Crops ›› 2024, Vol. 40 ›› Issue (1): 48-56.doi: 10.16035/j.issn.1001-7283.2024.01.007

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Transcriptome Analysis Reveals Mechanisms of Calycosin- 7-O-β-D-Glucoside Accumulation in Astragalus membranaceus Adventitious Roots through Hydrogen Peroxide

Dou Weize1(), Jiang Wen2, Feng Yichuan2, Jin Zhuo2, Quan Xueli2, Wu Songquan1,2()   

  1. 1College of Integration Science, Yanbian University, Yanji 133000, Jilin, China
    2Agricultural College, Yanbian University, Yanji 133000, Jilin, China
  • Received:2023-05-31 Revised:2023-09-01 Online:2024-02-15 Published:2024-02-20
  • Contact: Wu Songquan E-mail:1424327165@qq.com;arswsq@ybu.edu.cn

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Abstract:

Hydrogen peroxide (H2O2) significantly enhanced the content of calycosin-7-O-β-D-glucoside (CG) in Astragalus membranaceus adventitious roots (AMARs). To discover its biosynthetic mechanism of CG, AMARs were used to screen the differentially expressed genes (DEGs) through transcriptome sequencing after H2O2 treatment. The function and metabolic pathways were analyzed by using up-regulated genes, verified them by qRT-PCR method, and measured the relevant physiological and biochemical parameters. The results showed that AMARs activated ethylene signaling, induced all structural genes in the phenylpropane-isoflavone biosynthesis pathway, significantly increased the content of CG to counteract the oxidative stress caused by H2O2 treatment. Furthermore, H2O2 treatment also induced phenylalanine biosynthesis in order to increase the content of CG. These results revealed the accumulation mechanism of CG in AMARs which laid a theoretical basis for large-scale production of CG in the future.

Key words: H2O2, Oxidative stress, Astragalus membranaceus (Fisch.) Bunge, Transcriptome sequencing, Calycosin-7-O-β-D-glucoside

Table 1

Statistical of sequencing data quality evaluation"

样品
Sample		 Reads总数
Sum of
reads		 碱基总数
Sum of
base		 GC含量
GC
content
(%)		 碱基质量值
Quality score
of bases (%)
Q20 Q30
A1 21 235 913 6 331 453 200 42.92 98.00 94.15
A2 21 342 976 6 357 785 560 42.69 97.96 94.03
A3 22 438 643 6 700 654 314 42.65 97.80 93.64
B1 23 860 654 7 110 359 156 42.73 97.93 93.93
B2 21 448 660 6 403 522 654 42.69 97.71 93.38
B3 21 112 504 6 297 173 426 42.84 97.67 93.40
C1 21 096 613 6 286 682 948 42.62 97.66 93.33
C2 22 710 737 6 770 120 316 42.61 97.73 93.51
C3 21 934 340 6 540 593 324 42.74 98.05 94.20
D1 21 991 682 6 548 307 308 42.63 97.95 94.00
D2 23 063 182 6 885 014 356 42.80 97.84 93.77
D3 19 383 999 5 780 350 228 42.76 97.79 93.64

Table 2

The number of DEGs in H2O2 treatments"

分组
Group		 差异基因总数
Total number
of different
genes		 上调基因数
Number of
up-regulated
genes		 下调基因数
Number of
down-regulated
genes
B vs A 222 206 16
C vs A 468 347 121
D vs A 280 199 81
A vs B & C & D 135 127 8

Fig.1

Functional classification of GO for DEGs"

Fig.2

KEGG enrichment plots of DEGs (a) treated with H2O2 for 1 h, (b) treated with H2O2 for 3 h, and (c) treated with H2O2 for 6 h."

Fig.3

Relative gene expressions of isoflavone branching pathway Different letters indicate the significant difference (P < 0.05) in different treatmensts, “*”indicates a significant difference (P < 0.05) between control and H2O2 treatments, the same below."

Fig.4

Cluster heat map of isoflavone branching pathway differentially expressed genes Different color regions represent different clustering information, and the color from red to blue indicates the expression of differentially expressed genes from high to low."

Fig.5

Effects of the CG contents with H2O2 treatment"

Fig.6

Relative expressions of POD and its activity"

Fig.7

The relative expressions of ACS and ERF and the effects of H2O2 treatment on ethylene content"

Fig.8

Relative expressions of phenylalanine synthesis pathway of genes"

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