Biotechnology Bulletin ›› 2023, Vol. 39 ›› Issue (5): 192-204.doi: 10.13560/j.cnki.biotech.bull.1985.2022-1196
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LI Jing-rui(), WANG Yu-bo, XIE Zi-wei, LI Chang, WU Xiao-lei, GONG Bin-bin, GAO Hong-bo()
Received:
2022-09-27
Online:
2023-05-26
Published:
2023-06-08
Contact:
GAO Hong-bo
E-mail:yyljr@hebau.edu.cn;hongbogao@hebau.edu.cn
LI Jing-rui, WANG Yu-bo, XIE Zi-wei, LI Chang, WU Xiao-lei, GONG Bin-bin, GAO Hong-bo. Identification and Expression Analysis of PIN Gene Family in Melon Under High Temperature Stress[J]. Biotechnology Bulletin, 2023, 39(5): 192-204.
基因 Gene | 序列 Sequence(5'-3') | 退火温度 Annealing temperature Tm/℃ | GC含量 GC content/% | 产物长度 Product length/bp | |
---|---|---|---|---|---|
Actin3 | F | GGCAGTGGTGGTGAACATG | 59.04 | 57.89 | 149 |
R | TTCTGGTGATGGTGTGAGTC | 57.16 | 50.00 | ||
CmPIN3 | F | CTTCTCCTGTTTCTGATGTGT | 51.40 | 42.90 | 123 |
R | GTTCTGTCTTGGTGTCTCTTC | 52.00 | 47.60 | ||
CmPIN13 | F | CAGGTGATGAAAACTAAACGCCA | 59.75 | 43.48 | 172 |
R | CCGGTGCTCAAAATGTCAGG | 59.48 | 55.00 | ||
CmPIN18 | F | GAGTAGGAGGAACAGAGAAAG | 50.10 | 47.60 | 155 |
R | GACTAAGGACCAAGTGAGACC | 52.70 | 52.40 | ||
CmPIN2 | F | GGGTTTGGTGGTGGGTAAGA | 59.52 | 55.00 | 172 |
R | TTTTGAGTTTGACCGATGCAGG | 59.71 | 45.45 |
Table 1 Primers used in real-time fluorescence quantitative PCR
基因 Gene | 序列 Sequence(5'-3') | 退火温度 Annealing temperature Tm/℃ | GC含量 GC content/% | 产物长度 Product length/bp | |
---|---|---|---|---|---|
Actin3 | F | GGCAGTGGTGGTGAACATG | 59.04 | 57.89 | 149 |
R | TTCTGGTGATGGTGTGAGTC | 57.16 | 50.00 | ||
CmPIN3 | F | CTTCTCCTGTTTCTGATGTGT | 51.40 | 42.90 | 123 |
R | GTTCTGTCTTGGTGTCTCTTC | 52.00 | 47.60 | ||
CmPIN13 | F | CAGGTGATGAAAACTAAACGCCA | 59.75 | 43.48 | 172 |
R | CCGGTGCTCAAAATGTCAGG | 59.48 | 55.00 | ||
CmPIN18 | F | GAGTAGGAGGAACAGAGAAAG | 50.10 | 47.60 | 155 |
R | GACTAAGGACCAAGTGAGACC | 52.70 | 52.40 | ||
CmPIN2 | F | GGGTTTGGTGGTGGGTAAGA | 59.52 | 55.00 | 172 |
R | TTTTGAGTTTGACCGATGCAGG | 59.71 | 45.45 |
基因 Gene | 基因ID Gene ID | 氨基酸数量 Number of amino acids | 分子量 Molecular weight/kD | 理论等电点 Theoretical pI | 不稳定指数 Instability index | 脂溶指数 Aliphatic index | 总平均疏水指数 Grand average of hydropathicity | 跨膜结构域数目 Transmembrane domain number | 亚细胞定位 Subcellular location |
---|---|---|---|---|---|---|---|---|---|
CmPIN1 | MELO3C001206.2 | 135 | 15.15 | 6.69 | 24.33 | 111.93 | 0.795 | 4 | 液泡膜 Tonoplast membrane |
CmPIN2 | MELO3C017414.2 | 623 | 68.44 | 9.22 | 41.57 | 85.47 | -0.012 | 10 | 细胞质膜Cytomembrane |
CmPIN3 | MELO3C017357.2 | 591 | 64.52 | 8.5 | 43.16 | 88.76 | 0.064 | 10 | 细胞质膜Cytomembrane |
CmPIN4 | MELO3C009892.2 | 417 | 46.35 | 9.76 | 48.87 | 118.63 | 0.454 | 11 | 细胞质膜Cytomembrane |
CmPIN5 | MELO3C008726.2 | 402 | 43.62 | 6.23 | 34.38 | 113.26 | 0.501 | 9 | 液泡膜Tonoplast membrane |
CmPIN6 | MELO3C008744.2 | 51 | 5.25 | 6.51 | 30.81 | 139.61 | 1.208 | 0 | 细胞质Cytoplasm |
CmPIN7 | MELO3C031448.2 | 166 | 17.73 | 9.26 | 40.24 | 107.65 | 0.489 | 2 | 叶绿体Chloroplast |
CmPIN8 | MELO3C031243.2 | 51 | 5.19 | 7.98 | 30.81 | 141.57 | 1.275 | 0 | 细胞外基质Extracellular matrix |
CmPIN9 | MELO3C031244.2 | 109 | 12.42 | 7.65 | 27.69 | 102.02 | 0.693 | 3 | 液泡膜Tonoplast membrane |
CmPIN10 | MELO3C013710.2 | 297 | 32.35 | 7.11 | 45.14 | 114.81 | 0.405 | 6 | 内质网Endoplasmic reticulum |
CmPIN11 | MELO3C013711.2 | 428 | 46.24 | 5.02 | 40.85 | 117.78 | 0.576 | 10 | 细胞质膜Cytomembrane |
CmPIN12 | MELO3C017060.2 | 421 | 45.46 | 7.66 | 36.48 | 128.5 | 0.704 | 10 | 液泡膜Tonoplast membrane |
CmPIN13 | MELO3C019102.2 | 611 | 65.37 | 8.75 | 37.4 | 96.73 | 0.265 | 10 | 细胞质膜Cytomembrane |
CmPIN14 | MELO3C025264.2 | 417 | 45.67 | 8.86 | 34.71 | 123.72 | 0.748 | 10 | 细胞质膜Cytomembrane |
CmPIN15 | MELO3C005311.2 | 456 | 50.01 | 5.25 | 33.04 | 123.53 | 0.594 | 10 | 细胞质膜Cytomembrane |
CmPIN16 | MELO3C005326.2 | 636 | 69.50 | 7.15 | 36.26 | 91.42 | 0.111 | 10 | 细胞质膜Cytomembrane |
CmPIN17 | MELO3C018353.2 | 628 | 68.55 | 8.69 | 36.15 | 92.77 | 0.142 | 10 | 细胞质膜Cytomembrane |
CmPIN18 | MELO3C002132.2 | 642 | 70.30 | 9.01 | 36.31 | 83.99 | -0.029 | 10 | 细胞质膜Cytomembrane |
Table 2 Physicochemical property of CmPINs proteins in melon
基因 Gene | 基因ID Gene ID | 氨基酸数量 Number of amino acids | 分子量 Molecular weight/kD | 理论等电点 Theoretical pI | 不稳定指数 Instability index | 脂溶指数 Aliphatic index | 总平均疏水指数 Grand average of hydropathicity | 跨膜结构域数目 Transmembrane domain number | 亚细胞定位 Subcellular location |
---|---|---|---|---|---|---|---|---|---|
CmPIN1 | MELO3C001206.2 | 135 | 15.15 | 6.69 | 24.33 | 111.93 | 0.795 | 4 | 液泡膜 Tonoplast membrane |
CmPIN2 | MELO3C017414.2 | 623 | 68.44 | 9.22 | 41.57 | 85.47 | -0.012 | 10 | 细胞质膜Cytomembrane |
CmPIN3 | MELO3C017357.2 | 591 | 64.52 | 8.5 | 43.16 | 88.76 | 0.064 | 10 | 细胞质膜Cytomembrane |
CmPIN4 | MELO3C009892.2 | 417 | 46.35 | 9.76 | 48.87 | 118.63 | 0.454 | 11 | 细胞质膜Cytomembrane |
CmPIN5 | MELO3C008726.2 | 402 | 43.62 | 6.23 | 34.38 | 113.26 | 0.501 | 9 | 液泡膜Tonoplast membrane |
CmPIN6 | MELO3C008744.2 | 51 | 5.25 | 6.51 | 30.81 | 139.61 | 1.208 | 0 | 细胞质Cytoplasm |
CmPIN7 | MELO3C031448.2 | 166 | 17.73 | 9.26 | 40.24 | 107.65 | 0.489 | 2 | 叶绿体Chloroplast |
CmPIN8 | MELO3C031243.2 | 51 | 5.19 | 7.98 | 30.81 | 141.57 | 1.275 | 0 | 细胞外基质Extracellular matrix |
CmPIN9 | MELO3C031244.2 | 109 | 12.42 | 7.65 | 27.69 | 102.02 | 0.693 | 3 | 液泡膜Tonoplast membrane |
CmPIN10 | MELO3C013710.2 | 297 | 32.35 | 7.11 | 45.14 | 114.81 | 0.405 | 6 | 内质网Endoplasmic reticulum |
CmPIN11 | MELO3C013711.2 | 428 | 46.24 | 5.02 | 40.85 | 117.78 | 0.576 | 10 | 细胞质膜Cytomembrane |
CmPIN12 | MELO3C017060.2 | 421 | 45.46 | 7.66 | 36.48 | 128.5 | 0.704 | 10 | 液泡膜Tonoplast membrane |
CmPIN13 | MELO3C019102.2 | 611 | 65.37 | 8.75 | 37.4 | 96.73 | 0.265 | 10 | 细胞质膜Cytomembrane |
CmPIN14 | MELO3C025264.2 | 417 | 45.67 | 8.86 | 34.71 | 123.72 | 0.748 | 10 | 细胞质膜Cytomembrane |
CmPIN15 | MELO3C005311.2 | 456 | 50.01 | 5.25 | 33.04 | 123.53 | 0.594 | 10 | 细胞质膜Cytomembrane |
CmPIN16 | MELO3C005326.2 | 636 | 69.50 | 7.15 | 36.26 | 91.42 | 0.111 | 10 | 细胞质膜Cytomembrane |
CmPIN17 | MELO3C018353.2 | 628 | 68.55 | 8.69 | 36.15 | 92.77 | 0.142 | 10 | 细胞质膜Cytomembrane |
CmPIN18 | MELO3C002132.2 | 642 | 70.30 | 9.01 | 36.31 | 83.99 | -0.029 | 10 | 细胞质膜Cytomembrane |
基因 Gene | α-螺旋 Alpha helix/% | 延伸链 Extended strand/% | 无规则卷曲 Random coil/% | β-折叠 Beta turn/% |
---|---|---|---|---|
CmPIN1 | 51.11 | 22.22 | 25.19 | 1.48 |
CmPIN2 | 31.14 | 16.05 | 48.48 | 4.33 |
CmPIN3 | 30.12 | 15.91 | 49.24 | 4.74 |
CmPIN4 | 52.52 | 15.35 | 25.66 | 6.47 |
CmPIN5 | 46.02 | 16.67 | 31.59 | 5.72 |
CmPIN6 | 50.98 | 25.49 | 13.73 | 9.80 |
CmPIN7 | 24.70 | 27.71 | 39.76 | 7.83 |
CmPIN8 | 49.02 | 21.57 | 19.61 | 9.80 |
CmPIN9 | 46.79 | 25.69 | 26.61 | 0.92 |
CmPIN10 | 39.39 | 15.49 | 42.76 | 2.36 |
CmPIN11 | 40.42 | 18.93 | 36.45 | 4.21 |
CmPIN12 | 38.00 | 20.19 | 37.29 | 4.51 |
CmPIN13 | 31.75 | 14.24 | 49.75 | 4.26 |
CmPIN14 | 40.29 | 18.94 | 36.69 | 4.08 |
CmPIN15 | 41.45 | 17.11 | 38.38 | 3.07 |
CmPIN16 | 32.08 | 15.41 | 47.80 | 4.72 |
CmPIN17 | 30.89 | 16.40 | 47.13 | 5.57 |
CmPIN18 | 29.75 | 15.42 | 49.53 | 5.30 |
Table 3 Secondary structure of CmPINs proteins
基因 Gene | α-螺旋 Alpha helix/% | 延伸链 Extended strand/% | 无规则卷曲 Random coil/% | β-折叠 Beta turn/% |
---|---|---|---|---|
CmPIN1 | 51.11 | 22.22 | 25.19 | 1.48 |
CmPIN2 | 31.14 | 16.05 | 48.48 | 4.33 |
CmPIN3 | 30.12 | 15.91 | 49.24 | 4.74 |
CmPIN4 | 52.52 | 15.35 | 25.66 | 6.47 |
CmPIN5 | 46.02 | 16.67 | 31.59 | 5.72 |
CmPIN6 | 50.98 | 25.49 | 13.73 | 9.80 |
CmPIN7 | 24.70 | 27.71 | 39.76 | 7.83 |
CmPIN8 | 49.02 | 21.57 | 19.61 | 9.80 |
CmPIN9 | 46.79 | 25.69 | 26.61 | 0.92 |
CmPIN10 | 39.39 | 15.49 | 42.76 | 2.36 |
CmPIN11 | 40.42 | 18.93 | 36.45 | 4.21 |
CmPIN12 | 38.00 | 20.19 | 37.29 | 4.51 |
CmPIN13 | 31.75 | 14.24 | 49.75 | 4.26 |
CmPIN14 | 40.29 | 18.94 | 36.69 | 4.08 |
CmPIN15 | 41.45 | 17.11 | 38.38 | 3.07 |
CmPIN16 | 32.08 | 15.41 | 47.80 | 4.72 |
CmPIN17 | 30.89 | 16.40 | 47.13 | 5.57 |
CmPIN18 | 29.75 | 15.42 | 49.53 | 5.30 |
基因 Gene | 生长素相关 Auxin related | 赤霉素相关 Gibberellin related | 脱落酸相关 Abscisic acid related | 水杨酸相关 Salicylic acid related | 茉莉酸甲酯相关 Methyl jasmonate related | 光信号相关 Circadian- related | 分生组织相关 Meristem- related | 防御和应激相关 Defense related to stress | 低温反应相关 Low temperature related | 干旱诱导相关 Drought induced-related |
---|---|---|---|---|---|---|---|---|---|---|
CmPIN1 | 1 | - | 3 | - | 2 | 4 | 3 | 1 | - | - |
CmPIN2 | - | - | 3 | - | 4 | 3 | 1 | - | 1 | - |
CmPIN3 | 1 | - | 1 | - | - | 6 | - | 1 | - | 1 |
CmPIN4 | - | 1 | 2 | 1 | 2 | 3 | 2 | - | - | 1 |
CmPIN5 | - | 1 | 1 | - | 2 | 5 | - | - | - | - |
CmPIN6 | - | - | - | - | - | 1 | - | 3 | - | - |
CmPIN7 | - | - | - | - | - | 1 | - | 3 | - | - |
CmPIN8 | - | - | - | - | - | 1 | - | 3 | - | - |
CmPIN9 | 1 | - | 3 | - | 2 | 4 | 3 | 1 | - | - |
CmPIN10 | 1 | - | 3 | 3 | 2 | 5 | 1 | - | - | 1 |
CmPIN11 | 1 | - | 5 | 1 | 2 | - | - | 1 | - | - |
CmPIN12 | 1 | 1 | - | 1 | 4 | 7 | - | 2 | - | 1 |
CmPIN13 | - | - | 3 | - | 2 | 3 | - | - | - | - |
CmPIN14 | 1 | 1 | 1 | - | 4 | 4 | - | - | - | - |
CmPIN15 | - | - | 1 | - | 4 | 5 | - | 1 | 1 | - |
CmPIN16 | 1 | 1 | 2 | 1 | 4 | 2 | 1 | - | 2 | - |
CmPIN17 | 1 | 1 | 2 | - | 2 | 7 | - | 2 | - | - |
CmPIN18 | - | 1 | 4 | - | 6 | 2 | 1 | 1 | - | - |
总计Total | 9 | 7 | 34 | 7 | 42 | 63 | 12 | 19 | 4 | 4 |
Table 4 Predicted cis-elements in the promoter of the CmPINs genes
基因 Gene | 生长素相关 Auxin related | 赤霉素相关 Gibberellin related | 脱落酸相关 Abscisic acid related | 水杨酸相关 Salicylic acid related | 茉莉酸甲酯相关 Methyl jasmonate related | 光信号相关 Circadian- related | 分生组织相关 Meristem- related | 防御和应激相关 Defense related to stress | 低温反应相关 Low temperature related | 干旱诱导相关 Drought induced-related |
---|---|---|---|---|---|---|---|---|---|---|
CmPIN1 | 1 | - | 3 | - | 2 | 4 | 3 | 1 | - | - |
CmPIN2 | - | - | 3 | - | 4 | 3 | 1 | - | 1 | - |
CmPIN3 | 1 | - | 1 | - | - | 6 | - | 1 | - | 1 |
CmPIN4 | - | 1 | 2 | 1 | 2 | 3 | 2 | - | - | 1 |
CmPIN5 | - | 1 | 1 | - | 2 | 5 | - | - | - | - |
CmPIN6 | - | - | - | - | - | 1 | - | 3 | - | - |
CmPIN7 | - | - | - | - | - | 1 | - | 3 | - | - |
CmPIN8 | - | - | - | - | - | 1 | - | 3 | - | - |
CmPIN9 | 1 | - | 3 | - | 2 | 4 | 3 | 1 | - | - |
CmPIN10 | 1 | - | 3 | 3 | 2 | 5 | 1 | - | - | 1 |
CmPIN11 | 1 | - | 5 | 1 | 2 | - | - | 1 | - | - |
CmPIN12 | 1 | 1 | - | 1 | 4 | 7 | - | 2 | - | 1 |
CmPIN13 | - | - | 3 | - | 2 | 3 | - | - | - | - |
CmPIN14 | 1 | 1 | 1 | - | 4 | 4 | - | - | - | - |
CmPIN15 | - | - | 1 | - | 4 | 5 | - | 1 | 1 | - |
CmPIN16 | 1 | 1 | 2 | 1 | 4 | 2 | 1 | - | 2 | - |
CmPIN17 | 1 | 1 | 2 | - | 2 | 7 | - | 2 | - | - |
CmPIN18 | - | 1 | 4 | - | 6 | 2 | 1 | 1 | - | - |
总计Total | 9 | 7 | 34 | 7 | 42 | 63 | 12 | 19 | 4 | 4 |
Fig. 6 Effects of auxin inhibitors on the CmPIN2, CmPIN,CmPIN13, and CmPIN18 genes expression Error bars are the standard errors(SE). Different lowercase letters indicate significant differences(P < 0.05)
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