |
| SPL05 | 9.36 | 4.57 | 0 | 86.07 |
| SPL06 | 11.98 | 6.36 | 0 | 81.66 |
| SPL07 | 12.52 | 3.63 | 0 | 83.85 |
| SPL08 | 9.04 | 4.02 | 0 | 86.95 |
| SPL09 | 28.57 | 7.41 | 0 | 64.02 |
| SPL10 | 10.76 | 6.96 | 0 | 82.28 |
| SPL11 | 10.78 | 6.13 | 0 | 83.09 |
| SPL12 | 28.04 | 5.23 | 0 | 66.73 |
| SPL13 | 9.72 | 6.14 | 0 | 84.14 |
| SPL14 | 27.37 | 7.93 | 0 | 64.70 |
| SPL15 | 28.80 | 8.46 | 0 | 62.75 |
| SPL16 | 25.64 | 4.95 | 0 | 69.41 |
| SPL17 | 11.64 | 6.88 | 0 | 81.48 |
表3 ChSPL蛋白的二级结构
Table 3 Secondary structure of ChSPL proteins
蛋白 Protein | α-螺旋 Alpha helix (%) | 延伸链 Extended strand (%) | β-折叠 Beta turn (%) | 无规则卷曲 Random coil (%) |
|---|---|---|---|---|
| SPL01 | 31.48 | 5.56 | 0 | 62.96 |
| SPL02 | 9.40 | 4.96 | 0 | 85.64 |
| SPL03 | 11.05 | 6.05 | 0 | 82.89 |
| SPL04 | 31.48 | 5.56 | 0 | 62.96 |
| SPL05 | 9.36 | 4.57 | 0 | 86.07 |
| SPL06 | 11.98 | 6.36 | 0 | 81.66 |
| SPL07 | 12.52 | 3.63 | 0 | 83.85 |
| SPL08 | 9.04 | 4.02 | 0 | 86.95 |
| SPL09 | 28.57 | 7.41 | 0 | 64.02 |
| SPL10 | 10.76 | 6.96 | 0 | 82.28 |
| SPL11 | 10.78 | 6.13 | 0 | 83.09 |
| SPL12 | 28.04 | 5.23 | 0 | 66.73 |
| SPL13 | 9.72 | 6.14 | 0 | 84.14 |
| SPL14 | 27.37 | 7.93 | 0 | 64.70 |
| SPL15 | 28.80 | 8.46 | 0 | 62.75 |
| SPL16 | 25.64 | 4.95 | 0 | 69.41 |
| SPL17 | 11.64 | 6.88 | 0 | 81.48 |
图4 ChSPL基因的染色体定位(A)与物种间共线性分析(B)、种内共线性分析(C)
Fig. 4 Chromosome location (A) and interspecies collinearity analysis (B), intraspecific collinearity analysis (C) of ChSPL gene
图7 ChSPL在果实不同发育时期(A)和不同组织中(B)的表达量4-1:幼果期;4-2:硬核期;4-3:转色期;4-4:着色期;4-5:成熟期
Fig. 7 Expressions of ChSPL in different stages of fruit (A) and different tissues (B)4-1: Young fruit stage. 4-2: Hard core stage. 4-3: Turning stage. 4-4: Coloring stage. 4-5: Mature stage
图8 ChSPL在果实中不同发育时期的表达量变化4-1:幼果期(花后6周);4-2:硬核期(花后8周);4-3:着色期(花后16周);4-4:转色期(花后17周);4-5:成熟期(花后18周),不同字母表示不同发育时期表达量差异显著(P<0.05)
Fig. 8 Changes in the expressions of ChSPL over time under different development stages4-1: Young fruit stage (6 weeks after flowering). 4-2: Stone hardening stage (8 weeks after flowering). 4-3: Coloration stage (16 weeks after flowering). 4-4: Color turning stage (17 weeks after flowering). 4-5: Maturity stage (18 weeks after flowering). Different letters indicate significant differences in expressions at different developmental stages (P<0.05)
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