Biotechnology Bulletin ›› 2024, Vol. 40 ›› Issue (5): 203-214.doi: 10.13560/j.cnki.biotech.bull.1985.2023-1047
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WU Di(), YOU Xiao-feng, ZHENG Yi-zheng, LIN Nan, ZHANG Yan-yan, WEI Yi-cong()
Received:
2023-11-09
Online:
2024-05-26
Published:
2024-06-13
Contact:
WEI Yi-cong
E-mail:827820547@qq.com;2007047@fjtcm.edu.cn
WU Di, YOU Xiao-feng, ZHENG Yi-zheng, LIN Nan, ZHANG Yan-yan, WEI Yi-cong. Analysis of Endogenous Hormone Regulation Mechanism for Carotenoid Synthesis in Sarcandra glabra[J]. Biotechnology Bulletin, 2024, 40(5): 203-214.
基因 Gene | 正向引物 Forward primer(5'-3') | 反向引物Reverse primer(5'-3') |
---|---|---|
CAC | TCCGACAAATTGGAGGTTGC | TGCTGCTGACAACAATCACG |
DXS | GCCTTGAACGGACTCTTAGAC | AGGTGGAGAGCATCCTTGTT |
PSY | GGCGTCCTTGTTGTTGTACC | ATGAGTGCTCTAGTGTAGGCTATG |
NCED | CTCCAAGGCGTGTTCTATCG | CTCCAAGGCGTGTTCTATCG |
VDE | CACGACAACGAGTACCTTCAC | TGACAGCACCACCATATCCA |
PDS | ACGAGTACCAGGTGTTGTTCTA | AATCAGTTTGCCAGCATTCATTG |
LYCB | GGAAGATATACAGGAGAGGATGGT | TCTGAGGAAGCACTGGAAGT |
ZEP | GCTTGGAGGCAGAGTATTGAC | GCAGCCATTCTAGCCATTCC |
CYP97C1 | GATGAGAGCACAACAAGAGGTT | CGCCTTATTAACACTGGTGGAT |
Table 1 Primers used for real-time quantitative RT-PCR
基因 Gene | 正向引物 Forward primer(5'-3') | 反向引物Reverse primer(5'-3') |
---|---|---|
CAC | TCCGACAAATTGGAGGTTGC | TGCTGCTGACAACAATCACG |
DXS | GCCTTGAACGGACTCTTAGAC | AGGTGGAGAGCATCCTTGTT |
PSY | GGCGTCCTTGTTGTTGTACC | ATGAGTGCTCTAGTGTAGGCTATG |
NCED | CTCCAAGGCGTGTTCTATCG | CTCCAAGGCGTGTTCTATCG |
VDE | CACGACAACGAGTACCTTCAC | TGACAGCACCACCATATCCA |
PDS | ACGAGTACCAGGTGTTGTTCTA | AATCAGTTTGCCAGCATTCATTG |
LYCB | GGAAGATATACAGGAGAGGATGGT | TCTGAGGAAGCACTGGAAGT |
ZEP | GCTTGGAGGCAGAGTATTGAC | GCAGCCATTCTAGCCATTCC |
CYP97C1 | GATGAGAGCACAACAAGAGGTT | CGCCTTATTAACACTGGTGGAT |
化合物 Compound | 分子式 Formula | 离子模式 ESM | 质荷比 m/z | 保留时间 Retention/min | 二级鉴定分数 MS2 score |
---|---|---|---|---|---|
吲哚-3-羧酸 Indole-3-carboxylic acid | C9H7NO2 | - | 160.04 | 1.84 | 43.60 |
吲哚 Indole | C8H7N | - | 152.01 | 1.88 | 63.40 |
吲哚-5-羧酸 Indole-5-carboxylic acid | C9H7NO2 | - | 160.04 | 2.05 | 33.60 |
3-甲基羟基吲哚 3-Methyldioxyindole | C9H9NO2 | - | 162.06 | 2.27 | 38.70 |
5,6-二羟基吲哚 5,6-Dihydroxyindole | C8H7NO2 | - | 297.09 | 2.67 | 27.00 |
吲哚-3-甲醛 Indole-3-carboxaldehyde | C9H7NO | + | 110.04 | 3.88 | 0.00 |
5,6-吲哚醌-2-羧酸 5,6-Indolequinone-2-carboxylic acid | C9H5NO4 | + | 383.05 | 1.81 | 22.00 |
1-萘乙酸乙酯 Ethyl 1-naphthylacetic acid | C14H14O2 | + | 215.11 | 3.60 | 34.30 |
赤霉素3 Gibberellin A3 | C19H22O6 | - | 391.15 | 3.16 | 74.30 |
赤霉素4 Gibberellin A4 | C19H24O5 | - | 663.29 | 6.78 | 78.10 |
赤霉素36 Gibberellin A36 | C20H26O6 | + | 345.17 | 7.76 | 66.80 |
邻β-d-葡糖基玉米素 O-Beta-D-glucosylzeatin | C16H23N5O6 | - | 402.13 | 3.65 | 74.30 |
油菜素内酯 Brassinolide | C28H48O6 | + | 445.34 | 10.88 | 53.40 |
二氢茉莉酸 Dihydrojasmonic acid | C12H20O3 | - | 233.12 | 4.42 | 36.50 |
(-)-茉莉酸(-)-Jasmonic acid | C12H18O3 | + | 175.11 | 4.99 | 66.60 |
茉莉酸 Jasmonic acid | C12H18O3 | + | 175.11 | 7.34 | 69.00 |
5-甲氧基水杨酸 5-Methoxysalicylic acid | C8H8O4 | - | 167.04 | 1.35 | 86.90 |
赤霉素20 Gibberellin A20 | C19H24O5 | - | 353.15 | 7.05 | 47.80 |
赤霉素8 Gibberellin A8 | C19H24O7 | + | 387.13 | 4.17 | 66.10 |
赤霉素19 Gibberellin A19 | C20H26O6 | + | 385.17 | 6.84 | 70.20 |
赤霉素9 Gibberellin A9 | C19H24O4 | + | 317.18 | 8.95 | 42.20 |
(+)-脱落酸(+)-Abscisic acid | C15H20O4 | + | 247.13 | 5.70 | 81.20 |
油菜素甾酮 Castasterone | C28H48O5 | + | 447.34 | 10.77 | 60.70 |
褪黑素 Melatonin | C13H16N2O2 | - | 277.12 | 2.80 | 29.50 |
对水杨酸 P-salicylic acid | C7H6O3 | - | 137.03 | 1.56 | 28.90 |
茉莉酸甲酯 Methyl jasmonate | C13H20O3 | + | 207.14 | 3.00 | 45.30 |
二氢茉莉酮酸甲酯 Methyl dihydrojasmonate | C13H22O3 | - | 225.15 | 4.17 | 25.80 |
2-萘乙酸 2-Naphthylacetic acid | C12H10O2 | + | 187.08 | 8.05 | 33.90 |
玉米素 Zeatin | C10H13N5O | + | 220.12 | 1.57 | 26.50 |
玉米素核苷 cis-Zeatin riboside | C15H21N5O5 | + | 374.15 | 3.43 | 78.90 |
3-吲哚丙烯酸 3-Indoleacrylic acid | C11H9NO2 | - | 208.03 | 3.07 | 43.60 |
Table 2 Endogenous hormones in the leaves and roots of S. glabra
化合物 Compound | 分子式 Formula | 离子模式 ESM | 质荷比 m/z | 保留时间 Retention/min | 二级鉴定分数 MS2 score |
---|---|---|---|---|---|
吲哚-3-羧酸 Indole-3-carboxylic acid | C9H7NO2 | - | 160.04 | 1.84 | 43.60 |
吲哚 Indole | C8H7N | - | 152.01 | 1.88 | 63.40 |
吲哚-5-羧酸 Indole-5-carboxylic acid | C9H7NO2 | - | 160.04 | 2.05 | 33.60 |
3-甲基羟基吲哚 3-Methyldioxyindole | C9H9NO2 | - | 162.06 | 2.27 | 38.70 |
5,6-二羟基吲哚 5,6-Dihydroxyindole | C8H7NO2 | - | 297.09 | 2.67 | 27.00 |
吲哚-3-甲醛 Indole-3-carboxaldehyde | C9H7NO | + | 110.04 | 3.88 | 0.00 |
5,6-吲哚醌-2-羧酸 5,6-Indolequinone-2-carboxylic acid | C9H5NO4 | + | 383.05 | 1.81 | 22.00 |
1-萘乙酸乙酯 Ethyl 1-naphthylacetic acid | C14H14O2 | + | 215.11 | 3.60 | 34.30 |
赤霉素3 Gibberellin A3 | C19H22O6 | - | 391.15 | 3.16 | 74.30 |
赤霉素4 Gibberellin A4 | C19H24O5 | - | 663.29 | 6.78 | 78.10 |
赤霉素36 Gibberellin A36 | C20H26O6 | + | 345.17 | 7.76 | 66.80 |
邻β-d-葡糖基玉米素 O-Beta-D-glucosylzeatin | C16H23N5O6 | - | 402.13 | 3.65 | 74.30 |
油菜素内酯 Brassinolide | C28H48O6 | + | 445.34 | 10.88 | 53.40 |
二氢茉莉酸 Dihydrojasmonic acid | C12H20O3 | - | 233.12 | 4.42 | 36.50 |
(-)-茉莉酸(-)-Jasmonic acid | C12H18O3 | + | 175.11 | 4.99 | 66.60 |
茉莉酸 Jasmonic acid | C12H18O3 | + | 175.11 | 7.34 | 69.00 |
5-甲氧基水杨酸 5-Methoxysalicylic acid | C8H8O4 | - | 167.04 | 1.35 | 86.90 |
赤霉素20 Gibberellin A20 | C19H24O5 | - | 353.15 | 7.05 | 47.80 |
赤霉素8 Gibberellin A8 | C19H24O7 | + | 387.13 | 4.17 | 66.10 |
赤霉素19 Gibberellin A19 | C20H26O6 | + | 385.17 | 6.84 | 70.20 |
赤霉素9 Gibberellin A9 | C19H24O4 | + | 317.18 | 8.95 | 42.20 |
(+)-脱落酸(+)-Abscisic acid | C15H20O4 | + | 247.13 | 5.70 | 81.20 |
油菜素甾酮 Castasterone | C28H48O5 | + | 447.34 | 10.77 | 60.70 |
褪黑素 Melatonin | C13H16N2O2 | - | 277.12 | 2.80 | 29.50 |
对水杨酸 P-salicylic acid | C7H6O3 | - | 137.03 | 1.56 | 28.90 |
茉莉酸甲酯 Methyl jasmonate | C13H20O3 | + | 207.14 | 3.00 | 45.30 |
二氢茉莉酮酸甲酯 Methyl dihydrojasmonate | C13H22O3 | - | 225.15 | 4.17 | 25.80 |
2-萘乙酸 2-Naphthylacetic acid | C12H10O2 | + | 187.08 | 8.05 | 33.90 |
玉米素 Zeatin | C10H13N5O | + | 220.12 | 1.57 | 26.50 |
玉米素核苷 cis-Zeatin riboside | C15H21N5O5 | + | 374.15 | 3.43 | 78.90 |
3-吲哚丙烯酸 3-Indoleacrylic acid | C11H9NO2 | - | 208.03 | 3.07 | 43.60 |
Fig. 1 Heat map of the relative contents of endogenous hormones in the leaves and roots of S. glabra L1-6: Leaves(samples 1 to 6). R1-6: Root(samples 1 to 6). The same below
代谢酶Enzyme name | 缩写Abbreviation | KO 编码KO ID | unigene数量Number of unigene |
---|---|---|---|
1-脱氧-D-木酮糖5-磷酸合酶1-Deoxy-D-xylulose 5-phosphate synthase | DXS | K01662 | 3 |
15-cis-phytoene合成酶15-cis-Phytoene synthase | PSY | K02291 | 1 |
15 - cis - Phytoene去饱和酶15-cis-Phytoene desaturase | PDS | K02293 | 1 |
前番茄红素异构酶Prolycopene isomerase | CRTISO | K09835 | 1 |
番茄红素β-环化酶Lycopene beta-cyclase | LCYB | K06443 | 1 |
番茄红素环化酶Lycopene epsilon-cyclase | LCYE | K06444 | 1 |
胡萝卜素羟基Beta-ring hydroxylase | CHYB | K15747 | 2 |
类胡萝卜素ε-羟化酶Carotenoid epsilon hydroxylase | CYP97C1 | K09837 | 1 |
β -胡萝卜素异构酶Beta-carotene isomerase | DWARF27 | K17911 | 3 |
内酯合成酶Carlactone synthase | CCD8 | K17913 | 1 |
玉米黄质环氧化酶Zeaxanthin epoxidase | ZEP | K09838 | 3 |
紫黄质脱环氧化酶Violaxanthin de-epoxidase | VDE | K09839 | 1 |
9-顺式-环氧类胡萝卜素二加氧酶9-cis-Epoxycarotenoid dioxygenase | NCED | K09840 | 2 |
黄氧素脱氢酶Xanthoxin dehydrogenase | ABA2 | K09841 | 3 |
(+)- 脱落酸8’羟化酶(+)-Abscisic acid 8’-hydroxylase | CYP707A | K09843 | 1 |
脱落酸β-葡萄糖基转移酶Abscisate beta-glucosyltransferase | AOG | K14595 | 1 |
Table 3 Differential metabolic enzyme genes related to carotenoids in the leaves and roots of S. glabra
代谢酶Enzyme name | 缩写Abbreviation | KO 编码KO ID | unigene数量Number of unigene |
---|---|---|---|
1-脱氧-D-木酮糖5-磷酸合酶1-Deoxy-D-xylulose 5-phosphate synthase | DXS | K01662 | 3 |
15-cis-phytoene合成酶15-cis-Phytoene synthase | PSY | K02291 | 1 |
15 - cis - Phytoene去饱和酶15-cis-Phytoene desaturase | PDS | K02293 | 1 |
前番茄红素异构酶Prolycopene isomerase | CRTISO | K09835 | 1 |
番茄红素β-环化酶Lycopene beta-cyclase | LCYB | K06443 | 1 |
番茄红素环化酶Lycopene epsilon-cyclase | LCYE | K06444 | 1 |
胡萝卜素羟基Beta-ring hydroxylase | CHYB | K15747 | 2 |
类胡萝卜素ε-羟化酶Carotenoid epsilon hydroxylase | CYP97C1 | K09837 | 1 |
β -胡萝卜素异构酶Beta-carotene isomerase | DWARF27 | K17911 | 3 |
内酯合成酶Carlactone synthase | CCD8 | K17913 | 1 |
玉米黄质环氧化酶Zeaxanthin epoxidase | ZEP | K09838 | 3 |
紫黄质脱环氧化酶Violaxanthin de-epoxidase | VDE | K09839 | 1 |
9-顺式-环氧类胡萝卜素二加氧酶9-cis-Epoxycarotenoid dioxygenase | NCED | K09840 | 2 |
黄氧素脱氢酶Xanthoxin dehydrogenase | ABA2 | K09841 | 3 |
(+)- 脱落酸8’羟化酶(+)-Abscisic acid 8’-hydroxylase | CYP707A | K09843 | 1 |
脱落酸β-葡萄糖基转移酶Abscisate beta-glucosyltransferase | AOG | K14595 | 1 |
转录因子Transcription factor | 预测顺式作用元件Predicting cis-acting element | 差异内源激素变化Differential endogenous hormone changes(leaf/root) | 调控下游酶基因的表达趋势Regulating the expression trend of downstream enzyme genes(leaf/root) |
---|---|---|---|
MYB106 | SA、MeJA、GA、ABA | MeJA上调 | DXS、VDE、ZEP、CYP97C1、LCYB、PSY、CRTISO上调 |
SPL1 | MeJA、GA、ABA、Auxin | MeJA上调 | DXS、VDE、ZEP、CYP97C1、LCYB、PSY、CRTISO上调 |
NAC015 | SA、MeJA、GA | MeJA上调 | DXS、VDE、ZEP、CYP97C1、LCYB、PSY、CRTISO上调 |
WRKY44 | ABA、MeJA、SA | ABA下调、SA下调 | ABA2、AOG、DWARF27下调 |
BHLH16 | ABA、Zein、SA、GA、MeJA | ABA下调、SA下调、GA下调 | ABA2、AOG、DWARF27下调 |
EFR064 | ABA、Zein、SA、GA | ABA下调、SA下调、GA下调 | ABA2、AOG、DWARF27下调 |
Table 4 Regulation relationship of carotenoid biosynthetic pathway “endogenous hormone-transcription factor-metabolic enzyme gene expression” in S. glabra
转录因子Transcription factor | 预测顺式作用元件Predicting cis-acting element | 差异内源激素变化Differential endogenous hormone changes(leaf/root) | 调控下游酶基因的表达趋势Regulating the expression trend of downstream enzyme genes(leaf/root) |
---|---|---|---|
MYB106 | SA、MeJA、GA、ABA | MeJA上调 | DXS、VDE、ZEP、CYP97C1、LCYB、PSY、CRTISO上调 |
SPL1 | MeJA、GA、ABA、Auxin | MeJA上调 | DXS、VDE、ZEP、CYP97C1、LCYB、PSY、CRTISO上调 |
NAC015 | SA、MeJA、GA | MeJA上调 | DXS、VDE、ZEP、CYP97C1、LCYB、PSY、CRTISO上调 |
WRKY44 | ABA、MeJA、SA | ABA下调、SA下调 | ABA2、AOG、DWARF27下调 |
BHLH16 | ABA、Zein、SA、GA、MeJA | ABA下调、SA下调、GA下调 | ABA2、AOG、DWARF27下调 |
EFR064 | ABA、Zein、SA、GA | ABA下调、SA下调、GA下调 | ABA2、AOG、DWARF27下调 |
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