Comparative analysis of the difference in flavonoid metabolic pathway during coloring between red-yellow and red sweet cherry (Prunus avium L.).

The color of a fruit is an important contributor to the perception of its nutritional value. It is widely acknowledged that the color of sweet cherry changes obviously during ripening. Variations in anthocyanins and flavonoids account for the heterogeneous color of sweet cherries. In this study, we showed that anthocyanins but not carotenoids determine the color of sweet cherry fruits. The difference between red-yellow and red sweet cherry may be attributed to seven anthocyanins, including Cyanidin-3-O-arabinoside, Cyanidin-3,5-O-diglucoside, Cyanidin 3-xyloside, Peonidin-3-O-glucoside, Peonidin-3-O-rutinoside, Cyanidin-3-O-galactoside, Cyanidin-3-O-glucoside (Kuromanin), Peonidin-3-O-rutinoside-5-O-glucoside, Pelargonidin-3-O-glucoside and Pelargonidin-3-O-rutinoside. The content of 85 flavonols differed between red and red-yellow sweet cherries. The transcriptional analysis identified 15 key structural genes involved in the flavonoid metabolic pathway and four R2R3-MYB transcription factors. The expression level of Pac4CL, PacPAL, PacCHS1, PacCHS2, PacCHI, PacF3H1, PacF3H2, PacF3'H, PacDFR, PacANS1, PacANS2, PacBZ1 and four R2R3-MYB were positively correlated with anthocyanin content (ps < 0.05). PacFLS1, PacFLS2 and PacFLS3 expression was negatively correlated with anthocyanin content but positively correlated with flavonols content (ps < 0.05). Overall, our findings suggests that the heterogeneous expression of structural genes in the flavonoid metabolic pathway accounts for the variation in levels of final metabolites, leading to differences between red 'Red-Light' and red-yellow 'Bright Pearl'.

Phylogeny and pathogenicity of Fusarium spp. isolated from greenhouse melon soil in Liaoning Province.

Fungi of the Fusarium oxysporum are widely distributed around the world in all types of soils, and they are all anamorphic species. In order to investigate the relationships and differences among Fusarium spp., 25 Fusarium spp. were isolated from greenhouse melon soils in Liaoning Province, China. With these 25 strains, three positive control Fusarium strains were analyzed using universally primed PCR (UP-PCR). Seventy-three bands appeared after amplification using 6 primers, and 66 of these bands (90.4%) were polymorphic. All strains were clustered into eight groups, though 14 strains of F. oxysporum were clustered into a single group. We concluded that UP-PCR could reveal the genetic relationships and differences among Fusarium strains. Moreover, the UP-PCR results suggested that F. oxysporum is distinguishable from other Fusarium spp. Thus, UP-PCR is a useful method for Fusarium classification. The pathogenicity of 13 strains of F. oxysporum to muskmelon, cucumber and watermelon seedlings was studied by infecting the seedlings with a spore suspension after cutting the root. The results showed that the F. oxysporum strains were pathogenic to all three melon types, although the pathogenicity differed significantly among the 13 strains. In addition, all strains had the greatest pathogenicity toward watermelon. Since the factors affecting pathogenicity vary widely, they should be considered in future studies on Fusarium spp. The results of such studies may then yield an accurate description of the pathogenicity of Fusarium spp.

Construction of a new GFP vector and its use for Fusaruim oxysporum transformation.

In this study, the gfp fragment as a reporter gene had integrated into the form plasmid vector pBC-hygro which contains an expressive promoter of the fungus to facilitate the transformation of Fusarium oxysporum. The resultant plasmid pBC-hygro-GFP was identified by digestion with enzymes. Binary plasmids pBC-hygro-GFP were transformed into F. oxysporum by using the PEG-CaCl2 mediated transformation technique. Results show that the recombinant plasmid pBC-hygro-GFP was constructed correctly. The gfp gene was stably maintained and did not convey any significant loss of phenotype which would affect the survival and behaviour of the tagged strains. Introduction of the gfp gene into F. oxysporum provides a simple, specific and cost-effective method of strain identification for ecological studies. Transcriptional reporter vectors were constructed for using the green fluorescent protein (GFP) reporter.

Diversity analysis of type I ketosynthase in rhizosphere soil of cucumber.

Fusarium wilt [Fusarium oxysporum (Sch1.) f.sp. cucumerinum Owen.] is a major soil-borne disease of cucumber worldwide, and can cause huge yield losses. Biological control of Fusarium wilt of cucumber has received considerable attention. Many bacteria, particularly actinomycetes, are known to produce secondary metabolites synthesized by Polyketide synthases (PKSs) with a diverse range of biological activities. Ketosynthase (KS) gene diversity was analyzed in samples which were collected from rhizosphere soil of both diseased cucumber and healthy cucumber in Dalian, China. The phylogenetic analysis amino acid (AA) sequences indicated that the KS genes in the rhizosphere soil samples were clustered into diverse seven clades, including Sorangium cellulosum, Anabaena variabilis, Nostoc punctiforme, Xanthobacter autotrophicus, Streptomyces, myxobacteria and uncultured bacteria. Among seven major clades in the phylogenetic tree, two clades were peculiar to rhizosphere soil of diseased cucumber and one was peculiar to healthy cucumber. Among the 182 cloned KS genes, 147 KS genes were clustered with the uncultured bacteria group. Most of the KS genes showed about 80% similarity at the AA level to sequences known in GenBank. These results revealed the great diversity and novelty of KS genes in rhizosphere soil of cucumber.

[UP-PCR diversity analysis of Fusarium population isolated from greenhouse melon soils].

A total of 112 Fusarium isolates were obtained from 36 soil samples collected from the greenhouse melon fields of Liaoning Province, among which, 11 species were identified by traditional morphological classification and rDNA sequence analysis. Universally Primed PCR (UP-PCR) was conducted to analyze the 25 strains of test Fusarium isolates and 3 strains of positive control Fusarium isolates. The results indicated that a total of 73 bands appeared after amplification by using 6 primers, and 66 bands (90.4%) were polymorphic. The isolates were clustered into eight groups at the similarity of 0.736 by cluster analysis, among which, 14 isolates were clustered into one group. It was concluded that UP-PCR could present the genetic relationship and difference among Fusarium strains, being able to be used as an assistant method for Fusarium classification.