Biotechnology Bulletin ›› 2023, Vol. 39 ›› Issue (12): 158-168.doi: 10.13560/j.cnki.biotech.bull.1985.2023-0390
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YANG Yan1(), MO Yu-xing2, ZHOU Yi1, CHEN Hui-ming3, XIAO Lang-tao1(), WANG Ruo-zhong1()
Received:
2023-04-23
Online:
2023-12-26
Published:
2024-01-11
Contact:
XIAO Lang-tao, WANG Ruo-zhong
E-mail:976664175@qq.com;ltxiao@hunau.edu.cn;wangruozhong@hunau.edu.cn
YANG Yan, MO Yu-xing, ZHOU Yi, CHEN Hui-ming, XIAO Lang-tao, WANG Ruo-zhong. Effects of Cucumber Endocarp Juice on Seed Germination[J]. Biotechnology Bulletin, 2023, 39(12): 158-168.
时间 Time/min | 流动相A Mobile phase A/% | 流动相B Mobile phase B/% |
---|---|---|
0 | 90 | 10 |
5 | 98 | 2 |
8 | 85 | 15 |
14 | 73 | 27 |
15 | 70 | 30 |
20 | 65 | 35 |
25 | 5 | 95 |
30 | 5 | 95 |
30.1 | 98 | 2 |
35 | 98 | 2 |
Table 1 Basic situation of mobile phase and retention time
时间 Time/min | 流动相A Mobile phase A/% | 流动相B Mobile phase B/% |
---|---|---|
0 | 90 | 10 |
5 | 98 | 2 |
8 | 85 | 15 |
14 | 73 | 27 |
15 | 70 | 30 |
20 | 65 | 35 |
25 | 5 | 95 |
30 | 5 | 95 |
30.1 | 98 | 2 |
35 | 98 | 2 |
编号Code | 基因Gene | 引物序列Primer sequence(5'-3') |
---|---|---|
1 | CsNCED1 | F: CACGCTGTCAGCATCAATCATGG R: GAAGGCGTGATACGCACGTGG |
2 | CsNCED2 | F: AAGACGACGGCCACATCTTAGCC R: GACGTAAATTCACACTCATCCCGC |
3 | CsYP707A1 | F: CGAAGACCTCAAGGGATGCTACTAC R: GCAGCGAAGATGAGACCTATGACG |
4 | CsPYL2 | F: GATGCAGGATGCAGTTGAACCAATT R: AAACAGAACTCCACACTTCCCCACT |
5 | CsPP2C2 | F: AGTGGAACGGACATCGTGTGTTTG R: AGCAAGAGATGAAGCCCCGTGTT |
6 | CsSnRK2 | F: AGTTCACAGATGAAGGGTTGGAAGG R: GACAATGACTAGCACTCCAAGGCAC |
Table 2 Related genes and qPCR primers
编号Code | 基因Gene | 引物序列Primer sequence(5'-3') |
---|---|---|
1 | CsNCED1 | F: CACGCTGTCAGCATCAATCATGG R: GAAGGCGTGATACGCACGTGG |
2 | CsNCED2 | F: AAGACGACGGCCACATCTTAGCC R: GACGTAAATTCACACTCATCCCGC |
3 | CsYP707A1 | F: CGAAGACCTCAAGGGATGCTACTAC R: GCAGCGAAGATGAGACCTATGACG |
4 | CsPYL2 | F: GATGCAGGATGCAGTTGAACCAATT R: AAACAGAACTCCACACTTCCCCACT |
5 | CsPP2C2 | F: AGTGGAACGGACATCGTGTGTTTG R: AGCAAGAGATGAAGCCCCGTGTT |
6 | CsSnRK2 | F: AGTTCACAGATGAAGGGTTGGAAGG R: GACAATGACTAGCACTCCAAGGCAC |
试剂Reagent | 体积Volume/μL |
---|---|
2×ChamQ Universal SYBR qPCR Master Mix | 5 |
Forward primer | 0.2 |
Reverse primer | 0.2 |
cDNA | 0.4 |
ddH2O | Up to 10 |
Table 3 Reaction system
试剂Reagent | 体积Volume/μL |
---|---|
2×ChamQ Universal SYBR qPCR Master Mix | 5 |
Forward primer | 0.2 |
Reverse primer | 0.2 |
cDNA | 0.4 |
ddH2O | Up to 10 |
Fig. 1 Viviparous phenotypes of cucumber fruits A: C-L fruit; B: C-L split fruit; C: M-H fruit; D: M-H split fruit; E: seedlings taken from C-L fruit; F: germination phenotype of ungerminated seeds in C-L fruit after removal
Fig. 3 Germination rate of cucumber seeds treated with juice Different lowercase letters in the figure indicate significant differences in cucumber seed germination rates under different treatments(P<0.05). The same below
Fig. 4 Germination phenotypes of cucumber seeds treated with the extracts of juice A: CK(distilled water); B: EtOH layer; C: PET layer; D: EtOAc layer; E: n-BuOH layer; F: water layer
序号No | 保留时间tR/min | 分子式Formula | 化合物Compounds | 加和离子 Plus ion | 误差值Error/ppm | 实际值Experimental mass/(m/z) | 理论值Theoretical mass/(m/z) |
---|---|---|---|---|---|---|---|
1 | 1.16 | C12H20O4 | 延胡索酸丁二酯 | Na+ | -1.143 0 | 228.135 1 | 252.135 4 |
2 | 1.34 | C5H13NO | 缬氨醇 | H+ | -8.759 0 | 104.106 1 | 104.107 0 |
3 | 1.34 | C5H10O | 环戊醇 | NH4+ | -8.759 0 | 104.106 1 | 104.107 0 |
4 | 1.45 | C6H9N3O2 | 组氨酸 | H+ | -0.815 0 | 156.076 7 | 156.076 8 |
5 | 2.18 | C6H8O7 | 柠檬酸 | H+ | 6.500 0 | 193.035 5 | 193.034 3 |
6 | 2.28 | C5H7NO3 | D-5-Oxoproline | H+ | 9.141 0 | 130.051 1 | 130.049 9 |
7 | 9.95 | C12H26O7 | 六乙二醇 | NH4+ | -0.514 0 | 300.201 6 | 300.201 7 |
8 | 10.42 | C14H30O8 | 七聚乙二醇 | NH4+ | -7.189 0 | 344.225 4 | 344.257 9 |
9 | 10.42 | C12H28N3O7 | (1R,2R,3S,4R,6S)-4,6-Diammonio-2,3-dihydroxycyclohexyl2-ammonio-2-deoxy-α-D-glucopyranoside | NH4+ | -3.289 0 | 344.225 4 | 344.226 6 |
10 | 10.92 | C16H34O9 | 伸辛二醇 | NH4+ | -0.363 0 | 388.25 4 | 388.254 1 |
11 | 11.00 | C12H20O4 | 己二酸-3-甲基-1,5-戊二醇二酯 | Na+ | -1.143 0 | 251.125 1 | 251.125 4 |
12 | 11.33 | C12H19NO2 | DOM | NH4+ | -3.025 0 | 227.174 7 | 227.175 4 |
13 | 11.33 | C12H22N2O2 | 克罗乙胺 | H+ | -3.025 0 | 227.174 7 | 227.175 4 |
14 | 11.33 | C15H24 | 石竹烯 | Na+ | -10.145 0 | 227.174 7 | 227.177 0 |
15 | 11.43 | C18H38O10 | 酒甘醇 | NH4+ | 0.996 0 | 432.280 6 | 432.280 3 |
16 | 11.92 | C20H42O11 | 二甘醇 | NH4+ | -2.011 0 | 476.305 6 | 476.306 5 |
17 | 12.40 | C22H46O12 | 十一乙二醇 | NH4+ | 2.990 0 | 520.334 3 | 520.332 8 |
18 | 14.58 | C4H8O2 | 4-羟基-2-丁酮 | H+ | 5.896 0 | 89.060 2 | 89.059 7 |
19 | 14.62 | C8H18O3 | 原乙酸三乙酯 | H+ | 3.692 0 | 163.133 5 | 163.132 9 |
20 | 15.59 | C31H64O16 | 十六乙二醇单甲醚 | NH4+ | -1.671 0 | 710.452 1 | 710.453 3 |
21 | 16.68 | C6H12O2 | 己酸 | K+ | -8.091 0 | 155.045 6 | 155.046 9 |
22 | 16.68 | C9H8O | 4-甲氧基苯乙炔 | Na+ | -7.111 0 | 155.045 6 | 155.466 7 |
23 | 16.68 | C6H6N2O3 | 硝基酚 | H+ | 3.321 0 | 155.047 4 | 155.045 6 |
24 | 18.73 | C9H18N2O | 4-(4-piperidyl)morpholine | H+ | 6.252 0 | 171.150 3 | 171.149 2 |
25 | 23.49 | C16H32O2 | 棕榈酸 | NH4+ | -4.715 0 | 274.272 8 | 274.274 1 |
26 | 23.49 | C16H35NO2 | (2)- n-methylamino 3 7 11-trme- | H+ | -4.715 0 | 274.272 8 | 274.274 1 |
27 | 23.96 | C9H16O | 壬二烯醛类 | NH4+ | -7.653 0 | 158.152 7 | 158.153 9 |
28 | 23.96 | C9H19NO | 四甲基哌啶醇 | H+ | -7.953 0 | 158.152 7 | 158.153 9 |
29 | 23.96 | C7H17N4 | 1-(Diaminomethylene)-4-ethylpiperazin-1-ium | H+ | 0.837 0 | 158.152 7 | 158.152 6 |
30 | 25.86 | C14H28O2 | 肉豆蔻酸 | NH4+ | -0.777 0 | 246.242 8 | 246.242 6 |
31 | 27.36 | C19H47 N7O4 | 己二酸 | H+ | -3.471 0 | 483.374 7 | 483.376 2 |
32 | 31.85 | C6H12N4 | 乌洛托品 | H+ | -0.067 0 | 141.113 5 | 141.113 5 |
Table 4 Compounds of juice
序号No | 保留时间tR/min | 分子式Formula | 化合物Compounds | 加和离子 Plus ion | 误差值Error/ppm | 实际值Experimental mass/(m/z) | 理论值Theoretical mass/(m/z) |
---|---|---|---|---|---|---|---|
1 | 1.16 | C12H20O4 | 延胡索酸丁二酯 | Na+ | -1.143 0 | 228.135 1 | 252.135 4 |
2 | 1.34 | C5H13NO | 缬氨醇 | H+ | -8.759 0 | 104.106 1 | 104.107 0 |
3 | 1.34 | C5H10O | 环戊醇 | NH4+ | -8.759 0 | 104.106 1 | 104.107 0 |
4 | 1.45 | C6H9N3O2 | 组氨酸 | H+ | -0.815 0 | 156.076 7 | 156.076 8 |
5 | 2.18 | C6H8O7 | 柠檬酸 | H+ | 6.500 0 | 193.035 5 | 193.034 3 |
6 | 2.28 | C5H7NO3 | D-5-Oxoproline | H+ | 9.141 0 | 130.051 1 | 130.049 9 |
7 | 9.95 | C12H26O7 | 六乙二醇 | NH4+ | -0.514 0 | 300.201 6 | 300.201 7 |
8 | 10.42 | C14H30O8 | 七聚乙二醇 | NH4+ | -7.189 0 | 344.225 4 | 344.257 9 |
9 | 10.42 | C12H28N3O7 | (1R,2R,3S,4R,6S)-4,6-Diammonio-2,3-dihydroxycyclohexyl2-ammonio-2-deoxy-α-D-glucopyranoside | NH4+ | -3.289 0 | 344.225 4 | 344.226 6 |
10 | 10.92 | C16H34O9 | 伸辛二醇 | NH4+ | -0.363 0 | 388.25 4 | 388.254 1 |
11 | 11.00 | C12H20O4 | 己二酸-3-甲基-1,5-戊二醇二酯 | Na+ | -1.143 0 | 251.125 1 | 251.125 4 |
12 | 11.33 | C12H19NO2 | DOM | NH4+ | -3.025 0 | 227.174 7 | 227.175 4 |
13 | 11.33 | C12H22N2O2 | 克罗乙胺 | H+ | -3.025 0 | 227.174 7 | 227.175 4 |
14 | 11.33 | C15H24 | 石竹烯 | Na+ | -10.145 0 | 227.174 7 | 227.177 0 |
15 | 11.43 | C18H38O10 | 酒甘醇 | NH4+ | 0.996 0 | 432.280 6 | 432.280 3 |
16 | 11.92 | C20H42O11 | 二甘醇 | NH4+ | -2.011 0 | 476.305 6 | 476.306 5 |
17 | 12.40 | C22H46O12 | 十一乙二醇 | NH4+ | 2.990 0 | 520.334 3 | 520.332 8 |
18 | 14.58 | C4H8O2 | 4-羟基-2-丁酮 | H+ | 5.896 0 | 89.060 2 | 89.059 7 |
19 | 14.62 | C8H18O3 | 原乙酸三乙酯 | H+ | 3.692 0 | 163.133 5 | 163.132 9 |
20 | 15.59 | C31H64O16 | 十六乙二醇单甲醚 | NH4+ | -1.671 0 | 710.452 1 | 710.453 3 |
21 | 16.68 | C6H12O2 | 己酸 | K+ | -8.091 0 | 155.045 6 | 155.046 9 |
22 | 16.68 | C9H8O | 4-甲氧基苯乙炔 | Na+ | -7.111 0 | 155.045 6 | 155.466 7 |
23 | 16.68 | C6H6N2O3 | 硝基酚 | H+ | 3.321 0 | 155.047 4 | 155.045 6 |
24 | 18.73 | C9H18N2O | 4-(4-piperidyl)morpholine | H+ | 6.252 0 | 171.150 3 | 171.149 2 |
25 | 23.49 | C16H32O2 | 棕榈酸 | NH4+ | -4.715 0 | 274.272 8 | 274.274 1 |
26 | 23.49 | C16H35NO2 | (2)- n-methylamino 3 7 11-trme- | H+ | -4.715 0 | 274.272 8 | 274.274 1 |
27 | 23.96 | C9H16O | 壬二烯醛类 | NH4+ | -7.653 0 | 158.152 7 | 158.153 9 |
28 | 23.96 | C9H19NO | 四甲基哌啶醇 | H+ | -7.953 0 | 158.152 7 | 158.153 9 |
29 | 23.96 | C7H17N4 | 1-(Diaminomethylene)-4-ethylpiperazin-1-ium | H+ | 0.837 0 | 158.152 7 | 158.152 6 |
30 | 25.86 | C14H28O2 | 肉豆蔻酸 | NH4+ | -0.777 0 | 246.242 8 | 246.242 6 |
31 | 27.36 | C19H47 N7O4 | 己二酸 | H+ | -3.471 0 | 483.374 7 | 483.376 2 |
32 | 31.85 | C6H12N4 | 乌洛托品 | H+ | -0.067 0 | 141.113 5 | 141.113 5 |
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