Integrated Multi-Omics Analysis to Reveal the Molecular Mechanisms of Inflorescence Elongation in Medicago sativa.

The morphological architecture of inflorescence influences seed production. The regulatory mechanisms underlying alfalfa (Medicago sativa) inflorescence elongation remain unclear. Therefore, in this study, we conducted a comparative analysis of the transcriptome, proteome, and metabolome of two extreme materials at three developmental stages to explore the mechanisms underlying inflorescence elongation in alfalfa. We observed the developmental processes of long and short inflorescences and found that the elongation capacity of alfalfa with long inflorescence was stronger than that of alfalfa with short inflorescences. Furthermore, integrative analysis of the transcriptome and proteome indicated that the phenylpropanoid biosynthesis pathway was closely correlated with the structural formation of the inflorescence. Additionally, we identified key genes and proteins associated with lignin biosynthesis based on the differential expressed genes and proteins (DEGs and DEPs) involved in phenylpropanoid biosynthesis. Moreover, targeted hormone metabolome analysis revealed that IAA, GA, and CK play an important role in the peduncle elongation of alfalfa inflorescences. Based on omics analysis, we detected key genes and proteins related to plant hormone biosynthesis and signal transduction. From the WGCNA and WPCNA results, we furthermore screened 28 candidate genes and six key proteins that were correlated with lignin biosynthesis, plant hormone biosynthesis, and signaling pathways. In addition, 19 crucial transcription factors were discovered using correlation analysis that might play a role in regulating candidate genes. This study provides insight into the molecular mechanism of inflorescence elongation in alfalfa and establishes a theoretical foundation for improving alfalfa seed production.

Ectopic Expression of a Thellungiella salsuginea Aquaporin Gene, TsPIP1;1, Increased the Salt Tolerance of Rice.

Aquaporins play important regulatory roles in the transport of water and small molecules in plants. In this study, a ThellungiellasalsugineaTsPIP1;1 aquaporin was transformed into Kitaake rice, and three transgenic lines were evaluated by profiling the changes of the physiological metabolism, osmotic potential, and differentially expressed genes under salt stress. The TsPIP1;1 protein contains six transmembrane domains and is localized in the cytoplasm membrane. Overexpression of the TsPIP1;1 gene not only increased the accumulation of prolines, soluble sugars and chlorophyll, but also lowered the osmotic potential and malondialdehyde content in rice under salt stress, and alleviated the amount of salt damage done to rice organs by regulating the distribution of Na/K ions, thereby promoting photosynthetic rates. Transcriptome sequencing confirmed that the differentially expressed genes that are up-regulated in rice positively respond to salt stimulus, the photosynthetic metabolic process, and the accumulation profiles of small molecules and Na/K ions. The co-expressed Rubisco and LHCA4 genes in rice were remarkably up-regulated under salt stress. This data suggests that overexpression of the TsPIP1;1 gene is involved in the regulation of water transport, the accumulation of Na/K ions, and the translocation of photosynthetic metabolites, thus conferring enhanced salt tolerance to rice.

Expression of tomato SlTIP2;2 enhances the tolerance to salt stress in the transgenic Arabidopsis and interacts with target proteins.

Three independent transgenic Arabidopsis lines expressing SlTIP2;2 from Solanum lycopersicum L. cv. Lichun under the control of its endogenous promoter were used to analyze the expression of SlTIP2;2 and the salt stress tolerance under NaCl concentration gradient treatment. The expression patterns of SlTIP2;2 were shown to be tissue-specific and NaCl dose-dependent under salt stress. SlTIP2;2-transformed Arabidopsis plants exhibited enhanced salt stress tolerance, and the physiological parameters suggested that SlTIP2;2 has close links with the ion homeostasis and antioxidant enzymes activities in salt-stressed transgenic Arabidopsis. Moreover, SlTIP2;2 expression significantly affected the Na(+) and K(+) fluxes from the root meristematic zones and resulted in remarkable changes in the morphology of the pith ray cells in the inflorescence stems of transgenic Arabidopsis. Based on the yeast growth assay, ß-galactosidase activity testing and bimolecular fluorescence complementation, SlTIP1;1, SlTIP2;1 and an UDP-galactose transporter were confirmed to interact with SlTIP2;2, which may greatly broaden our understanding of the physiological functions of aquaporins.

Integrated Metabolomic and Transcriptomic Profiles Provide Insights into the Mechanisms of Anthocyanin and Carotenoid Biosynthesis in Petals of Medicago sativa ssp. sativa and Medicago sativa ssp. falcata.

The petals of Medicago sativa ssp. sativa and M. sativa ssp. falcata are purple and yellow, respectively. Free hybridization between M. sativa ssp. sativa and M. sativa ssp. falcata has created hybrids with various flower colors in nature. Moreover, the flower colors of alfalfa are closely correlated with yield, nutritional quality, stress tolerance and other agronomic characteristics. To elucidate the underlying mechanisms of flower color formation in M. sativa ssp. sativa and M. sativa ssp. falcata, we conducted an integrative analysis of the transcriptome and metabolome of alfalfa with three different petal colors (purple, yellow and cream). The metabolic profiles suggested that anthocyanins and carotenoids are the crucial pigments in purple and yellow flowers, respectively. A quantitative exploration of the anthocyanin and carotenoid components indicated that the accumulations of cyanidin, delphinidin, peonidin, malvidin, pelargonidin and petunidin derivatives are significantly higher in purple flowers than in cream flowers. In addition, the content of carotenes (phytoene, α-carotene and ß-carotene) and xanthophylls (α-cryptoxanthin, lutein, ß-cryptoxanthin, zeaxanthin, antheraxanthin and violaxanthin derivatives) was markedly higher in yellow flowers than in cream flowers. Furthermore, we found that delphinidin-3,5-O-diglucoside and lutein were the predominant pigments accumulated in purple and yellow flowers, respectively. The transcriptomic results revealed that twenty-five upregulated structural genes (one C4H, three 4CL, twelve CHS, two CHI, one F3H, one F3'H, one F3'5'H and four DFR) are involved in the accumulation of anthocyanins in purple flowers, and nine structural genes (two PSY, one ZDS, two CRTISO, two BCH, one ZEP and one ECH) exert an effect on the carotenoid biosynthesis pathway in yellow flowers. The findings of this study reveal the underlying mechanisms of anthocyanin and carotenoid biosynthesis in alfalfa with three classic flower colors.

Integrative Metabolomic and Transcriptomic Analysis Elucidates That the Mechanism of Phytohormones Regulates Floral Bud Development in Alfalfa.

Floral bud growth influences seed yield and quality; however, the molecular mechanism underlying the development of floral buds in alfalfa (Medicago sativa) is still unclear. Here, we comprehensively analyzed the transcriptome and targeted metabolome across the early, mid, and late bud developmental stages (D1, D2, and D3) in alfalfa. The metabolomic results revealed that gibberellin (GA), auxin (IAA), cytokinin (CK), and jasmonic acid (JA) might play an essential role in the developmental stages of floral bud in alfalfa. Moreover, we identified some key genes associated with GA, IAA, CK, and JA biosynthesis, including CPS, KS, GA20ox, GA3ox, GA2ox, YUCCA6, amid, ALDH, IPT, CYP735A, LOX, AOC, OPR, MFP2, and JMT. Additionally, many candidate genes were detected in the GA, IAA, CK, and JA signaling pathways, including GID1, DELLA, TF, AUX1, AUX/IAA, ARF, GH3, SAUR, AHP, B-ARR, A-ARR, JAR1, JAZ, and MYC2. Furthermore, some TFs related to flower growth were screened in three groups, such as AP2/ERF-ERF, MYB, MADS-M-type, bHLH, NAC, WRKY, HSF, and LFY. The findings of this study revealed the potential mechanism of floral bud differentiation and development in alfalfa and established a theoretical foundation for improving the seed yield of alfalfa.

Microstructure Evolution and Improved Permeability of Ceramic Waste-Based Bricks.

The resource and large-scale utilization of waste ceramic materials, magnesium slag, and coal gangue are one of the important ways for the sustainable development in metallurgy, coal, and other related enterprises. In this paper, waste ceramic materials were used as aggregates; coal gangue and magnesium slag were used as mixed binder; and the all solid-waste-based permeable bricks with excellent performance were prepared by forming pressure at 5 MPa. The mechanical properties and water permeability of the all-solid-waste-based permeable bricks were evaluated. The results proved that the porous channel of permeable brick is mainly composed of waste ceramic materials with a particle size of 2-3 mm. Pore structures below 200 µm were mainly composed of fine aggregate and mixed binder. Using 60% coarse aggregate, 20% fine aggregate, 10% coal gangue, and 10% magnesium slag as raw materials, the all-solid-waste-based permeable bricks were obtained by pressing at 6 MPa and sintering at 1200 °C, which exhibited the best performance, and its water permeability, compressive strength, and apparent porosity were 1.56 × 10-2 cm/s, 35.45 MPa, and 13.15%, respectively. Excellent water permeability, compressive strength, and apparent porosity of the all solid-waste-based permeable bricks were ascribed to the high content of connecting open pores, and closely adhesive force were ascribed to the porous microstructure constructed by the grading of waste ceramic materials and the tight conjoined points of the liquid phases in coal gangue and magnesium slag at a high sintering temperature.

N2O emissions and source partitioning using stable isotopes under furrow and drip irrigation in vegetable field of North China.

N2O emissions have increased significantly over the last decades, with much of the increase being ascribed to the expansion in agricultural land. Agricultural water management has significant influence on N2O emissions, hence the investigation of N2O emissions and the underlying production mechanisms under different irrigation managements would provide insights for N2O emission reduction and rational water usage. Here, we used the stable isotope method to evaluate the N2O isotopic signatures and the site preference, to clarify the N2O emission dynamics and the N2O source partitioning under different irrigation managements. We applied a furrow irrigation system and a drip irrigation system to maintain two different soil water conditions, allowance of dry-wet cycles and relatively stable water conditions, respectively. We found that the N2O emission was significantly decreased under drip irrigation compared to furrow irrigation with cumulated N2O flux of 526.3 mg m-2 and 571.0 mg m-2, respectively. In general, furrow irrigation with its dry-wet alternations promoted N2O emissions, while drip irrigation created a relatively stable environment that reduced N2O emissions. The intramolecular 15N isotopic composition of N2O was used to partition the relative contribution of denitrification and nitrification. Nitrification dominated the processes driving N2O production under both treatments, nearly accounting for 76% up to 100% during the initial N2O peaks. Effective measure for mitigating N2O emissions from the investigated vegetable field could be obtained by replacing the traditional furrow irrigation with drip irrigation.