Effects of tandem fermentation of edible mushroom and L. plantarum on sensory, polysaccharide, vitamin C, and γ-aminobutyric acid of Rosa roxburghii Tratt and coix seed beverage.

A new Rosa roxburghii Tratt (RRT) and coix seed (CS) beverage rich in multi-active ingredients was developed. Edible mushrooms and L. plantarum were selected for fermentation in stages. Some physicochemical properties, γ-aminobutyric acid (GABA), polysaccharides and sensory were studied during the T. versicolor and L. plantarum fermentation. T. versicolor increased the free amino acid through enzymatic protein digestion in the early growth stage and used these amino acids to synthesize its bacteriophage protein. T. versicolor and L. plantarum increased the polysaccharide and GABA of the fermentation broth. Vitamin C was retained as much as possible, with a slight loss occurring mainly in the aerobic fermentation stage of T. versicolor. Its less loss in exchange was for a higher value of T. versicolor polysaccharide, protein enhancement, and bitterness reduction. This study provides a reference for the deep processing of Guizhou's unique agricultural products and edible mushroom fermented beverage.

Correlation between Inflorescence Architecture and Floral Asymmetry-Evidence from Aberrant Flowers in Canna L. (Cannaceae).

Floral symmetry studies often focus on the development of monosymmetric and polysymmetric flowers, whereas asymmetric flowers and their position and function within the inflorescence structure are largely neglected. Cannaceae is one of the few families that possesses truly asymmetric flowers, serving as a model to study the characters and mechanisms involved in the development of floral asymmetry and its context within the developing and mature inflorescence. In this study, inflorescence structure and floral morphology of normal asymmetric flowers and 16 aberrant flower collections from Canna indica L. and C. glauca L. were photographed, analyzed, and compared with attention to stamen petaloidy, floral symmetry, and inflorescence branching patterns anterior and posterior to the aberrant flower. In comparison with normal flowers, the aberrant flowers are arranged into abnormal partial florescences, and vary in floral symmetry, orientation, and degree of androecial petaloidy. The appendage of the fertile stamen is universally located distal from the higher order bract, indicating an underlying influence of inflorescence architecture. A synthetic model is proposed to explain the relationship between floral symmetry and inflorescence structure. Data from the observation of aberrant phenotypes strongly support the hypothesis that irregular petaloidy of the stamens is correlated with an asymmetric morphogenetic field within the inflorescence that contributes to the overall floral asymmetry in Canna flowers.

Effect of Adding Bifidobacterium animalis BZ25 on the Flavor, Functional Components and Biogenic Amines of Natto by Bacillus subtilis GUTU09.

Natto is a high-value fermented soybean produced by B. subtilis. However, B. subtilis produces a pungent amine odor. This study compared the volatile organic compounds (VOCs), free amino acids (FAAs) and biogenic amines (BAs), nattokinase (NK) of natto made by two-strain fermentation with Bifidobacterium animalis BZ25 and Bacillus subtilis GUTU09 (NMBB) and that of natto made by single-strain fermentation with Bacillus subtilis GUTU09 (NMB). Compared with NMB, volatile amine substances disappeared, ketones and aldehydes of NMBB were reduced, and alcohols increased. Besides that, the taste activity value of other bitter amino acids was lowered, and BA content was decreased from 255.88 mg/kg to 238.35 mg/kg but increased NK activity from 143.89 FU/g to 151.05 FU/g. Correlation analysis showed that the addition of BZ25 reduced the correlation between GUTU09 and BAs from 0.878 to 0.808, and pH was changed from a positive correlation to a negative one. All these results showed that the quality of natto was improved by two-strain co-fermentation, which laid a foundation for its potential industrial application.

Effect of Fermentation Parameters on Natto and Its Thrombolytic Property.

Natto is a popular food because it contains a variety of active compounds, including nattokinase. Previously, we discovered that fermenting natto with the combination of Bacillus subtilis GUTU09 and Bifidobacterium animalis subsp. lactis BZ25 resulted in a dramatically better sensory and functional quality of natto. The current study further explored the effects of different fermentation parameters on the quality of natto fermented with Bacillus subtilis GUTU09 and Bifidobacterium BZ25, using Plackett-Burman design and response surface methodology. Fermentation temperature, time, and inoculation amount significantly affected the sensory and functional qualities of natto fermented with mixed bacteria. The optimal conditions were obtained as follows: soybean 50 g/250 mL, triangle container, 1% sucrose, Bacillus subtilis GUTU09 to Bifidobacterium BZ25 ratio of 1:1, inoculation 7%, fermentation temperature 35.5 °C, and fermentation time 24 h. Under these conditions, nattokinase activity, free amino nitrogen content, and sensory score were increased compared to those before optimization. They were 144.83 ± 2.66 FU/g, 7.02 ± 0.69 mg/Kg and 82.43 ± 5.40, respectively. The plate thrombolytic area and nattokinase activity both increased significantly as fermentation time was increased, indicating that the natto exhibited strong thrombolytic action. Hence, mixed-bacteria fermentation improves the taste, flavor, nattokinase activity, and thrombolysis of natto. This research set the groundwork for the ultimate manufacturing of natto with high nattokinase activity and free amino nitrogen content, as well as good sensory and thrombolytic properties.

Why pendulum symmetry is absent from the cymose partial inflorescences of Cannaceae? Insights into the essential characteristic of cincinni.

In a typical cincinnus, the neighboring two flowers are generally enantiomorphic, which leads to the pendulum symmetry of the entire cyme. While in a two-flowered Cannaceae cincinnus, the flowers develop the same chirality. In this study, we observed several abnormal cincinni of Canna indica that extended longer than their normal form, which presented a second enantiomorphic flower, thus reflecting a typical pendulum symmetry. The chirality change of the second flower was strongly associated with the position of the lateral cincinnus meristem, which determines the angle size of the cincinnus zigzag shift and may serve as a key factor controlling the formation of pendulum symmetry. We propose that alternating floral chirality and the concomitant pendulum symmetry are the essential characteristics of a typical cincinnus. Accordingly, Canna flowers with the same chirality are arranged in modified cincinni.

Isolation of Bacillus spp. with High Fibrinolytic Activity and Performance Evaluation in Fermented Douchi.

ABSTRACT: Fibrinolytic enzymes are effective and highly safe in treating cardiovascular and cerebrovascular diseases. Therefore, screening fibrinolytic enzyme-producing microbial strains with excellent fermentation performance is of great value to industrial applications. The fibrin plate method was used in screening strains with high yields of fibrinolytic enzymes from different fermented food products, and the screened strains were preliminarily identified using molecular biology. Then, the strains were used for solid-state fermentation of soybeans. Moreover, the fermentation product douchi was subjected to fibrinolytic activity measurement, sensory evaluation, and biogenic amine content determination. The fermentation performance of each strain was comprehensively evaluated through principal component analysis. Finally, the target strain was identified based on strain morphology, physiological and biochemical characteristics, 16S rDNA sequence, and phylogenetic analysis results. A total of 15 Bacillus species with high fibrinolysin activity were selected. Their fibrinolytic enzyme-producing activity levels were higher than 5,500 IU/g. Through molecular biology analysis, we found 4 strains of Bacillus subtilis, 10 strains of Bacillus amyloliquefaciens, and 1 strain of Bacillus velezensis. The principal component analysis results showed that SN-14 had the best fermentation performance and reduced the accumulation of histamine and total amine, the fibrinolytic activity of fermented douchi reached 5,920.5 ± 107.7 IU/g, and the sensory score was 4.6 ± 0.3 (out of 5 points). Finally, the combined results of physiological and biochemical analyses showed SN-14 was Bacillus velezensis. The high-yield fibrinolytic and excellent fermentation performance strain Bacillus velezensis SN-14 has potential industrial application.

Expression and Function Studies of CYC/TB1-Like Genes in the Asymmetric Flower Canna (Cannaceae, Zingiberales).

The asymmetric flower, lacking any plane of symmetry, is rare among angiosperms. Canna indica L. has conspicuously asymmetric flowers resulting from the presence of a half-fertile stamen, while the other androecial members develop as petaloid staminodes or abort early during development. The molecular basis of the asymmetric distribution of fertility and petaloidy in the androecial whorls remains unknown. Ontogenetic studies have shown that Canna flowers are borne on monochasial (cincinnus) partial florescences within a racemose inflorescence, with floral asymmetry likely corresponding to the inflorescence architecture. Given the hypothesized role of CYC/TB1 genes in establishing floral symmetry in response to the influence of the underlying inflorescence architecture, the spatiotemporal expression patterns of three Canna CYC/TB1 homologs (CiTBL1a, CiTBL1b-1, and CiTBL1b-2) were analyzed during inflorescence and floral development using RNA in situ hybridization and qRT-PCR. In the young inflorescence, both CiTBL1a and CiTBL1b-1 were found to be expressed in the bracts and at the base of the lateral florescence branches, whereas transcripts of CiTBL1b-2 were mainly detected in flower primordia and inflorescence primordia. During early flower development, expression of CiTBL1a and CiTBL1b-1 were both restricted to the developing sepals and petals. In later flower development, expression of CiTBL1a was reduced to a very low level while CiTBL1b-1 was detected with extremely high expression levels in the petaloid androecial structures including the petaloid staminodes, the labellum, and the petaloid appendage of the fertile stamen. In contrast, expression of CiTBL1b-2 was strongest in the fertile stamen throughout flower development, from early initiation of the stamen primordium to maturity of the ½ anther. Heterologous overexpression of CiTBL genes in Arabidopsis led to dwarf plants with smaller petals and fewer stamens, and altered the symmetry of mature flowers. These data provide evidence for the involvement of CYC/TB1 homologs in the development of the asymmetric Cannaceae flower.

Irregular adaxial-abaxial polarity rearrangement contributes to the monosymmetric-to-asymmetric transformation of Canna indica stamen.

In flowering plants, lateral organs including stamens develop according to the precise regulation of adaxial-abaxial polarity. However, the polarity establishment process is poorly understood in asymmetric stamens. Canna indica (Zingiberales: Cannaceae) is a common ornamental plant with an asymmetric stamen comprising a one-theca anther and a petaloid appendage. In this study, we depicted the monosymmetric-to-asymmetric morphogenesis of C. indica stamen, and the morphogenesis of the monosymmetric stamen of a sister species was used as a contrast. We chose a HD-ZIP III gene family member and a YABBY family member as the adaxial and abaxial polarity marker genes, respectively, and tested their expression using mRNA in situ hybridization. The expression patterns of the two genes changed dynamically and asymmetrically during the stamen development process. Compared with their homologues in Arabidopsis thaliana, these two genes exhibited some specific expression patterns. We hypothesize that the distinctive adaxial-abaxial polarity participates in the irregular morphogenesis of C. indica stamen, which mediates the putative stamen-to-petaloid staminode conversion in this species.

RNA-Seq analysis reveals the distinctive adaxial-abaxial polarity in the asymmetric one-theca stamen of Canna indica.

Proper establishment of adaxial-abaxial polarity is essential for the development of lateral organs, while former researches were mostly focused on the polarity regulation in leaves, and little is known in stamens, especially in the asymmetric ones. Canna indica (Zingiberales: Cannaceae) is a widely cultivated ornamental plant and the representative species to study the evolutionary development of Zingiberales. The androecium of Canna indica comprises 3-4 petaloid staminodes and a fertile stamen (FS), which consists of a one-theca anther and a petaloid appendage. The partially petaloid stamen is considered as an intermediate state organ from a two-thecae stamen to a completely petaloid staminode. Using RNA-Seq, we quantified the expressions of the transcripts in anther and petaloid appendage, and detected 64,430 and 57,041 unigenes in these two organs, respectively. 4574 unigenes were down-regulated, and 3525 were up-regulated in petaloid appendage compared with those in anther. GO enrichment analysis indicated that the function of cytokinin is more related to cell differentiation in anther, while auxin is more to cell division in petaloid appendage. B- and C-class floral homeotic genes were expressed in these two androecium parts. Most of the class III HD-ZIP family members, which specify adaxial identity, were expressed lower in petaloid appendage than in anther; while KANADIs and YABBYs, which promote abaxial identity, exhibited opposite expression patterns. In situ hybridization showed that the adaxial marker gene was mainly expressed in the region between the two protrusions of the anther, while the abaxial marker was mainly expressed in petaloid appendage. We hypothesize that the adaxial-abaxial polarity participates in the distinctive anther-petaloid appendage patterning within the asymmetric FS of Canna indica.

Corrigendum: Temporal-Spatial Transcriptome Analyses Provide Insights into the Development of Petaloid Androecium in Canna indica.

[This corrects the article on p. 1194 in vol. 7, PMID: 27582744.].

Temporal-Spatial Transcriptome Analyses Provide Insights into the Development of Petaloid Androecium in Canna indica.

Canna indica (Zingiberales) is one of the most important ornamental species characterized with beautiful petaloid staminodes, which are considered to evolve from stamens. However, the genetic basis for the development of petaloid staminodes remains unclear largely because the genomic sequences are not available. By using RNA-Seq, we sequenced the transcripts in the flower of C. indica, and quantified the temporal gene expressions in flower primordium and differentiated flower, as well as the spatial gene expressions in petal and petaloid staminode. In total, 118,869 unigenes were assembled, among which 67,299 unigenes were annotated. Quantification analysis identified the differentially expressed genes in the temporal and spatial two comparisons, based on which, Gene Ontology enrichment analysis highlighted the representative terms in each sample, such as specification of organ number in flower primordium, growth in differentiated flower, secondary cell wall biogenesis in petal and cell division in petaloid staminode. Among the 51 analyzed MADS-box unigenes, 37 were up-regulated in differentiated flower compared with those in flower primordium. A-class unigenes were expressed higher in petal than in petaloid staminode, and C-class unigenes were expressed oppositely, whereas B-class unigenes demonstrated close expression levels in these two organs, indicating that petaloid staminode retains stamen identity to some degree. In situ hybridization provided more detailed expression patterns of these unigenes, and revealed the extended expression of B-class to the carpel at later stages when the style turned flat. These results constitute a preliminary basis for the study of flower development in C. indica and can be applied in further study of the evolution of Zingiberales.