基于转录组分析油菜素内酯对高温胁迫下酿酒葡萄花色苷合成及果实品质的调控机制

doi:10.3864/j.issn.0578-1752.2023.06.010

Abstract

Abstract:

【Objective】 The aims of the study were to analyze the genes involved in the regulation of grape anthocyanin accumulation and fruit quality by 2,4-Epibrassinolide (EBR) under high-temperature stress, and to explore the molecular mechanism of EBR regulation anthocyanin accumulation in grapes under high-temperature stress. 【Method】 Cabernet Sauvignon grapes were treated with high-temperature stress using infrared emitter, and sprayed 0.6 mg∙L^-1 of EBR before the veraison. The content of total anthocyanins, total sugar, reducing sugar and sucrose were quantified using the ultraviolet visible spectrophotometer. The mechanism of EBR-mediated accumulation of anthocyanin under high-temperature stress was analyzed by transcriptome sequencing. 【Result】 Starting from veraison, the anthocyanin content increased gradually under various treatments. At maturity, the total anthocyanin content in the high temperature group (HT) was significantly lower than that in the control group (CK), and the anthocyanin content in the high temperature and EBR group (HTE) was higher than that in the HT group, but lower than CK group. Under HT treatment, the accumulation pattern of total sugar, reducing sugar and sucrose was similar to that of anthocyanins and lower than those of CK group at maturity stage. Compared with HT group, the contents of various sugars in HTE group were increased. The differences in transcriptome levels of Cabernet Sauvignon fruits under the three treatments were analyzed. Through GO and KEGG enrichment, 14 differential genes related to sucrose and starch metabolic pathways, among which 10 genes were significantly up-regulated and 4 genes were significantly down-regulated under HT and HTE treatments. The expressions of 11 genes were different in the phenylpropane metabolic pathway. Seven genes involved in anthocyanin synthesis were up-regulated under the HT treatment, and 4 genes involved in lignin synthesis were significantly up-regulated under the HT treatment, indicating that high temperature might promote lignin synthesis and reduce the accumulation of anthocyanins. In the endogenous hormone signaling pathway, the expression of the ABA signaling receptor genes PP2C and SnRK2 was significantly increased under high-temperature stress, and might be involved in regulating the synthesis of grape anthocyanin under high-temperature stress together with EBR. The expression patterns of some differential genes were verified by qRT-PCR, which confirmed the accuracy of transcriptome data. 【Conclusion】 EBR alleviated the inhibitory effect of high temperature stress on grapevine anthocyanin accumulation, probably due to the fact that EBR reduced the expression of lignin-related genes and changed the expression pattern of grape endogenous hormone signal transduction genes.

Key words: Vitis vinifera L. cv Cabernet Sauvignon, 2,4-Epibrassinolide, high-temperature, anthocyanin, RNA-seq

WANG YueNing, DAI HongJun, HE Yan, WEI Qiang, GUO XueLiang, LIU Yan, YIN MengTing, WANG ZhenPing. Regulation Mechanism of Brassinolide on Anthocyanins Synthesis and Fruit Quality in Wine Grapes Under High Temperature Stress Based on Transcriptome Analysis[J].Scientia Agricultura Sinica, 2023, 56(6): 1139-1153.

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基因名称 Gene name	正向引物 Forward primer (5′-3′)	反向引物 Reverse Primer (5′-3′)	NCBI登录号 Accession number
VvActin	CTTGCATCCCTCAGCACCTT	TCCTGTGGACAATGGATGGA	NB000705
VvF3′5′H	ACTAAGCCACAGGAAACTAA	AAACCGCTCAGACCAAAACC	LOC100243414
VvSS	CCAGCCAACGTCATTAGCCT	CCATGACTTGTGGCCTTCCT	LOC100252799
VvLDOX	GCCTAAGACACCAAGCGACTACG	CAACCCAAGCGATAGCACCGATAG	LOC100233142
VvLAR	TGCTTTTGTGATTTTGTTAGAGG	CCCTTCCCCGATTGAGAGTA	LOC100232982
VvINV	CCAGAAAACTTGTAGAAGCACC	GTTGACGCATTCCTTAAGGATC	LOC100232951
VvBZR1	GCACGCCATCTTGCTCCAT	AAGCTCCAGATCATCCAAACCTA	LOC100262472
VvF3′H	GGTGATGTTAGGCAGGAGAGTGTTC	GCTCCACCACCATCTCTTTGAACTC	LOC100232896
VvPP2C	CCACGCTTCCCTCAAACCCTCCCA	CATCCACCACTCCATCATCCCCGC	LOC100242943
VvSnRK2	CAATGCATGCTTTGAAAGTGTG	GCCAAACTGCAGTCTGATTTTA	LOC100260262

时间 Time	处理 Treatment	花色苷 Anthocyanins (mg∙L^-1)	总糖 TotalSugar (mg∙g^-1)	还原糖 Reducing sugar (mg∙g^-1)	蔗糖 Sucrose (mg∙g^-1)
7.20	CK	0.2±0.51a	14.61±1.92a	8.90±0.02a	7.12±0.15a
	HT	0.2±0.14a	14.56±2.02a	9.13±0.18a	7.35±0.26a
	HTE	0.2±0.28a	14.22±1.80a	9.14±0.74a	7.12±.028a
8.10	CK	12.91±0.86c	143.72±5.14a	20.71±1.61a	15.64±0.65b
	HT	14.30±0.79ab	124.72±3.52b	29.88±1.14a	16.56±0.48ab
	HTE	15.09±0.24a	137.35±6.55a	36.30±3.02a	16.92±0.54a
8.30	CK	35.24±2.11ab	199.21±4.55a	127.68±2.36a	23.85±5.48a
	HT	32.61±2.59c	174.96±0.52b	118.02±2.18b	19.92±2.61c
	HTE	36.04±2.98a	199.30±6.63a	124.29±9.47ab	22.34±0.69ab
9.20	CK	49.10±3.07a	225.87±4.19a	128.04±11.85a	26.67±6.93bc
	HT	39.33±2.75c	216.42±4.57b	108.17±15.81c	27.07±6.21b
	HTE	41.19±2.64b	222.41±5.16a	121.40±10.70b	29.43±5.35a

样品名称 Sample	原始序列数据 Raw reads (M)	原始序列长度 Raw bases (G)	过滤后数据 Clean reads (M)	过滤后序列长度 Clean bases (G)	比对率 Valid bases (%)	Q30 (%)	GC (%)
CK1	50.55	7.58	49.83	7.26	95.71	91.50	47.66
CK2	47.78	7.17	47.06	6.86	95.75	92.75	47.52
CK3	47.30	7.10	46.61	6.79	95.68	91.75	47.55
HT1	48.07	7.21	47.37	6.88	95.37	89.64	47.94
HT2	49.42	7.41	48.66	7.08	95.49	92.00	47.56
HT3	49.86	7.48	49.12	7.16	95.69	91.16	47.17
HTE1	48.84	7.33	48.09	7.01	95.71	92.44	47.45
HTE2	51.05	7.66	50.31	7.31	95.52	91.53	47.74
HTE3	50.46	7.57	49.75	7.25	95.78	92.67	47.37

处理 Treatment	差异转录因子个数 Transcription factor number	上调 Up	下调 Down	转录因子家族 Transcription factor family
HT vs CK	95	74	21	AP2、bHLH、bZIP、C2H2、Dof、ERF、HD-ZIP、LBD、MIKC-MADS、MYB、NAC、WRKY
HTE vs CK	57	44	13	bHLH、bZIP、Dof、ERF、HD-ZIP、LBD、MYB、NAC、WRKY
HT vs HTE	6	1	5	MYB

Regulation Mechanism of Brassinolide on Anthocyanins Synthesis and Fruit Quality in Wine Grapes Under High Temperature Stress Based on Transcriptome Analysis

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