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As a common mutation trait in plants, leaf colour mutation is related to the degree of chlorophyll and anthocyanin changes and the destruction of chloroplast structure. This study summarizes the latest research progress in leaf colour mutation mechanism, including the metabolic basis of plant leaf colour mutation, leaf colour mutation caused by gene mutation in the chlorophyll metabolism pathway, leaf colour mutation caused by blocked chloroplast development, leaf colour mutation controlled by key transcription factors and noncoding RNAs, leaf colour mutation caused by environmental factors, and leaf colour mutation due to the involvement of the mevalonate pathway. These results will lay a theoretical foundation for leaf colour development, leaf colour improvement, and molecular breeding for leaf colour among tree species. [ABSTRACT FROM AUTHOR]

Chamaemelum nobile is a traditional Chinese herbal medicine, whose secondary metabolites used in the pharmacology of Chinese medicine. Among them, the flavonoids have great research value. Flavanone 3-hydroxylase (F3H) is one of the core enzymes in the early steps of flavonoid biosynthesis. This study aimed to elucidate the structures, functions, and expression levels of F3H families from C. nobile. Four members of the F3H family were screened from C. nobile transcriptome data and performed bioinformatics analysis. Results showed that CnF3H1~4 had a high similarity with the other F3H plants, and all genes contained two conserved isopenicillin N synthase-like and oxoglutarate/iron-dependent dioxygenase domains. Further analysis revealed that the four CnF3H proteins contained some differences in binding sites. The results of secondary and 3-D structures displayed that the composition and proportion of the four CnF3H secondary structures were basically the same, and their 3D structures were consistent with the secondary structures. The phylogenetic tree displayed that CnF3H2, CnF3H3, and CnF3H4 were grouped with Asteraceae. The expression patterns of CnF3Hs in the roots, stems, leaves, and flowers of C. nobile were evaluated using the value of RPKM. The results indicated that CnF3Hs had significant difference in the expression of different tissues. Especially, CnF3H1~3 and CnF3H4 had the highest expression levels in the flowers and roots, respectively. Hence, CnF3Hs played a significant role in the flavonoid metabolism. [ABSTRACT FROM AUTHOR]

Ginkgo biloba is widely planted, and the extracts of leaves contain flavonoids, terpene esters and other medicinal active ingredients. WRKY proteins are a large transcription factor family in plants, which play an important role in the regulation of plant secondary metabolism and development, as well as the response to biotic and abiotic stress. In our study, we identified 40 genes with conserved WRKY motifs in the G. biloba genome and classified into groups I (groups I-N and -C), II (groups IIa, b, c, d, and e), and III, which include 12, 26, and 2 GbWRKY genes, respectively. Meanwhile, the expression patterns of 10 GbWRKY (GbWRKY2, GbWRKY3, GbWRKY5, GbWRKY7, GbWRKY11, GbWRKY15, GbWRKY23, GbWRKY29, GbWRKY31, GbWRKY32) under different tissue and abiotic stress conditions were analyzed. Under stress treatment, the expression patterns of 10 WRKY genes were changed. 10 ginkgo WRKY transcription factors were induced by ETH and SA, but there are two different induced response modes. The expression of 10 WRKY genes was inhibited under low temperature, high temperature and MeJA hormone induction. Most WRKY genes were up-regulated under the induction of high salt and ABA. GbWRKYs were differentially expressed in various tissues after abiotic stress and plant hormone treatments, thereby indicating their possible roles in biological processes and abiotic stress tolerance and adaptation. Our results provided insight into the genome-wide identification of GbWRKYs, as well as their differential responses to stresses and hormones. These data can also be utilized to identify potential molecular targets to confer tolerance to various stresses in G. biloba. [ABSTRACT FROM AUTHOR]

The contents of flavonoids and other related components, in addition to the activities of related enzymes, such as phenylalanine ammonia- lyase (PAL), peroxidase (POD), chalcone synthase (CHS), and UDP-glucose:flavonoid 3-O- glucosyltransferase (UFGT), were determined in Ginkgo biloba L. leaves during leaf development. The results indicated that POD activity increased gradually in the early stages and reached its first activity peak in the beginning of Jul., the second one in late Aug., and the third one (the highest peak) in the middle of Oct. The changes in POD activity were irregular, and its activity was maintained at a low level in the early stages. However, it increased rapidly from the middle of Sep., and reached its highest value in the middle of Oct. Three peaks were obtained for PAL activity. The first peak, with the highest value was in early to middle Jul. The second one, with the lowest value, was in late Aug., and the last one, with a higher value and the longest period, was after the middle of Sep. UFGT activity had a speedy increase in the early stages and continued to increase after May. However, the net increase remained low. CHS activity was enhanced during all stages and showed two obvious peaks. The first time was in early Jul., and the other, with a higher value, was in the middle of Nov. Although PAL activity had a close relationship with the flavonoid content, no notable correlation could be identified between them. A significant correlation between CHS activity and the flavonoid content showed that CHS was one of the key enzymes involved in flavonoid biosynthesis. The relationship between flavonoid content and other related enzymes is discussed in this study. [ABSTRACT FROM AUTHOR]

The flavonoid content determines the quality of Ginkgo biloba that can be increased by using of plant growth regulators. The objective of study was to observe the effect of 5-aminolevulinic acid (ALA), a key precursor in the biosynthesis of porphyrins and a new plant growth regulator, on photosynthetic rate, chlorophyll and soluble sugar content, flavonoid accumulation, and flavonoid enzyme activity in G. biloba leaves. The ginkgo seedlings were grown in greenhouse conditions under low levels (10 and 100 mg l-1 ) of foliar application of ALA. Photosynthetic rates of leaves increased significantly at day 4 in response to both ALA concentrations and remained elevated as compared to control for further 12 days. Chlorophyll and soluble sugar contents were significantly increased by day 4 and continued to increase by day 16; however, Chl a/b ratio remained unchanged. Total polyphenols, flavonoids, and anthocyanins, phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS) and chalcone isomerase (CHI) activities were increased from day 4 to 16 after ALA treatment. The increase in chlorophyll and soluble sugar contents, and activities of flavonoid enzymes (PAL, CHS and CHI) were likely to be closely associated with improvement of the accumulation of total polyphenols, flavonoids, anthocyanins and advance of leaf quality by ALA treatment. Foliar treatment with a low concentration of ALA therefore, might provide a useful means of improving pharmacological properties of G. biloba leaves. [ABSTRACT FROM AUTHOR]

Ginkgolide is one of the important secondary metabolites of Ginkgo biloba. The synthesis of plant secondary metabolites is influenced by exogenous phytohormones. In this study, ten-year-old ginkgo grafted seedlings were used as the test material, and the effects of 0, 2, 4, 6, 8, and 10 mmol L-1 gibberellin (GA3) on endogenous indole acetic acid (IAA), abscisic acid (ABA), GA3, and ginkgolide contents in ginkgo leaves were investigated. Results showed that exogenous GA3 treatment inhibited endogenous GA3, changed the contents of endogenous IAA and ABA in ginkgo leaves, and affected the accumulation of secondary metabolite ginkgolides. Among the given concentrations, the 4 mmol L-1 GA3 treatment could remarkably increase ginkgolide content. We found that the best harvesting period of ginkgo leaves were in late August and late September, in which the content of ginkgolides was the highest. [ABSTRACT FROM AUTHOR]

Fungal diseases of plants continue to contribute to heavy crop losses in spite of the best control efforts of plant pathologists. Breeding for disease-resistant varieties and the application of synthetic chemical fungicides are the most widely accepted approaches in plant disease management. CMN1308 strain of Bacillus amyloliquefaciens was isolated from healthy Chinese chestnut fruit, which has antibiosis and induced resistance to the prevention mechanism of pathogenic bacterium after picking. In order to evaluate the antibiosis mechanism, CMN1308 was fostered with the method of confront culture. The antimicrobial components were also isolated from the culture of CMN1308, and their stability and antimicrobial activity was tested under different treatments such as temperature, pH and UV. The results showed that CMN1308 displays advantages in regard to spatial competition against the major pathogens of chestnut, Rhizopus stolonifer, Fusarium solani, Stachybotrys chartarum, Cryphonectria parasitica, Lasiodiplodia theobromae, Penicillium expansum and Aspergillus niger. Among this, CMN1308 had the best antimicrobial activity against P. expansum, with the inhibition zone diameter of 27.1 mm. The antimicrobial material isolated from CMN1308 culture showed a strong inhibition to the growth of P. expansum hyphal and also had a good stability to high temperature, alkali media and UV, but was sensitive to acidic conditions. Furthermore, CMN1308 increased the peroxidase, polyphenol oxidase activity and reduced the MDA content in chestnuts after infecting by pathogenic fungi. Thus, producing antibiotic compounds and inducible resistance are the main factors that may explain the antibacterial mechanism of CMN1308 on chestnut pathogenic. The results of this study might help to optimize the practical use of CMN1308 in the biological control of chestnut rot or other fruit rot infected by pathogenic fungi. [ABSTRACT FROM AUTHOR]

Terpene trilactones (TTL) is a pharmacological ingredient in Ginkgo biloba and its content has become one of the key indices for medicinal value evaluation of ginkgo. 1-Deoxy-D-xylulose 5-phosphate reductoisomerase (DXR) catalyzes the first step specific for isopentenyl diphosphate production in methylerythritol phosphate pathway, which provide the basic structure required for TTLs biosynthesis. To understand the mechanism controlling the GbDXR gene expression, the GbDXR promoter sequence was isolated and subjected to transient expression with the green fluorescent protein (GFP) in tobacco plants. Characteristic analysis revealed various cis -acting elements that related to light-regulated transcription, hormone signaling (auxin, ethylene), adversity stress and defense signaling (heat/dehydration stress) in the GbDXR promoter region. In transient expression assay, deletion of different portions of the upstream GbDXR promoter identified that the promoter region -3230bp to -865bp conserve the positive regulation function, which could promote the expression of GFP in the cytoplasm of tobacco leaf epidermal cells. The regulation function of the promoter region -865bp to -262bp remained to be elucidated. EMSA analysis suggested possible interactions of GbERF10 and GbERF17 with the ERF-binding elements in the upstream of GbDXR promoter. For abiotic stresses treatment, the expression of GbDXR gene could be significantly induced by UVB and drought stress. In general, the GbDXR gene expressed differently in different ginkgo tissues but exhibited the highest transcriptional level in the root, with the maximum TTLs content simultaneously. The positive relationship between gene expression level and TTLs content indicated that the GbDXR is responsible for TTLs biosynthesis in G. biloba. [ABSTRACT FROM AUTHOR]

Four separation methods of antimicrobial substances produced by CMN1308 (Bacillus amyloliquefaciens) were evaluated and selected according to number of antimicrobial substances and its activity in vitro. The results showed that extraction by acid precipitation of the fermentation supernatant of CMN1308 was the best with a diameter of inhibition zone of pathogen fungi P. expansum of 12.3 mm in a laboratory bioassay. Applying a silica thin layer chromatography (TLC), SDS-PAGE and other separation technologies we isolate antimicrobial substances, and the separated band were cut off for mass spectrometry analysis. The TLC of crude extract of CMN1308 show a topical band corresponding with the surfactin standard (Rf value =0.75), proved that the strain CMN1308 can produce this surface active compound. The mycoprotein extracted from CMN1308 was separated by Tricine-SDS-PAGE modified with the addition of urea in the separation gel. After mass spectrometric analysis and protein characterization, the isolated mycoprotein showed a maximum ion peak at M/Z of 2679 and molecular weight of 29.5 kDa, matching with protein flagellin. The extracellular antimicrobial protein of strain CMN1308 display four bands after urea-Tricine-SDS-PAGE, but after mass spectrometry analysis only two bands were identified. Band "A" with a maximum ion peak at M/Z of 1926 and molecular weight of 49.8 kDa, aligned with NCBI database, matching with DLDH (dihydrolipoamide dehydrogenase enzyme). Band "D" show the maximum ion peak at M/Z of 2936 and molecular weight of 22.4 kD, matching with a chitin binding protein. Thus, the strain CMN1308 has the potential to be developed as a commercial biological control agent for chestnut common pathogenic fungi. [ABSTRACT FROM AUTHOR]

The terpene trilactones (TTLs) are believed to be important for the pharmacological properties of Ginkgo biloba leaves extract. 3-Hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) is a critical enzyme involved in the biosynthetic pathway of TTLs. In this study, an 1.22kb fragment of 5' flanking region of the HMGR gene (GbHMGR), was isolated from G. biloba by genome walking. Extensive sequence analysis revealed the presence of evolutionarily conserved and over-represented putative cis-acting elements in light-regulated transcription, hormone signaling (gibberellic acid, jasmonate and salicylic acid), elicitor and stress responses (cold/dehydration responses), and plant defense signaling (W-box/WRKY) that are common to the promoter region of GbHMGR. EMSA analysis suggested possible functionality of W-box in GbHMGR promoter region. The behavior of gene transcripts in ginkgo callus upon light, low temperature, MeJA and SA treatments further verified the regulatory function of GbHMGR promoter. A significant positive relationship between gene expression level and total TTL contents suggested that GbHMGR might be one of key genes involved in TTL biosynthesis in G. biloba. [ABSTRACT FROM AUTHOR]

As a kind of transcription factors gene family, MADS-box genes play an important role in plant development processes. To find genes involved in the floral transition of Ginkgo biloba, a MADS-box gene, designated as GbMADS2, was cloned from G. biloba based on EST sequences by RT-PCR. Sequence analysis results showed that the cDNA sequence of GbMADS2 contained a 663 bp length ORF encoding 221 amino acids protein, which displayed typical structure of plant MADS-box protein including MADS, I, and K domains and C terminus. The sequence of GbMADS2 protein was highly homologous to those of MADS-box proteins from other plant species with the highest homologous to AGAMOUS (CyAG) from Cycas revoluta. The phylogenetic tree analysis revealed that GbMADS2 belonged to AGAMOUS clade genes. Real-time PCR analysis indicated that expression levels of GbMADS2 gene in female and male flower were significantly higher than those in root, stem, and leaves, and that GbMADS2 expression level increased along with time of flower development. The spatial and time-course expression profile of GbMADS2 implied that GbMADS2 might be involved in development of reproductive organs. The isolation and expression analysis of GbMADS2 provided basis for further studying the molecular mechanism of flower development in G. biloba. [ABSTRACT FROM AUTHOR]

Recently, there has been an increasing interest among researchers in using combinations of biological control agents to exploit potential synergistic effects among them. In the present study, there were investigated commercially acceptable formulations of Bacillus cereus CE3 wetting powder with long storage life and retained efficacy to control chestnut and other fruit rot caused by Endothia parasitica (Murr) and Fusarium solani. The study sought to develop a new B. cereus formulation that would be more effective and better suited to the conditions of field application. By a series of experiments, the formulation was confirmed as follows: 60% B. cereus freeze5dried powder, 28.9% diatomite as carrier, 4% sodium lignin sulfonate as disperser, 6% alkyl naphthalene sulfonate as wetting agent, 1% K2HPO4 as stabilizer, 0.1% β-cyclodextrin as ultraviolet protectant. The controlling experiments showed that the diluted 100 times of 60% B. cereus wetting powder had 79.47% corrosion rate to chestnut pathogens; and this result is comparable to the diluted 1,000 times of 70% thiophanate5 methyl. Safety evaluation results showed that rats acute oral lethal dose 50% was 5,000.35, therefore application of B. cereus wettable powder could not cause a person or animal poisoning. This work illustrated that 60% B. cereus wetting powder had commercial potential; however, to apply this formulation as a biological pesticide in the field, masses production processes need to be further studied. [ABSTRACT FROM AUTHOR]

The flavonoids content determines the quality characteristics of Ginkgo biloba extract that could be increased by using of plant growth regulators. The objective of study was to investigate the effect of chlorocholine chloride (CCC), an anti-gibberellin growth retardant, on photosynthesis, chlorophyll, soluble sugar, total amino acids and phenylalanine contents, flavonoid accumulation, and flavonoids enzyme activity in G. biloba leaves. The ginkgo seedlings were grown in the greenhouse conditions with foliar applications of 0 (control), 0.5, 1.0 and 2.0 g l-1 CCC. Results showed that 0.5, 1.0 and 2.0 g l-1 CCC treatments significantly increased photosynthetic rates of leaves, the contents of chlorophyll, soluble sugar, total amino acids and phenylalnine in ginkgo leaves. Total polyphenols, flavonoids, anthocyanins content, phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS) and chalcone isomerase (CHI) activities were all significantly increased by 1.0 and 2.0 g l-1 CCC treatments. Foliar treatment with CCC therefore might be a useful means of improving pharmacological properties of G. biloba leaves. [ABSTRACT FROM AUTHOR]