Optimizing techno-functionality of germinated whole wheat flour steamed bread via glucose oxidase(Gox)and pentosanase(Pn)enzyme innovation

Germination,a powerful biofortification technique,holds immense potential in bolstering the micronutrient profile of essential staple grains,thereby paving the way for optimal nutritional enhancement.The primary goal of this study was to improve the technological functionality of germinated wheat flour by incorporating pentosanase(Pn)and glucose oxidase(Gox)enzymes,with particular emphasis on the evolutionary changes in its components.The inclusion of Gox did not produce any substantial impact on the volumetric characteristics of the steamed bread.The incorporation of Pn and Gox has been seen to enhance the overall excellence of steamed bread by optimizing loaf volume and textural characteristics while also improving the thermal stability of the dough.The existence of two endothermic peaks could be attributed to bound water or alterations in the granules within the starch crystallization region.Adding Pn and Gox reduced and increased the formation and stability time of the dough,respectively.A certain ratio was employed to assess alternations in the crystallinity of starch granules over a limited range.After steaming,a significant decrease in IR1047/1022 was observed,indicating that the elevated temperature partially disrupted the internal starch crystal structure,leading to a gelatinization reaction with water.The ratio of tensile resistance(R)and elongation(E)of dough increased significantly compared to the control.The results obtained from this study indicate that the simultaneous inclusion of enzymes(Pn+Gox)holds significant promise for expanding the technological functionality of germinated wheat flour dough and improving the quality attributes of steamed bread.

Germinated brown rice relieves hyperlipidemia by alleviating gut microbiota dysbiosis

Hyperlipidemia is a frequent metabolic disorder that is closely associated with diet. It is believed that brown rice, containing the outer bran layer and germ, is beneficial for the remission of hyperlipidemia. This study established a rat model of hyperlipidemia by feeding a high-fat diet. The hypolipidemic potential of germinated brown rice(Gbrown) and germinated black rice(a germinated black-pigmented brown rice, Gblack) were explored in the model rats, mainly in the aspects of blood lipids, lipases, apolipoproteins, and inflammation. The gut microbiota in hyperlipidemic rats receiving diverse dietary interventions was determined by 16S rDNA sequencing. The results showed that the intervention of Gbrown/Gblack alleviated the hyperlipidemia in rats, evidenced by decreased TC, TG, LDL-C, and apolipoprotein B, and increased HDL-C, HL, LPL, LCAT, and apolipoprotein A1. Gbrown/Gblack also weakened the inflammation in hyperlipidemia rats, evidenced by decreased TNF-α, IL-6, and ET-1. In addition, 16S rDNA sequencing revealed that the diet of Gbrown/Gblack elevated the abundance and diversity of gut microbiota in hyperlipidemia rats. At the phylum level, Gbrown/Gblack decreased Firmicutes, increased Bacteroidetes, and decreased the F/B ratio in hyperlipidemia rats. At the genus level, Gbrown/Gblack decreased Streptococcus and increased Ruminococcus and Allobaculum in hyperlipidemia rats. Some differential microbial genera relating to lipid metabolism were also determined, such as the Lachnospira and Ruminococcus in the Gblack group, and the Phascolarctobacterium, Dorea, Turicibacter, and Escherichia-Shigella in the Gbrown group. Notably, the beneficial effect of Gblack was stronger than Gbrown. To sum up, the dietary interventions of Gbrown/Gblack contributed to the remission of hyperlipidemia by alleviating the dysbiosis of gut microbiota.

Identification of an anticancer compound against HT-29 cells from Phellinus linteus grown on germinated brown rice

Objective:To isolate and identify the anticancer compound against proliferation of human colon cancer cells from ethyl acetate(EtOAc)extract ol Phellinus linteus grown on germinated brown rice(PB).Methods:EtOAc extract of PB was partitioned with n-hexane,EtOAc,and water-saturated n-butanol.Anticancer compound of n-hexane layer was isolated and identified by HPLC and NMR,respectively.Cytotoxicity against HT-29 cells was tested by SRB assay.Results:The n-hexane layer obtained after solvent fractionation of PB EtOAc extracts showed a potent anticancer activity against the HT-29 cell line.Atractylenolide I,a eudesmane-type sesquiterpene lactone,a major anticancer substance of PB,was isolated from the n-hexane layer by silica gel column chromatography and preparative-HPLC.This structure was elucidated by one-and two-dimensional NMR spectroscopic data.Atractylenolide I has not been reported in mushrooms or rice as of yet.The isolated compound dose-dependently inhibited the growth of HT-29 human colon cancer cells.Conclusions:Atractylenolide I might contribute to the anticancer effect of PB.

Study on evaluation of gamma oryzanol of germinated brown rice by near infrared spectroscopy

Germinated brown rice(GBR)is rich in gamma oryzanol which increase its consumption popularity,particularly in the health food market.The objective of this research was to apply the near infraredspectroscopy(NIRS)for evaluation of gamma oryzanol of the germinated brown rice.The germinated brown rice samples were prepared from germinated rough rice(soaked for 24 and 48 h,incubated for 0,6,12,18,24,30 and 36 h)and purchased from local supermar kets.The germinated brown rice sampleswere subjected to NIR scanning before the evaluation of gamma oryzanol by using partial extractionmet hodology.The prediction model was established by partial least square regression(PLSR)andvalidated by full cross validation method.The NIRS model established from various varieties of germinated brown rice bought from diferent markets by first derivatives+vector normalizationpretreated spectra showed the optimal prediction with the correlation of determination(R?),root mean squared error of cross validation(RMSECV),and bias of 0.934,8.84×10^(-5) mg/100 g drymatter and 1.06×10^(-5) mg/100 g dry matter,respectively.This is the first report on the application of NIRS in the evaluation of gamma oryzanol of the germinated brown rice.This information is veryuseful to the germinated brown rice production factory and consumers.

Variation in Antioxidants, Bioactive Compounds and Antioxidant Capacity in Germinated and Ungerminated Grains of Ten Rice Cultivars

Present study was conducted to characterise ten rice(Oryza sativa L.) cultivars viz. IET-23466, Dhan-201, IET-23448, MAS-946, IET-23445, IET-23463, IET-23455, PR-123, PR-115 and IET-23449 based on antioxidants(total phenolics, flavonols and tannins), bioactive compounds(phytic acid, gamma amino butyric acid, tocopherol and reduced ascorbate) and antioxidant activity(1,1-diphenyl-2-picrylhydrazyl radical scavenging activity, hydroxyl radical scavenging activity, free radical antioxidant power and total reducing power) with an aim to identify cultivars containing higher health promoting components after germination. Three cultivars IET-23466, IET-23463 and PR-123 performed better as revealed by higher level of antioxidants, bioactive compounds and antioxidant activity before and after germination. Three cultivars MAS-946, IET-23445 and IET-23449 had moderate level of antioxidants, bioactive compounds and antioxidant activity. Four cultivars Dhan-201, IET-23448, IET-23455 and PR-115 performed intermediately. Strong positive correlation was observed among total phenolics and the antioxidant activity. Phytic acid was found to be negatively correlated to the antioxidant activity. Our results highlighted that cultivars IET-23466, IET-23463 and PR-123 hold great potential after germination and would open up a useful opportunity for the functional food industry, and consumption of these cultivars after germination would afford health benefits to consumers since they contain higher level of antioxidants.

Rheological, Organoleptical and Quality Characteristics of Gluten-Free Rice Cakes Formulated with Sorghum and Germinated Chickpea Flours

Gluten-free bakery products are produced for people suffering from celiac disease due to sensitivity to gluten. So, the aim of this work was to study the effect of partial substitution of rice flour at 20%, 30% and 40% with sorghum and germinated chickpea flours on rheological, physical, sensory properties and staling rate of prepared gluten-free cake. Obtained results showed that germinated chickpea flour had the highest contents of crude protein (23.62%), lipids (4.89%) and crude fibers (5.76%) as compared to other grain flours. Gradual increase in gelatinization temperature and decrease in maximum viscosity and setback were fond of cake batters with increasing the substituting levels of rice flour from 20% to 40% with sorghum or germinated chickpea flours in comparison to rice flour batter. No significant difference was observed in specific volume between rice cake and cake containing 20% of germinated chickpea and sorghum flours which given 2.71, 2.62 and 2.56 cm3/g, respectively. Data also revealed that gluten-free cake lightness (L*) and total intensity significantly increased with 40% of sorghum flour followed by 30 and 20% of sorghum flour. Substitution of 40% rice flour with germinated chickpea flour resulted in significant high in redness (a*), yellowness (b*) and chroma of gluten-free cake being 11.95, 33.72 and 35.77, respectively followed by substituted levels 30% and 20% which attributed to their natural yellow pigments compared with other investigated samples. Overall acceptability of gluten-free cakes was improved after adding 20, 30% of sorghum flour and 20% of germinated chickpea flour which being 90.7, 88.6 and 87.5, respectively compared to rice flour cake (88.4). Rice flour cake was more freshness than wheat flour cake and other treatments during storage duration. After 6 days, rice cake had significant high freshness (306.2%) followed by gluten-free cake samples with 20% of sorghum or germinated chickpea flours which given 294.5% and 289.4%, respectively. Generally, it could be concluded that the quality and nutritional properties of gluten-free rice cakes could be improved when using 20%, 30% of sorghum flour and 20% of germinated chickpea flour as substitution levels of rice flour.

The metabolomics variations among rice, brown rice, wet germinated brown rice, and processed wet germinated brown rice

Germination and processing are always accompanied by significant changes in the metabolic compositions of rice. In this study, polished rice(rice), brown rice, wet germinated brown rice(WGBR), high temperature and pressure-treated WGBR(WGBR-HTP), and low temperature-treated WGBR(WGBR-T18) were enrolled. An untargeted metabolomics assay isolated 6 122 positive ions and 4 224 negative ions(multiple difference ≥1.2 or ≤0.8333, P<0.05, and VIP≥1) by liquid chromatography-mass spectrum. These identified ions were mainly classified into three categories, including the compounds with biological roles, lipids, and phytochemical compounds. In addition to WGBR-T18 vs. WGBR, massive differential positive and negative ions were revealed between rice of different forms. Flavonoids, fatty acids, carboxylic acids, and organoxygen compounds were the dominant differential metabolites. Based on the Kyoto Encyclopedia of Genes and Genomes(KEGG) database, there 7 metabolic pathways(phenylalanine/tyrosine/tryptophan biosynthesis, histidine metabolism, betalain biosynthesis, C5-branched dibasic acid metabolism, purine metabolism, zeatin biosynthesis, and carbon metabolism) were determined between brown rice and rice. Germination changed the metabolic pathways of porphyrin and chlorophyll, pyrimidine, and purine metabolisms in brown rice. In addition, phosphonate and phosphinate metabolism, and arachidonic acid metabolism were differential metabolic pathways between WGBR-HTP and WGBR-T18. To sum up, there were obvious variations in metabolic compositions of rice, brown rice, WGBR, and WGBR-HTP. The changes of specific metabolites, such as flavonoids contributed to the antioxidant, anti-inflammatory, anti-cancer, and immunomodulatory effects of GBR. HTP may further improve the nutrition and storage of GBR through influencing specific metabolites, such as flavonoids and fatty acids.

Optimization of Germinated Conditions for Somatic Embryos in Liriodendron Hybrids

The development of somatic embryos is a key technology in plant breeding. To gain the best cultural conditions,the factors influencing on germination and transplanting survival rate of the somatic embryos in Liriodendron hybrids were optimized,including the development period of somatic embryos,macro-element,Vitamin C,genotypes,plant hormones and transplant medium in this study. The results showed that the optimal development phase of germination was torpedo-shaped embryo,and the base cultural medium of 3/4 MS with vitamin C can keep normal form from somatic embryos. The germination rate of somatic embryo had obvious genotype difference,among which the hybrid 1 × 5002 was the most sensitive( 82. 46%),and the hybrid 1 × 4088 was the most insensitive( 13. 24%). The medium with 0. 1 mg/L of KT + 0. 1 mg/L of IBA can help promote the germination of somatic embryogenesis. In addition,the yellow soil was more suitable than mixed peat soil for transplanting Liriodendron hybrids seedlings. Therefore,the obtainment of the optimal conditions has important guidance for production practices of Liriodendron hybrids.

Dietary Fibre and Fatty Acid Evaluation of Boiled, Roasted, Germinated and Fermented Breadnut (Ukwa bekee) Seed Flours

Evaluation of dietary fiber and fatty acid composition of boiled, roasted, fermented and germinated breadnut seed flour was investigated. The seeds were dehulled and washed. It was divided into four equal parts for different processing techniques: boiled, roasted, germinated and fermented. They were analyzed for dietary fibre and fatty acid composition using standard methods. Data were subjected to statistical analysis using Statistical Product for Service Solution (SPSS) version 23.0. Values were expressed as means and standard deviation, Duncan multiple range test was used in separating the means at 95% confidence interval. Dietary fibre composition showed that breadnut seed flours range from 4.94%(fermented breadnut seed flour) to 5.42%(roasted breadnut seed flour). Fatty acid profile showed that breadnut seed flours contain a greater amount of non-essential fatty acids which were highly significant (p < 0.05) in oleic acid (57.93%) for roasted breadnut seed flour followed by linoleic acid (25.76%) in fermented breadnut flour. Stearic acid was the only prominent non-essential fatty acid in the breadnut seed flour and ranged from 4.40%(fermented breadnut seed flour) to 6.27%(germinated breadnut seed flour). The study revealed that all the processed breadnut seed flours have appreciable quantities of dietary fibre and essential fatty acids than non-essential fatty acids. Breadnut seed should be recommended in wheat substitution in bakery industry and culinary uses. It is also recommended for weight loss program.

Effects of germination and aeration treatment following segmented moisture conditioning on theγ-aminobutyric acid accumulation in germinated brown rice

Brown rice was treated by segmented moisture conditioning to reach the suitable water content and aerated with air for germination.The effects of germination and aeration treatment on theγ-aminobutyric acid(GABA)content in germinated brown rice were studied.The results showed that the germination rate,glutamic acid,glutamate decarboxylase activity and GABA content in germinated brown rice increased significantly and then decreased with the increase of germination and aeration treatment parameters.Correlation analysis also revealed that there was a significant positive correlation between GABA accumulation and glutamic acid content,glutamate decarboxylase activity.These results suggested that the aeration treatment during germination following segmented moisture conditioning could contribute to a high GABA content of germinated brown rice.

OsbZIP01 Affects Plant Growth and Development by Regulating OsSD1 in Rice

As the‘Green Revolution’gene,SD1(encoding GA20ox2),has been widely applied to improve yield in rice breeding.However,research on its transcriptional regulation is limited.Here,we identified a transcription factor OsbZIP01,which can suppress the expression of SD1 and regulate gibberellin(GA)biosynthesis in rice.Knockout mutants of OsbZIP01 exhibited increased plant height,while the overexpression lines showed a semi-dwarf phenotype and diminished germination rate.Furthermore,the semi-dwarf phenotype of OE-bZIP01,was caused by the reduced internode length,which was accompanied by a thin stem width.The predominant expression of OsbZIP01 was observed in leaves and sheaths.OsbZIP01 protein was localized in the nucleus and showed transcriptional repression activity.In addition,OsbZIP01 could directly bind to the promoter of the OsSD1 gene,and inhibit its transcription.The semi-dwarf phenotype of OE-bZIP01 could be rescued by exogenous GA_(3).Meanwhile,the bzip01 sd1 double mutant showed a shorter shoot length compared with the wild type,indicating that OsbZIP01 regulated plant growth mainly through the GA biosynthesis pathway.Collectively,OsbZIP01 negatively regulates GA biosynthesis by restraining SD1 transcription,thereby affecting plant growth and development.

Relationship between Seed Priming Mediated Seedling Vigor and Yield Performance of Spring Wheat

Seed priming is a pre-germinated technique that can enhance seed germination percentage,faster and synchro-nized germination,better seedling growth,and yield under stress conditions.To ascertain the most effective seed priming method that would ensure the potential yield of wheat in Bangladesh,two experiments were carried out from December 2021 to March 2022 at the Department of Agronomy,Bangladesh Agricultural University.Two wheat varieties namely BARI Gom-28 and BWMRI Gom-1 were subjected to a range of priming chemicals in both lab and pot tests.These compounds included the following:control(no priming),hydropriming(distilled water),10000 ppm KNO_(3),15000 ppm KNO_(3),40000 ppm Mannitol,60000 ppm Mannitol,10000 ppm NaCl,20000 ppm NaCl,100 ppm PEG,150 ppm PEG,500 ppm NaOCl,1000 ppm NaOCl,10000 ppm CaCl_(2),20000 ppm CaCl_(2),10000 ppm KCl and 20000 ppm KCl.A complete randomized design(CRD)with three repli-cations was used to set up the experiments.The results showed that BARI Gom-28 and BWMRI Gom-1 responded best to KCl priming in terms of rapid seed germination and strong seedling development.On the other hand,the best priming agents for plant growth and productivity turned out to be CaCl_(2) and KCL.The results of this study support the possibility of using seed priming as a technique to improve wheat plant development and output by raising seed emergence and survival rates.

Effect of biochar on the metabolome of soybean seedlings

The use of biochar can have several effects on plant germination,depending on raw material,preparation method and application dose.However,the molecular mechanisms that lead to those results have yet to be elucidated.The aim of this research was to improve the understanding of these mechanisms by characterizing the metabolic effects of sugarcane bagasse biochar on soybean germination.Three types of biochars were prepared by pyrolysis at 300℃(SCB300),400℃(SCB400)and 600℃(SCB600).Then,each one was mixed into sand at 1%,3%,5%(w/w)dose,respectively.The experiment was performed in 8 days of incubation,when the number of germinated seeds and the average radicle length were determined.To evaluate the metabolome,the dry biomass(DB)was subjected to extraction with a mixture of methanol-d4 and D2O(1:1 v/v).The extracts were submitted to metabolomics analysis by Proton Nuclear Magnetic Resonance.The Relative Germination,Relative Average Radicle Growth and Germination Index increased in all treatments compared to control.On the other hand,the DB increased in all treatments,except for SCB300,at doses of 1%and 3%w/w.Seven metabolites(alanine,asparagine,acetic acid,citric acid,glycerol,fatty acids and sucrose)were identified and quantified in DB extracts as the most influential finding for the separation of treatments.Taken together,these results strongly suggested that biochars accelerated the catabolism of triacylglycerols to sucrose and induced a slight osmotic stress.

Preparation of Tartary Buckwheat Seed Coating Agent and Its Effect on Germination

To mitigate the wastage of seed resources and reduce the usage of pesticides and fertilizers, seed coating agentshave gained popularity. This study employs single-factor and multi-index orthogonal experimental design methodsto investigate the seed coating formula and physical properties of Tartary buckwheat. The specific effects ofeach component on Tartary buckwheat seed germination are analyzed. The findings reveal that the seed coatingagent formulated with 1.5% polyvinyl alcohol, 0.15% sodium alginate, 0.2% op-10, 0.1% polyacrylamide, 8% colorant,3% ammonium sulfate, 1% potassium dihydrogen phosphate, and 0.15% carbendazim exhibits the mosteffective coating. It demonstrates optimal physical properties and promotes seed germination efficiently. The suspensionrate of this seed coating agent reaches 91.12%, with a mere 2.13% coating shedding rate and 2.5% coatingseed rot rate. Furthermore, it achieves a germination percentage of 99.17%, which is 20.84% higher than the lowestgroup. The germination potential and index are also significantly higher than the lowest group, with anincrease of 20.84% and 26.56%, respectively. Additionally, the vitality index is 553.08, a 15.75% increase comparedto the lowest group. The application of seed coating agents helps reduce seed resource loss, increase plant numbers,and ultimately enhance agricultural yields. This finding holds practical significance in agriculturalproduction.

Effects of Low Temperature Stress on Germination and Physiological Characteristics of Different Sweet Maize Varieties

[Objectives]The paper was to explore the effects of low temperature stress on germination and physiological characteristics of different sweet maize varieties.[Methods]Taking Taitian 264,Zhexuetian 1 and Chaotian 4 as the research objects,the changes in germination potential,germination index,plant height,biomass,and antioxidant enzyme activity of maize seeds were studied under optimal temperature conditions(25℃)and low temperature stress conditions(10℃).[Results]Under 10℃stress,the germination rate and germination index of Taitian 264 were higher than that of Zhexuetian 1 and Chaotian 4.Under low temperature stress,Taitian 264 exhibited the least reduction in height and biomass,while Zhexuetian 1 had the most reduction.Additionally,the SOD and POD activities of Taitian 264 were higher than that of Zhexuetian 1 and Chaotian 4 under both temperature conditions,while the MDA content of Taitian 264 was lower.Taitian 264 showed strong germination ability against low temperature stress.[Conclusions]This study provides a basis for timely sowing practices of sweet maize in agricultural production.

The Toxic Effects of Strong Chlorin Disinfectant on Mangroves and Emission Thresholds

In this study, based on the simulated discharge results of chemical disinfectants, hypocotyl germination concentration gradient pre-test and concentration gradient determination experiment were set up respectively. Laboratory cultivation was conducted to compare and analyze the root germination and germination indexes, three mangrove hypocotyls of Kandelia candel (Linn.) Druce, Ceriopstagal C.B. Rob. and Bruguiera sexangula var. Rhynchopetalas’ efficiency of cumulative root germination, cumulative germination and the cumulative expansion of the second pair of leaves, one-way analysis of variance was used to obtain the tolerance threshold of three mangrove hypocotyls to strong chlorin disinfectant. The study determined that the by-products of strong chlorin disinfectant, the toxic threshold concentrations of Kandelia candel (Linn.) Druce, Ceriopstagal C.B. Rob. and Bruguiera sexangula var. rhynchopetala are close to 0.55 mg/L, 0.55 mg/L and 0.25 mg/L, respectively. This concentration range is lower than the average concentration of 1.183 mg/L of active chlorine emitted from strong chlorine concentrate during pond clearing in high-level shrimp ponds, indicating that transient emissions of strong chlorine concentrate during pond clearing can have a toxic effect on mangrove plants. The strength of tolerance of the embryonic axes of the three mangrove species to effective chlorine contamination was, Ceriopstagal C.B. Rob. stronger than Bruguiera sexangula var. rhynchopetala, and Kandelia candel (Linn.) Druce is the weakest.

Nondestructive determination of GABA in germinated brown rice with near infrared spectroscopy based on wavelet transform denoising

The objective of this study was to analyze the content ofγ-aminobutyric acid(GABA)in germinated brown rice(GBR)by using near-infrared spectroscopy(NIRS)and the pretreatment method of wavelet de-noising(WD).The prediction accuracy of the NIRS model established by the Daubechies5 wavelet basis function at 3 level denoising treatment is the highest,the correlation coefficient for calibration(rc)is 0.931,the root mean square error of calibration(RMSEC)is 0.4038 mg/100 g,the Bias of calibration is 0.006,the correlation coefficient for prediction(rp)is 0.916,the root mean square error of prediction(RMSEP)is 0.4329 mg/100 g,the Bias of prediction is 0.010,and the ratio of performance to deviation(RPD)is 4.911.Results showed that the predicted and actual values had high correlation.Therefore,these results indicate that the NIRS model treated by WD is feasible to detect GABA content in GBR rapidly and nondestructively.

Effect of Forest Litter on the Regeneration of Larix sibirica: Insight from Aqueous Extract and Litter Coverage

The effect of litter on forest regeneration depends on the characteristics of regional climate and also shows community specificity. The influences of plant litter on seed germination and seedling growth of Larch Siberian forest in the Altai Mountains were investigated through two simulated experiments including litter coverage and litter aqueous extracts. In the litter coverage experiment, three litter coverage methods including above (D), below (S) and in the middle (Z) of litter were set with the litter coverage thickness of 0, 1, 2, and 4 cm, while two aqueous extract obtained methods using the air-dried litter and litter ash after fir were used with the concentration of 10%, 40%, 80% and 100% in the present study. Results showed that: the aqueous extracts obtained using the air-dried litter restrained the seed germination, while the aqueous extracts obtained using litter ash improved the seed germination. Compared with other litter concentration, the influences of 100% concentration reach highest. The seed germination rate, seed germination potential and vital index under the treatment of seeds above the litter coverage were highest, which were significantly higher than other treatments. The above-ground biomass was significantly higher and the inhibition index of below-ground bio-mass was significantly lower under the treatment of seed above the litter with thin litter cover-age (S1) compared to other litter coverage treatments. These results indicated that the litter aqueous extract and the litter coverage had a combined effect on the seed germination and seedling growth of Siberian larch forest. Fire disturbance could promote seed germination by modifying the adverse effects of litter aqueous extracts and litter coverage, and thus plays an important role in the regeneration of Siberian larch in the Altai Mountains.

Whole lifecycle observation of single‐spore germinated Streptomyces using a nanogap‐stabilized microfluidic chip

Streptomyces is a model bacterium to study multicellular differentiation and the major reservoir for antibiotics discovery.However,the cellular‐level lifecycle of Streptomyces has not been well studied due to its complexity and lack of research tools that can mimic their natural conditions.In this study,we developed a simple microfluidic chip for the cultivation and observation of the entire lifecycle of Streptomyces development from the single‐cell perspective.The chip consists of channels for loading samples and supplying nutrients,microwell arrays for the seeding and growth of single spores,and air chambers beside the microwells that facilitate the development of aerial hyphae and spores.A unique feature of this chip is that each microwell is surrounded by a 1.5µm nanogap connected to an air chamber,which provides a stabilized water–air interface.We used this chip to observe the lifecycle development of Streptomyces coelicolor and Streptomyces griseus germinated from single spores,which revealed differentiation of aerial hyphae with progeny spores at micron‐scale water–air interfaces and air chambers.Finally,we demonstrated the applicability of this chip in phenotypic assays by showing that the microbial hormone A‐Factor is involved in the regulatory pathways of aerial hyphae and spore formation.The microfluidic chip could become a robust tool for studying multicellular differentiation,single‐spore heterogeneity,and secondary metabolism of single‐spore germinated Streptomyces.

Observation of Flowering Process of Grape(Vitis vinifera L.)—Insight into the Starting of Pollination of Flower Hiding in Calyptras(Cap)

It is generally agreed that many Vitis vinifera L.cultivars are self-fertile,where self-pollination often occurs before capfall in a process called cleistogamy.Therefore,it is difficult to identify the right time to remove stamens before self-pollination during the cross-breeding of grape.For this paper,we observed the process of grape flowering and measured the pollen viability and stigma receptivity of grape flowers of‘Shine Muscat’in order to identify the starting time of self-pollination before capfall and to provide useful information for improving the efficiency of cross-breeding.The results demonstrate that the anther is not cracked during the visible clusters and separated clusters stages.Meanwhile,in the separated floral buds,flowering begins,and full bloom stages,the pollen viability is 60.7%,73.2%and 80.3%,respectively;however,at the berry set stage,pollen viability drops to zero.The top of the mature stigma is composed of a layer of nearly cylindrical papillary cells,and the stigma receptivity for pollen changes with the development of flowers:in particular,no reaction was observed in the visible clusters stage;weak positive reaction at the separated clusters stage;strong positive reaction at the separated floral buds,flowering begins,and full bloom stages;and no reaction at the berry set stage.In the separated floral buds stage,pollen tubes were seen germinating in the style.In the flowering begins stage,more pollen tubes were observed at the entry of the ovary.During the full bloom stage,most pollen tubes elongated into the ovary base and some entered the pearl hole.At the berry set stage,newborn endosperm nucleus could be seen in the ovule.From the above,we can conclude that the initiation time of closed fertilization for‘Shine Muscat’grape can be judged as the separated floral buds stage,and it is best to discard the stamen before the separated floral buds stage when conducting cross-breeding.