Unveiling the impact of nitrogen deficiency on alkaloid synthesis in konjac corms (Amorphophallus muelleri Blume).

BACKGROUND: Konjac corms are known for their alkaloid content, which possesses pharmacological properties. In the primary cultivation areas of konjac, nitrogen deficiency is a common problem that significantly influences alkaloid synthesis. The impact of nitrogen deficiency on the alkaloids in konjac corms remains unclear, further complicated by the transition from mother to daughter corms during their growth cycle. RESULTS: This study examined 21 alkaloids, including eight indole alkaloids, five isoquinoline alkaloids, and eight other types of alkaloids, along with the associated gene expressions throughout the development of Amorphophallus muelleri Blume under varying nitrogen levels. Nitrogen deficiency significantly reduced corm diameter and fresh weight and delayed the transformation process. Under low nitrogen conditions, the content of indole alkaloids and the expression of genes involved in their biosynthesis, such as tryptophan synthase (TRP) and tryptophan decarboxylase (TDC), exhibited a substantial increase in daughter corms, with fold changes of 61.99 and 19.31, respectively. Conversely, in the mother corm, TDC expression was markedly reduced, showing only 0.04 times the expression level observed under 10 N treatment. The patterns of isoquinoline alkaloid accumulation in corms subjected to nitrogen deficiency were notably distinct from those observed for indole alkaloids. The accumulation of isoquinoline alkaloids was significantly higher in mother corms, with expression levels of aspartate aminotransferase (GOT), chorismate mutase (CM), tyrosine aminotransferase (TAT), and pyruvate decarboxylase (PD) being 4.30, 2.89, 921.18, and 191.40 times greater, respectively. Conversely, in daughter corms, the expression levels of GOT and CM in the 0 N treatment were markedly lower (0.01 and 0.83, respectively) compared to the 10 N treatment. CONCLUSIONS: The study suggests that under nitrogen deficiency, daughter corms preferentially convert chorismate into tryptophan to synthesize indole alkaloids, while mother corms convert it into tyrosine, boosting the production of isoquinoline alkaloids. This research provides valuable insights into the mechanisms of alkaloid biosynthesis in A. muelleri and can aid in developing nitrogen fertilization strategies and in the extraction and utilization of alkaloids.

Gamma-aminobutyric acid as a regulator of astaxanthin production in Haematococcus lacustris under salinity: Exploring physiology, signaling, autophagy, and multi-omics landscape.

Haematococcus lacustris-derived natural astaxanthin has significant commercial value, but stressful conditions alone impair cell growth and reduce the total productivity of astaxanthin in industrial settings. This study used gamma-aminobutyric acid (GABA) to increase biomass, astaxanthin productivity, and tolerance to salinity. GABA under NaCl stress enhanced the biomass to 1.76 g/L, astaxanthin content to 30.37 mg g-1, and productivity to 4.10 mg/L d-1, outperforming the control. Further analysis showed GABA enhanced nitrogen assimilation, Ca2+ level, and cellular GABA content, boosting substrate synthesis, energy metabolism, osmoregulation, autophagy, and antioxidant defenses. GABA also activated signaling pathways involving phytohormones, cAMP, cGMP, and MAPK, aiding astaxanthin synthesis. The application of biomarkers (ethylene, salicylic acid, trans-zeatin) and an autophagy inhibitor cooperated with GABA to further enhance the total astaxanthin productivity under NaCl stress. Combining GABA with 25 µM salicylic acid maximized astaxanthin yield at 4.79 mg/L d-1, offering new strategies for industrial astaxanthin production.

Complete chloroplast genome and phylogenetic analysis of Amorphophallus paeoniifolius (Araceae).

Amorphophallus paeoniifolius (Dennst.) Nicolson, 1885, often known as elephant foot yam, is a tropical tuber crop that originates from south-east Asia and belongs to the Araceae family. It is known for its high production potential and popularity as a medicinal plant. However, the phylogeny and genes for this species are still unavailable. In this study, the first complete chloroplast genome of A. paeoniifolius was reported and phylogenetic analysis was conducted with Araceae species. The chloroplast genome was 176,258 bp in length with 34.80% overall GC content and includes a large single-copy (LSC) region (93,951 bp), a small single-copy (SSC) region (15,013 bp), and a pair of inverted repeat (IRs) regions (33,647 bp). The chloroplast genome has 130 genes, which include 85 protein-coding genes, 37 tRNA genes, and eight rRNA genes. A maximum-likelihood (ML) phylogenetic analysis indicated that all Amorphophallus species formed a single monophyletic clade with a high bootstrap value and A. paeoniifolius was closely related to A. konjac, A. albus, A. krausei, and A. titanum. The chloroplast genome reported in this study will be useful for further taxonomic and evolutionary studies of Amorphophallus.

The endophytic fungal community plays a crucial role in the resistance of host plants to necrotic bacterial pathogens.

Konjac species (Amorphophallus spp.) are the only plant species in the world that are rich in a large amount of konjac glucomannan (KGM). These plants are widely cultivated as cash crops in tropical and subtropical countries in Asia, including China. Pectobacterium carotovorum subsp. carotovorum (Pcc) is one of the most destructive bacterial pathogens of konjac. Here, we analyzed the interactions between Pcc and susceptible and resistant konjac species from multiple perspectives. At the transcriptional and metabolic levels, the susceptible species A. konjac and resistant species A. muelleri exhibit similar molecular responses, activating plant hormone signaling pathways and metabolizing defense compounds such as phenylpropanoids and flavonoids to resist infection. Interestingly, we found that Pcc stress can lead to rapid recombination of endophytic microbial communities within a very short period (96 h). Under conditions of bacterial pathogen infection, the relative abundance of most bacterial communities in konjac tissue decreased sharply compared with that in healthy plants, while the relative abundance of some beneficial fungal communities increased significantly. The relative abundance of Cladosporium increased significantly in both kinds of infected konjac compared to that in healthy plants, and the relative abundance in resistant A. muelleri plants was greater than that in susceptible A. konjac plants. Among the isolated cultivable microorganisms, all three strains of Cladosporium strongly inhibited Pcc growth. Our results further elucidate the potential mechanism underlying konjac resistance to Pcc infection, highlighting the important role of endophytic microbial communities in resisting bacterial pathogen infections, especially the more direct role of fungal communities in inhibiting pathogen growth.

The complete chloroplast genome sequence of Amorphophallus konjac (Araceae) from Yunnan, China and its phylogenetic analysis in the family Araceae.

This work determined and analyzed the complete chloroplast genome sequence of Amorphophallus konjac K. Koch ex N.E.Br 1858 from Yunnan, China. The genome size was 167,470 bp, of which contains a large single-copy region (LSC 93,443 bp), a small single-copy region (SSC 21,575 bp), and a pair of inverted repeat regions (IR 26,226 bp). The chloroplast genome has 131 genes, including 86 protein-coding genes, 37 tRNAs, and eight rRNAs. A previous study reported deletion of accD, psbE, and trnG-GCC genes in the A. konjac chloroplast genome. Our study supports the conservative structure of A. konjac and does not support the gene deletion mentioned above. Phylogenetic analysis indicated that A. konjac shares a close relationship with another A. konjac (collected from Guizhou) and A. titanium by forming a clade in the genus Amorphophallus. Our results provide some useful information to the evolution of the family Araceae.

Melatonin, a phytohormone for enhancing the accumulation of high-value metabolites and stress tolerance in microalgae: Applications, mechanisms, and challenges.

High-value metabolites, such as carotenoids, lipids, and proteins, are synthesized by microalgae and find applications in various fields, including food, health supplements, and cosmetics. However, the potential of the microalgal industry to serve these sectors is constrained by low productivity and high energy consumption. Environmental stressors can not only stimulate the accumulation of secondary metabolites in microalgae but also induce oxidative stress, suppressing cell growth and activity, thereby resulting in a decrease in overall productivity. Using melatonin (MT) under stressful conditions is an effective approach to enhance the productivity of microalgal metabolites. This review underscores the role of MT in promoting the accumulation of high-value metabolites and enhancing stress resistance in microalgae under stressful and wastewater conditions. It discusses the underlying mechanisms whereby MT enhances metabolite synthesis and improves stress resistance. The review also offers new perspectives on utilizing MT to improve microalgal productivity and stress resistance in challenging environments.

Comprehensive understanding of the regulatory mechanism by which selenium nanoparticles boost CO2 fixation and cadmium tolerance in lipid-producing green algae under recycled medium.

Recycled medium plus cadmium is a promising technique for reducing the cultivation cost and enhancing the yield of microalgae lipids. However, oxidative stress and cadmium toxicity significantly hinder the resulting photosynthetic efficiency, cell growth and cell activity. Herein, selenium nanoparticles (SeNPs) were used to increase the total biomass, biolipid productivity, and tolerance to cadmium. Wide-ranging analyses of photosynthesis, energy yield, fatty acid profiles, cellular ultrastructure, and oxidative stress biomarkers were conducted to examine the function of SeNPs in CO2 fixation and cadmium resistance in Ankistrodesmus sp. EHY. The application of 15 µM cadmium and 2 mg L-1 SeNPs further enhanced the algal biomass productivity and lipid productivity to 500.64 mg L-1 d-1 and 301.14 mg L-1 d-1, respectively. Moreover, the rates of CO2 fixation, chlorophyll synthesis and total nitrogen removal were similarly increased by the application of SeNPs. Exogenous SeNPs strengthened cell growth and cadmium tolerance by upregulating photosynthesis, the TCA cycle and the antioxidant system, reducing the uptake and translocation of cadmium, and decreasing the levels of reactive oxidative stress (ROS), extracellular polymeric substances (EPSs) and cellular Cd2+ level in EHY under recycled medium and cadmium stress conditions. Additionally, a maximum energy yield of 127.40 KJ L-1 and a lipid content of 60.15% were achieved in the presence of both SeNPs and cadmium stress. This study may inspire the efficient disposal of recycled medium and biolipid production while also filling the knowledge gaps regarding the mechanisms of SeNP functions in carbon fixation and cadmium tolerance in microalgae.

Synergistic effects of gamma-aminobutyric acid and melatonin on seed germination and cadmium tolerance in tomato.

The effects of exogenous γ-aminobutyric acid (GABA) and melatonin (MT) on tomato seed germination and shoot growth exposed to cadmium stress were investigated. On the one hand, treatment with MT (10-200 µM) or GABA (10-200 µM) alone could significantly relieve cadmium stress in tomato seedlings, which is reflected in increasing the germination rate, vigor index, fresh weight, dry weight and radicle lengths of tomato seeds, as well as the soluble content compared to the absence of exogenous treatment, and the alleviating effect reached the peak in the 200 µM GABA or 150 µM MT alone. On the other hand, exogenous MT and GABA showed synergistic effects on the germination of tomato seed under cadmium stress. Moreover, the application of 100 µM GABA combined with 100 µM MT markedly decreased the contents of Cd and MDA by upregulating the activities of antioxidant enzymes, thereby alleviating the toxic effect of cadmium stress on tomato seeds. Collectively, the combinational strategy showed significant positive effects on seed germination and cadmium stress resistance in tomato.

Exogenous prolinebooststheco-accumulation ofastaxanthin and biomassin stress-induced Haematococcus pluvialis.

This paper aims to explore the role of proline (Pro) in the production of biomass and astaxanthin (AST) in stress-induced Haematococcus pluvialis. The astaxanthin content and productivity were 24.02 mg g-1 and 2.22 mg/L d-1 under abiotic stresses, respectively. After 100 µM Pro supplementation, the biomass, AST and lipid contents reached 1.43 g/L, 29.91 mg g-1 and 56.79 %, which were enhanced by 19.16 %, 33.52 % and 11.08 %, respectively, compared to the control. Pro-treated regulated chlorophyll, carbohydrate and protein accumulation and upregulated carotenogenic, lipogenic and antioxidant enzymes-associated gene levels; as well as increased endogenous Pro content, but reduced ROS (Reactive oxygen species) and MDA (Malondialdehyde) levels and alleviated oxidative stress, which might be involved in AST biosynthesis. Further data showed Pro has a positive role in biomass and AST coaccumulation in different H. pluvialis species, suggesting application of Pro was an effective strategy to improve AST productivity of H. pluvialis.

A low-cost technique for biodiesel production in Ankistrodesmus sp. EHY by using harvested microalgal effluent.

The present study aims to use Ankistrodesmus sp. EHY to develop a viable and economic lipid production strategy using recycling of harvested microalgal effluent. In comparison to the control, the highest lipid content (52.4 %) and productivity (250.72 mg L-1 d-1) were achieved when 40 % recycled medium was used. Consistent with the trend of lipid accumulation, the six key lipogenetic genes were upregulated, as well as reactive oxygen species (ROS), glutathione (GSH) and genes encoding antioxidant enzymes during cultivation in recycled medium. Moreover, the consumption of dissolved organic carbon (DOC) and the increased humic acid (HA) in the recycled medium might also be associated with lipid biosynthesis. The biodiesel parameters of alga biomass-derived lipids were fitted to the standard of commercial biodiesel. In conclusion, this study offers an economically viable strategy for microalgal biofuel production and wastewater treatment using recycling of harvested microalgal effluent.

Myo-inositol facilitates astaxanthin and lipid coproduction in Haematococcus pluvialis by regulating oxidative stress and ethylene signalling.

In the present study, exogenous myo-inositol (MI) was applied to induce natural astaxanthin and biolipid accumulation in Haematococcus pluvialis. Under 200 µM MI, algal cells exhibited 62.11 % and 34.67 % increases in astaxanthin and lipid content, respectively, compared to the control. The carotenogenesis and lipogenesis genes were upregulated by induction of MI. Interestingly, MI addition elevated the ethylene (ETH) content and activated antioxidant enzyme-associated gene levels, which could be involved in alleviating oxidative stress. Further data showed that the ETH signal played a positive function in stimulating astaxanthin biosynthesis under MI induction. Supplementation with ethephon plus MI boosted the astaxanthin content to 33.08 ± 0.03 mg g-1 by further upregulating astaxanthin biosynthesis genes and blocking reactive oxidative species (ROS) levels, and vice versa under ETH inhibition. This study provides a potential induction approach for natural astaxanthin production and explains the role of ethylene signalling in regulating astaxanthin synthesis by H. pluvialis.

Promoting lutein production from the novel alga Acutodesmus sp. by melatonin induction.

In the current research, a novel microalgae strain was isolated from Yajiageng Red Rock Beach and identified as Acutodesmus sp. HLGY. To obtain high-efficiency production of lutein from algae, the feasibility of using melatonin (MT) to increase lutein yield of Acutodesmus sp. HLGY was evaluated. Under the 7.5 µM MT treatment, the lutein content and lutein productivity were 17.44 mg g-1 and 46.50 mg L-1 d-1, which were 1.53 times those of the control. Furthermore, exogenous MT increased the transcripts of key lutein synthesis- and antioxidant enzyme-related genes. Simultaneously, the carbohydrate, protein, and cellular reactive oxygen species (ROS) levels and lipid content were suppressed. More importantly, the ethylene and γ-aminobutyric acid contents were markedly increased by MT, which may be linked to the increase in lutein biosynthesis. This study proposes a valuable biotechnological approach for lutein production via a novel Acutodesmus sp. strain using MT induction and provides insights into the role of MT in promoting lutein biosynthesis.

Phytohormone-like small biomolecules for microalgal biotechnology.

Microalgae are highly adaptable to abiotic stress and produce valuable metabolites, but microalgal commercialization is still difficult because of minimal yields. The application of phytohormone-like small biomolecules is effective in simultaneously improving the productivity of valuable microalgal biomass-derived metabolites and stress tolerance. This represents a significant opportunity for microalgal biotechnology.

Enhancing astaxanthin and lipid coproduction in Haematococcus pluvialis by the combined induction of plant growth regulators and multiple stresses.

Coupling chemical induction and abiotic stresses is a beneficial strategy for astaxanthin (Asta) induction in Haematococcus pluvialis. The combined application of melatonin (MT) and putrescine (Put) induced Asta and lipid biosynthesis in H. pluvialis under adverse conditions. Under MT and Put inductions, the highest Asta and lipid contents were 3.64% and 55.84%, which were 1.71- and 1.17-times higher than the control group, respectively. The combination of MT and Put also enhanced the expression of carotenogenic, lipogenic and antioxidant enzyme genes. Additionally, this combined treatment increased the endogenous Put content while decreasing the reactive oxygen species (ROS) and γ-aminobutyric acid (GABA) levels. Further results proved that endogenous Put promoted Asta production and alleviated oxidative stress by regulating carotenogenesis and GABA and ROS signaling. This study describes a potential process for stimulating Asta and lipid coproduction and highlights the connections among MT, Put, signaling molecules, Asta and lipid synthesis in H. pluvialis.

Coupling of myo-inositol with salinity regulates ethylene-induced microalgal lipid hyperproduction in molasses wastewater.

With the goal of cost-effective and high-efficient microalgae-based biodiesel production, this study evaluated the feasibility of the joint strategy concerning myo-inositol (MI) and salinity stress on lipid productivity of Monoraphidium sp. QLY-1 in molasses wastewater (MW). The maximal lipid productivity (147.79 mg L-1 d-1) was obtained under combined 0.5 g L-1 MI and 10 g L-1 NaCl treatment, which was 1.40-fold higher than the control. Meanwhile, the nutrients removal from MW was markedly increased under MI-NaCl treatment. Moreover, exogenous MI upregulated key lipogenic genes' expressions, activated autophagic activity and ethylene (ET) signaling, and ultimately alleviated the salinity-induced damage via reactive oxygen species (ROS) signaling. Further pharmacologic experiment confirmed the indispensable role of ET in the lipogenesis progress under the combined treatment. These data demonstrated the combined salinity stress and MI treatment to be capable for lipid hyperproduction and wastewater nutrients removal, which contributes to practically integrating the microalgae cultivation with wastewater treatment.

A joint strategy comprising melatonin and 3-methyladenine to concurrently stimulate biomass and astaxanthin hyperaccumulation by Haematococcus pluvialis.

Haematococcus pluvialis is a commercial microalgae used for natural astaxanthin production. This study aims to investigate the roles of melatonin (MT) and 3-methyladenine (3-MA) in regulating the cell growth and biosynthesis of astaxanthin and fatty acids under adverse conditions by H. pluvialis. Upon the dual treatments, the maximum astaxanthin concentration (46.78 mg L-1) was 2.39- and 1.35-fold higher compared with the control and MT treatments, respectively. Concomitantly, the combined application of MT and 3-MA suppressed autophagy but promoted the production of biomass and lipids and upregulated carotenogenesis, lipogenesis and antioxidant enzyme-related genes at the transcriptional level, which were linked with astaxanthin and lipid biosynthesis and oxidative stress. Additionally, astaxanthin exhibited a noticeable increase under MT coupled with 3-MA in the other two strains of H. pluvialis. This study proposed a potential method for astaxanthin induction and provided insights into the function of autophagy in modulating cell growth and astaxanthin synthesis.

A fed-batch feeding with succinic acid strategy for astaxanthin and lipid hyper-production in Haematococcus pluviualis.

Effects of succinic acid (SA) in fed-batch feeding mode on astaxanthin and lipids biopoduction of Haematococcus pluvialis against abiotic stresses were explored. By comparison with the control, the initial addition of SA on day 0 increased the production of astaxanthin by 71.61%. More importantly, the maximum values of astaxanthin (35.88 mg g-1) and lipid (54.79%) contents were obtained after supplementation of SA on day 7. Meanwhile, under SA treatment, the chlorophyll, carbohydrate, and protein levels were reduced, but the intracellular levels of SA and reactive oxygen species (ROS), the transcription levels of astaxanthin and fatty acids biosynthesis-, and antioxidant system-related genes were increased. Furthermore, scaling-up cultivation in bioreactor further enhanced the astaxanthin productivity from H. pluvialis. Generally, this study proved the intermittent SA feeding method in fed-batch culture as a potent strategy that facilitated massive astaxanthin and lipids production in algae.

Enhancement of Lipid Productivity and Self-flocculation by Cocultivating Monoraphidium sp. FXY-10 and Heveochlorella sp. Yu Under Mixotrophic Mode.

To maintain high microalgae lipid productivity and flocculation efficiency simultaneously and reduce the production cost of microalgae lipids, Monoraphidium sp. FXY-10 with high lipid-producing capacity and Heveochlorella sp. Yu with strong self-flocculation ability were cocultivated and studied. Cocultivated microalgae lipid productivity and flocculation efficiency were increased to 203.8 mg L-1 day-1 and 70.55%, respectively, which is potentially related to the excessive competitive depletion of nitrogen sources and the upregulation of correlative key genes in lipid anabolic metabolism. Under cocultivation conditions, microalgae cells could enter the stationary phase 2 days earlier than that under monocultivation conditions, thus reducing the culture time. Relative expression of the accD, ME, and rbcL genes was upregulated to varying degrees, and the enzyme activities of ACCase, ME, and RuBisCO were also significantly increased compared with those in monocultivation. Moreover, fatty acid composition showed that microalgae lipids in cocultivation exhibited potential as a feedstock for biodiesel.

Role of copper in the enhancement of astaxanthin and lipid coaccumulation in Haematococcus pluvialis exposed to abiotic stress conditions.

This study investigated the effects of copper (Cu) on astaxanthin and lipid biological synthesis in unicellular alga Haematococcus pluvialis under high-light (HL) and nitrogen-deficiency (ND) conditions. During a 15-day cultivation period, the astaxanthin and lipid contents reached the peak values (3.32% and 47.72%) under 6 µM Cu treatment, which were increased by 66.87% and 34.99% compared to nontreated group, respectively. The application of Cu also increased the transcriptional expression of biosynthesis genes and antioxidant enzyme-related genes, as well as increased the intracellular calcium (Ca2+) level but led to a decrease in reactive oxygen species (ROS) levels. Additionally, Cu treatment induced the activation of calcium-dependent protein kinases (CDPKs) and mitogen-activated protein kinases (MAPKs). This approach simultaneously facilitated astaxanthin and lipid production, and the role of Cu were elucidated on the regulation of signal transduction (e.g., Ca2+, CDPK, MAPK and ROS) in the carotenogenesis and lipogenesis in H. pluvialis.