Transcriptome Analysis of Lycium ruthenicum Murr. Shoots in Anthocyanin Biosynthesis Response to Salt Stress

doi:10.13560/j.cnki.biotech.bull.1985.2021-1576

Abstract

Abstract:

In order to analyze the process of Lycium ruthenicum Murr.（L. ruthenicum）shoots in anthocyanin biosynthesis response to salt stress，L. ruthenicum seedlings were treated with 300 mmol/L NaCl，and the shoots were collected at 3 d after treatment，and the anthocyanin content and transcriptome were analyzed by LC-MS/MS and RNA-seq. Combined with the analysis of species and contents of anthocyanin and differential expressed genes（DEGs）in GO and KEGG pathway，DEGs related to anthocyanin biosynthesis induced by salt stress were mined，and the results of transcriptome data were verified by RT-qPCR. The results showed delphinidin-3-O-rutinoside in L. ruthenicum shoots was the highest upregulated anthocyanin（11.9 times），and the content of petunia-3-O-rutinoside was the highest anthocyanin（5.313±0.286）μg/g. Total 1 416 DEGs（P<0.01）were screened，including 867 up-regulated and 549 down-regulated DEGs，which were classified into GO terms，such as catalytic activity，photosystem and single-organism metabolic process. DEGs were significantly enriched in 14 KEGG pathways. Meanwhile，there were 7 DEGs involved in anthocyanin biosynthesis and all of them were up-regulated，and27 DEGs were involved in phytohormone signal transduction，in which 15 DEGs were related to ABA signal transduction pathway. The expressions of 11 MYB and 7 bHLH transcription factors were significantly changed. Sum up，the results of this study suggested that DEGs related to ABA pathway，MYB，bHLH transcription factor and anthocyanin biosynthesis play important role in anthocyanin biosynthesis in L.ruthenicum shoot responses to NaCl stress.

Key words: Lycium ruthenicum Murr., shoots, NaCl, anthocyanin, molecular modulation

GUO Ai, JIANG Mu-yan, Ha Li-ma-ti·Ba He-tai-li, LIU Yu-yuan, WANG Jing. Transcriptome Analysis of Lycium ruthenicum Murr. Shoots in Anthocyanin Biosynthesis Response to Salt Stress[J]. Biotechnology Bulletin, 2022, 38(10): 173-183.

Figures/Tables 11

References 32

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基因ID Gene ID	序列Sequence（5'-3'）
LrH2B1	F：AGTGCTTCCTGGTGAATTGG
LrH2B1	R：TGGATAATACCTAGCCCTAGTTTCC
Cluster-45.236520	F：TTTGCCTCTGGCTTTGTCTACC
Cluster-45.236520	R：TGGATACTGCTGTTGCCTTCAT
Cluster-45.192144	F：TGCACTGTTTGTCTCCTAGCCTCC
Cluster-45.192144	R：ACGCCATTCTTCTCGTCTCCATC
Cluster-45.35768	F：AAAACAGGACATGGCCTTTATG
Cluster-45.35768	R：CTGAGTTTGTAGGCAAGGTGGA
Cluster-45.156739	F：GCACAAAGTCTCCGAGAAGG
Cluster-45.156739	R：GGGGGCTTCATATTTCTTGTT
Cluster-45.33457	F：GAAGCCAACAGGTGAAACGACG
Cluster-45.33457	R：TCTTCATCAAAGCTCCCAAATGCT
Cluster-45.168632	F：GGTTTGATGAAAGGGGTGAAGAA
Cluster-45.168632	R：CCTGACGTGGCAGTATCGCTGAG，
Cluster-45.207146	F：TGAAGAATTAAGAGAGTCGAGGTCG
Cluster-45.207146	R：AAGTTGGCAATGGTGATGGTGTC.
Cluster-45.136887	F：TCAGATGAACCCAATTCACCAA
Cluster-45.136887	R：ACCCTAAAGAGCTGTCGGATGT
Cluster-45.146553	F：GGCACAGTAGCTGAATCCAAGA
Cluster-45.146553	R：CCTTTACCAACTTCTGCCCATA
Cluster-45.150170	F：GAGCAAGGCAGCAATAGCAGAT
Cluster-45.150170	R：AAGGTGGTGACATTGAGGTGAAG
Cluster-45.146155	F：GGTCAAGCCACTGATAGCCACA
Cluster-45.146155	R：GCATTCCTGCTTTCCCTGTGAT
Cluster-45.117764	F：TATCTTTCTTAGGCCCTTCCCTC
Cluster-45.117764	R：GCGTTATTGTTATTATTGTCGGTGT

基因ID Gene ID	序列Sequence（5'-3'）
LrH2B1	F：AGTGCTTCCTGGTGAATTGG
LrH2B1	R：TGGATAATACCTAGCCCTAGTTTCC
Cluster-45.236520	F：TTTGCCTCTGGCTTTGTCTACC
Cluster-45.236520	R：TGGATACTGCTGTTGCCTTCAT
Cluster-45.192144	F：TGCACTGTTTGTCTCCTAGCCTCC
Cluster-45.192144	R：ACGCCATTCTTCTCGTCTCCATC
Cluster-45.35768	F：AAAACAGGACATGGCCTTTATG
Cluster-45.35768	R：CTGAGTTTGTAGGCAAGGTGGA
Cluster-45.156739	F：GCACAAAGTCTCCGAGAAGG
Cluster-45.156739	R：GGGGGCTTCATATTTCTTGTT
Cluster-45.33457	F：GAAGCCAACAGGTGAAACGACG
Cluster-45.33457	R：TCTTCATCAAAGCTCCCAAATGCT
Cluster-45.168632	F：GGTTTGATGAAAGGGGTGAAGAA
Cluster-45.168632	R：CCTGACGTGGCAGTATCGCTGAG，
Cluster-45.207146	F：TGAAGAATTAAGAGAGTCGAGGTCG
Cluster-45.207146	R：AAGTTGGCAATGGTGATGGTGTC.
Cluster-45.136887	F：TCAGATGAACCCAATTCACCAA
Cluster-45.136887	R：ACCCTAAAGAGCTGTCGGATGT
Cluster-45.146553	F：GGCACAGTAGCTGAATCCAAGA
Cluster-45.146553	R：CCTTTACCAACTTCTGCCCATA
Cluster-45.150170	F：GAGCAAGGCAGCAATAGCAGAT
Cluster-45.150170	R：AAGGTGGTGACATTGAGGTGAAG
Cluster-45.146155	F：GGTCAAGCCACTGATAGCCACA
Cluster-45.146155	R：GCATTCCTGCTTTCCCTGTGAT
Cluster-45.117764	F：TATCTTTCTTAGGCCCTTCCCTC
Cluster-45.117764	R：GCGTTATTGTTATTATTGTCGGTGT

物质Compound	CK含量CK content/（μg·g^-1）	Na含量Na content/（μg·g^-1）	P值P -value	倍数变化Fold change
飞燕草素-3-O-葡萄糖苷 Delphinidin-3-O-glucoside	0.151±0.0409	1.13±0.0283	0.000027	7.5
飞燕草素-3-O-芸香糖苷 Delphinidin-3-O-rutinoside	0.432±0.0581	5.15±0.134	0.0000655	11.9
矮牵牛素-3-O-芸香糖苷 Petunidin-3-O-rutinoside	2.33±0.323	5.313±0.286	0.000659	2.28
柚皮素-7-O-葡萄糖苷 Naringenin-7-O-glucoside	0.112±0.0189	0.339±0.0457	0.0015	3.02

物质Compound	CK含量CK content/（μg·g^-1）	Na含量Na content/（μg·g^-1）	P值P -value	倍数变化Fold change
飞燕草素-3-O-葡萄糖苷 Delphinidin-3-O-glucoside	0.151±0.0409	1.13±0.0283	0.000027	7.5
飞燕草素-3-O-芸香糖苷 Delphinidin-3-O-rutinoside	0.432±0.0581	5.15±0.134	0.0000655	11.9
矮牵牛素-3-O-芸香糖苷 Petunidin-3-O-rutinoside	2.33±0.323	5.313±0.286	0.000659	2.28
柚皮素-7-O-葡萄糖苷 Naringenin-7-O-glucoside	0.112±0.0189	0.339±0.0457	0.0015	3.02

样品Sample	原始测序数据Raw reads	过滤后测序数据Clean reads	过滤后数据Clean bases	Error/%	Q20/%	Q30/%	GC content/%
CK1	55 697 476	54 900 826	8.24 G	0.02	98.48	95.29	42.65
CK2	60 551 930	59 757 658	8.96 G	0.03	97.2	91.95	42.48
CK3	55 743 892	54 933 528	8.24 G	0.03	97.25	92.05	42.53
Na1	61 583 804	61 024 066	9.15 G	0.03	96.83	91.12	42.21
Na2	46 367 542	45 803 662	6.87 G	0.02	98.39	95.06	42.32
Na3	47 635 904	46 871 318	7.03 G	0.02	98.48	95.32	42.31