Analyzing Morphology, Metabolomics, and Transcriptomics Offers Invaluable Insights into the Mechanisms of Pigment Accumulation in the Diverse-Colored Labellum Tissues of Alpinia.

Alpinia plants are widely cherished for their vibrant and captivating flowers. The unique feature of this genus lies in their labellum, a specialized floral structure resulting from the fusion of two non-fertile staminodes. However, the intricate process of pigment formation, leading to distinct color patterns in the various labellum segments of Alpinia, remains a subject of limited understanding. In this study, labellum tissues of two Alpinia species, A. zerumbet (yellow-orange flowers) and A. oxyphylla (white-purple flowers), were sampled and analyzed through morphological structure observation, metabolite analysis, and transcriptome analyses. We found that hemispherical/spherical epidermal cells and undulate cell population morphology usually display darker flower colors, while flat epidermal cells and cell populations usually exhibit lighter flower colors. Metabolomic analysis identified a high concentration of anthocyanins, particularly peonidin derivatives, in segments with orange and purple pigments. Additionally, segments with yellow pigments showed significant accumulations of flavones, flavanols, flavanones, and xanthophylls. Furthermore, our investigation into gene expression levels through qRT-PCR revealed notable differences in several genes that participated in anthocyanin and carotenoid biosynthesis among the four pigmented segments. Collectively, these findings offer a comprehensive understanding of pigmentation in Alpinia flowers and serve as a valuable resource for guiding future breeding efforts aimed at developing Alpinia varieties with novel flower colors.

Exploring the Heat Shock Transcription Factor (HSF) Gene Family in Ginger: A Genome-Wide Investigation on Evolution, Expression Profiling, and Response to Developmental and Abiotic Stresses.

Ginger is a valuable crop known for its nutritional, seasoning, and health benefits. However, abiotic stresses, such as high temperature and drought, can adversely affect its growth and development. Heat shock transcription factors (HSFs) have been recognized as crucial elements for enhancing heat and drought resistance in plants. Nevertheless, no previous study has investigated the HSF gene family in ginger. In this research, a total of 25 ZoHSF members were identified in the ginger genome, which were unevenly distributed across ten chromosomes. The ZoHSF members were divided into three groups (HSFA, HSFB, and HSFC) based on their gene structure, protein motifs, and phylogenetic relationships with Arabidopsis. Interestingly, we found more collinear gene pairs between ZoHSF and HSF genes from monocots, such as rice, wheat, and banana, than dicots like Arabidopsis thaliana. Additionally, we identified 12 ZoHSF genes that likely arose from duplication events. Promoter analysis revealed that the hormone response elements (MEJA-responsiveness and abscisic acid responsiveness) were dominant among the various cis-elements related to the abiotic stress response in ZoHSF promoters. Expression pattern analysis confirmed differential expression of ZoHSF members across different tissues, with most showing responsiveness to heat and drought stress. This study lays the foundation for further investigations into the functional role of ZoHSFs in regulating abiotic stress responses in ginger.

Combined Metabolome and Transcriptome Analyses of Young, Mature, and Old Rhizome Tissues of Zingiber officinale Roscoe.

Ginger (Zingiber officinale Roscoe) is known for its unique pungent taste and useability in traditional Chinese medicine. The main compounds in ginger rhizome can be classified as gingerols, diarylheptanoids, and volatile oils. The composition and concentrations of the bioactive compounds in ginger rhizome might vary according to the age of the rhizome. In this regard, the knowledge on the transcriptomic signatures and accumulation of metabolites in young (Y), mature (M), and old (O) ginger rhizomes is scarce. This study used HiSeq Illumina Sequencing and UPLC-MS/MS analyses to delineate how the expression of key genes changes in Y, M, and O ginger rhizome tissues and how it affects the accumulation of metabolites in key pathways. The transcriptome sequencing identified 238,157 genes of which 13,976, 11,243, and 24,498 were differentially expressed (DEGs) in Y vs. M, M vs. O, and Y vs. O, respectively. These DEGs were significantly enriched in stilbenoid, diarylheptanoid, and gingerol biosynthesis, phenylpropanoid biosynthesis, plant-hormone signal transduction, starch and sucrose metabolism, linoleic acid metabolism, and α-linoleic acid metabolism pathways. The metabolome profiling identified 661 metabolites of which 311, 386, and 296 metabolites were differentially accumulated in Y vs. M, Y vs. O, and M vs. O, respectively. These metabolites were also enriched in the pathways mentioned above. The DEGs and DAMs enrichment showed that the gingerol content is higher in Y rhizome, whereas the Y, M, and O tissues differ in linoleic and α-linoleic acid accumulation. Similarly, the starch and sucrose metabolism pathway is variably regulated in Y, M, and O rhizome tissues. Our results showed that ginger rhizome growth slows down (Y > M > O) probably due to changes in phytohormone signaling. Young ginger rhizome is the most transcriptionally and metabolically active tissue as compared to M and O. The transitioning from Y to M and O affects the gingerol, sugars, linoleic acid, and α-linoleic acid concentrations and related gene expressions.

Functional characterization of an Indian sandalwood (Santalum album L.) dual-localized bifunctional nerolidol/linalool synthase gene involved in stress response.

Essential oils extracted from the heartwood of Indian sandalwood (Santalum album L.) contain linalool and nerolidol as minor components. However, nerolidol/linalool synthase (NES/LIS), which produce linalool and nerolidol, have yet to be characterized in sandalwood. Using a transcriptomic-based approach, a terpene synthase gene was screened from unigenes of transcriptome data derived from S. album seedlings exposed to low temperature (4 °C). The enzyme encoded by these complementary DNAs belongs to the TPS-b clade. Recombinant SaNES/LIS is a bifunctional enzyme that can catalyze the formation of (E)-nerolidol from farnesyl diphosphate and linalool from geranyl diphosphate, respectively. Whereas SaNES/LIS was primarily localized in chloroplastids, both as granular fluorescence and as diffuse fluorescence, it was also detected in the cytosol of a limited number of cells. Agrobacterium tumefaciens-mediated transient gene expression in planta produced the same terpene products as those obtained in vitro. Real-time PCR analysis showed the highest expression of SaNES/LIS in fruits, with about a three-fold higher level than in leaves, followed by flowers, heartwood and roots. SaNES/LIS transcripts were differentially activated in different tissues in response to methyl jasmonate, cold, high temperature, strong illumination, and drought stress. Our results provide novel insight into the role of sandalwood terpenoids in response to various environmental stresses.

Electrochemical/Fe3+/peroxymonosulfate system for the degradation of Acid Orange 7 adsorbed on activated carbon fiber cathode.

Acid Orange 7 (AO7), as a most common and widely used synthetic dyes in the printing and dyeing industry, was hardly degradable by traditional wastewater treatment methods. Here, activated carbon fiber (ACF) as an in-situ regenerated cathodic adsorbent in the electrochemical/Fe3+/peroxymonosulfate process (EC/ACF/Fe3+/PMS) was firstly investigated for AO7 removal and compared with several different processes. The results indicated that the effective adsorption of AO7 on ACF can be enhanced under electrolytic conditions, while the adsorbed AO7 on ACF can be completely degraded and mineralized in EC/ACF/Fe3+/PMS process resulting in the in-situ regeneration of ACF. Besides, the electrical energy per order values were investigated, which showed an apparent reduction of electrical energy consumption from 0.42831 to 0.09779 kWh m-3 when ACF-cathode replaced Pt-cathode. Further study revealed that higher conversion rate of Fe2+ from Fe3+ was observed with ACF-cathode. It deserved to be mentioned that the removal efficiency of AO7 was satisfactory and stable even after reusing ACF cathode for 10 times. Furthermore, structure and elements of ACF surface were investigated, which indicated the structure of ACF was intact in EC/ACF/Fe3+/PMS due to inhibition of ACF corrosion by electron migration at cathode. In addition, the total iron content of the effluent in EC/ACF/Fe3+/PMS was lower than that of EC/Fe3+/PMS due to the deposition of iron on ACF-cathode surface. Therefore, advantages of EC/ACF/Fe3+/PMS for AO7 degradation were not only a much higher oxidation efficiency and in-situ regenerated cathodic adsorbent, but also a lower electrical energy consumption and lesser iron ions contents in the effluent.

Backwashing performance of self-cleaning screen filters in drip irrigation systems.

The self-cleaning screen filter is one of the most common types used in drip irrigation systems. Backwashing pressure difference and backwashing time for two screen filters with one geometry and two different screens (178 µm and 124 µm) using two water qualities (tap water and sand-water mixture) were studied in a total of 88 runs (42 runs for tap water, 22 and 24 filtration cycles for sand-water mixture and backwashing, respectively). The backwashing pressure difference and backwashing time were calculated using the experimental data, and the results were largely in the range of measured values. Three constraint conditions (flowrate, sand condition and filtration time) of backwashing pressure difference were analysed, and the optimal values of backwashing pressure difference were given as 60.0 and 70.0 kPa for 178 µm and 124 µm filters, respectively. The backwashing time of the screen filter should be an optimal value that ensures that the pressure difference between the internal and external surfaces of the screen decreased to the initial value, and the sand concentration of the backwashed outlet decreased to a small, stable value. Based on the results of the backwashing experiment and prototype observation, the optimal backwashing time was given as 30 to 45 s for both screen filters.

Analytical equation for outflow along the flow in a perforated fluid distribution pipe.

Perforated fluid distribution pipes have been widely used in agriculture, water supply and drainage, ventilation, the chemical industry, and other sectors. The momentum equation for variable mass flow with a variable exchange coefficient and variable friction coefficient was developed by using the momentum conservation method under the condition of a certain slope. The change laws of the variable momentum exchange coefficient and the variable resistance coefficient along the flow were analyzed, and the function of the momentum exchange coefficient was given. According to the velocity distribution of the power function, the momentum equation of variable mass flow was solved for different Reynolds numbers. The analytical solution contains components of pressure, gravity, friction and momentum and reflects the influence of various factors on the pressure distribution along the perforated pipe. The calculated results of the analytical solution were compared with the experimental values of the study by Jin et al. 1984 and Wang et al. 2001 with the mean errors 8.2%, 3.8% and 2.7%, and showed that the analytical solution of the variable mass momentum equation was qualitatively and quantitatively consistent with the experimental results.

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.

Molecular basis of floral petaloidy: insights from androecia of Canna indica.

Floral organs that take on the characteristics of petals can occur in all whorls of the monocot order Zingiberales. In Canna indica, the most ornamental or 'petaloid' parts of the flowers are of androecial origin and are considered staminodes. However, the precise nature of these petaloid organs is yet to be determined. In order to gain a better understanding of the genetic basis of androecial identity, a molecular investigation of B- and C-class genes was carried out. Two MADS-box genes GLOBOSA (GLO) and AGAMOUS (AG) were isolated from young inflorescences of C. indica by 3' rapid amplification of cDNA ends polymerase chain reaction (3'-RACE PCR). Sequence characterization and phylogenetic analyses show that CiGLO and CiAG belong to the B- and C-class MADS-box gene family, respectively. CiAG is expressed in petaloid staminodes, the labellum, the fertile stamen and carpels. CiGLO is expressed in petals, petaloid staminodes, the labellum, the fertile stamen and carpels. Expression patterns in mature tissues of CiGLO and CiAG suggest that petaloid staminodes and the labellum are of androecial identity, in agreement with their position within the flower and with described Arabidopsis thaliana expression patterns. Although B- and C-class genes are important components of androecial determination, their expression patterns are not sufficient to explain the distinct morphology observed in staminodes and the fertile stamen in C. indica.

Development and characterization of microsatellite markers for Alpinia oxyphylla (Zingiberaceae).

PREMISE OF THE STUDY: Microsatellite loci were isolated and characterized for population genetic studies of Alpinia oxyphylla (Zingiberaceae), a perennial rhizomatous herbaceous plant often used medicinally in China. • METHODS AND RESULTS: A total of 85 loci were identified using a magnetic bead enrichment method, of which 23 were polymorphic. The level of polymorphism was characterized in 32 individuals from two populations; the number of alleles per locus ranged from 1 to 13; and observed heterozygosity and expected heterozygosity varied from 0 to 1 (mean: 0.6441) and 0 to 0.887 (mean: 0.6241), respectively. • CONCLUSIONS: The polymorphic microsatellite markers generated from this study will be useful for genetic diversity and structure analysis of A. oxyphylla.