Competitive Biosynthesis of Bacterial Alginate Using Azotobacter vinelandii 12 for Tissue Engineering Applications

This study investigated the effect of various cultivation conditions (sucrose/phosphate concentrations, aeration level) on alginate biosynthesis using the bacterial producing strain Azotobacter vinelandii 12 by the full factorial design (FFD) method and physicochemical properties (e.g., rheological properties) of the produced bacterial alginate. We demonstrated experimentally the applicability of bacterial alginate for tissue engineering (the cytotoxicity testing using mesenchymal stem cells (MSCs)). The isolated synthesis of high molecular weight (Mw) capsular alginate with a high level of acetylation (25%) was achieved by FFD method under a low sucrose concentration, an increased phosphate concentration, and a high aeration level. Testing the viscoelastic properties and cytotoxicity showed that bacterial alginate with a maximal Mw (574 kDa) formed the densest hydrogels (which demonstrated relatively low cytotoxicity for MSCs in contrast to bacterial alginate with low Mw). The obtained data have shown promising prospects in controlled biosynthesis of bacterial alginate with different physicochemical characteristics for various biomedical applications including tissue engineering.

A Vacuolar Invertase CsVI2 Regulates Sucrose Metabolism and Increases Drought Tolerance in Cucumis sativus L.

Vacuolar invertase (VI) can irreversibly degrade sucrose into glucose and fructose and involve in plants abiotic-stress-tolerance. Cucumber (Cucumis sativus L.) is susceptible to drought stress, especially during the seedling stage. To date, the involvement of VI in drought tolerance in cucumber seedlings is in urgent need of exploration. In the present study, a cucumber vacuolar invertase gene, CsVI2, was isolated and functionally characterized. The results showed that (1) CsVI2 showed vacuolar invertase activity both in vivo and in vitro; (2) the transcript level of CsVI2, along with VI activity, was significantly induced by drought stress. Moreover, the expression of sucrose synthase 3 (CsSUS3) was increased and that of sucrose phosphate synthase 1 (CsSPS1) was decreased after exposure to drought stress, which was followed by an increase in sucrose synthase activity and a decrease in sucrose phosphate synthase activity; (3) CsVI2-overexpressing transformed cucumber seedlings showed enhanced vacuolar invertase activity and drought tolerance and 4) protein–protein interaction modelling indicated that a cucumber invertase inhibitor, CsINVINH3, can interact with CsVI2. In summary, the results indicate that CsVI2 as an invertase can regulate sucrose metabolism and enhance drought stress in cucumber seedlings.

Genotypic variation in root morphology, cotton subtending leaf physiology and fiber quality against nitrogen

Background Nitrogen (N) is important for improving various morphological and physiological processes of cotton but their contribution to fiber quality is still lacking. Aims The current study aimed to explore the relationship between root morphology, subtending leaf physiology, and fiber quality of contrasting N-efficient cotton genotypes in response to N. Methods We analyzed the above parameters of CCRI 69 (N-efficient) and Xinluzao-30 (XLZ-30, N-inefficient) under control (2.5 mmol·L−1) and high N (5 mmol·L−1) conditions. Results The results showed that root morphological traits were increased in CCRI-69 under control conditions than high N. Subtending leaf morphology, chlorophyll and carotenoid contents, free amino acids, and soluble proteins were higher under high N as compared with the control. However, soluble sugars, fructose, sucrose contents, and sucrose phosphate synthase were higher under control conditions than high N across the growth stages. Irrespective of the N conditions, all morphological and physiological traits of cotton subtending leaf were higher in CCRI-69 than XLZ-30. Except for fiber uniformity, fiber quality traits like fiber length, strength, micronaire, and elongation were improved under control conditions than high N. Between the genotypes, CCRI-69 had significantly higher fiber length, strength, micronaire, and elongation as compared with XLZ-30. Strong positive correlations were found between root morphology, soluble sugars, sucrose content, and sucrose phosphate synthase activity with fiber quality traits, respectively. Conclusions These findings suggest that CCRI-69 performed better in terms of growth and fiber quality under relatively low N condition, which will help to reduce fertilizer use, the cost of production, and environmental pollution.

The Involvement of Glucose in Hydrogen Gas-Medicated Adventitious Rooting in Cucumber

Hydrogen gas (H2) and glucose (Glc) have been reported as novel antioxidants and signal molecules involved in multiple biological processes in plants. However, the physiological roles and relationships of H2 and Glc in adventitious rooting are less clear. Here, we showed that the effects of different concentrations Glc (0, 0.01, 0.05, 0.10, 0.50 and 1.00 mM) on adventitious rooting in cucumber were dose-dependent, with a maximal biological response at 0.10 mM. While, the positive roles of hydrogen rich water (HRW, a H2 donor)-regulated adventitious rooting were blocked by a specific Glc inhibitor glucosamine (GlcN), suggesting that Glc might be responsible for H2-regulated adventitious root development. HRW increased glucose, sucrose, starch and total sugar contents. Glucose-6-phosphate (G6P), fructose-6-phosphate (F6P) and glucose-1-phosphate (G1P) contents were also increased by HRW. Meanwhile, the activities of sucrose-related enzymes incorporating sucrose synthase (SS) and sucrose phosphate synthase (SPS) and glucose-related enzymes including hexokinase (HK), pyruvate kinase (PK) and adenosine 5′-diphosphate pyrophosphorylase (AGPase) were increased by HRW. Moreover, HRW upregulated the expression levels of sucrose or glucose metabolism-related genes including CsSuSy1, CsSuSy6, CsHK1, CsHK3, CsUDP1, CsUDP1-like, CsG6P1 and CsG6P1-like. However, these positive roles were all inhibited by GlcN. Together, H2 might regulate adventitious rooting by promoting glucose metabolism.

EFFECT OF SALT (NaCl) ON PHOTOSYNTHESIS AND CARBOHYDRATES IN LEAVES OF TWO RAGI (ELEUSINE CORACANA (L.) GAERTN) VARIETIES

Salinity is one of the most widespread environmental threats to global crop production, especially in arid and semi-arid regions. Photosynthesis and carbohydrates were determined in two ragi (Eleusine coracana (L.) Gaertn) varieties (CO13 and PAIYUR-1), subjected to salt stress of different concentrations (0, 40, 80 and 120mM). Salinity was given as a basal dose and sampling was done in leaves on 30th Days. After Treatment (DAT). There was a marked variation in the photosynthetic rates and ribulose-1, 5-bisphosphate carboxylase activity between the two ragi varieties subjected to salt stress. Photosystem II (PSII) and sucrose phosphate synthase activities were also significantly reduced as measured by salt stressed conditions. The quantity of glucose and sucrose decreased with increasing salt stress while starch showed a reverse trend under salt-stressed conditions. The results revealed that CO-13 exhibits higher photosynthetic rates and activities of ribulose-1,5-bisphosphate carboxylase, sucrose phosphate synthase with photochemical efficiency of PSII compared to PAIYUR-1

Structure and Expression Analysis of Sucrose Phosphate Synthase, Sucrose Synthase and Invertase Gene Families in Solanum lycopersicum

Sucrose phosphate synthase (SPS), sucrose synthase (SUS) and invertase (INV) are all encoded by multigene families. In tomato (Solanum lycopersicum), a comprehensive analysis of structure characteristics of these family genes is still lacking, and the functions of individual isoforms of these families are mostly unclear under stress. Here, the structure characteristics of the three families in tomato were analyzed; moreover, as a first step toward understanding the functions of isoforms of these proteins under stress, the tissue expression pattern and stress response of these genes were also investigated. The results showed that four SPS genes, six SUS genes and nineteen INV genes were identified in tomato. The subfamily differentiation of SlSPS and SlSUS might have completed before the split of monocotyledons and dicotyledons. The conserved motifs were mostly consistent within each protein family/subfamily. These genes demonstrated differential expressions among family members and tissues, and in response to polyethylene glycerol, NaCl, H2O2, abscisic acid or salicylic acid treatment. Our results suggest that each isoform of these families may have different functions in different tissues and under environmental stimuli. SlSPS1, SlSPS3, SlSUS1, SlSUS3, SlSUS4, SlINVAN5 and SlINVAN7 demonstrated consistent expression responses and may be the major genes responding to exogenous stimuli.

Comparative transcriptome analysis reveals key genes potentially related to organic acid and sugar accumulation in loquat

Organic acids and sugars are the primary components that determine the quality and flavor of loquat fruits. In the present study, major organic acids, sugar content, enzyme activities, and the expression of related genes were analyzed during fruit development in two loquat cultivars, ’JieFangZhong’ (JFZ) and ’BaiLi’ (BL). Our results showed that the sugar content increased during fruit development in the two cultivars; however, the organic acid content dramatically decreased in the later stages of fruit development. The differences in organic acid and sugar content between the two cultivars primarily occured in the late stage of fruit development and the related enzymes showed dynamic changes in activies during development. Phosphoenolpyruvate carboxylase (PEPC) and mNAD malic dehydrogenase (mNAD-MDH) showed higher activities in JFZ at 95 days after flowering (DAF) than in BL. However, NADP-dependent malic enzyme (NADP-ME) activity was the lowest at 95 DAF in both JFZ and BL with BL showing higher activity compared with JFZ. At 125 DAF, the activity of fructokinase (FRK) was significantly higher in JFZ than in BL. The activity of sucrose synthase (SUSY) in the sucrose cleavage direction (SS-C) was low at early stages of fruit development and increased at 125 DAF. SS-C activity was higher in JFZ than in BL. vAI and sucrose phosphate synthase (SPS) activities were similar in the two both cultivars and increased with fruit development. RNA-sequencing was performed to determine the candidate genes for organic acid and sugar metabolism. Our results showed that the differentially expressed genes (DEGs) with the greated fold changes in the later stages of fruit development between the two cultivars were phosphoenolpyruvate carboxylase 2 (PEPC2), mNAD-malate dehydrogenase (mNAD-MDH), cytosolic NADP-ME (cyNADP-ME2), aluminum-activated malate transporter (ALMT9), subunit A of vacuolar H+-ATPase (VHA-A), vacuolar H+-PPase (VHP1), NAD-sorbitol dehydrogenase (NAD-SDH), fructokinase (FK), sucrose synthase in sucrose cleavage (SS-C), sucrose-phosphate synthase 1 (SPS1), neutral invertase (NI), and vacuolar acid invertase (vAI). The expression of 12 key DEGs was validated by quantitative reverese transcription PCR (RT-qPCR). Our findings will help understand the molecular mechanism of organic acid and sugar formation in loquat, which will aid in breeding high-quality loquat cultivars.