黄瓜内果皮汁液对种子萌发的影响

doi:10.13560/j.cnki.biotech.bull.1985.2023-0390

摘要/Abstract

摘要：

明确黄瓜内果皮汁液在种子萌发中的作用、化学成分及机制，为基于黄瓜来源的种子萌发抑制剂研发提供一定的理论依据。以易胎萌黄瓜（C-L）种子为供试材料，研究难胎萌黄瓜（M-H）内果皮汁液（质量分数为100%、50%）及其不同提取液（质量分数为3%）对C-L种子萌发的影响，并分析汁液化学成分。针对质量分数为100%的M-H汁液和蒸馏水（对照）处理的种子生理生化指标动态变化进行分析。质量分数为100%、50%的M-H黄瓜内果皮汁液对C-L种子萌发均表现出显著的抑制作用，20 h抑制率分别为100%和73%。乙醇相、正丁醇相在种子整个萌发过程中均发挥抑制作用，与对照相比，26 h其抑制率分别提高12.22%和47.77%。在汁液中鉴定到影响种子萌发的化合物：肉豆蔻酸、棕榈酸、柠檬酸和组氨酸。质量分数为100%汁液处理的种子可溶性糖含量变化不大。植物激素ABA含量一直高于对照，ABA/GA₃ 0-20 h显著高于对照。ABA合成酶基因CsNCED1 10-15 h表达量增加，CsNCED2 15-25 h表达量显著增加；ABA信号转导基因CsPYL2、CsPP2C2、CsSnRK2和代谢基因CsCYP707A1 10 h后表达量均显著增加，20 h时表达量均最高。黄瓜内果皮汁液中的肉豆蔻酸、棕榈酸、柠檬酸、组氨酸可能通过促使ABA合成基因CsNCED2及信号转导基因CsPYL2、CsPP2C2、CsSnRK2表达量增加，从而使得ABA含量提高，同时阻碍可溶性糖代谢，进而有效抑制种子萌发。

关键词: 黄瓜, 内果皮汁液, 种子萌发, 化学成分, 植物激素, ABA

Abstract:

The roles, chemical composition and mechanisms of endocarp juice on seed germination were studied, which may provide a theoretical basis for the research of cucumber seed germination inhibitors from cucumber source. Viviparous cucumber（C-L）seed was used as materials and cultivated by not viviparous（M-H）endocarp juice（mass fractions was 100% and 50%）and its various extracts（mass fractions was 3%）. The chemical composition of juice was analyzed. During seed germination, the physiological and biochemical factors were analyzed when the culture medium was a mass fraction of 100% juice and distilled water（control）. Cucumber endocarp juice with a mass fraction of 100% and 50% effectively inhibited cucumber seed germination, with inhibition rates of 100% and 73%, respectively, at 20 h. Throughout the process, the EtOH and n-BuOH layers inhibited the seed germination, at 26 h, the inhibition rates were 12.22% and 47.77% higher than those of the control, respectively. Myristic acid, palmitic acid, citric acid, and histidine were identified in the juice that affected seed germination. The soluble sugar content of the seeds was insignificant change after the 100% juice treatment process. From 0 to 20 h, ABA content was always higher than that of the control, and ABA/GA₃ was significantly higher than that of the control. The expressions of the ABA synthase genes CsNCED1 and CsNCED2 rose from 10 to 15 h and from 15 to 25 h, respectively. After 10 h, the expressions of ABA signal transduction genes CsPYL2, CsPP2C2, CsSnRK2, and catabolic gene CsCYP707A1 increased significantly, with the highest expression of four genes detected at 20 h. Cucumber endocarp juice contains myristic acid, palmitic acid, citric acid, and histidine, which may decrease seed germination by increasing the expressions of ABA synthesizing CsNCED2 and signal transduction genes CsPYL2, CsPP2C2, and CsSnRK2, as well as inhibiting soluble sugar metabolism.

Key words: cucumber, endocarp juice, seed germination, chemical composition, phytohormone, ABA

杨艳, 莫雨杏, 周祎, 陈惠明, 肖浪涛, 王若仲. 黄瓜内果皮汁液对种子萌发的影响[J]. 生物技术通报, 2023, 39(12): 158-168.

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.

图/表 14

参考文献 24

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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

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

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

序号No	保留时间t_R/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