基于转录组学分析辣椒对磷营养逆境的响应

doi:10.3864/j.issn.0578-1752.2024.14.011

Abstract

Abstract:

【Objective】 This study aimed to explore the changes in the transcriptional level and physiological responses of pepper (Capsicum annuum L.) seedlings under different phosphorus stress gradients, to analyze the key pathways, and to integrate related experiments to elucidate the physiological mechanisms of the pepper response to nutritional stress of phosphorus. This study also identified the transcription factors and core genes that regulate the nutritional stress of phosphorus, so as to provide a theoretical basis and genetic resources for the breeding of peppers. 【Method】 This study used the roots of the pure CA#8 seedlings of pepper as experimental material, grown by Hoagland hydroponics in a single four-leaf stage and subjected to four different gradient phosphorus stress treatments, namely control group (CK, 200 μmol·L^-1 NH₄H₂PO₄) and phosphorus deficiency group (DP, 0 μmol·L^-1 NH₄H₂PO₄), low phosphorus stress group (LP, 20 μmol·L^-1 NH₄H₂PO₄), and high phosphorus stress group (HP, 1 000 μmol·L^-1 NH₄H₂PO₄). Transcriptome sequencing (RNA-Seq) was performed on pepper roots systems 2 days after treatment, and the endogenous hormone levels and antioxidant enzyme activities were measured after 0, 2, 4, and 6 days of treatment, respectively. 【Result】 Compared with the CK group, the DP, LP, and HP groups exhibited 626, 107, and 171 differentially expressed genes (DEG), respectively. 10 DEGs related to the phosphorus signaling pathway, 4 DEGs related to the plant hormone signal transduction pathway, and 7 DEGs related to antioxidant enzyme activities were identified by GO, KEGG enrichment analysis, and WGCNA analysis. Real-time fluorescence quantitative (qRT-PCR) was used to validate the accuracy of transcriptome data. Quantitative measurements of endogenous hormones in the roots of pepper seedlings showed that with increasing duration phosphorus stress, the levels of various growth-promoting endogenous hormones such as gibberellin (GA), brassinolide (BR), cytokinin (CTK), indole acetic acid (IAA), strigolactone (SL), and jasmonic acid (JA) decreased in the DP, LP and HP groups, while the levels of growth-inhibiting hormones such as ethylene (ETH) and abscisic acid (ABA) increased, especially under phosphorus deficiency and low phosphorus stress, where growth inhibition of the seedling roots was the most significant. Various phosphorus stress-induced increased activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) in pepper root tissue, while the activities of SOD decreased at the later stage of stress, and POD and CAT tended to be stable too. 【Conclusion】 During various phosphorus nutritional stress, the pepper seedlings mitigated the effects of phosphorus stress on growth by responding to phosphorus signaling pathways, plant hormone signal transduction pathways, and differentially expressed genes related to antioxidant enzyme activity, thus improving the tolerance to phosphorus stress of pepper seedlings.

Key words: Capsicum annuum L., phosphorus stress, phosphorus starvation response, transcriptome, gene mining

XU MingRui, WANG XiaoJuan, YANG YaLi, MA YueFei, LIU WanMao, SUN Ying. Transcriptomics-Based Analysis of Pepper Responses to Phosphorus Nutritional Stress[J].Scientia Agricultura Sinica, 2024, 57(14): 2827-2846.

Figures/Tables 11

Table 1

Real-time fluorescent quantitative PCR primer sequence"

基因分类 Gene classification	基因名称 Gene name	基因ID Gene ID	正向引物 Forward primer (5´-3´)	反向引物 Reverse primer (5´-3´)
内参基因 Internal reference gene	CaActin	LOC107850541	AGTGGTCGTACAACTGGTATTG	GCAAGATCCAAACGAAGAATGG
磷信号相关基因 Phosphorus signaling related genes	CaSPX3	LOC107857225	AAGAGTGTGAAAGCACCATAGA	CTTCACCTGCAGCTCCTATTT
	CaSPX2	LOC107840775	AGAGCACCATATCCGCTTTAC	CCAGGTATCTTCGAGTCCACTA
	CaSPX1	LOC107863810	ACTCAGTGGCGAGCTTATTC	TGGCTCAGTTTGGTACAAGAG
	CaPHO1;H1	LOC107848491	ACACGCGGAGAAGATGATTC	AGCATGTTCAAGGTCCGATAAG
	CaPHO1;H10	LOC107848334	CTCATGGAAGACAGGGTTCTAC	GTGAATCAGTCTCTCCGTTCTC
	CaPht1;9	LOC107866766	CATCGCAGGTATGGGCTTATT	CCGTTGGCGATCTGTAGTAAAT
	CaPht1;4	LOC107868451	TCCGGACTCTCCTTTGGTAATA	CGTGGCCGATAAAGGGTAAT
	CaPAP2	LOC107866942	GGAGATGGAGGAAATCAAGAAGG	TCCAATATACCGTGCCCAAAG
	CaPAP15	LOC107864505	GGACCTGGCTAATGTTGATAGG	CGATAGTGTGCCGTGTAAGTAG
	CaPAP16	LOC107872241	GGGAGCTAGGATTCTGGAGATA	GCTTGCCGTAGAGCTCAATA
抗氧化酶活性相关基因 Antioxidant enzyme activity related genes	CaPER44	LOC107847017	GTACGAGGATACGAGCTGATTG	CCTTCCTGTGGGTATGTTGTAG
	CaPER41	LOC107850917	AGGAGATGGTTGTGTTGACTG	GGTTCATTGTTGGATCAGCATTAG
	CaPER55	LOC107860772	GCTGAAGGGCAACTAGAAGAG	AGACGCAAAGTAGCTGGAATAG
	CaPER7	LOC107861283	TGTCCGACGTTGGAAAGTATAG	CTAACGAAGCAGTCATGGAAATG
	CaSODB	LOC107873354	TGCTCACGATAGACGTTTGG	ATGACACGAGCTTCTCCATAAA
	CaGRXC6	LOC107871022	GGTGGTGGTATGGATGAGATG	GGAACAAGTCCACCACTTAGAT
	CaGRXS3	LOC107874941	TTTCTGCTCTCGGCTGTAATC	TGATCTCACTTTCTCCACCAAC
植物激素信号转导相关基因 Phytohormone signaling related genes	CaAUX22D	LOC107863844	GCCACCAGTTCGATCTTACA	AAGGTGCTCCATCCATACTAAC
	CaPIF4	LOC107878077	GTGGATCGGGCAGTAGTTATG	TACTCTGGCATTCTGGTTCTTC
	CaCYP707A1	LOC107859996	GTACCAGGGAATCTGGGAATAC	GCCGGAGTCTAACAAAGTCATA
	CaSAUR36	LOC107857408	GTCGGGTCGAAATTGATGGA	TGAAGTAAACCACAGGCACTAA

Table 1

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样品名称 Sample	原始序列数 Raw reads (M)	原始序列长度 Raw bases (G)	高质量序列数 Clean reads (M)	高质量序列长度 Clean bases (G)	比对到参考基因组 Mapped reads (%)	Q30 (%)	GC (%)
CK1	51.10	7.36	48.96	7.05	95.80	96.58	42.98
CK2	50.57	7.31	48.63	7.03	96.17	96.89	43.10
CK3	50.69	7.27	48.35	6.93	95.38	96.54	43.09
DP1	50.25	7.24	48.13	6.93	95.76	96.75	43.15
DP2	50.63	7.28	48.37	6.95	95.54	96.85	43.25
DP3	50.37	7.22	47.98	6.87	95.25	96.90	43.24
HP1	50.65	7.31	48.62	7.01	96.00	96.87	43.14
HP2	50.99	7.28	48.35	6.90	94.82	95.27	43.27
HP3	51.53	7.38	49.02	7.02	95.15	96.82	43.14
LP1	48.92	7.04	46.85	6.75	95.77	96.80	43.20
LP2	51.66	7.37	49.00	6.99	94.85	96.70	43.18
LP3	50.80	7.32	48.66	7.01	95.79	96.59	43.21

Transcriptomics-Based Analysis of Pepper Responses to Phosphorus Nutritional Stress

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