用茉莉酸甲酯诱导促进龙胆草不定根培养物中龙胆内酯的产生和生物合成基因的转录反应

IF 5.4 Q1 PLANT SCIENCES Current Plant Biology Pub Date : 2024-10-17 DOI:10.1016/j.cpb.2024.100397
Jingwen Hao , Jian Zang , Dan Liu , Ning Hao , Rujun Zhou
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引用次数: 0

摘要

非生物诱导剂在调控植物生长、发育和特殊代谢的各个方面发挥着至关重要的作用。本研究旨在通过筛选诱导剂类型、优化诱导条件以及估测秦艽不定根中生物合成基因的转录反应,进一步提高秦艽苷的含量。结果表明,在所测试的诱导剂中,茉莉酸甲酯(MeJA)对葶苈不定根生物量积累的诱导效果最好,其鲜重(FW)和干重(DW)分别为 13.26 ± 0.57 g flask-1 和 1.31 ± 0.25 g flask-1。研究了诱导时间和 MeJA 浓度对葶苈子不定根生物量和龙胆内酯含量的影响。在含有 3.0 mg L-1 1-naphthlcetic acid (NAA) 和 1.0 mg L-1 kinetin (KT) 的 MS 液体培养基中,当 MeJA 浓度为 100 μM L-1 时,培养 6 天的根系可获得最大的 FW(15.73 ± 0.41 g flask-1)和 DW(1.51 ± 0.19 g flask-1);当 MeJA 浓度为 100 μM L-1 时,培养 6 天的根系可获得最大的 FW(15.73 ± 0.41 g flask-1)和 DW(1.51 ± 0.19 g flask-1)。同时,龙胆甙的含量也明显增加到 62.62 ± 0.27 mg g-1 DW,是未处理对照的 2.49 倍。为了进一步了解龙胆草对 MeJA 诱导的转录响应,研究人员估算了 12 个与龙胆草甙生物合成相关的候选基因在葶苈不定根中的表达水平,这些基因涉及甲羟戊酸(MVA)、赤藓糖醇磷酸甲酯(MEP)和仲呋喃类途径。结果表明,与未处理的对照组相比,葶苈不定根中有 10 个基因(ACCT1、HMGR1、MCK1、MVD1、GPPS4、G10H、IS3、DL7H1、DXS5 和 ISPH5)上调,而 DXR1 和 IDI1 基因下调,差异显著(阈值 P ≤0.05)。转录分析表明,12 个候选基因是龙胆内酯生物合成途径中的关键调控基因。总之,研究结果为利用MeJA诱导提高珍贵龙胆内酯的产量提供了一种前景广阔、可行且稳定的方法。此外,这些研究还为今后通过代谢工程策略在葶苈子不定根中进行龙胆草甙生物合成奠定了基础。
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Promotion of gentiopicroside production and transcriptional responses of biosynthetic genes in adventitious root cultures of Gentiana scabra Bunge by elicitation with methyl jasmonate
Abiotic elicitors play a crucial role in regulating various aspects of plant growth, development, and specialized metabolism. This study aimed to further increase the gentiopicroside content by screening elicitor types, optimizing elicitation conditions, and estimating transcriptional responses of biosynthetic genes in the adventitious roots of Gentiana scabra. The results showed that methyl jasmonate (MeJA) was the most effective inducer for biomass accumulation in the adventitious roots of G. scabra among tested elicitors, with fresh weight (FW) and dry weight (DW) of 13.26 ± 0.57 g flask−1 and 1.31 ± 0.25 g flask−1, respectively. The effects of the induction time and concentration of MeJA on the biomass and gentiopicroside content in the adventitious roots of G. scabra were investigated. The maximum FW (15.73 ± 0.41 g flask−1) and DW (1.51 ± 0.19 g flask−1) were obtained when the roots were cultured for 6 days in MS liquid medium containing 3.0 mg L−1 1-naphthlcetic acid (NAA) and 1.0 mg L−1 kinetin (KT) at MeJA concentration of 100 μM L−1. Also, the gentiopicroside content significantly increased to 62.62 ± 0.27 mg g−1 DW, and was 2.49 times higher than that for the nontreated control. The expression levels of 12 candidate gentiopicroside biosynthesis–related genes involved in the mevalonic acid (MVA), methyl erythritol phosphate (MEP), and secoiridoid pathways were estimated in the adventitious roots of G. scabra to further understand the transcriptional response to MeJA elicitation. Among these, 10 genes (ACCT1, HMGR1, MCK1, MVD1, GPPS4, G10H, IS3, DL7H1, DXS5, and ISPH5) were upregulated whereas DXR1 and IDI1 genes were downregulated in the adventitious roots of G. scabra compared with nontreated control, with significant differences having threshold P value ≤0.05. The transcriptional analyses revealed that 12 candidate genes were the key regulated genes in the gentiopicroside biosynthetic pathway. Overall, the findings provided a promising, feasible, and stable approach to utilizing MeJA elicitation to increase the production of valuable gentiopicroside. Additionally, they provided a foundation for future gentiopicroside biosynthesis through metabolic engineering strategies in the adventitious roots of G. scabra.
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来源期刊
Current Plant Biology
Current Plant Biology Agricultural and Biological Sciences-Plant Science
CiteScore
10.90
自引率
1.90%
发文量
32
审稿时长
50 days
期刊介绍: Current Plant Biology aims to acknowledge and encourage interdisciplinary research in fundamental plant sciences with scope to address crop improvement, biodiversity, nutrition and human health. It publishes review articles, original research papers, method papers and short articles in plant research fields, such as systems biology, cell biology, genetics, epigenetics, mathematical modeling, signal transduction, plant-microbe interactions, synthetic biology, developmental biology, biochemistry, molecular biology, physiology, biotechnologies, bioinformatics and plant genomic resources.
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