Fuliang Cao

Effect of feeding fermented Ginkgo biloba leaves on growth performance, meat quality, and lipid metabolism in broilers

Aspergillus niger-fermented Ginkgo biloba leaves (FR) and its comparative effect with vitamin E (VE) and nonfermented (NF) Ginkgo leaves on growth, lipid metabolism, antioxidant capacity, and meat quality of broiler chicks were investigated. In total, 360 one-day-old broiler chicks were randomly allocated into 6 dietary treatments, which were then denoted as control group (basal diet), VE group (containing respectively 15 and 30 IU/kg of all-rac-α-tocopherol acetate in the starter and grower phase), NF group (containing respectively 0.35% and 0.7% NF in the starter and grower phase), and FR1, FR2, and FR3 groups containing respectively 0.2, 0.35, and 0.5% FR in the starter and 0.4, 0.7, and 1.0% FR in the grower phase. The results on performance showed that a significant (P < 0.05) reduction of feed:gain ratio of birds in the FR2 group (22-42 d and 1-42 d) was observed when compared with that of the control and NF groups. With dietary FR increasing, the serum α-tocopherol concentration increased linearly (P = 0.001). Compared with the control, broilers had higher (P < 0.05) serum high-density lipoprotein concentration, total superoxide dismutase activities, and total antioxidant capacity when they were provided with the FR2 and FR3 diet. Whereas the low-density lipoprotein and triglyceride concentrations were lower (P < 0.05 or P < 0.01) in broilers from FR2 or FR3 groups. As the dietary FR increased, abdominal fat (P = 0.002) and muscle malondialdehyde (P = 0.001) concentrations decreased. Furthermore, 24-h pH, 24-h drip loss, and cooking loss were greatly improved (P < 0.05) as the levels of FR increased. Birds fed with FR had a lower (P < 0.05) C16:0 and C18:0 concentrations but a greater (P = 0.001) concentration of C18:2, C18:3, and C20:4 than that of the control. In conclusion, FR can improve the growth performance and lipid metabolism of broilers with decreased abdominal fat deposition. Also, the antioxidant capacity and meat quality improving effects observed in broilers fed FR products might result from the increased retention of α-tocopherol and reduction in lipid peroxidation, as evidenced by the decrease in malondialdehyde and the increase in total superoxide dismutase activities.

Effects of feeding fermented Ginkgo biloba leaves on small intestinal morphology, absorption, and immunomodulation of early lipopolysaccharide-challenged chicks

Aspergillus niger-fermented-Ginkgo biloba leaves (FG) and the comparative effect with nonfermented (NF)-Ginkgo leaves were investigated on morphology, absorption, and immunity of small intestine after lipopolysaccharide (LPS)-challenge in chicks at an early age. Broiler chicks (180 d of age) were divided into 3 treatment groups and were fed 1 of 3 diets: basal diet or basal diet supplemented with 0.5% NF or FG (control, NF, and FG group, respectively). Half of the birds from each treatment group were injected intraperitoneally with LPS (500 μg/kg of BW) or 0.9% NaCl solution (the sham control groups) at 10, 12, 15, 17, 19, and 21 d of age. The results indicated that when LPS-challenged birds were pretreated with FG, the decrease of ADG, ADFI, duodenal and jejunal relative weights, villus height, crypt depth, alkaline phosphatase activity, and plasma d-xylose were dramatically attenuated (P < 0.05 or P < 0.01). Meanwhile, a significant decrease (P < 0.05 or P < 0.01) of duodenal and jejunal interferon-γ, interleukin (IL)-4, IL-13, IL-18, inducible nitric oxide synthase, and duodenal sodium glucose co-transporter 1 mRNA expressional levels were found in LPS-challenged birds pretreated with FG. In conclusion, FG-supplemented diets minimized the deleterious effects of LPS and improved intestinal development, absorption, and immunity in immune-stressed chickens.

Expression of Selected Ginkgo biloba Heat Shock Protein Genes After Cold Treatment Could Be Induced by Other Abiotic Stress

Heat shock proteins (HSPs) play various stress-protective roles in plants. In this study, three HSP genes were isolated from a suppression subtractive hybridization (SSH) cDNA library of Ginkgo biloba leaves treated with cold stress. Based on the molecular weight, the three genes were designated GbHSP16.8, GbHSP17 and GbHSP70. The full length of the three genes were predicted to encode three polypeptide chains containing 149 amino acids (Aa), 152 Aa, and 657 Aa, and their corresponding molecular weights were predicted as follows: 16.67 kDa, 17.39 kDa, and 71.81 kDa respectively. The three genes exhibited distinctive expression patterns in different organs or development stages. GbHSP16.8 and GbHSP70 showed high expression levels in leaves and a low level in gynoecia, GbHSP17 showed a higher transcription in stamens and lower level in fruit. This result indicates that GbHSP16.8 and GbHSP70 may play important roles in Ginkgo leaf development and photosynthesis, and GbHSP17 may play a positive role in pollen maturation. All three GbHSPs were up-regulated under cold stress, whereas extreme heat stress only caused up-regulation of GbHSP70, UV-B treatment resulted in up-regulation of GbHSP16.8 and GbHSP17, wounding treatment resulted in up-regulation of GbHSP16.8 and GbHSP70, and abscisic acid (ABA) treatment caused up-regulation of GbHSP70 primarily.

Competitive interactions in Ginkgo and crop species mixed agroforestry systems in Jiangsu, China

Intercropping Ginkgo and crop species in southern China is receiving increasing attention because it offers potential advantages for resource utilization, higher economic income to farmers and increased sustainability in crop production, We carried out a 2-year field intercropping system composed of Ginkgo with wheat, broad bean, and rapeseed, respectively, to determine the competitive interactions between the different species, and productivity and the economic yield of each intercropping system. The density of Ginkgo and crop species was varied systematically in a two-way density matrix composed of three monoculture densities and nine intercropping of all possible pairwise combinations of monoculture densities. Intercropping systems were assessed on the basis of several intercropping indices such as land equivalent ratio, relative crowding coefficient, relative competition intensity and vector competition analysis. The results showed that the combined biomass production of the component crop species was significantly greater in the Ginkgo/crop mixtures than in monocultures crops (Ginkgo, broad bean, wheat, and rapeseed). Ginkgo: rapeseed ratio 24:12, Ginkgo: bread bean ratio 24:5, and Ginkgo: wheat ratio 24:200 had the best total biomass production. Ginkgo: rapeseed (and broad bean) ratio 24:5 and Ginkgo: wheat ratio 24:200 in respective Ginkgo/crop mixtures had the maximum economic yield. Vector competition analysis showed that Ginkgo/rapeseed mixture exhibited an antagonistic interaction type and therefore is not suitable for intercropping. Ginkgo/broad bean mixture demonstrated the most beneficial effects among the three intercropping systems.

Effects of dietary supplementation with fermented ginkgo leaves on antioxidant capacity, intestinal morphology and microbial ecology in broiler chicks

Abstract The purpose of this study was to evaluate the effects of supplementing diets with three types of fermented Ginkgo-leaves (FGL) on growth, antioxidant capacity, intestinal morphology and microbial ecology in broiler chicks. A total of 300 d-old broilers were randomly allocated to 4 dietary treatments with 6 replications of 10 birds each. Birds were fed on basal diets (Control) or basal diets supplemented with 0.5% FGL with Candida utilis (CF group), Aspergillus niger (AF group) or their combined fermentation (CAF group), respectively, for a 42 d feeding trial. AF and CAF supplementation improved body weight gain (BWG) (22–42 d) and feed conversion ratio (22–42 d and 1–42 d). Concentrations of serum α-tocopherol in CAF group, as well as hepatic α-tocopherol in the three FGL groups were increased, while hepatic reactive oxygen species (ROS) levels were greatly decreased in group AF and CAF. Chickens in AF and CAF groups had decreased hepatic protein carbonyls and malondialdehyde (MDA), as well as jejunal and ileal protein carbonyls. The total superoxide dismutase (T-SOD) activities and glutathione (GSH) of both jejunum and ileum of the CAF group were higher than the other groups. Duodenal and jejunal villous height of birds fed on the AF and CAF diets were increased, while jejunal crypt depth (CD) was decreased. Furthermore, birds fed on AF and CAF supplemented diets had increased ileal lactobacilli populations. Decreased ileal and caecal Escherichia coli and Salmonellas populations was found for the birds fed on CAF supplemented diets. The present study may indicate that the improved feed efficiency and intestinal functions in the group supplemented with AF and CAF are directly connected with the improved antioxidant capacity and intestinal microbial ecology.

Improving Flavonoid Extraction from Ginkgo biloba Leaves by Prefermentation Processing

This paper presents a prefermentation treatment method involving fungi to improve flavonoid extraction from the leaves of Ginkgo biloba . The fungi employed for this treatment were screened from the soil present under an ancient ginkgo tree. Seventy-six strains belonging to 23 genera were isolated and identified by a molecular identification method employing 18S rDNA sequences. Thirty-three strains grew well using ginkgo leaves as the growth medium. One strain, Gyx086, with higher extracted yield of flavonoids and more similar to the control, was finally selected for prefermentation processing. The major fermentation factors were optimized by response surface methodology. The optimal conditions for the highest total falvonoid yield were 27.8 °C for temperature, 64.2% for moisture content, and 61 h for fermentation time. Under the optimal condition, a actual total flavonoid yield of 27.59 ± 0.52 mg/g dry weight culture sample was obtained, which was about 70% higher than that of unfermented gingko leaf samples.

Comparative Proteomic and Physiological Analysis Reveals the Variation Mechanisms of Leaf Coloration and Carbon Fixation in a Xantha Mutant of Ginkgo biloba L.

Yellow-green leaf mutants are common in higher plants, and these non-lethal chlorophyll-deficient mutants are ideal materials for research on photosynthesis and plant development. A novel xantha mutant of Ginkgo biloba displaying yellow-colour leaves (YL) and green-colour leaves (GL) was identified in this study. The chlorophyll content of YL was remarkably lower than that in GL. The chloroplast ultrastructure revealed that YL had less dense thylakoid lamellae, a looser structure and fewer starch grains than GL. Analysis of the photosynthetic characteristics revealed that YL had decreased photosynthetic activity with significantly high nonphotochemical quenching. To explain these phenomena, we analysed the proteomic differences in leaves and chloroplasts between YL and GL of ginkgo using two-dimensional gel electrophoresis (2-DE) coupled with MALDI-TOF/TOF MS. In total, 89 differential proteins were successfully identified, 82 of which were assigned functions in nine metabolic pathways and cellular processes. Among them, proteins involved in photosynthesis, carbon fixation in photosynthetic organisms, carbohydrate/energy metabolism, amino acid metabolism, and protein metabolism were greatly enriched, indicating a good correlation between differentially accumulated proteins and physiological changes in leaves. The identifications of these differentially accumulated proteins indicates the presence of a specific different metabolic network in YL and suggests that YL possess slower chloroplast development, weaker photosynthesis, and a less abundant energy supply than GL. These studies provide insights into the mechanism of molecular regulation of leaf colour variation in YL mutants.

An efficient approach to identify Ginkgo biloba cultivars by using random amplified polymorphic DNA markers with a manual cultivar identification diagram strategy.

Cultivar identification is a key step to avoid the formation of homonyms and synonyms of Ginkgo biloba. In this study, a new approach based on combinational utilization of polymorphic bands produced from 6 different random amplified polymorphic DNA (RAPD) primers was developed for identifying 42 Ginkgo cultivars, and a manual cultivar identification diagram that consisted of polymorphic bands produced from different RAPD primers was reported. To check the reliability and efficiency of the cultivar identification diagram, 5 randomly chosen cultivars were further tested, and the workability of the diagram was verified. This new approach will be very helpful for Ginkgo cultivar discrimination and protection, and will also be beneficial for the nursery industry for early identification of Ginkgo seedlings.

Deep eutectic solvents as green media for efficient extraction of terpene trilactones from Ginkgo biloba leaves

ABSTRACT Deep eutectic solvents (DESs)-based ultrasonic extraction of terpene trilactones (TTLs) from Ginkgo biloba leaves was efficiently developed. Sixteen DESs were prepared, and DESs composed of choline chloride-urea (ChCl-U) and betaine-ethylene glycol (BE-EG) gave higher TTL extraction yields than the present, most efficient solvent 70% ethanol. The extraction conditions were further optimized, and the optimum conditions were as follows: taking BE-EG containing 40% (w/w) water as the extraction solvent, 1:10 of G. biloba leaves powder-to-solvent ratio, and ultrasonic treatment at 45°C and 100 W for 20 min. A total extraction yield of 1.94 ± 0.03 mg/g was obtained under the optimum conditions, which indicated that 99.37% of TTLs could be extracted from the G. biloba leaves powder by a single extraction. Moreover, the polyamide resin was used to recover the TTLs in DES extracting solution, and recovery yield of 95.1% was attained. Therefore, BE-EG containing 40% (w/w) water was a potential alternative solvent for TTLs extraction from G. biloba leaves. GRAPHICAL ABSTRACT

Role of Temperature and Soil Moisture Conditions on Flavonoid Production and Biosynthesis-Related Genes in Ginkgo (Ginkgo biloba L.) Leaves

The environmental factors affecting flavonoid biosynthesis and accumulation from Ginkgo (Ginkgo biloba L.) plants in production plantation settings is investigated to maximize their production. Objectives: To develop an understanding of the environmental factors (temperature and available soil moisture and their inter-relationship) affecting flavonoids biosynthesis and accumulation in Ginkgo leaves for selection, development, and production purposes for commercial production. Methods: A factorial experiment (temperature: 15/5, 25/15 and 35/25°C (day/night) and soil moisture: 55-60, 40-45 and 30-35% of field capacity) using 2-year-old Ginkgo seedlings to estimated flavonoids content and their expression level in leaves sampled 10, 20, 30, 40 and 50days using high performance liquid chromatography and quantitative real-time PCR (qRT-PCR) of seven flavonoid biosynthesis-related genes, respectively. Results: Flavonoid accumulation was significantly higher under lower temperature (15/5°C) and available soil moisture (40-45 and 30-35%) while it was severely suppressed under high temperature. qRT-PCR indicated that flavonoid biosynthesis differed greatly among the studied genes with some genes being up-regulated under lower or higher temperature and/or moisture. Conclusion: Total flavonoid content was greatly affected by temperature and soil moisture content and their biosynthesis varied among the studied genes suggesting the presence of a synergistic effect on the expression of genes within the flavonoid biosynthesis pathway. The results contributed to the understanding of the environmental factors needed for the successful production of flavonoid from Ginkgo leaves harvest plantations.