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Acta Agronomica Sinica ›› 2020, Vol. 46 ›› Issue (12): 1894-1904.doi: 10.3724/SP.J.1006.2020.04050

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Mechanism exploration on alkaloids accumulation and TAs metabolic pathways regulation in Atropa belladonna L. treated with exogenous methyl jasmonate under UV-B stress

Yu-Si SHAN1,2(), Zheng-Qi XIN1,2, Xiao HE1,2, Huan-Huan DAI1,2, Neng-Biao WU1,2,*()   

  1. 1School of Life Science, Southwest University, Chongqing 400715, China
    2Key Laboratory of Eco-environments in Three Gorges Region, Ministry of Education, Chongqing 400715, China
  • Received:2020-02-28 Accepted:2020-06-02 Online:2020-12-12 Published:2020-07-28
  • Contact: Neng-Biao WU E-mail:shanyusi3354@foxmail.com;wunb@swu.edu.cn
  • Supported by:
    National Natural Science Foundation of China(30500041)

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Abstract:

In order to preliminarily explore the mechanism of TAs metabolic pathway in A. belladonna treated with exogenous methyl jasmone under UV-B stress, the effects on the contents of hyoscyamine and scopolamine, the upstream products in alkaloid synthesis, signal molecule and the expression level of key enzyme genes of secondary metabolism were studied under different concentrations of exogenous MeJA and different treatment time under using A. belladonna as materials. UV-B stress treatment significantly reduced the contents of hyoscyamine and scopolamine, inhibited the accumulation of precursors in the TAs synthesis pathway, which harmful to TAs synthesis. The content of TAs in A. belladonna increased to some extent, the contents of precursor amino acids (ornithine, arginine), polyamine content and key enzymes activities in the synthesis of putrescine in the secondary metabolic pathway all increased to some extent after treatment with the appropriate concentration of MeJA. The concentrations of signal molecule NO firstly increased and then decreased with the rising MeJA concentration, and reached the highest when MeJA concentration was 250 μmol L -1. The expression of key enzyme genes in TAs synthesis pathway showed that exogenous MeJA could increase the relative gene expression levels of TR I, PPAR, H6H to some extent. Those indicated that exogenous MeJA could induce the increase in the contents of upstream products in TAs synthesis by stimulating the burst of NO resulted in more precursor materials for the TAs synthesis pathway and affect the high expression of TR I, PPAR and H6H. It alleviated the inhibiton of UV-B stress on TAs of A. belladonna and increased the contents of hyoscyamine and scopolamine effectively. The results provided a theoretical basis for further studying the mechanism of exogenous elicitors to regulate the TAs secondary metabolic pathway of A. belladonn under stress, and effectively improved the stress resistance of A. belladonn and the accumulation of medicinal ingredients in actual production.

Key words: methyl jasmonate, UV-B stress, Atropa belladonna L., tropane alkaloids, secondary metabolism

Table 1

Primer for quantitative RT-PCR"

基因
Gene		 上游引物
Forward primer (5′-3′)		 下游引物
Reverse primer (5′-3′)
PGK TCGCTCTTGGAGAAGGTTGAC CTTGTCCGCAATCACTACATCAG
PMT CCTACTTACCCTACTGGTGTTATC GCGAAAGATGGCAAAATAAAAGC
TR I TTCTTTGCTTCCCTGCTGCTTC CAGGCCAACCTTAGTATCACACAG
PPAR GATTGGAGTTGTTGGATTGG CCTTGAAGTGTAAGAGATGGA
H6H TTCCACTTGAGCAGAAAGCAAAGC CCTCATGGTCAACTTCCTCACTTCC

Table 2

Effects on the hyoscyamine and scopolamine contents under exogenous methyl jasmonate with different concentrations in A. belladonna leaves under UV-B stress (μg g-1 DW)"

指标
Indicator		 处理组
Treatment group		 处理时间Treatment time
4 d 8 d 12 d 16 d
莨菪碱
Hyoscyamine		 CK1 477.988±30.032 d 464.993±39.364 a 510.286±29.327 a 489.386±33.957 a
CK2 514.323±36.595 c 403.706±14.900 c 350.179±35.316 d 295.220±30.438 d
T1 527.659±38.739 bc 423.530±36.157 bc 375.603±19.982 c 312.172±19.933 cd
T2 542.565±35.559 bc 468.808±27.539 a 392.611±37.487 bc 361.054±22.242 bc
T3 581.520±17.200 a 447.993±24.571 b 425.491±47.568 b 384.890±44.869 b
T4 564.200±33.989 ab 430.305±15.919 bc 369.667±40.042 c 332.059±26.077 c
东莨菪碱
Scopolamine		 CK1 267.673±24.260 a 261.248±22.428 a 294.230±42.998 a 282.621±28.477 a
CK2 221.892±20.219 c 198.795±34.491 c 174.729±14.562 d 121.804±15.205 d
T1 243.889±28.790 ab 217.900±17.821 bc 193.653±21.253 cd 130.244±24.996 cd
T2 237.625±28.217 ab 236.775±13.461 ab 221.873±19.211 b 150.869±26.035 c
T3 248.261±20.375 ab 234.134±26.418 b 229.996±29.857 b 185.867±23.527 b
T4 225.705±14.739 bc 220.173±26.487 bc 208.852±24.722 bc 164.412±30.129 c

Fig. 1

Effects on the contents of Ornithine and Arginine under exogenous methyl jasmonate with different concentrations in A. belladonna leaves under UV-B stress Bars superscripted by different lowercase letters show significant differences among different treatment groups at P < 0.05. Treatments are the same as those given in Table 2."

Fig. 2

Effects on ODC and ADC activities under exogenous methyl jasmonate with different concentrations in A. belladonna leaves under UV-B stress Bars superscripted by different lowercase letters show significant differences among different treatment groups at P < 0.05. Treatments are the same as those given in Table 2."

Fig. 3

Effects on Polyamine contents (A), PAs contents (B) and (Spm+Spd)/Put (C) under exogenous methyl jasmonate with different concentrations in A. belladonna leaves under UV-B stress Bars superscripted by different lowercase letters show significant differences among different treatment groups at P < 0.05. Treatments are the same as those given in Table 2."

Fig. 4

Effects on NO concentration under exogenous methyl jasmonate with different concentrations in A. belladonna leaves under UV-B stress Bars superscripted by different lowercase letters show significant differences among different treatment groups at P < 0.05. Treatments are the same as those given in Table 2."

Fig. 5

Effects on relative gene expression level of PMT, TR I, PPAR, H6H under exogenous methyl jasmonate with different concentrations in A. belladonna leaves (A) and roots (B) under UV-B stress Bars superscripted by different lowercase letters show significant differences among different treatment groups at P < 0.05. Treatments are the same as those given in Table 2."

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