不同品种蝴蝶兰黄酮代谢途径分析

doi:10.3969/j.issn.1001-1803.2022.12.005

Abstract

Abstract:

High performance liquid chromatography （HPLC） was used to study the flavonoid metabolism pathways of 18 Phalaenopsis. A HPLC technology was established to analyze the pathway of flavonoid metabolism （regulate color）. Fluorescence quantitative PCR （Real-Time PCR） was used to further analyze the expression level of key genes in Phalaenopsis flavonoid synthesis pathway, and an Agrobacterium-mediated transient transformation of Phalaenopsis was used to obtain transgenic strains. Phalaenopsis HPLC separation technology can successfully separate 29 components, which is quite different from other species of grape and hyacinth. Real-Time PCR analysis of the key genes in the flavonoid synthesis pathway of different species of Phalaenopsis finds that the temporal and spatial expression patterns of Chalcone synthase （CHS）, Flavonol-3-hydroxylase （F3H） and Chalcone isomerase （CHI） of different species are basically the same. The key enzyme gene RNAi interference vector is constructed by Agrobacterium-mediated method to transform Phalaenopsis transiently. The interference expression of key enzyme genes in the flavonoid synthesis pathway will reduce the color of Phalaenopsis. In conclusion, the HPLC research method of Phalaenopsis flavonoid metabolism pathway is established, and its molecular mechanism is preliminary studied, which provids a theoretical and practical basis for revealing the color regulation mechanism of Phalaenopsis orchids.

Key words: Phalaenopsis, flavonoid metabolism, high performance liquid chromatography, molecular mechanism

CLC Number:

TQ658

Cheng Ling,Wang Jianping. Analysis of flavonoid metabolic pathways of different varieties of Phalaenopsis and their molecular mechanism[J].China Surfactant Detergent & Cosmetics, 2022, 52(12): 1293-1299.

Figures/Tables 8

Tab. 1

Fig. 1

Fig. 2

Tab. 2

Tab. 3

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Fig. 5

References 13

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基因	序列	功能
PhF3'5'H-F	CGAACTCCAATGGCTTCTCC	Real-Time PCR
PhF3'5'H-R	CCCAACTCCTCTCAGCATACCT	Real-Time PCR
PhDFR-F	TTCAATCCAAAGACCCTGAGAAC	Real-Time PCR
PhDFR-R	TTACGCTGCCTGACCTCTTG	Real-Time PCR
PhF3'H-F	ATCGGACTTAACCGACTCGC	Real-Time PCR
PhF3'H-R	CAACACGTGGAAGAGAGAGC	Real-Time PCR
Tub-F	GCCTAAGGTCTCGGATACTGTT	Real-Time PCR
Tub-R	GCTCATGGTGGCAGATATGA	Real-Time PCR

组分	时间/min	分子结构	分子及碎片分子量
1	1.532 5	—	—
2	3.871	—	—
3	5.046	飞燕草素-3-O-芸香糖苷-7-O-葡萄糖苷	773/303
4	5.698	飞燕草素-3-O-葡萄糖苷	465/303
5	6.276	—	—
6	7.683	—	—
7	9.052	—	—
8	11.047	飞燕草素-3-O-咖啡酰葡萄糖苷-7-O-葡萄糖苷	789/627/465/303
9	12.032	—	—
10	13.684	—	—
11	15.673	天竺葵素-3-O-桑布糖苷	565/271
12	16.176	—	—
13	17.591	矢车菊素-3-O-丙二酰葡萄糖苷-7-O-葡萄糖苷-3’-O-葡萄糖苷	859/773/287
14	19.158	飞燕草素	303
15	20.464	矢车菊素-3-O-苹果酰葡萄糖苷-7-O-葡萄糖苷	—
16	21.389	矢车菊素-3-O-咖啡酰葡萄糖苷	611/449/287
17	22.648	矢车菊素-3-O-丙二酰槐糖苷-7-O-阿魏酰葡萄糖苷-3’-O-丙二酰葡萄糖苷	1283/1197/949/287
18	22.967	矢车菊素-3-O-咖啡酰阿拉伯糖苷	581
19	24.057	矢车菊素-3-O-槐糖苷-7-O-芥子酰葡萄糖苷-3’-O-对香豆酰葡萄糖苷	1287
20	25.685	天竺葵素-3-O-咖啡酰葡萄糖苷-7-O-对香豆酰葡萄糖苷	903/433/271
21	27.486	矢车菊素-3-O-葡萄糖苷-7-O-阿魏酰葡萄糖苷-3’-O-芥子酰葡萄糖苷	1155
22	30.562	天竺葵素-3-O-咖啡酰葡萄糖苷	595/433/271
23	34.018	矢车菊素-3-O-乙酰葡萄糖苷-7-O-芥子酰葡萄糖苷-3’-O-芥子酰葡萄糖苷	1227
24	31.281	—	—
25	31.893	矢车菊素-3-O-丙二酰葡萄糖苷-7-O-芥子酰葡萄糖苷-3’-O-芥子酰葡萄糖苷	1271
26	32.182	天竺葵素-3-O-葡萄糖苷-7-O-芥子酰葡萄糖苷	801/595/271
27	39.425	矢车菊素-3-O-丙二酰葡萄糖苷-7-O-阿魏酰葡萄糖苷	873/697/611
28	42.183	天竺葵素-3-O-对香豆酰葡萄糖苷-7-O-芥子酰葡萄糖苷	947/741/595/271
29	46.837	天竺葵素-3-O-丙二酰葡萄糖苷-7-O-芥子酰葡萄糖苷	887/681/433/271

品种	花青素含量/%			总花青素含量/%	总黄酮含量/%
品种	天竺葵素	矢车菊素	飞燕草素	总花青素含量/%	总黄酮含量/%
聚宝红玫瑰	34.57±0.11	60.23±1.14	3.63±0.18	0.82±0.09	3.15±0.08
红梅	39.39±0.35	59.52±1.22	0.57±0.04	1.15±0.07	2.68±0.12
花F1	33.64±0.28	65.34±1.37	0.65±0.05	0.71±0.04	1.42±0.06
翔凤	27.74±0.25	71.53±1.45	0.37±0.03	1.19±0.07	1.86±0.02
红彤	23.19±0.24	75.17±1.58	0.49±0.07	1.18±0.05	2.14±0.11
大富贵	29.31±0.28	70.46±1.78	0.00±0.00	0.48±0.02	1.27±0.05
金尚美人	34.40±0.39	62.16±1.27	1.87±0.23	0.86±0.05	2.15±0.07
红珍珠	21.08±0.14	76.36±0.45	1.12±0.24	0.82±0.09	2.28±0.06
新红龙	25.87±0.85	73.05±1.32	0.43±0.08	0.81±0.07	1.64±0.14
红钻石	28.37±1.34	69.62±2.57	0.74±0.05	1.06±0.10	2.02±0.06
S083	26.22±0.39	72.02±2.74	0.63±0.08	0.86±0.06	1.26±0.09
A4656	17.73±0.46	69.06±1.53	3.76±0.32	1.27±0.10	1.39±0.07
眉红	19.29±0.58	77.71±2.47	1.65±0.26	0.84±0.05	1.63±0.06
红天使	17.35±0.29	79.10±3.63	2.37±0.24	0.68±0.05	1.57±0.08
大辣椒	34.05±1.42	65.82±3.47	0.00±0.00	0.97±0.07	1.41±0.09
眉白	0.00±0.00	0.00±0.00	0.00±0.00	0.00±0.00	3.15±0.08
A5744	0.00±0.00	0.00±0.00	0.00±0.00	0.00±0.00	1.76±0.09
P48	0.00±0.00	0.00±0.00	0.00±0.00	0.00±0.00	1.07±0.04

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Analysis of flavonoid metabolic pathways of different varieties of Phalaenopsis and their molecular mechanism

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