Genome-Wide Analysis of the PHT Gene Family and Its Response to Mycorrhizal Symbiosis in Tomatoes under Phosphate Starvation Conditions.

Phosphate is one of the essential mineral nutrients. Phosphate transporter genes (PHTs) play an important role in Pi acquisition and homeostasis in tomato plants. However, basic biological information on PHT genes and their responses of symbiosis with arbuscular mycorrhizal in the genome remains largely unknown. We analyzed the physiological changes and PHT gene expression in tomatoes (Micro-Tom) inoculated with arbuscular mycorrhizal (AM) fungi (Funneliformis mosseae) under different phosphate conditions (P1: 0 µM, P2: 25 µM, and P3: 200 µM Pi). Twenty-three PHT genes were identified in the tomato genomics database. Protein sequence alignment further divided the 23 PHT genes into three groups, with similar classifications of exons and introns. Good colonization of plants was observed under low phosphate conditions (25 µM Pi), and Pi stress and AM fungi significantly affected P and N accumulation and root morphological plasticity. Moreover, gene expression data showed that genes in the SlPHT1 (SlPT3, SlPT4, and SlPT5) gene family were upregulated by Funneliformis mosseae under all conditions, which indicated that these gene levels were significantly increased with AM fungi inoculation. None of the analyzed SlPHT genes in the SlPH2, SlPHT3, SlPHT4, and SlPHO gene families were changed at any Pi concentration. Our results indicate that inoculation with AM fungi mainly altered the expression of the PHT1 gene family. These results will lay a foundation for better understanding the molecular mechanisms of inorganic phosphate transport under AM fungi inoculation.

The Roles of Phosphorus and Nitrogen Nutrient Transporters in the Arbuscular Mycorrhizal Symbiosis.

More than 80% of land plant species can form symbioses with arbuscular mycorrhizal (AM) fungi, and nutrient transfer to plants is largely mediated through this partnership. Over the last few years, great progress has been made in deciphering the molecular mechanisms underlying the AM-mediated modulation of nutrient uptake progress, and a growing number of fungal and plant genes responsible for the uptake of nutrients from soil or transfer across the fungal-root interface have been identified. In this review, we outline the current concepts of nutrient exchanges within this symbiosis (mechanisms and regulation) and focus on P and N transfer from the fungal partner to the host plant, with a highlight on a possible interplay between P and N nutrient exchanges. Transporters belonging to the plant or AM fungi can synergistically process the transmembrane transport of soil nutrients to the symbiotic interface for further plant acquisition. Although much progress has been made to elucidate the complex mechanism for the integrated roles of nutrient transfers in AM symbiosis, questions still remain to be answered; for example, P and N transporters are less studied in different species of AM fungi; the involvement of AM fungi in plant N uptake is not as clearly defined as that of P; coordinated utilization of N and P is unknown; transporters of cultivated plants inoculated with AM fungi and transcriptomic and metabolomic networks at both the soil-fungi interface and fungi-plant interface have been insufficiently studied. These findings open new perspectives for fundamental research and application of AM fungi in agriculture.

LMWP (S3-3) from the Larvae of Musca domestica Alleviate D-IBS by Adjusting the Gut Microbiota.

Diarrhea-based Irritable Bowel Syndrome (D-IBS) and diarrhea are both associated with ecological imbalance of the gut microbiota. Low Molecular Weight Peptides (LMWP) from the larvae of Musca domestica have been shown to be effective in the treatment of diarrhea and regulation of gut microbiota. Meanwhile, the single polypeptide S3-3 was successfully isolated and identified from LMWP in our previous studies. It remains unclear exactly whether and how LMWP (S3-3) alleviate D-IBS through regulating gut microbiota. We evaluated the gut microbiota and pharmacology to determine the regulation of gut microbiota structure and the alleviating effect on D-IBS through LMWP (S3-3). The rates of loose stools, abdominal withdrawal reflex (AWR) and intestinal tract motility results revealed that LMWP (S3-3) from the larvae of Musca domestica had a regulating effect against diarrhea, visceral hypersensitivity and gastrointestinal (GI) dysfunction in D-IBS model mice. Additionally, 16S rRNA gene sequencing was utilized to examine the gut microbiota, which suggests that LMWP induce structural changes in the gut microbiota and alter the levels of the following gut microbiota: Bacteroidetes, Proteobacteria and Verrucomicrobia. LMWP putatively functioned through regulating 5-HT, SERT, 5-HT2AR, 5-HT3AR and 5-HT4R according to the results of ELISA, qRT-PCR and IHC. The findings of this study will contribute to further understanding how LMWP (S3-3) attenuate the effects of D-IBS on diarrhea, visceral hypersensitivity and GI dysfunction.

Anticancer Effects of Five Biflavonoids from Ginkgo Biloba L. Male Flowers In Vitro.

Ginkgo biloba L., an ancient dioecious gymnosperm, is now cultivated worldwide for landscaping and medical purposes. A novel biflavonoid-amentoflavone 7''-O-ß-D-glucopyranoside (1)-and four known biflavonoids were isolated and identified from the male flowers of Ginkgo. The anti-proliferative activities of five biflavonoids were evaluated on different cancer lines. Bilobetin (3) and isoginkgetin (4) exhibited better anti-proliferative activities on different cancer lines. Their effects were found to be cell-specific and in a dose and time dependent manner for the most sensitive HeLa cells. The significant morphological changes validated their anticancer effects in a dose-dependent manner. They were capable of arresting the G2/M phase of the cell cycle, inducing the apoptosis of HeLa cells dose-dependently and activating the proapoptotic protein Bax and the executor caspase-3. Bilobetin (3) could also inhibit the antiapoptotic protein Bcl-2. These might be the mechanism underlying their anti-proliferation. In short, bilobetin (3) and isoginkgetin (4) might be the early lead compounds for new anticancer agents.

Overlapping Root Architecture and Gene Expression of Nitrogen Transporters for Nitrogen Acquisition of Tomato Plants Colonized with Isolates of Funneliformis mosseae in Hydroponic Production.

Understanding the impact of arbuscular mycorrhizal fungi (AMF) upon the nitrogen (N) uptake of tomato (Lycopersicum esculentum L.) plants is crucial for effectively utilizing these beneficial microorganisms in industrial hydroponic tomato production. Yet it remains unknown whether, besides fungal delivery, the AMF also affects N uptake via altered plant root growth or whether, together with changed N transporters expression of hosts, this impact is isolate-specific. We investigated tomato root architecture and the expression of LeAMT1.1, LeAMT1.2, and LeNRT2.3 genes in roots inoculated with five isolates of Funneliformis mosseae, these collected from different geographical locations, under greenhouse conditions with nutritional solution in coconut coir production. Our results revealed that isolate-specific AMF inoculation strongly increased the root biomass, total root length, surface area, and volume. Linear relationships were found between the total root length and N accumulation in plants. Furthermore, expression levels of LeAMT1.1, LeAMT1.2, and LeNRT2.3 were significantly up-regulated by inoculation with F. mosseae with isolate-specific. These results implied N uptake greater than predicted by root growth, and N transporters up-regulated by AMF symbiosis in an isolate-specific manner. Thus, an overlap in root biomass, architecture and expression of N transporters increase N acquisition in tomato plants in the symbiosis.

Non-targeted metabolomic analysis of variation of volatile fractions of ginseng from different habitats by HS-SPME-GC-MS coupled with chemometrics.

Cultivated ginseng (CG), transplanted ginseng (TG) and mountain cultivated ginseng (MCG) classified by the habitat type all belong to Panax ginseng and were reported to have similar types of secondary metabolites. Nonetheless, owing to the distinctly diverse habitats in which these ginseng types grow, their pharmacological effects differ. In the present study, an emerging analytical approach involving headspace solid-phase microextraction-gas chromatography-mass spectrometry (HS-SPME-GC-MS) was established to effectively distinguish among CG, TG and MCG. First, the volatile components were analysed and identified by using the NIST library combined with measured retention indices (Kovats', RI), and a total of 78 volatile components were finally characterized, which included terpenes, alcohols, esters, aldehydes and alkynols. Furthermore, multivariate statistical approaches, principal component analysis (PCA) and orthogonal partial least-squares discrimination analysis (OPLS-DA) were subsequently utilized to screen for compounds of significance. Under optimized HS-SPME-GC-MS conditions, 12, 16, and 16 differential markers were screened in the CG-TG, CG-MCG and TG-MCG groups, respectively. Our study suggested that HS-SPME-GC-MS analysis combined with metabolomic analytical methods and chemometric techniques can be applied as potent tools to identify chemical marker candidates to distinguish CG, TG and MCG.

Beneficial Effects of the Five Isolates of Funneliformis mosseae on the Tomato Plants Were Not Related to Their Evolutionary Distances of SSU rDNA or PT1 Sequences in the Nutrition Solution Production.

The symbiosis and beneficial effects of arbuscular mycorrhizal fungi (AM fungi) on plants have been widely reported; however, the effects might be unascertained in tomato industry production with coconut coir due to the nutrition solution supply, or alternatively with isolate-specific. Five isolates of AM fungi were collected from soils of differing geographical origins, identified as Funneliformis mosseae and evidenced closing evolutionary distances with the covering of the small subunit (SSU) rDNA regions and Pi transporter gene (PT1) sequences. The effects of these isolates on the colonization rates, plant growth, yield, and nutrition uptake were analyzed in tomato nutrition solution production with growing seasons of spring-summer and autumn-winter. Our result indicated that with isolate-specific effects, irrespective of geographical or the SSU rDNA and PT1 sequences evolution distance, two isolates (A2 and NYN1) had the most yield benefits for plants of both growing seasons, one (E2) had weaker effects and the remaining two (A2 and T6) had varied seasonal-specific effects. Inoculation with effective isolates induced significant increases of 29.0-38.0% (isolate X5, T6) and 34.6-36.5% (isolate NYN1, T6) in the plant tissues respective nitrogen and phosphorus content; the plant biomass increased by 18.4-25.4% (isolate T6, NYN1), and yields increased by 8.8-12.0% (isolate NYN1, A2) compared with uninoculated plants. The maximum root biomass increased by 28.3% (isolate T6) and 55.1% (isolate E2) in the autumn-winter and spring-summer growing seasons, respectively. This strong effect on root biomass was even more significant in an industry culture with a small volume of substrate per plant. Our results reveal the potential benefits of using selected effective isolates as a renewable resource that can overcome the suppressing effects of sufficient nutrient availability on colonization rates, while increasing the yields of industrially produced tomatoes in nutrition solution with coconut coir.