Yao Deheng, Chen Yukun, Xu Xiaoping, Lin Yuling, Lai Zhongxiong
Institute of Horticultural Biotechnology, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
College of Food and Biological Engineering, Fujian Polytechnic Normal University, Fuqing 350300, China.
Int J Mol Sci. 2022 Mar 28;23(7):3704. doi: 10.3390/ijms23073704.
Our aim in the experiment was to study the effects of methyl jasmonates (MeJA) on the active compounds of rosemary suspension cells, the metabolites' change of contents under different concentrations of MeJA, including 0 (CK), 10 (M10), 50 (M50) and 100 μM MeJA (M100). The results demonstrated that MeJA treatments promoted the accumulation of rosmarinic acid (RA), carnosic acid (CA), flavonoids, jasmonate (JA), gibberellin (GA), and auxin (IAA); but reduced the accumulations of abscisic acid (ABA), salicylic acid (SA), and aspartate (Asp). In addition, 50 and 100 μM MeJA promoted the accumulation of alanine (Ala) and glutamate (Glu), and 50 μM MeJA promoted the accumulation of linoleic acid and alpha-linolenic acid in rosemary suspension cells. Comparative RNA-sequencing analysis of different concentrations of MeJA showed that a total of 30, 61, and 39 miRNAs were differentially expressed in the comparisons of CKvsM10, CKvsM50, CKvsM100, respectively. The analysis of the target genes of the differentially expressed miRNAs showed that plant hormone signal transduction, linoleic acid, and alpha-linolenic acid metabolism-related genes were significantly enriched. In addition, we found that miR160a-5p target , miR171d_1 and miR171f_3 target , miR171b-3p target , and miR156a target , which played a key role in rosemary suspension cells under MeJA treatments. qRT-PCR of 12 differentially expressed miRNAs and their target genes showed a high correlation between the RNA-seq and the qRT-PCR result. Amplification culture of rosemary suspension cells in a 5 L stirred bioreactor showed that cell biomass accumulation in the bioreactor was less than that in the shake flask under the same conditions, and the whole cultivation period was extended to 14 d. Taken together, MeJA promoted the synthesis of the active compounds in rosemary suspension cells in a wide concentration range via concentration-dependent differential expression patterns. This study provided an overall view of the miRNAs responding to MeJA in rosemary.
我们进行该实验的目的是研究茉莉酸甲酯（MeJA）对迷迭香悬浮细胞活性化合物的影响，以及在不同浓度MeJA（包括0（CK）、10（M10）、50（M50）和100 μM MeJA（M100））作用下代谢产物含量的变化。结果表明，MeJA处理促进了迷迭香酸（RA）、肌醇六磷酸（CA）、类黄酮、茉莉酸（JA）、赤霉素（GA）和生长素（IAA）的积累；但降低了脱落酸（ABA）、水杨酸（SA）和天冬氨酸（Asp）的积累。此外，50和100 μM MeJA促进了丙氨酸（Ala）和谷氨酸（Glu）的积累，50 μM MeJA促进了迷迭香悬浮细胞中亚油酸和α-亚麻酸的积累。不同浓度MeJA的比较RNA测序分析表明，在CKvsM10、CKvsM50、CKvsM100的比较中，分别有30、61和39个miRNA差异表达。对差异表达miRNA的靶基因分析表明，植物激素信号转导、亚油酸和α-亚麻酸代谢相关基因显著富集。此外，我们发现miR160a - 5p靶基因、miR171d_1和miR171f_3靶基因、miR171b - 3p靶基因和miR156a靶基因在MeJA处理下的迷迭香悬浮细胞中起关键作用。对12个差异表达miRNA及其靶基因进行qRT-PCR分析，结果显示RNA测序与qRT-PCR结果高度相关。在5 L搅拌式生物反应器中对迷迭香悬浮细胞进行放大培养，结果表明在相同条件下，生物反应器中的细胞生物量积累低于摇瓶中的，且整个培养周期延长至14天。综上所述，MeJA通过浓度依赖性差异表达模式在较宽浓度范围内促进了迷迭香悬浮细胞中活性化合物的合成。本研究提供了迷迭香中响应MeJA的miRNA的整体情况。
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