Sucrose Concentration Research Articles

24-epibrassinolide enhances drought tolerance in grapevine (Vitis vinifera L.) by regulating carbon and nitrogen metabolism.

Exogenous application of 24-epibrassinolide can alleviate oxidative damage, improve photosynthetic capacity, and regulate carbon and nitrogen assimilation, thus improving the tolerance of grapevine (Vitis vinifera L.) to drought stress. Brassinosteroids (BRs) are a group of plant steroid hormones in plants and are involved in regulating plant tolerance to drought stress. This study aimed to investigate the regulation effects of BRs on the carbon and nitrogen metabolism in grapevine under drought stress. The results indicated that drought stress led to the accumulation of superoxide radicals and hydrogen peroxide and an increase in lipid peroxidation. A reduction in oxidative damage was observed in EBR-pretreated plants, which was probably due to the improved antioxidant concentration. Moreover, exogenous EBR improved the photosynthetic capacity and sucrose phosphate synthase activity, and decreased the sucrose synthase, acid invertase, and neutral invertase, resulting in improved sucrose (190%) and starch (17%) concentrations. Furthermore, EBR pretreatment strengthened nitrate reduction and ammonium assimilation. A 57% increase in nitrate reductase activity and a 13% increase in glutamine synthetase activity were observed in EBR pretreated grapevines. Meanwhile, EBR pretreated plants accumulated a greater amount of proline, which contributed to osmotic adjustment and ROS scavenging. In summary, exogenous EBR enhanced drought tolerance in grapevines by alleviating oxidative damage and regulating carbon and nitrogen metabolism.

The Impact of the Growth Regulators and Cultivation Conditions of Temporary Immersion Systems (TISs) on the Morphological Characteristics of Potato Explants and Microtubers

Potatoes (Solanum tuberosum L.) constitute one of the most economically important annual crops. In terms of tissue culture, potato microtubers (MTs) have a number of advantages over conventional plants. These advantages include their small size, which greatly facilitates storage, transport, and germplasm exchange compared to in vitro plants. One effective solution for the production and mass propagation of healthy MTs is the use of temporary immersion systems (TISs). In this study, in a SETISTM system containing kinetin/gibberellic acid (GA)/indole-3-butyric acid (IBA) hormones, we investigated the effects of different nutrient media on the morphological characteristics of potato explants and MTs. We determined the optimal cycling duration (3 h) with an immersion frequency of 2 min. The results revealed that the optimal nutrient medium for culturing single-node potato explants in a SETISTM bioreactor was the M7 medium containing kinetin (2 mg/L), GA (0.5 mg/L), and IBA (0.5 mg/L). The optimal nutrient medium for obtaining potato MTs was the M1 medium (hormone-free) with a high concentration of sucrose (9%) at 18 °C under dark growing conditions. Thus, a universal nutrient medium, employed in a bioreactor, was selected for the mass propagation of potato MTs for both domestic and foreign potato varieties.

Phosphorus application under continuous wheat-cotton straw retention enhanced cotton root productivity and seedcotton yield by improving the carbohydrate metabolism of root

ContextStraw retention could reduce phosphorus (P) application without decreasing seedcotton yield but related physiological mechanisms were unclear. Cotton root is the first organ to sense the changes in soil environment, and its growth and development, especially carbohydrate metabolism, significantly affected the formation of seedcotton yield. Therefore, it is necessary to explore the responses of cotton root carbohydrate metabolism to straw retention combined with P application. ObjectiveThis study was to investigate the effects of straw retention combined with P application on root carbohydrate metabolism and its relationship with seedcotton yield. MethodsA field experiment (2020–2022) was conducted to evaluate the changes in seedcotton yield, cotton canopy apparent photosynthesis rate (CAP), biomass, root productivity, root carbohydrate contents and related enzyme activities under different straw management [removal (S0), retention (S1)] and P rates (0, 100, 200 kg P2O5 ha−1). ResultsStraw retention and P application both increased seedcotton yield and had a significant interaction effect on it. In the 5th-7th year of straw retention, straw retention reduced 24 %-26 % P application to reach a similar yield of S0 with 100 kg P2O5 ha−1. Analyzing the reasons: (1) Straw retention combined with P application increased cotton CAP, providing adequate assimilates for cotton development; (2) Straw retention combined with P application promoted the utilization of root carbohydrates, thereby increasing root productivity and seedcotton yield. Specifically, compared with S0 without P, S1 combined with P application increased the enzyme activities related to carbohydrate metabolism, especially sucrose synthase (7.6 %-59.4 %) and acid invertase (3.9 %-58.6 %), which reduced sucrose (3.3 %-32.2 %) and starch (4.3 %-28.7 %) contents at the peak boll setting stage. Efficient utilization of sucrose and starch enhanced root productivity, the ratio of root to shoot (R/S) reduced by 2.7 %-21.6 %, while the boll loading of root system (BLR) and boll capacity of root system (BCR) increased by 0.6 %-39.2 % and 3.1 %-50.6 % at the boll opening stage, respectively, favoring the formation of seedcotton yield. Furthermore, when the R/S, BLR, and BCR reached 0.12, 0.81 boll g−1, and 4.33 g g−1, cotton harvested the maximum theoretical yield of 3780, 3774, and 4069 kg ha−1, respectively. ConclusionsStraw retention combined with P application increased root productivity and seedcotton yield by promoting the utilization of root carbohydrates. ImplicationsThis study revealed the internal mechanism for straw retention combined with P application to affect seedcotton yield from the angle of root carbohydrate metabolism and provided a theoretical basis for rationally reducing P fertilizer under straw retention.

Physicochemical and textural properties of gummy candies prepared with fruit vinegar

Foods with additional functional value derived from natural sources are becoming more and more in demand these days, including confectionary products. Therefore, the study aimed to develop a gummy candy recipe, enriched with different fruit vinegars and determine the components' effect on the products' mineral, acid, sugar, antioxidant, and polyphenol content, and texture of the confectionery products with analytical methods. In terms of macroelement content, potassium was present in the largest amount in the tested candies (167.87 and 178.83 mg kg-1) and liquid ingredients (57,71–306.93 mg kg-1). In the case of microelements, iron (4.81, 4.79 mg kg-1) and boron (4.62, 4.49 mg kg-1) were significant in the products, iron is also found in gelatin in significant quantities (3.51 mg kg-1), however, boron came exclusively from fruit-derived components (1.02–9.40 mg kg-1). Among the tested organic acids, malic acid (21.67–46.12 mg g-1), and acetic acid (47.74–50.06 mg g-1) were significant in both the raw materials and products, while the fructose content (2.44–439.35 mg g-1) was prominent among the sugars, but the glucose (75.99–163.44 mg g-1) and sucrose (73.61–157.00 mg g-1) content were also significant. Unfortunately, the polyphenol (1.37–1.46 mg GAE/g) and antioxidant (0.29–0.75 mg AAE/g) content of the products was not significant, maybe because of their heat sensitivity. In terms of the texture of the candies, the two candies prepared with different fruit vinegars had similar textural properties, however, in terms of hardness (3523.10 N/m2), gumminess (3266.25 N/m2), and chewiness (3425.33 N/m2) the gummy candy with apple cider vinegar proved to be better. Unlike many other confectionery products, the sugar content of the developed product is provided only by the used the apple concentrate, it does not contain added sugar or sweeteners. The addition of fruit vinegars has been proven to have a positive effect on the nutritional values of candies. By replacing gelatin with another gelling agent, the product can also be prepared in a vegan form.

Foliar application of sodium selenite affects the growth, antioxidant system, and fruit quality of strawberry.

Selenium (Se) plays a vital role in various physiological processes in plants and is regarded as an essential micronutrient for human health as well. In this study, sodium selenite solution at 10, 40, 70, and 100 mg·L-1 concentrations was foliar sprayed, and the strawberry plant growth, antioxidant system, and fruit quality with an emphasis on sugar and acid content were assessed. The results showed that 10 mg·L-1 of sodium selenite treatment promoted plant growth, while all the treated concentrations could enhance photosynthesis, the antioxidant system in leaves, the content of Se, and ascorbic acid in fruits. More importantly, 40 mg·L-1 sodium selenite treatment significantly increased fruit weight, total soluble solid, total phenolic content, and anthocyanins, as well as improved the shape index. Furthermore, it decreased the total flavonoid and proanthocyanidin content. Particularly, sodium selenite treatment at 40 and 70 mg·L-1 largely increased the ratio of soluble sugars to titratable acid. The changes of predominant sugars and organic acids during fruit development were further investigated. The sucrose, fructose, and glucose content was upregulated by sodium selenite treatment through upregulating the activities of sucrose phosphate synthase (SPS) and acid invertase, as well as the FaSPS expression. In addition, sodium selenite treatment inhibited the activity of citrate synthase and phosphoenolpyruvate carboxylase, rather than modulating their transcript levels to reduce the citric acid content. This work presented a potentially efficient approach to enhance plant growth and fruit quality and supplement Se in strawberry, providing insights into the mechanism of regulating sugar and acid metabolism by Se.

Engineering UDP-Glycosyltransferase UGTPg29 for the Efficient Synthesis of Ginsenoside Rg3 from Protopanaxadiol.

Rare ginsenosides Rg3 and Rh2, which exhibit diverse pharmacological effects, are derivatives of protopanaxadiol (PPD). UDP-glycosyltransferases, such as the M315F variant of Bs-YjiC (Bs-YjiCm) from Bacillus subtilis and UGTPg29 from Panax ginseng, can efficiently convert PPD into Rh2 and Rh2 into Rg3, respectively. In the present study, the N178I mutation of Bs-YjiCm was introduced, resulting in an increase in Rh2 production. UDP-glycosyltransferase UGTPg29 was then engineered to improve its robustness through semi-rational design. The variant R91M/D184M/A287V/A342L, which indicated desirable stability and activity, was utilized in coupling with the N178I variant of Bs-YjiCm and sucrose synthase AtSuSy from Arabidopsis thaliana to set up a "one-pot" three-enzyme reaction for the biosynthesis of Rg3. The influential factors, including the ratio and concentration of UDP-glycosyltransferases, pH, and the concentrations of UDP, sucrose, and DMSO, were optimized. On this basis, a fed-batch strategy was adopted to achieve a Rg3 yield as high as 12.38 mM (9.72 g/L) with a final yield of 68.78% within 24 h. This work may provide promising UDP-glycosyltransferase candidates for ginsenoside biosynthesis.

The underlying mechanisms by which boron mitigates copper toxicity in Citrus sinensis leaves revealed by integrated analysis of transcriptome, metabolome and physiology.

Both copper (Cu) excess and boron (B) deficiency are often observed in some citrus orchard soils. The molecular mechanisms by which B alleviates excessive Cu in citrus are poorly understood. Seedlings of sweet orange (Citrus sinensis (L.) Osbeck cv. Xuegan) were treated with 0.5 (Cu0.5) or 350 (Cu350 or Cu excess) μM CuCl2 and 2.5 (B2.5) or 25 (B25) μM HBO3 for 24wk. Thereafter, this study examined the effects of Cu and B treatments on gene expression levels revealed by RNA-Seq, metabolite profiles revealed by a widely targeted metabolome, and related physiological parameters in leaves. Cu350 upregulated 564 genes and 170 metabolites, and downregulated 598 genes and 58 metabolites in leaves of 2.5μM B-treated seedlings (LB2.5), but it only upregulated 281 genes and 100 metabolites, and downregulated 136 genes and 40 metabolites in leaves of 25μM B-treated seedlings (LB25). Cu350 decreased the concentrations of sucrose and total soluble sugars and increased the concentrations of starch, glucose, fructose and total nonstructural carbohydrates in LB2.5, but it only increased the glucose concentration in LB25. Further analysis demonstrated that B addition reduced the oxidative damage and alterations in primary and secondary metabolisms caused by Cu350, and alleviated the impairment of Cu350 to photosynthesis and cell wall metabolism, thus improving leaf growth. LB2.5 exhibited some adaptive responses to Cu350 to meet the increasing need for the dissipation of excessive excitation energy (EEE) and the detoxification of reactive oxygen species (reactive aldehydes) and Cu. Cu350 increased photorespiration, xanthophyll cycle-dependent thermal dissipation, nonstructural carbohydrate accumulation, and secondary metabolite biosynthesis and abundances; and upregulated tryptophan metabolism and related metabolite abundances, some antioxidant-related gene expression, and some antioxidant abundances. Additionally, this study identified some metabolic pathways, metabolites and genes that might lead to Cu tolerance in leaves.

Micropropagation and genetic transformation of Byblis liniflora

AbstractByblis, a small genus of carnivorous plants predominantly found in Australia, is characterized by its passive trapping mechanism and unique floral features. The chemical composition of Byblis, including identified phenylethanoid glycosides, particularly acteoside, highlights its pharmacological potential with various biological activities. In vitro culture techniques have been established for propagation, with micropropagation protocols developed for different Byblis species. However, information on genetic transformation, vital for trait modification and enhanced pharmacological interest, remains limited. This study focuses on optimizing micropropagation, adventitious regeneration, and genetic transformation methods for Byblis liniflora. Adventitious regeneration rates were highest in medium with reduced Murashige and Skoog salts (MS/10) and sucrose (3 gL−1) concentrations. Zeatin supplementation (1 mgL−1) further improved regeneration rates and bud development with 100% of regenerated root explants and 8.8 shoots per explant. Liquid MB3 medium supplemented with indole-3-acetic acid (IAA) 5 mgL−1 facilitated efficient rooting and acclimatization. The establishment of an efficient Rhizobium-mediated genetic transformation method yielded transgenic plants expressing green fluorescent protein (GFP). Molecular analysis confirmed transgene integration, marking the first successful genetic transformation in the Byblis genus. These advancements pave the way for exploring gene function and enhancing pharmacological properties, thereby broadening our understanding and utilization of carnivorous plants like Byblis.

In Vitro Propagation and Conservation of Lavandula stoechas subsp. luisieri and Pterospartum tridentatum, Two Important Medicinal and Aromatic Species from Portugal.

Lavandula stoechas subsp. luisieri and Pterospartum tridentatum are two valuable aromatic and medicinal plants. Their biometric and morphological parameters, such as the number of new shoots, length of the longest shoot, multiplication rate, and fresh weight, were evaluated using the multiplication MS medium protocol. The rooting protocols involved immersing the explants in IBA (1 g L-1) and a commercial IBA (3.3 g L-1) preparation (Clonex®). Slow-growth conservation assays were carried out using two different sucrose concentrations (15 g L-1 and 30 g L-1), and a control, with the cultures kept at 4 °C for 12 months. The multiplication rate for L. stoechas subsp. luisieri was 6.8, and that of P. tridentatum was 13.3, achieved using the MS medium supplemented with 0.2 mg L-1 BAP, 1 mg L-1 BAP, and 0.5 mg L-1 IBA. The application of Clonex® showed the best ex vitro rooting results in L. stoechas subsp. luisieri (77%) and P. tridentatum (90%). In the slow-growth conservation assays, at 4 °C, in darkness for 12 months, an excellent survival rate was achieved in L. stoechas subsp. luisieri (>80%) and P. tridentatum (>90%), even at the reduced sucrose concentration. This study demonstrates the effectiveness of in vitro multiplication and ex vitro rooting protocols for two valuable aromatic and medicinal plants. These findings are significant for the ex situ conservation of these species, as they provide effective long-term preservation and utilization strategies.

Biochemical composition and volatile profile analysis of three varieties of Coffea arabica and their correlation with the microclimate of Mount Tangkuban Perahu, West Java, Indonesia

Abstract. Firnabillah RA, Mustari E, Marwani E. 2024. Biochemical composition and volatile profile analysis of three varieties of Coffea arabica and their correlation with the microclimate of Mount Tangkuban Perahu, West Java, Indonesia. Biodiversitas 25: 3264-3276. The quality of coffee is determined by various biochemical compounds, including chlorogenic acid, caffeine, trigonelline, sucrose, and oil, as well as the volatile metabolite profile in coffee beans. Genetic factors and environmental conditions at the coffee cultivation site influence the composition of these compounds. This study aimed to analyze the biochemical composition and volatile profile of various coffee samples, including Ateng, Tim-Tim, and Sigarar Utang, and investigate their correlation with microclimate factors. Different analytical methods were used to examine the green beans, including High-Performance Liquid Chromatography-Mass Spectrometry (HPLC-MS) for caffeine, chlorogenic acids, sucrose, and trigonelline, Soxhlet methods for oil content determination, and Gas Chromatography-Mass Spectrometry (GC-MS) for volatile compound analysis. The results showed distinct biochemical components and metabolite profiles among the three arabica coffee varieties. The Ateng variety had the highest concentrations of chlorogenic acid, while the Tim-Tim variety had the lowest content of these compounds. Trigonelline and caffeine content was similar across all varieties. Sucrose content was highest in the Sigarar Utang variety, while oil content was higher in the Ateng and Tim-Tim varieties compared to Sigarar Utang. Principal Component Analysis (PCA) showed significant metabolite variation among the three varieties, with PC1 and PC2 accounting for 69.2% of the variance. Furthermore, Partial Least Squares-Discriminant Analysis (PLS-DA) successfully distinguished the three varieties and identified potential biomarker compounds for classification. Additionally, the study discovered that the environmental factor, specifically light intensity affected the sucrose and oil compounds in arabica green beans coffee. The study concluded that the three varieties' biochemical composition and volatile metabolites differed. Certain compounds were found to correlate with measured microclimatic factor.

Honeybee (Apis mellifera L.) pollination enhances the yield and flavor quality of kiwifruit.

Pollination is essential for achieving high yields and enhancing the quality of kiwifruit cultivation, both of which significantly influence growers' interests and consumers' preferences. However, compared to studies on yield, there are fewer studies exploring the impact of pollination methods on the flavor of kiwifruit Actinidia chinensis Planchon. This study examined the effects of bee (Apis mellifera L.) pollination and artificial pollination on the yield and flavor of kiwifruit in the main producing areas of China. Compared with those pollinated artificially, bee-pollinated kiwifruit exhibited a greater fruit set rate, heavier fruit weight, and greater number of seeds. Notably, the number of seeds was positively correlated with fruit weight in bee-pollinated kiwifruit, whereas no such correlation was detected in artificially pollinated fruit. Bee pollination not only enhanced the yield but also improved the flavor of kiwifruit. Specifically, bee-pollinated kiwifruit contained higher levels of sucrose and lower concentrations of glucose and fructose, while the acid content was less affected by pollination methods. Furthermore, significant differences were observed in the volatile organic compound (VOC) levels in kiwifruit subjected to different pollination treatments, with bee-pollinated fruit exhibiting a superior flavor. Our findings provide new insights into the beneficial role of bee pollination in enhancing kiwifruit yield and quality, underscoring the crucial importance of bees in kiwifruit pollination.

Examining the impact of dry climates temperature on citrus fruit internal ripening

High temperatures alter the ripening process of citrus fruit, affecting quality, flavor, harvest time and marketing period. For example, citrus fruits ripen faster and are sweeter in tropical hot-humid climates than in Mediterranean warm-dry climates due to higher sugar accumulation and organic acid catabolism. Considering that: 1) over 60 % of the world's citrus exports come from countries with warm Mediterranean climates, and 2) the Mediterranean region is warming 20 % faster than the global average, the citrus industry will face significant challenges in the coming years. The range of high temperatures and the timing at which they determine sugar and acid content in dry climates have not been studied under field conditions. Therefore, a study was undertaken over 6 years to determine the relationship between high temperature and citrus ripening in two dry macroclimates, hot (Arid, in Morocco) and warm (Mediterranean, in Spain), and 5 microclimates of the Mediterranean region.Heat stress in these dry climates correlates with changes in fruit quality (lower juice and sugar content, and higher organic acids), depending on the maximum temperature threshold and the time of onset and duration of heat. In the arid climate, the heat stress threshold was found to be a significant number of days above 35 °C. This was particularly important at the beginning of summer (July in the northern hemisphere). However, in the current Mediterranean climate of Spain, the percentage of summer days above 35 °C is still low indicating that other microclimatic conditions are involved in determining fruit quality. In particular, an increase in the number of days with temperatures between 27 °C and 33 °C in late summer and early autumn was found to correlate with reduced citric acid concentration. Microclimate did not determine sucrose, fructose, and glucose concentrations.

A novel high-sensitivity sensor with asymmetric elliptical shaped exposed core and gold coating for sucrose and chemical concentration detection

The article introducing an innovative exposed core SPR biosensor employing an asymmetrical elliptic air hole PCF known for remarkable sensitivity. Gold-plated fiber sensor detects changes in refractive index. This sensor effectively covers a RI scale of 1.28 to 1.42 for analytes, showcasing its versatility in simultaneous detection. Taking into account the RI change at the outer surface, attractive sensing implementations such as optimal wavelength sensitivity of 97,000 nm/RIU and optimal amplitude sensitivity of 529.20 RIU−1 are attained. Furthermore, with a resolution of 9.09 × 10−6 RIU, a figure in merit of 170 RIU−1, an FWHM of 570 nm, and a detection accuracy of 0.0166 nm−1, the suggested sensor is impressive. This suggested sensor finds application in monitoring sucrose solutions across a spectrum ranging from 0% to 45% chemical concentration over time. It has an ideal amplitude sensitivity of 530.95 RIU−1 and an ideal wavelength sensitivity of 10,000 nm/RIU for sucrose solutions at the 40% concentration level. Furthermore, the sensor shows an ideal wavelength sensitivity of 35,000 nm/RIU and an ideal amplitude sensitivity of 793.80 RIU−1 for the detection of 2-propanol. Nevertheless, beyond sucrose noticing auspicious sensing qualities by the suggested sensor its feasibility for impact fully identifying a range of biochemical and organic samples. As a result, the proposed sensor holds promise as an exemplary choice the realms in biomedical sensing, the detection of lower RI analyses as well as chemical analysis. Streamlining practical application, the sensor's structure incorporates eight elliptical air holes, uncomplicated readily manufactural with existing technologies.

The chloroplast pentatricopeptide repeat protein RCN22 regulates tiller number in rice by affecting sugar levels via the TB1–RCN22–RbcL module

As an important yield component, rice tiller number controls panicle number and determines grain yield. Regulation of rice tiller number by chloroplast pentatricopeptide repeat (PPR) proteins has not been reported previously. Here, we report the rice reduced culm number22 (rcn22) mutant, which produces few tillers owing to suppressed tiller bud elongation. Map-based cloning revealed that RCN22 encodes a chloroplast-localized P-type PPR protein. We found that RCN22 specifically binds to the 5′ UTR of RbcL mRNA (encoding the large subunit of Rubisco) and enhances its stability. The reduced abundance of RbcL mRNA in rcn22 leads to a lower photosynthetic rate and decreased sugar levels. Consequently, transcript levels of DWARF3 (D3) and TEOSINTE BRANCHED1 (TB1) (which encode negative regulators of tiller bud elongation) are increased, whereas protein levels of the positive regulator DWARF53 (D53) are decreased. Furthermore, high concentrations of sucrose can rescue the tiller bud growth defect of the rcn22 mutant. On the other hand, TB1 directly binds to the RCN22 promoter and downregulates its expression. The tb1/rcn22 double mutant shows a tillering phenotype similar to that of rcn22. Our results suggest that the TB1–RCN22–RbcL module plays a vital role in rice tiller bud elongation by affecting sugar levels.

Research on the rapid tissue breeding technology of the endangered plant Cibotium barometz (L.) J. Sm.

Cibotium barometz (L.) J. Sm., a fern, holds significant value in traditional Chinese medicine. This research aimed to establish a rapid tissue breeding technology for Cibotium barometz (L.) and provide a theoretical basis for its artificial breeding and industrial planting. The optimum conditions for spores germination were selected, including both the best sterilization time and the ideal spore germination medium. Subsequently, the optimal medium for proliferation and differentiation were chosen, and the optimal concentration of plant hormones was screened to determine the optimal medium. The rooting experiment was performed in two single-factor experiments by using differentiated sporophytes as materials. These experiments showed that the optimal sterilization time was 11 min and the best medium for germination was 1/8MS + 0.02 mg/L IAA. MS medium was suitable for proliferation, while WPM medium was suitable for sporophyte differentiation. The best medium for growth was MS + 1.0 mg/L IAA + 0.5 mg/L 6-BA. In the absence of sucrose, the seedlings barely grew and failed to differentiate into sporophytes. They showed optimal growth and differentiation at a sucrose concentration of 20 g/L. The plant growth regulators plant growth regulators used, including IAA, NAA, 6-BA, and GA3, showed more effective differentiation, wherease KT had a slightly weaker effect. The rooting rate reached the maximum when the concentrations of ABT and IBA were at 1.5 mg•L− 1 and IAA was at 1.0 mg•L− 1. The IBA had a significant effect on the average and the longest root length, while the three PGRs had no obvious effect on the average or maximum diameter. Plant tissue culture seedlings still have good genetic stability after subsequent generation. In this study, the optimal cultivation conditions for rapid breeding technology in tissue culture, covering stages from germination and growth to differentiation and rooting, were obtained. Moreover, sporophytes were successfully transplanted to the external environment. This study provided a theoretical reference for the industrial planting of C. barometz.

In-vitro Regeneration of An Endangered Medicinal Plant Anodendron paniculatum (Apocynaceae)

Objectives: The present study aims to establish a standardized and efficient regeneration protocol for Anodendron paniculatum A. DC. (Apocynaceae) by optimizing the effect of various plant growth regulators as it is categorized globally as an endangered medicinal plant. Methods: MS medium fortified with 3% sucrose (w/v) and different concentrations of cytokinins 6-benzylamino purine (BAP), kinetin, thidiazuron either alone or in combination with auxins, indole 3-acetic acid (IAA), naphthalene acetic acid (NAA) and GA3. For the in vitro root induction, individual and healthy shoots (3-4 cm in length) were excised in the 7th week and were cultured onto basal and half-strength MS medium supplemented with various concentrations of auxins and incubated in the culture room. Findings: MS medium supplemented with thidiazuron (2.5 𝜇M) in combination with indole 3-acetic acid (2.5 𝜇M) and GA3 (1.25 𝜇M) was most effective, providing multiple shoot regeneration for 79.33% of cotyledonary leaf explants associated with a high number of shoots per explant (20.00 ± 1.15) and length of shoot (2.66 ± 0.08 cm). Whereas, the frequency of response, mean number of shoots, and respective lengths were varied with different concentrations of 6-benzylamino purine when used in combination with IAA or NAA and BAP (5.0 𝜇M) along with IAA (2.5 𝜇M) at the frequency of 47.66%. Whereas, the addition of GA3 (1.25 𝜇M) in combination with BAP (5.0 𝜇M) and IAA (2.5 𝜇M) has shown a maximum number of multiple shoot formation/explant (12.33) with length of shoot (1.53 ± 0.08 cm). MS medium supplemented with 10 𝜇M 6-benzyl amino purine and 2.5 𝜇M naphthalene acetic acid recorded the highest response (53.66%) with more number of the shoot (9.66) and length of the shoot (1.83 ± 0.03 cm) per explant. The maximum percentage of response (84.33%) was recorded at 1 2 strength MS medium with 5.0 𝜇M NAA, whereas the highest number of roots per shoot (6.66 ± 0.33) was induced at MS + 5.0 𝜇M NAA. The highest length of roots (3.66 cm) was recorded at basal MS medium fortified with 5.0 𝜇M NAA and 7.5 𝜇M NAA. Novelty: The standardized protocol method is of first of its kind, a quick and suitable approach for the high quantity production of Anodendron paniculatum with appropriate concentrations of plant growth regulators that meet thecommercial demand of endangered medicinal plants. Keywords: Anodendron paniculatum; Endangered plant; Plant growth regulators; Regeneration; Protocol standardization

The Influence of Sucrose on Biomass and Glycosides Content of Callus Cultured from the Leaves of Stevia rebaudiana Bertoni

Stevia rebaudiana Bertoni contains diterpenoid steviol glycosides that have no adverse impact on blood sugar levels despite being 300 times sweeter than sugar. This study aimed to investigate the rate of callus induction from stevia leaves and the content of glycosides when changing the sucrose percentage in the culture medium.. Murashige and Skoog (MS) culture medium supported by 4.0 mg/l naphthalene acetic acid (NAA) and 1.0 mg/l benzyl adenine (BA) was used, and different concentrations of sucrose (2, 3, 4, 5 and 6%) were tested .The extraction of glycosides from leaf and callus tissues was performed by using methanol. Extracted glycosides were analyzed by high-performance liquid chromatography (HPLC). The results showed significant influences of sucrose on callus initiation. The concentration of 3% sucrose had the highest fresh and dry weight. No callus was induced in the MS medium with a high concentration of sucrose (5% and 6%). However, the highest glycoside content, stevioside and rebaudioside were obtained from callus treated with 4% concentration followed by 3% sucrose the treatment . The highest fresh and dry weight average was obtained with a 3% concentration of sucrose, this treatment also increased the concentration of glycosides in the callus twice as much as in the leaves, and concentrations of 3% and 4% of sucrose gave very similar concentrations of glycosides in callus in terms of being double what was found in the leaves something which may aid in the development of a stevia glycosides based industry.

Transcriptome and weighted gene co-expression network analyses reveal key genes and pathways involved in early fruit ripening in Citrus sinensis

BackgroundThe fruit ripening period is an important target trait in fruit tree crop breeding programs. Thus, citrus tree breeders seek to develop extreme early ripening cultivars that allow optimization of citrus maturation periods. In this study, we explored the regulatory network involved in fruit ripening in Citrus sinensis using the ‘Newhall’ navel orange variety and its early-ripening mutant, ‘Gannanzao’. This research will provide a basis for further research on important signaling pathways, gene functions and variety breeding of Citrus sinensis related to fruit ripening period.ResultsPhysiological analyses suggested that early fruit ripening in ‘Gannanzao’ is regulated by early accumulation of abscisic acid (ABA), persistently high levels of jasmonic acid (JA), and higher sucrose content in the pericarp. Pericarp samples from ‘Gannanzao’ and ‘Newhall’ navel oranges were sampled for RNA sequencing analysis at 180, 200, and 220 days after flowering; 1430 differentially expressed genes (DEGs) were identified. Functional enrichment analysis indicated that these DEGs were mainly enriched in the plant hormone signal transduction and sugar metabolism pathways, as well as other pathways related to fruit ripening. Important DEGs associated with fruit ripening in ‘Gannanzao’ included genes involved in ABA and JA metabolism and signal transduction, as well as sugar metabolism. Weighted gene co-expression network analysis showed that the deep pink module had the strongest correlations with ABA content, JA content, and early ripening. Based on gene functionality and gene expression analyses of 37 genes in this module, two candidate hub genes and two ethylene response factor 13 (ERF13) genes (Cs_ont_5g000690 and Cs_ont_5g000700) were identified as key genes regulated by ABA and JA signaling. These findings will help to clarify the mechanisms that underlie early citrus fruit ripening and will lead to the development of excellent genetic resources for further breeding of extreme early-ripening varieties.ConclusionsThrough analyses of the ‘Newhall’ navel orange cultivar and its early-ripening mutant ‘Gannanzao’, we identified genes involved in ABA and JA metabolism, signal transduction, and sugar metabolism that were related to fruit ripening. Among these, two ERF13 genes were inferred to be key genes in the regulation of fruit ripening. These findings provide insights into the genetic architecture related to early fruit ripening in C. sinensis.

Carbohydrate digestion in the stomach of horses grazed on pasture, fed hay or hay and oats

Concentrations of starch, mono- and disaccharides, fructans, hemicellulose and cellulose were analysed in feed and gastric digesta of horses in relation to acid insoluble ash as a marker indigestible in the stomach. Twenty-four horses were allocated to pasture 24 h/d (PST; n = 4), hay ad libitum (HAY; n = 8), hay ad lib. and oats at 1 g starch/kg body weight (BWT)/meal (OS1; n = 6) and hay ad lib. and oats at 2 g starch/kg BWT/meal (OS2; n = 5). One horse was excluded from the analysis. The horses were fed the ration a minimum of 34 days. Following euthanasia and dissection, digesta was sampled from Pars nonglandularis (PNG) and Pars glandularis (PG). Oat starch concentration in gastric digesta decreased from 309 to 174 g/kg dry matter (DM) in OS1 (44 %-reduction) and from 367 to 261 g/kg DM in OS2 (29 %-reduction) (P < 0.001). Glucose, fructose and sucrose disappeared from gastric digesta distinctly more in PST, HAY and OS1 than in OS2. In PST and HAY, sucrose concentration was completely cleared (P < 0.001). The concentration of fructans was reduced predominantly in PST (84 %-reduction) and HAY (54 %-reduction), mainly in the PNG (P < 0.05). Fructan degradation did not occur in the high-starch diet (OS2). Some evidence for fibre degradation was observed in PST (P < 0.01). Soluble carbohydrates disappear from the stomach dependent on the type of ration, which may lead to changes in the composition of the gastric microbial community and the endogenous response.

Mechanisms and Applications of Pseudomonas monteilii SX001: A Promising Agent for Improving Cucumber Tolerance to Salt Stress

To investigate the effects of Pseudomonas monteilii SX001 on various parameters of cucumber plants under salt stress, the salt-sensitive cucumber variety “Jinyou No. 4” was used as the test material, and coconut bran was used to simulate salt stress by applying NaCl solution. The results indicated that salt stress significantly reduced the morphological structure, relative growth rate, root morphology, and photosynthetic parameters of the cucumber plants. Leaf starch, soluble sugar, and sucrose contents significantly increased, whereas their levels in roots decreased. Cell membrane damage leads to the accumulation of reactive oxygen species and malondialdehyde, with notable increases in the activities of major antioxidant enzymes such as SOD, CAT, and POD. Nitrogen metabolism was disrupted, as evidenced by a significant decrease in nitrate nitrogen content and an increase in ammonium nitrogen content, as well as a significant reduction in the activity of NR enzymes involved in nitrogen metabolism. The enzyme activity in the cucumber rhizosphere soil decreased. However, Pseudomonas monteilii SX001 significantly enhanced the growth of cucumber seedlings under salt stress, improved photosynthetic efficiency, and facilitated sugar transformation and transport via glucose metabolism. Additionally, Pseudomonas monteilii SX001 reduced the reactive oxygen content and increased antioxidant enzyme activity. It also increased the activity of substrate enzymes and decreased the diversity of rhizosphere soil microorganisms but also increased the abundance of Asticcacaulis, Acinetobacter, Brevundimonas, Pseudomonas, and Enterobacter. These findings demonstrate that Pseudomonas monteilii SX001 is a promising bioinoculant for alleviating salt stress in cucumber production and improving soil health.