Global Metabolic Changes by Bacillus Cyclic Lipopeptide Extracts on Stress Responses of Para Rubber Leaf

Changing environmental conditions can generate abiotic stress, such as the scarcity of water and exposure to chemicals. This includes biotic stress like Phytophthora palmivora infection, which causes leaf fall disease and inhibits the growth rate of para rubber seedlings, resulting in economic loss. To prevent abiotic and biotic stresses, biocontrol agents such as cyclic lipopeptides (CLPs) from Bacillus spp. have been introduced to reduce the use of chemically synthesized fungicides and fertilizers. This study aimed to use Bacillus CLP extracts as a biological agent to stimulate the plant growth system in para rubber seedlings under stress conditions compared with the exogenous plant hormone (salicylic acid, SA). CLP extracts obtained from B. subtilis PTKU12 and exogenous SA were applied to the leaves of para rubber seedlings. The extracted metabolites from each treatment were analyzed by untargeted metabolomics for metabolite identification and metabolic networks under stress responses. In both treatments, 1,702 and 979 metabolites were detected in the positive and negative ion modes of electrospray ionization, respectively. The differential analysis revealed that the accumulation of up-regulated metabolites in the treatment of CLP extracts was higher than in the exogenous SA treatment, belonging to 56 metabolic pathways. The analysis of metabolic pathways indicated that CLP extracts employed alanine, aspartate, and glutamate metabolisms for stress responses leading to plant growth promotion. These findings revealed that the metabolic network for plant growth promotion induced by BacillusCLP extracts could be considered a protective option for para rubber plantations. Doi: 10.28991/ESJ-2023-07-03-022 Full Text: PD

การแสดงออกของยีนที่เกี่ยวข้องกับการต้านทานโรคในผลส้มเหนี่ยวนำโดยสารไลโปเปปไทด์จากเชื้อแบซิลลัส ซับทิลลิส อิลิซิเตอร์และเชื้อราก่อโรค

Thesis (Ph.D., Biochemistry)--Prince of Songkla University, 2018Cyclic lipopeptides (CLPS) obtained from Bacillus subtilis ABS-S14 had ability to activate some important genes, proteins, and metabolites which are involved in plant defensive pathways to stress responses and infection caused by Penicillium digitatum in mandarin fruit. The effects of CLP extract, fengycin, iturin A and surfactin in enhancement of the expression of PAL, LOX, ACS1, ACO, CHI, GLU, POD and PR1 genes in postharvest mandarin fruit were revealed. Also the proteins relating to Ca2+ pathway, ABA signaling pathway, reactive oxygen species pathway, and ubiquitin- proteasome system including energy production for plant resistance were significantly accumulated. During stress responses (non-fungal infection), fengycin activated plant production of a protein that is involved in plant development and the ubiquinone biosynthetic process while iturin A and surfactin were shown to be involved in auxin and abscisic acid modulating signaling pathways. In addition, a comparative elicitor-protein binding assay of each CLP illustrated that iturin A attached to 12-oxophytodienoate reductase 2 that plays role in oxylipin biosynthetic process required for jasmonic acid production led to inducing systemic resistance (ISR). Furthermore, metabolomic analysis revealed that CLPs elicited a high accumulation of some amino acids; aspartic acid, homoserine, threonine, serine, glycine and tryptophan, which are primary metabolites via the metabolism of glycine, serine and threonine, and the secondary metabolites; serotonin and tyramine, which are active in the metabolism of tryptophan and tyrosine, respectively in postharvest mandarin fruit.สารไลโปเปปไทด์จากเชื้อแบคทีเรียแบซิลัส ซับทิลลิสมีประสิทธิภาพในการกระตุ้นการแสดงออกของยีน โปรตีน และสารเมแทบอไลท์ที่เกี่ยวข้องกับระบบภูมิต้านทานโรคพืช เพื่อตอบสนองต่อสภาวะเครียดและการติดเชื้อราเขียวในผลส้มโชกุน โดยสารสกัดไลโปเปปไทด์ รวมถึง fengycin iturin A และ Surface สามารถกระตุ้นการแสดงออกของยีนที่เกี่ยวข้องกับการ ต้านทานโรคพืชได้ เช่น ยีน PAL LOX ACS1 ACO CHI GLU POD และ PR1 เป็นต้น ทั้งนี้สาร สกัดไลโปเปปไทด์ยังสามารถเพิ่มปริมาณของโปรตีนที่เกี่ยวข้องกับวิถีแคลเซียม กรดแอบไซซิกอนุพันธ์ออกซิเจนที่ว่องไว ระบบยูบิควิโน-โปรติเอโซม รวมไปถึงผลผลิตของสารที่ให้พลังงานเพื่อ การต้านทานโรคพืช อีกทั้ง fengycin ยังเพิ่มปริมาณของโปรตีนที่เกี่ยวข้องกับการพัฒนาของพืช และการสังเคราะห์ยูบิควิโนนในกลุ่มที่ไม่มีการติดเชื้อราเขียว ในขณะที่โปรตีนที่เกี่ยวข้องกับ ฮอร์โมนพืช เช่น ออกซินและกรดแอบไซซิกนั้นถูกเพิ่มปริมาณโดยการกระตุ้นจาก iturin A และ surfactin การทดสอบการจับระหว่างสารไลโปเปปไทด์กับโปรตีนที่เกี่ยวข้องกับการต้านทานโรค พืชสะท้อนให้เห็นว่า iturin A สามารถจับกับโปรตีน 12-oxophytodienoate reductase 2 ซึ่ง เกี่ยวข้องกับกระบวนการสังเคราะห์ oxylipin ที่นําไปสู่การกระตุ้นระบบการต้านทานโรคพืช induced systemic resistance (ISR) การวิเคราะห์ทางด้านเมแทโบโลมิกส์โดยวิธี dansylation isotope labeling LC-MS ชี้ชัดว่า สารกลุ่มไลโปเปปไทด์สามารถกระตุ้นการเพิ่มปริมาณของกรดอะมิโนบางชนิดได้ เช่นกรดแอสพาร์ติก โฮโมเซอรีน ทรีโอนีน เซอรีน ไกลซีน และ ทริปโตเฟน ซึ่งถือเป็นเมแทโบไลท์ชนิดปฐมภูมิ รวมทั้ง เซโรโทนิน และทริปตามีน ซึ่งถือเป็นสารเมแทโบไลท์ ชนิดทุติยภูมิที่เกี่ยวข้องกับกระบวนการเมแทบิลิซึมของทริปโตเฟนและไทโรซีน ตามลําดับ ในผล ส้มโชกุนหลังการเก็บเกี่ยวเพื่อตอบสนองต่อสภาะเครีย

Supplementary data

There are the supplementary data.</p

Targeted transcriptional and proteomic studies explicate specific roles of Bacillus subtilis iturin A, fengycin, and surfactin on elicitation of defensive systems in mandarin fruit during stress.

Application of Bacillus cyclic lipopeptides (CLPs); fengycin, iturin A and surfactin has shown a great potential in controlling the spread of green mold pathogen invasion (Penicillium digitatum) in wounded mandarin fruit during postharvest period. The limited defensive protein profiles followed specific expression of pivotal genes relating to plant hormone mediating signaling pathways of the CLPs' action on stimulating host plant resistance have been exhibited. The present study aimed to elucidate the specific effect of individual CLP obtained from Bacillus subtilis ABS-S14 as elicitor role on activation of plant defensive system at transcriptional and proteomic levels with and without P. digitatum co-application in mandarin fruit. Fengycin and iturin A elevated the gene expression of PAL, ACS1, ACO, CHI, and GLU while significantly stimulating plant POD transcription was only detected in the treatments of surfactin both with and without following P. digitatum. An increase of LOX and PR1 gene transcripts was determined in the treatments of individual CLP with fungal pathogen co-application. Fengycin activated production of unique defensive proteins such as protein involved in ubiquinone biosynthetic process in treated flavedo without P. digitatum infection. Proteins involved in the auxin modulating pathway were present in the iturin A and surfactin treatments. CLP-protein binding assay following proteome analysis reveals that iturin A attached to 12-oxophytodienoate reductase 2 involved in the oxylipin biosynthetic process required for jasmonic acid production which is implicated in induced systemic resistance (ISR). This study suggests specific elicitor action of individual CLP, particularly iturin A showed the most powerful in stimulating the ISR system in response to stresses in postharvest mandarins

Exploring metabolic pathway alterations in obese fermented feces mediated by individual fruit extracts of Triphala components using untargeted metabolomics

Objective: This work aimed to explore the modified pathways impacted by changes in fecal metabolites among female obese adults during the human gut model period, both in the presence and absence of individual Triphala constituent fruit extract. Methods and results: The human gut model employed individual fruit extracts from Phyllanthus emblica, Terminalia bellerica, and T. chebula, comparing them to a control group. Fermentation used fecal samples from female obese adults over 24 h. Metabolite extraction, untargeted metabolomics, and pathway analysis identified metabolic changes. Treatments with P. emblica extract, T. bellerica extract, and T. chebula extract revealed the statistical detection of 128, 734, and 757 up-regulated metabolites, respectively, while 31, 90, and 92 down-regulated metabolites were identified. Pathway analysis revealed that P. emblica extract primarily influenced vitamin B6 metabolism, whereas the treatments with T. bellerica extract and T. chebula extract predominantly engaged glycine, serine, and threonine metabolism for metabolic regulation within the human gut model. Conclusion: By examining obese fecal metabolite changes and their association with metabolic pathway modulation through individual fruit extracts of Triphala constituents in a human gut model, this study provides a comprehensive understanding of Triphala components’ potential for managing obesity and its applications in the food industry

Metaproteomic Analysis of Gut Resistome in the Cecal Microbiota of Fattening Pigs Raised without Antibiotics

ABSTRACT Improper use of antibiotics in swine could reduce commensal bacteria and possibly increase pathogen infections via the gut resistome. This study aimed to compare the metaproteomic profiles of the gut resistome and related metabolism in the cecal microbiota of fattening pigs raised under antibiotic-free (ABF) conditions with those of ordinary industrial pigs (controls [CTRL]). The top three relatively abundant microbes in both groups were Escherichia coli, Ruminococcus, and Lactobacillus, followed by Bacteroides and Bifidobacterium. E. coli, Lactobacillus, and Bacteroides were found to be increased in the CTRL group, whereas Ruminococcus and Clostridium were greater in the ABF group. The highest abundances of antibiotic resistance proteins (log2 expression levels [ELs] of >10) were found to be for tetracycline resistance (Tetr) and aminoglycoside resistance (AMGr) proteins found in Bacteroides, with a significant increase in the CTRL group. High Tetr (ELs of 5.32) was found in Ruminococcus in the CTRL group, although pigs in both groups had never received tetracycline, possibly reflecting the influence of environments in farms. In E. coli, AMGr and β-lactamase family proteins were observed in both groups (ELs of 3 to 6), whereas multidrug resistance protein MdtL was significantly expressed in the CTRL group (ELs of around 3). In the ABF group, CRISPR-associated endonucleases Cas1 and Cas9, which function to defend against viruses, were markedly observed in Ruminococcus and Lactobacillus, respectively, with ELs of 8.6 and 4.15, respectively. In conclusion, this study demonstrated that CRISPR-associated endonucleases were markedly observed in the ABF group, whereas higher levels of Tetr, AMGr, and multidrug resistance protein MdtL was markedly observed in dominant bacterial species in the CTRL group. IMPORTANCE In order to control and reduce antibiotic use in animals, the Department of Livestock Development, Thailand, has launched a campaign for antibiotic-free livestock production. The present study has shown for the first time that CRISPR-associated endonucleases Cas1 and Cas9, which function to defend against viruses, were markedly observed in Ruminococcus and Lactobacillus, respectively, in ceca of pigs raised without antibiotics (ABF). The highest abundances of antibiotic resistance proteins were for tetracycline (Tetr) and aminoglycoside resistance (AMGr) proteins found in Bacteroides, with a significant increase in the controls. In E. coli, the microbe with the highest relative abundance, AMGr and β-lactamase family proteins were observed in both groups, whereas multidrug resistance protein MdtL was significantly expressed in the controls. Pigs in both ABF and control groups had never received tetracycline, possibly reflecting the influence of farm environments. We suggest that pigs raised without antibiotics may have more beneficial microorganisms for the gut than pigs raised with antibiotics

Dynamics of the fecal microbiome and antimicrobial resistome in commercial piglets during the weaning period

10.1038/s41598-021-97586-9Scientific Reports1111809