Transcriptome Analysis of the Response of Tree Peony‘Hu Hong’Under High Temperature Stress

doi:10.16420/j.issn.0513-353x.2023-0941

Acta Horticulturae Sinica ›› 2024, Vol. 51 ›› Issue (12): 2800-2816.doi: 10.16420/j.issn.0513-353x.2023-0941

• Genetic & Breeding·Germplasm Resources·Molecular Biology • Previous Articles Next Articles

Transcriptome Analysis of the Response of Tree Peony‘Hu Hong’Under High Temperature Stress

BU Wenxuan¹, YAO Yiping¹, HUANG Yu², YANG Xingyu¹, LUO Xiaoning¹, ZHANG Minhuan¹^,²(), LEI Weiqun², WANG Zheng³, TIAN Jianing¹, CHEN Lujie¹, QIN Liping²

¹College of Landscape Architecture，Central South University of Forestry and Technology，Changsha 410004，China
²College of Art and Design，Nanning University，Nanning 530200，China
³College of Landscape Architecture and Art，Henan Agricultural University，Zhengzhou 450046，China

Received:2024-03-01 Revised:2024-09-03 Online:2024-12-25 Published:2024-12-13
Contact: ZHANG Minhuan

Abstract

Abstract:

To analyze the molecular regulation mechanism of tree peony（Paeonia suffruticosa）in southern China，‘Hu Hong’was selected as the test material，the leaves treated at 20 ℃ and 40 ℃ were collected after 0，2，6，12 and 24 h，respectively. Transcriptomics and physiological and biochemical indicators were used to explore the differences in gene expression at different treatment times under high temperature stress. The result showed that the leaves showed water loss and biofilm damage，the relative conductivity of leaves and the proline content increased significantly with the prolongation of high temperature stress time. Free proline，relative conductivity and soluble protein were identified as important indicators to measure the changes of physiological mechanism in the response of peony to high temperature stress. A total of 492 693 unigene and 840 509 transcripts were generated by RNA-seq. 36 174 differentially expressed genes（DEGs）were annotated in 5 databases including GO and KEGG. The GO analysis showed that the main enrichment pathways were the reaction pathway to heat and the cycle pathway of reduced pentose phosphate. KEGG enrichment analysis showed that DEG is involved in signal transduction，photosynthesis and metabolism play a major role in responding to high temperature stress. Among the DEGs，multiple candidate genes and transcription factors involved in heat stress have been screened out，such as heat shock transcription factors PSHSFA2b，PSHSFA3，PSHSFB2a，PSHSFB2b，PSHSFB4，PSHSFC1 and some NAC，MYB，bHLH family genes. The expression of 11 selected genes under high temperature stress was detected by qRT-PCR，and the results were similar to RNA seq.

Key words: peony, high temperature stress, transcriptome, physiological and biochemical indicators

BU Wenxuan, YAO Yiping, HUANG Yu, YANG Xingyu, LUO Xiaoning, ZHANG Minhuan, LEI Weiqun, WANG Zheng, TIAN Jianing, CHEN Lujie, QIN Liping. Transcriptome Analysis of the Response of Tree Peony‘Hu Hong’Under High Temperature Stress[J]. Acta Horticulturae Sinica, 2024, 51(12): 2800-2816.

Figures/Tables 14

Table 1 Genes and primers used for qRT-PCR

基因名称（编号） Gene name（number）	底物名称 Substrate name	引物序列（5′-3′） Primer sequence
HSP20（TRINITY_DN57811_c1_g1_i1）	S539-1	F：TGCCCAAAGAGGAAGAGAA；R：CCCATGCCATCAGAAATTAA
WRKY20（TRINITY_DN58753_c6_g1_i1）	S539-4-2	F：CTAATGGCATCACAACCGTC；R：ACTAGTTTTGACGCGTTTTGT
HSPA1s（TRINITY_DN58827_c1_g2_i1）	S539-3	F：TTGTCAGCCGGAGAAAGC；R：GCAGGAGGCGGAGAAGTA
HSFC1（TRINITY_DN58986_c1_g1_i2）	S539-4	F：AAGGCGGTGGTCTTGGTT；R：TGGGGGTGTTGAGTTGGA
HSFA3（TRINITY_DN61524_c1_g1_i2）	S539-5	F：CATTTTGGTGGTGCTTGACA；R：AGACGGTTTCGTTCTTGGC
CRF4（TRINITY_DN63834_c0_g1_i1）	S539-6	F：TTCAACAATCAACACTACGGG；R：AGAGTTAAGCACACGGGGTT
HSFB2b（TRINITY_DN64862_c0_g3_i2）	S539-7	F：GACTACTTTCGCAAGGGGA；R：TTACCGTCGTTGAAGGAGC
NAC55（TRINITY_DN66043_c1_g1_i3）	S539-8	F：ACAATACCAGTGCCGCTTC；R：CCCCTAATACCCCTAACGAC
MCU3（TRINITY_DN67541_c3_g1_i1）	S539-9	F：ATTAGGGAATTATTTTGGGGG；R：TGGAAGGTGATTTTTTGTGAA
HSP90B（TRINITY_DN71090_c1_g1_i1）	S539-10	F：AGGTCCTTGATTATGGGATGAT；R：TGGATGGTCTGCTAATATGGAA
HSP70（TRINITY_DN74057_c3_g1_i1）	S539-11-2	F：CTGGGACTTGCCGATGAC；R：ACGACACTGAGAAGGCCCT
内参 Reference	S539-DN59422	F：CGAATCTTGTCTTGACCCCC；R：ATTGTCACCACCATCCCTACC

Fig. 1 Changes of leaf phenotype，relative water content and relative conductivity of‘Hu Hong’peony under high temperature（40 ℃）treatment t-test，* α = 0.05.

Fig. 2 Changes of chlorophyll a，chlorophyll b content，chlorophyll a/b and malondialdehyde content in leaves of‘Hu Hong’peony under high temperature（40 ℃）treatment t-test，* α = 0.05.

Fig. 3 Changes of soluble sugar，soluble protein and free proline content in leaves of‘Hu Hong’peony under high temperature（40 ℃）treatment t-test，* α = 0.05.

Fig. 4 Changes of superoxide dismutase（SOD），peroxidase（POD）and catalase（CAT）activities in leaves of Paeonia suffruticosa‘Hu Hong’under high temperature（40 ℃）treatment t-test，* α = 0.05.

Table 2 Correlation analysis of physiological indexes of‘Hu Hong’peony under high temperature stress

指标 Index	RWC	Rec	Chl	SS	SP	MDA	Pro	SOD	POD	CAT
RWC	1
Rec	-0.916^*	1
Chl	0.881^*	-0.852	1
SS	-0.924^*	0.922^*	-0.857	1
SP	-0.909^*	0.974^**	-0.764	0.947^*	1
MDA	-0.871	0.788	-0.943^*	0.917^*	0.763	1
Pro	-0.922^*	0.995^**	-0.812	0.911^*	0.983^**	0.751	1
SOD	0.748	-0.430	0.595	-0.635	-0.482	-0.731	-0.447	1
POD	-0.817	0.892^*	-0.557	0.771	0.929^*	0.504	0.931^*	-0.359	1
CAT	-0.690	0.914^*	-0.604	0.755	0.902^*	0.508	0.916^*	-0.066	0.897^*	1

Table 3 Principal component analysis results of physiological indexes of‘Hu Hong’peony under high temperature stress

生理指标 Physiologic indexs	特征向量 Eigenvector
生理指标 Physiologic indexs	主成分1 Principal component 1	主成分2 Principal component 2
RWC	-0.968	0.180
Rec	0.981	0.173
Chl	-0.880	0.273
SS	0.968	-0.108
SP	0.975	0.174
MDA	0.865	-0.426
Pro	0.978	0.196
SOD	-0.594	0.719
POD	0.866	0.378
CAT	0.830	0.550
特征值 Characteristic value	8.058	1.361
方差贡献率/% Variance contribution rate	80.578	13.612
累积贡献率/% Cumulative contribution rate	80.578	94.191

Fig. 5 Volcano map of differentially expressed genes of‘Hu Hong’peony under high temperature（40 ℃）and normal temperature（20 ℃）treatment

Fig. 6 Differentially expressed gene expression profiles of‘Hu Hong’peony leaves under high temperature stress Above is the ID of the trend，and below is the number of genes in the trend.

Fig. 7 GO enrichment analysis bubble diagram of differentially expressed genes between high temperature（40 ℃）treatment and control（20 ℃）at the same time period of‘Hu Hong’peony

Fig. 8 GO enrichment analysis bubble diagram of differentially expressed genes in different time periods of high temperature （40 ℃）treatment and control（20 ℃）of‘Hu Hong’peony

Fig. 9 Heat map of differentially expressed transcription factors between high temperature（40 ℃）treatment and control（20 ℃）at the same time period of‘Hu Hong’peony

Fig. 10 Differentially expressed transcription factor gene expression profiles of‘Hu Hong’peony at different temperatures

Fig. 11 qRT-PCR verification of transcriptome data of‘Hu Hong’peony in response to high temperature stress

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Transcriptome Analysis of the Response of Tree Peony‘Hu Hong’Under High Temperature Stress

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