• Journal of Plant Biotechnology
• /
• v.43 no.1
• /
• pp.37-48
• /
• 2016

The transcriptional profiles of 'Tamnara' grapevine (Vitis labruscana L.) to Rhizobium vitis were determined using 12,000 gene oligonucleotide microarray chips constructed with 6,776 unigenes based on the EST sequencing. Among them, 95 clones were up-regulated more than three times and 90 were down-regulated more than 5-times in the R. vitis-inoculated grapevines relative to the control vines. Treatment of salicylic acid showed that 337 clones were upregulated and 52 clones were down regulated in grapevines. Microarray analysis, reverse transcription-polymer chain reaction, and slot blot hybridization analysis revealed that 5, 14, and 64 clones were up-regulated and 10, 12, and 61 clones were down-regulated in wounded, salicylic acid-treated, and R. vitis-inoculated 'Tamnara' grapevine leaves, respectively. The expression patterns of ${\beta}$-1,3-glucanase, proline-rich protein, and lipoxygenase genes of 'Tamnara' moderately resistant to R. vitis were similar to those of resistant 'Concord' and 'Delaware' grapevines. However, chalcone synthase genes in 'Tamnara' grapevines showed similar expression patterns to susceptible grapevines 'Neomuscat' and 'Rizamat'. Further expression studies with various clones for each gene should be conducted to elucidate their roles in resistant responses against pathogens or other stimuli in grapevines. These results could provide better resources for understanding the mechanism of defense responses against crown gall disease and clues for identifying new genes that may play a role in defense against R. vitis in grapevines.

• The Plant Pathology Journal
• /
• v.32 no.4
• /
• pp.347-356
• /
• 2016

Plants protect themselves from pathogen attacks via several mechanisms, including hypersensitive cell death. Recognition of pathogen attack by the plant resistance gene triggers expression of carboxylesterase genes associated with hypersensitive response. We identified six transcripts of carboxylesterase genes, Vitis flexuosa carboxylesterase 5585 (VfCXE5585), Vf-CXE12827, VfCXE13132, VfCXE17159, VfCXE18231, and VfCXE47674, which showed different expression patterns upon transcriptome analysis of V. flexuosa inoculated with Elsinoe ampelina. The lengths of genes ranged from 1,098 to 1,629 bp, and their encoded proteins consisted of 309 to 335 amino acids. The predicted amino acid sequences showed hydrolase like domains in all six transcripts and contained two conserved motifs, GXSXG of serine hydrolase characteristics and HGGGF related to the carboxylesterase family. The deduced amino acid sequence also contained a potential catalytic triad consisted of serine, aspartic acid and histidine. Of the six transcripts, Vf-CXE12827 showed upregulated expression against E. ampelina at all time points. Three genes (VfCXE5585, VfCXE12827, and VfCXE13132) showed upregulation, while others (VfCXE17159, VfCXE18231, and VfCXE47674) were down regulated in grapevines infected with Botrytis cinerea. All transcripts showed upregulated expression against Rhizobium vitis at early and later time points except VfCXE12827, and were downregulated for up to 48 hours post inoculation (hpi) after upregulation at 1 hpi in response to R. vitis infection. All tested genes showed high and differential expression in response to pathogens, indicating that they all may play a role in defense pathways during pathogen infection in grapevines.

• Journal of Plant Biotechnology
• /
• v.44 no.2
• /
• pp.125-134
• /
• 2017

Enhanced disease susceptibility1 (EDS1) is a regulator of basal defense responses required for resistance mediated by TIR-NBS-LRR containing R proteins. We identified three transcripts of EDS1-like genes encompassing diverse/separate expression patterns, based on the transcriptome analysis by Next Generation Sequencing (NGS) of V. flexuosa inoculated with Elsinoe ampelina. These genes were designated VfEDL1 (Vitis flexuosa Enhanced Disease Susceptibility1-like1), VfEDL2 and VfEDL3, and contained 2464, 1719 and 1599 bp, with 1791, 1227 and 1599 bp open reading frames (ORFs), encoding proteins of 596, 408 and 532 amino acids, respectively. The predicted amino acid sequences of all three genes showed the L-family lipase-like domain (class 3 lipase domain), and exhibited a potential lipase catalytic triad, aspartic acid, histidine and serine in the conserved G-X-S-X-G. All three VfEDL genes were upregulated at 1 hpi against the bacterial and fungal pathogens Rizhobiumvitis and E. ampelina, respectively, except VfEDL1, which was downregulated against E. ampelina at all time points. Against E. ampelina, VfEDL2 and VfEDL3 showed downregulated expression at later time points. When evaluated against R. vitis, VfEDL1 showed downregulated expression at all time points after 1 hpi, while VfEDL3 showed upregulation up to 24 hpi. Based on the expression response, all three genes may be involved in plant resistant responses against R. vitis, and VfEDL2 and VfEDL3 show additional resistant responses against E. ampelina infection.

• Horticultural Science & Technology
• /
• v.34 no.4
• /
• pp.537-548
• /
• 2016

The objective of this study was to evaluate the antimicrobial activities of 9 kinds of medicinal plants against crown gall in grapevine. The medicinal plants extracted with several solvent systems were screened for in vitro antibacterial activity by the disc diffusion method. The ethanol and ethyl acetate extracts from magic lily flowers, tachys roots, asian plantain flowers and seeds, sweet wormwood leaves, stems and flowers, immature bitter melon fruits, cockscomb flowers, and peach tree resin showed in vitro antimicrobial activities against Rhizobium vitis with growth inhibition zones ranging from 10 to 27 mm in diameter. The minimum inhibitory concentration values of extracts against R.vitis ranged from 10,000 in Asian plantain flower and 50,000 fold diluted extracts in sweet wormwood flowers, stems, leaves, cockscomb leaves and immature bitter melon fruits. The active fractions of ethyl acetate and ethanol extracts from the medicinal plants were partially separated through silica gel column chromatography and thin layer chromatography (TLC). The active fractions were separated at Rf 0.36, 0.69, 0.75, 0.84, and 0.94 in sweet wormwood extracts, Rf 0.96 and 0.99 in cockscomb flower extracts, Rf 0.92 and 0.97 in cockscomb leaf extracts, and Rf 0.85 in immature bitter melon fruit extracts in TLC analysis developed with hexane:ethyl acetate (20:80, v/v) and methanol:chloroform (20:80, v/v). Among extracts from plants with in vitro antimicrobial activities, sweet wormwood, cockscomb leaves, and immature bitter melon fruits showed in vivo antimicrobial activities with inhibition activity of 100, 67, and 83.3%, respectively, in 'Kyoho' grapevine inoculated with R. vitis compared with the untreated control. These findings indicate that extracts of medicinal plants could be used as sustainable candidates to control crown gall disease caused by R. vitis in grapevines.

• Korean Journal of Soil Science and Fertilizer
• /
• v.35 no.1
• /
• pp.12-26
• /
• 2002

The phylogenetic relationships for 33 strains belonging to the genera Azorhizobium, Bradyrhizobium, Mesorhizobium, Rhizobium and Sinorhizobium were conducted by the sequence analyses of the ITS regions. The sequence homologies of these strains showed the high variations(28.0 - 94.9%). According to the phylogenetic analysis of ITS regions. 37 ITS clones from 33 strains of 32 species were classified into four groups. Group I included all strains of the genus Sinorhizobium as core members and R. giardinii as a peripheral member. The genus Rhizobium strains were clustered into group II which was very heterogeneous and the tree toplogy of this group were very unstable. Among the members of group II. the taxonomic position of R. radiobacter and R. rubi was not clearly identified on the basis of ITS I regions. R. undicola and R. vitis were remotely related with other Rhizobium strains including R. leguminosarum, R. galegae, R. gallicum, R. mongolense, R. tropici, R. hainanense, R. rhizogense and R. huautlense of group II were supposed to be loosely related to R. leguminosarum. While the stains of the genera Bradyrhizobium constituted group III with Azorhizobium caulindans, the strains of the genus Mesorhizobium formed group IV on the relatively high sequence homology level.