Integrated Transcriptomics and Metabolomics Reveal Key Insights into Iridoid Biosynthesis in Gentiana crassicaulis Seeds during Germination.

Background:Gentiana crassicaulis Duthie ex Burk., a key species used in traditional Chinese medicine for treating rheumatic pain and stroke, contains iridoids as its primary active component. However, the biosynthetic mechanisms underlying iridoid production are not fully understood. Methods: This study focused on iridoid biosynthesis during the germination of G. crassicaulis seeds, integrating metabolomic and transcriptomic analyses to uncover the underlying pathways and key candidate genes. Results: 196,132 unigenes and 10 iridoid compounds were identified through RNA-seq and ultra performance liquid chromatography-quadrupole time of flight-mass spectrometer (UPLC-Q-TOF-MS), respectively. The intersection of results from Pearson correlation analysis and weighted gene co-expression network analysis (WGCNA) revealed a significant correlation between 26 genes and iridoid levels, suggesting their potential role in the iridoid metabolism. Notably, six highly expressed candidate genes (DL7H, SLS, CYP76, CYP72A2, CYP84A1, and 13-LOX3) and five iridoids (loganic acid, sweroside, swertiamarin, gentiopicroside, and 6'-O-ß-D-glucosyl-gentiopicroside) responded to methyl jasmonate stimulation in G. crassicaulis seedlings. Conclusions: by combining the known functions of candidate gene families, It is hypothesized that the CYP716 and LOX families exert indirect influences on iridoid metabolism, while the CYP71, CYP81, CYP72, CYP76, CYP710 families, 2OG-FeII family, and the glucosyltransferase family are likely to play direct roles in the biosynthetic transformations of the five iridoids. This study provides a theoretical basis for further functional gene validation and metabolic engineering aimed at enhancing iridoid production. The insights gained could lead to improved iridoid production efficiency in medicinal plants, ultimately benefiting the quality and efficacy of medicinal materials.

Aluminum and UV-C light on seed germination and initial growth of white oats.

Aluminum (Al) may be beneficial to crops, but in excess becomes detrimental to the germination and initial development of seedlings. The main determining indicators are the type of crop and exposure duration. The aim of this study was to examine the influence of Al and of UV-C light on the germination and initial growth of white oats. Seeds were sown on germitest paper in a solution of 100, 200, 300, 400, or 500 mg/L of aluminum chloride and kept in a germination chamber at 20°C for a 12-hr photoperiod. Germination and seedling growth parameters were determined after 5 and 10 days. The seeds were also exposed to two doses of UV-C (0.85 and 3.42 kJ m-2) under aluminum chloride stress (200 mg/L). Data demonstrated that treatment with aluminum chloride significantly decrease in germination at 200 mg/L and total seedling length at 100 mg/L. Exposure of seeds to UV-C light under excess Al (200 mg/L) did not show a significant effect on germination and growth compared to control (non-irradiated). Results indicated that exposure to high concentration of Al in the medium adversely altered germination and initial growth of white oat seedlings. Although UV-C light alone was not detrimental to the germination process, treatment with UV-C light also failed to mitigate the toxic effects of Al.

Andean Crops Germination: Changes in the Nutritional Profile, Physical and Sensory Characteristics. A Review.

Andean crops such as quinoa, amaranth, cañihua, beans, maize, and tarwi have gained interest in recent years for being gluten-free and their high nutritional values; they have high protein content with a well-balanced essential amino acids profile, minerals, vitamins, dietary fiber, and antioxidant compounds. During the germination bioprocess, the seed metabolism is reactivated resulting in the catabolism and degradation of macronutrients and some anti-nutritional compounds. Therefore, germination is frequently used to improve nutritional quality, protein digestibility, and availability of certain minerals and vitamins; furthermore, in specific cases, biosynthesis of new bioactive compounds could occur through the activation of secondary metabolic pathways. These changes could alter the technological and sensory properties, such as the hardness, consistency and viscosity of the formulations prepared with them. In addition, the flavor profile may undergo improvement or alteration, a critical factor to consider when integrating sprouted grains into food formulations. This review summarizes recent research on the nutritional, technological, functional, and sensory changes occur during the germination of Andean grains and analyze their potential applications in various food products.

Shrimp injection with dsRNA targeting the microsporidian EHP polar tube protein reduces internal and external parasite amplification.

The microsporidian Enterocytozoon hepatopenaei (EHP) is a major threat to shrimp health worldwide. Severe EHP infections in shrimp cause growth retardation and increase susceptibility to opportunistic infections. EHP produces spores with a chitin wall that enables them to survive prolonged environmental exposure. Previous studies showed that polar tube extrusion is a prerequisite for EHP infection, such that inhibiting extrusion should prevent infection. Using a proteomic approach, polar tube protein 2 of EHP (EhPTP2) was found abundantly in protein extracts obtained from extruded spores. Using an immunofluorescent antibody against EhPTP2 for immunohistochemistry, extruded spores were found in the shrimp hepatopancreas (HP) and intestine, but not in the stomach. We hypothesized that presence of EhPTP2 might be required for successful EHP spore extrusion. To test this hypothesis, we injected EhPTP2-specific double-stranded RNA (dsRNA) and found that it significantly diminished EHP copy numbers in infected shrimp. This indicated reduced amplification of EHP-infected cells in the HP by spores released from previously infected cells. In addition, injection of the dsRNA into EHP-infected shrimp prior to their use in cohabitation with naïve shrimp significantly (p < 0.05) reduced the rate of EHP transmission to naïve shrimp. The results revealed that EhPTP2 plays a crucial role in the life cycle of EHP and that dsRNA targeting EHP mRNA can effectively reach the parasite developing in host cells. This approach is a model for future investigations to identify critical genes for EHP survival and spread as potential targets for preventative and therapeutic measures in shrimp.

Graphene oxide decreases the effects of salt stress on Persian clover seed germination.

Among biotic and abiotic stresses, the most restrictive for plant distribution is salt stress, where different concentrations might exert harmful effects on seed germination. Recently, nanomaterials were used successfully to mitigate these stresses, indicating that plants may be able to develop normally in adverse conditions. The aim of this study was to examine the effects of graphene oxide (GO) on the germination of Persian clover seedlings under salt stress conditions. Following sown on substrate paper, seeds were tested after exposure to different concentrations of graphene oxide (0, 125, 250, or 500 mg L-1 GO), sodium chloride (0; -0.1; -0.2; -0.3, or -0.4 MPa NaCl) and/or GO + salt concomitantly, and then stored for 7 days in a germination chamber at 20°C in the presence of light. Seed germination and growth parameters of seedlings were determined. Graphene oxide demonstrated protective effect against salt stress as evident by no marked adverse effects on seed germination where GO blocked the salt-induced reduction in germination. The results obtained provide references for the safe application of nanomaterials and emphasize the significance of GO as a promising material for reducing the toxicity of salts in agriculture.

Germination of pecan seeds changes the microbial community.

Endophytes are core of the plant-associated microbiome, and seed endophytes are closely related to the plant growth and development. Seed germination is an important part of pecan's life activities, but the composition and changes of microbes during different germination processes have not yet been revealed in pecan seeds. In order to deeply explore the characteristics of endophytes during the germination process of pecan, high-throughput sequencing was performed on seeds at four different germination stages. Findings of present study was found that the diversity and composition of microorganisms were different in different germination stages, and the microbial richness and diversity were highest in the seed endocarp break stage. It was speculated that the change of endophytes in pecan seeds was related to the germination stage. By evaluating the relationship between microbial communities, the core microbiota Cyanobacteria, Proteobacteria and Actinobacteria (bacterial) and Anthophyta and Ascomycota (fungal) core microbiota were identified in germinating pecan seeds. Finally, biomarkers in different germination processes of pecan seeds were identified by LEfSe analysis, among which Proteobacteria, Gamma proteobacteria and, Cyanobacteria and Ascomycota and Sordariomycetes were most abundant. Thus, this study will help to explore the interaction mechanism between pecan seeds and endophytes in different germination processes, and provide materials for the research and development of pecan seed endophytes.

Modification on phenolic profiles and enhancement of antioxidant activity of proso millets during germination.

Changes in phenolic profiles and antioxidant activity of three varieties of proso millet during germination were investigated. Total phenolic content (TPC) and total flavonoid content (TFC) increased significantly with prolongation in germination period. After germination for 6 days, TPC of the free and bound fractions increased 6.30-8.66-fold and 77.65-116.18%, respectively. The free and bound phenolic compounds identified by UPLC-MS/MS, displayed significant variations. Feruloylquinic acid and N,N'-bis-(p-coumaroyl)-putrescine biosynthesized during germination, are reported for the first time in proso millets. Other phenolics including trans- and cis-ferulic, trans-p-coumaric, vanillic acid and ferulic acid dimers (DFAs) were increased significantly along with a new DFA (8,5'-DFA) seemingly produced during germination. The germinated proso milllets displayed superior antioxidant activity than the corresponding ungerminated samples indicating that germination could be one applicable method for improving phenolic profiles and antioxidant capacity of proso millets. Thus germinated proso millet could be exploited as a functional ingredient in several products.

Chia (Salvia hispanica L.) Seed Germination: a Brief Review.

Chia (Salvia hispanica L.) is a seed native to northern Mexico and southern Guatemala that has started to be consumed in recent years in other regions of the world owing to its nutritional and functional properties. Germination of chia seeds seems to be able to further improve these properties, and it has been the subject of some studies. In general, germination has proven to be a simple and inexpensive process capable of improving the content of phenolic compounds and the antioxidant capacity of foods, as well as reducing antinutritional factors that interfere with nutrient absorption. A particular characteristic of chia seeds is that they produce mucilage when they are hydrated. For this reason, the germination conditions of the seed need to be adapted. The nutritional guidelines of some countries, such as Brazil, Germany and Sweden, recommend that the diet of the population should be more plant-based, thus encouraging the consumption of foods with a high content of bioactive compounds and nutrients, e.g., germinated seeds. This review briefly explored the germination conditions of chia seeds as well as the changes in phytonutrient content and antinutritional factors after their germination process. The main information available in the literature is that germination of chia seeds can increase the contents of protein, fiber, and total phenolic compounds. As a conclusion, germination of chia seeds is favorable for increasing their health benefits and nutritional value. However, chia germination parameters should be adjusted and microbiological risks should be properly evaluated.

Effect of heat-moisture treatment of germinated black rice on the physicochemical properties and its utilization by lactic acid bacteria.

The heat susceptibility of starch granule structure has been considering as significant limitation of germinated black rice (GBR) using in food processing industry. Therefore, this study aimed to improve the physicochemical and antioxidation property as well as its effect on the probiotics of GBR by heat moisture treatment (HMT). Black rice germinated at 37.5 °C for 12, 24, and 36 h were studied. Ultrastructural image of each sample was visualized through scanning electron microscope. The results illustrated 24 h-GBR retain its former shape with rough surface. Subsequently, 24 h-GBR was structurally modified by HMT with moisture levels of 20% and 25% for 1 and 2 h. The results showed that pasting properties of HMT-treated GBR were improved. This was particularly on, GBR using HMT condition of 25% moisture for 2 h decrease in breakdown viscosity was shown. Moreover, phenolic content of HMT-treated GBR was higher than those of GBR. Besides, the number of the Lactobacillus paracasei TOKAI 13 was increased in GBR and HMT-treated GBR with counts of 10.08 ± 0.83 Log CFU/ml and 9.31 ± 0.33 Log CFU/ml, respectively, with significant increases in antioxidant property. Therefore, the HMT-GBR could be utilized as an alternative functional ingredient in food processing products.

Oilseed rape (Brassica napus): the importance of aminopeptidases in germination under normal and heavy metals stress conditions.

BACKGROUND: Oilseed rape is one of the most important oilseed crops worldwide, crucial in the food and feed industries. Different environment and climatic conditions can influence its sustainable cultivation and crop yield. Aminopeptidases are crucial enzymes in many physiological processes in all organisms, including humans, so it is important to learn their behavior in food and feed sources. This study presents, for the first time, a detailed discussion on the importance of aminopeptidases, during the oilseed rape germination process, under standard and stress conditions. RESULTS: During the germination of oilseed rape under standard conditions, a significant increase in aminopeptidases activity toward N-terminal amino acids - phenylalanine (Phe), alanine (Ala), glycine (Gly), leucine (Leu), proline (Pro), methionine (Met) - was observed. The change was substrate specific, with the highest increase being observed for Gly (3.2-fold), followed by Ala (2.9-fold), Pro (2.5-fold), Met (1.5-fold), and Phe (1.3-fold). Generally, N-terminal Phe was preferentially cleaved. Germination under stress conditions, caused by several heavy metal ions (e.g. divalent copper, zinc, cadmium, and lead ions), negatively influenced the plants' growth and quality, but significantly enhanced the expression of genes encoding aminopeptidases (or potentially activated aminopeptidases precursors), which was related to the dramatic increase of their activity. CONCLUSIONS: The activity/concentration of aminopeptidases in plants is adjusted to the needs at each stage of development and stress factors occurrence. The most significant increase of activity toward N-terminal Gly and Pro proved the key role of aminopeptidases in the defense mechanisms, by supplying the plants with osmoprotectants and organic nitrogen. The results provide new concepts of oilseed rape growth and cultivation under different conditions. © 2021 Society of Chemical Industry.

Methods for Assessment of Viability and Germination of Plasmodiophora brassicae Resting Spores.

Clubroot caused by the obligate biotrophic parasite Plasmodiophora brassicae is a destructive soil borne disease of cruciferous crops. Resting spores of P. brassicae can survive in the soil for a long period without hosts or external stimulants. The viability and germination rate of resting spores are crucial factors of the inoculum potential in the field. The accurate assessment of viability and germination rate is the foundation to evaluate the effect of control methods. In this study, we evaluated several methods for the assessment of viability and germination rate of P. brassicae resting spores. Dual staining with calcofluor white-propidium iodide (CFW-PI) or single stain with Evans blue showed reliable accuracy in estimating viability. CFW-PI was capable of reliably determining the viability within 10 min, while Evans blue required overnight incubation to obtain accurate results. Due to DNA degradation of heat treatments, acetone was selected to evaluate the efficiency of propidium monoazide (PMA)-quantitative PCR (qPCR) used for the quantification of DNA from viable cells. The staining with 4,6-Diamidine-2-phenylindole dihydrochloride (DAPI) and the use of differential interference contrast microscopy were suitable for the determination of resting spore germination rates. The latter method also allowed recording individual germination states of spores. Alternatively, dual staining with CFW-Nile red was successfully used to assess the germination rate of resting spores with a lethal pre-treatment. This study evaluates and confirms the suitability of various microscopic and molecular genetic methods for the determination of viability and germination of P. brassicae resting spores. Such methods are required to study factors in the soil regulating survival, dormancy and germination of P. brassicae resting spores causing clubroot disease in Brassicaceae hosts and therefore are fundamental to develop novel strategies of control.

Genome-wide analyses of late pollen-preferred genes conserved in various rice cultivars and functional identification of a gene involved in the key processes of late pollen development.

BACKGROUND: Understanding late pollen development, including the maturation and pollination process, is a key component in maintaining crop yields. Transcriptome data obtained through microarray or RNA-seq technologies can provide useful insight into those developmental processes. Six series of microarray data from a public transcriptome database, the Gene Expression Omnibus of the National Center for Biotechnology Information, are related to anther and pollen development. RESULTS: We performed a systematic and functional study across the rice genome of genes that are preferentially expressed in the late stages of pollen development, including maturation and germination. By comparing the transcriptomes of sporophytes and male gametes over time, we identified 627 late pollen-preferred genes that are conserved among japonica and indica rice cultivars. Functional classification analysis with a MapMan tool kit revealed a significant association between cell wall organization/metabolism and mature pollen grains. Comparative analysis of rice and Arabidopsis demonstrated that genes involved in cell wall modifications and the metabolism of major carbohydrates are unique to rice. We used the GUS reporter system to monitor the expression of eight of those genes. In addition, we evaluated the significance of our candidate genes, using T-DNA insertional mutant population and the CRISPR/Cas9 system. Mutants from T-DNA insertion and CRISPR/Cas9 systems of a rice gene encoding glycerophosphoryl diester phosphodiesterase are defective in their male gamete transfer. CONCLUSION: Through the global analyses of the late pollen-preferred genes from rice, we found several biological features of these genes. First, biological process related to cell wall organization and modification is over-represented in these genes to support rapid tube growth. Second, comparative analysis of late pollen preferred genes between rice and Arabidopsis provide a significant insight on the evolutional disparateness in cell wall biogenesis and storage reserves of pollen. In addition, these candidates might be useful targets for future examinations of late pollen development, and will be a valuable resource for accelerating the understanding of molecular mechanisms for pollen maturation and germination processes in rice.

Malting process optimization for protein digestibility enhancement in finger millet grain.

Finger millet (Eleusine coracana) is a nutritious, gluten-free, and drought resistant cereal containing high amounts of protein, carbohydrate, and minerals. However, bio-availability of these nutrients is restricted due to the presence of an excessive level of anti-nutrient components, mainly phytic acid, tannin, and oxalate. It has been shown that a well-designed malting/germination process can significantly reduce these anti-nutrients and consequently enhance the nutrient availability. In the present study, the effects of two important germination factors, duration and temperature, on the enhancement of in-vitro protein digestibility of finger millet were thoroughly investigated and optimized. Based on a central composite design, the grains were germinated for 24, 36, and 48 h at 22, 26, and 30 °C. For all factor combinations, protein, peptide, phytic acid, tannin, and oxalate contents were evaluated and digestibility was assessed. It was shown that during the malting/germinating process, both temperature and duration factors significantly influenced the investigated quantities. Germination of finger millet for 48 h at 30 °C increased protein digestibility from 74 % (for native grain) up to 91 %. Besides, it notably decreased phytic acid, tannin, and oxalate contents by 45 %, 46 %, and 29 %, respectively. Linear correlations between protein digestibility and these anti-nutrients were observed.