TMT-Based Proteomic Analysis of Continuous Cropping Response in Codonopsis tangshen Oliv.

The growth and development of Codonopsis tangshen, an important herb used in Chinese traditional medicine, have been seriously affected by continuous cropping obstacles. Therefore, understanding the molecular responses of C. tangshen to continuous cropping is imperative to improve its resistance to continuous cropping obstacles. Here, physiological and biochemical results showed that the levels of chlorophyll and malonaldehyde (MDA) were higher in the continuous cropping (LZ) group compared with those of the non-continuous cropping (FLZ) group, while superoxide dismutase (SOD) content was lower in the LZ group than in the FLZ group. Tandem mass tag (TMT)-based proteomic analysis was performed to investigate the response mechanism to continuous cropping obstacles in C. tangshen. A total of 70 differentially expressed proteins (DEPs) were significantly involved in relevant pathways, including photosynthesis, oxidative phosphorylation, ribosome activity, and secondary metabolites. The results suggest that these DEPs in C. tangshen might play a critical role in response to continuous cropping. These findings could provide scientific basis for improving C. tangshen's resistance to continuous cropping obstacles.

Transcriptomic Analysis Provides Novel Insights into the Heat Stress-Induced Response in Codonopsis tangshen.

Codonopsis tangshen Oliv (C. tangshen) is a valuable traditional Chinese medicinal herb with tremendous health benefits. However, the growth and development of C. tangshen are seriously affected by high temperatures. Therefore, understanding the molecular responses of C. tangshen to high-temperature stress is imperative to improve its thermotolerance. Here, RNA-Seq analysis was performed to investigate the genome-wide transcriptional changes in C. tangshen in response to short-term heat stress. Heat stress significantly damages membrane stability and chlorophyll biosynthesis in C. tangshen, as evidenced by pronounced malonaldehyde (MDA), electrolyte leakage (EL), and reduced chlorophyll content. Transcriptome analysis showed that 2691 differentially expressed genes (DEGs) were identified, including 1809 upregulated and 882 downregulated. Functional annotations revealed that the DEGs were mainly related to heat shock proteins (HSPs), ROS-scavenging enzymes, calcium-dependent protein kinases (CDPK), HSP-HSP network, hormone signaling transduction pathway, and transcription factors such as bHLHs, bZIPs, MYBs, WRKYs, and NACs. These heat-responsive candidate genes and TFs could significantly regulate heat stress tolerance in C. tangshen. Overall, this study could provide new insights for understanding the underlying molecular mechanisms of thermotolerance in C. tangshen.

Transcriptome analysis provides novel insights into the soil amendments induced response in continuously cropped Codonopsis tangshen.

Codonopsis tangshen Oliv (C. tangshen) is an important Chinese traditional medicinal plant with various health benefits. However, the growth of C. tangshen are seriously affected by continuous cropping, which led to the decrease of the yield and quality. A field experiment was conducted to learn the effects of soil amendments on the growth of C. tangshen under continuous cropping condition, and the biological events which occurred at molecular level were investigated. The results indicated that the content of chlorophyll a (Chl a), chlorophyll b (Chl b), and carotenoid (Car) was significantly higher in SCPM (silicon-calcium-potassium-magnesium fertilizer), SCPMA (SCPM combined with azoxystrobin) and SCPMAOM (SCPM combined with azoxystrobin and organic manure) treatments. Moreover, the yield and the levels of alkaloid, polysaccharide, flavone and total protein in the treatments of SCPM, SCPMA and SCPMAOM were significantly higher than those in the control, and these indexes were all highest in the SCPMAOM treatment. RNA-sequencing (RNA-Seq) is an economical and efficient method to obtain genetic information for species with or without available genome data. In this study, RNA-Seq was performed to understand how continuously cropped C. tangshen responded to the soil amendments at the transcriptome level. The number of differentially expressed genes (DEGs) were as follows: CK vs. SCPM (719 up- and 1456 down-), CK vs. SCPMA (1302 up- and 1748 down-), CK vs. SCPMAOM (1274 up- and 1678 down-). The soil amendments affected the growth of C. tangshen mainly by regulating the genes involved in pathways of 'photosynthesis,' 'plant hormone signal transduction,' 'biosynthesis of unsaturated fatty acids,' 'phenylpropanoid biosynthesis,' and 'starch and sucrose metabolism,' etc. qRT-PCR was performed to validate the expressions of 10 target genes such as CP26, PsaF, and POX, etc., which verified the reliability of RNA-Seq results. Overall, this study revealed the roles and underlying mechanisms of the soil amendments in regulating the growth of continuously cropped C. tangshen at transcriptome level. These findings are beneficial for improving the continuous cropping tolerance and may be valuable for future genetic improvement of C. tangshen.

Exploration of habitat-related chemomarkers for Magnoliae officinalis cortex applying both global and water-soluble components-based metabolomics method.

BACKGROUND: The quality of traditional Chinese medicines (TCMs) has been closely related to their growth regions. The geo-herbalism of TCMs is just like the protected destination of origin on foodstuffs and wines, telling us the specific geographic regions could yield TCMs with superior quality. However, the impact of habitat on TCMs could hardly been indicated in current quality evaluation, defects were as follows: (1) few studies involved the effect of environmental factors, (2) more attentions were paid to several abundant compounds, while global components especially water-soluble compounds were prone to be ignored. PURPOSE: A new integrated metabolomics analysis based on global and water-soluble components was proposed aiming to explore habitat-related chemomarkers for TCMs combined with correlation analysis to environmental factors. The geo-herbalism of Magnoliae officinalis cortex (MOC) was studied as an example. METHODS: Multi-metabolomics approach based on UPLC/Q-TOF-MS and GC-MS combined with LC-2ECD were employed to analyze global components and accurately quantified water-soluble compounds, respectively. Meanwhile, decision tree, partial least squares discriminant analysis (PLS-DA) as well as hierarchical clustering analysis (HCA) heat map was applied to classify different samples and explore habitat-related chemomarkers. In addition, support vector machines model was used to verify the importance of screened out chemomarkers in predicting sample classification, and the impact of environmental factors on the markers were also demonstrated by correlation analysis. RESULTS: By analyzing 148 batches of MOC samples from 21 habitats, 238 variables were picked and 84 of them were identified by UNIFI, meanwhile, seven water-soluble compounds were accurately quantified. Among them, thirteen markers including Var.1, magnolignan E, magnoloside N isomer, α-agarofuran, γ-eudesmol, ß-eudesmol, magnolosides A, B, D, F, H, L and M were suggested importance in grouping Chuan-po and the other MOC samples. Support vector machines model also indicated well prediction performance with an accuracy of 96.97%. Most markers belong to water-soluble compounds and temperature and precipitation contributed to such chemical differences. CONCLUSIONS: The proposed strategy based on multi-metabolomics analysis could aid exploration of habitat-related chemomarkers for TCMs. Meanwhile, the screened out water-soluble compounds could perform equivalent functions in recognition of Daodi medicinal materials (DMMs) and non-DMM samples compared to the global components to some extent.

Transcriptome analysis reveals novel insights into the continuous cropping induced response in Codonopsis tangshen, a medicinal herb.

Codonopsis tangshen Oliv. (C. tangshen Oliv.), a famous medicinal herb in China, is seriously affected by continuous cropping (C-cro). The physiological and biochemical results indicated that C-cro significantly affected the malonaldehyde (MDA) and chlorophyll content, as well as activities of catalase (CAT) and superoxide dismutase (SOD) when compared with the non-continuous cropping (NC-cro) group. Transcriptome profiling found 762 differentially expressed genes, including 430 up-regulated and 332 down-regulated genes by C-cro. In addition, pathway enrichment analysis revealed that genes related to 'Tyrosine degradation I', 'Glycogen synthesis' and 'Phenylalanine and tyrosine catabolism' were up-regulated, and genes associated with 'Signal transduction', 'Immune system', etc. were down-regulated by C-cro. The expression of target genes was further validated by Q-PCR. In this study, we demonstrated the effects of C-cro on C. tangshen at the transcriptome level, and found possible C-cro responsive candidate genes. These findings could be further beneficial for improving the continuous cropping tolerance.

[Effects of primary processing on quality of cortex Magnolia officinalis].

In this paper, the changes of volatile oil and non-volatile ingredients in Cortex Magnolia Officinalis before and after primary processing were determined by an HPLC and a GC-MS method. The method is based on quantitative determination of three index ingredients, beta-eudesmol, honokiol and magnolol, and on qualitative fingerprinting analysis using HPLC and GC. Big differences were observed between processed and unprocessed samples according to their chromatographic fingerprinting data calculated by statistic software. Compared with unprocessed samples, the contents of honokiol and magnolol in processed samples increased, whereas the contents of beta-eudesmol and magnoloside A in processed samples decreased. Magnoloside A was isolated from this plant for the first time.

[Effects of tree ages and geographic area on quality of bark of Magnolia officinalis and M. officinalis var. biloba].

The effects of tree ages and geographic area on quality of bark of Magnolia officinalis (MO) and M. officinalis var. biloba (MOB) were studied. The volatile oils of samples extracted by a water stream distillation method were analyzed using a GC/MS method. The non-volatile ingredients of samples were determined by an HPLC method. Seven and twenty characteristic peaks were detected in the HPLC and GC fingerprint of MO, respectively, and the similar HPLC and GC fingerprint were showed in MOB. For samples collected in the same geographic area with different ages, the concentrations of active ingredients varied barely, whereas for those samples collected from different geographic area, differences on the concentrations of active ingredients were observed, for example, the concentrations of the amount of magnolol and honokiol in samples collected in Hubei province were higher than those collected in other areas. Big differences on the concentrations of active ingredients were found between MO and MOB, for MO, the ratio of honokiol/ magnolol was more than 0.7, whereas the ratio-was less than 0.7 in the case of MOB, and the concentrations of honokiol in MO were higher than those in MOB.