Effects of Temperature on Systemic Infection and Symptom Expression of Turnip mosaic virus in Chinese cabbage (Brassica campestris).

Using the Chinese cabbage (Brassica campestris) cultivar 'Chun-goang' as a host and turnip mosaic virus (TuMV) as a pathogen, we studied the effects of ambient temperature (13°C, 18°C, 23°C, 28°C and 33°C) on disease intensity and the speed of systemic infection. The optimal temperature for symptom expression of TuMV was 18-28°C. However, symptoms of viral infection were initiated at 23-28°C and 6 days post infection (dpi). Plants maintained at 33°C were systemically infected as early as 6 dpi and remained symptomless until 12 or 22 dpi, depending on growth stage at the time of inoculation. It took 45 days for infection of plants grown at 13°C. Quantitative real-time polymerase chain reaction (q-PCR) results showed that the accumulation of virus coat protein was greater in plants grown at 23-28°C. The speed of systemic infection increased linearly with rising ambient temperature, up to 23°C. The zero-infection temperature was 10.1°C. To study the effects of abruptly elevated temperatures on systemic infection, plants inoculated with TuMV were maintained at 10°C for 20 d; transferred to a growth chamber at temperatures of 13°C, 18°C, 23°C, 28°C, or 33°C for 1, 2, or 3 d; and then moved back to 10°C. The numbers of plants infected increased as duration of exposure to higher temperatures and dpi increased.

Gibberellin application at pre-bloom in grapevines down-regulates the expressions of VvIAA9 and VvARF7, negative regulators of fruit set initiation, during parthenocarpic fruit development.

Fruit set is initiated only after fertilization and is tightly regulated primarily by gibberellins (GAs) and auxins. The application of either of these hormones induces parthenocarpy, fruit set without fertilization, but the molecular mechanism underlying this induction is poorly understood. In the present study, we have shown that the parthenocarpic fruits induced by GA application at pre-bloom result from the interaction of GA with auxin signaling. The transcriptional levels of the putative negative regulators of fruit set initiation, including Vitis auxin/indole-3-acetic acid transcription factor 9 (VvIAA9), Vitis auxin response factor 7 (VvARF7), and VvARF8 were monitored during inflorescence development in seeded diploid 'Tamnara' grapevines with or without GA application. Without GA application, VvIAA9, VvARF7, and VvARF8 were expressed at a relatively high level before full bloom, but decreased thereafter following pollination. After GA application at 14 days before full bloom (DBF); however, the expression levels of VvIAA9 and VvARF7 declined at 5 DBF prior to pollination. The effects of GA application on auxin levels or auxin signaling were also analyzed by monitoring the expression patterns of auxin biosynthesis genes and auxin-responsive genes with or without GA application. Transcription levels of the auxin biosynthesis genes Vitis anthranilate synthase ß subunit (VvASB1-like), Vitis YUCCA2 (VvYUC2), and VvYUC6 were not significantly changed by GA application. However, the expressions of Vitis Gretchen Hagen3.2 (VvGH3.2) and VvGH3.3, auxin-responsive genes, were up-regulated from 2 DBF to full bloom with GA application. Furthermore, the Vitis GA signaling gene, VvDELLA was up-regulated by GA application during 12 DBF to 7 DBF, prior to down-regulation of VvIAA9 and VvARF7. These results suggest that VvIAA9 and VvARF7 are negative regulators of fruit set initiation in grapevines, and GA signaling is integrated with auxin signaling via VvDELLA during parthenocarpic fruit development in grapevines.

Transcriptional changes of gibberellin oxidase genes in grapevines with or without gibberellin application during inflorescence development.

The concept that gibberellin (GA) application on seeded grapevines induces seedlessness has been known for decades in viticulture. GA was applied to inflorescence clusters of seeded diploid grapevine cultivar 'Tamnara' (Vitis spp.) at 14 days before full bloom (DBF). Morphological and molecular effects of GA application were examined on the induction of parthenocarpic fruit development. With GA application, ovaries were enlarged and pollen tube growth was completely inhibited. Vitis GA oxidase enzymes, key determinants for GA level, were characterized through phylogenetic analysis with Arabidopsis GA oxidase enzymes. Five VvGA 20-oxidase (VvGA20ox), three VvGA 3-oxidase (VvGA3ox), and nine VvGA 2-oxidase (VvGA2ox) family proteins, and one VvGA methyltransferase (VvGAMT) and one Vitis cytochrome P450 714A1 proteins were identified, and their expression patterns were analyzed during inflorescence development from 14 DBF to 5 days after full bloom (DAF). VvGA2ox1, VvGA20ox3, and VvGA3ox2 were the most abundantly expressed genes in each gene family at 7, 5, and 2 DBF, respectively. Following GA application at 14 DBF inducing seedlessness, GA catabolic genes such as VvGAMT2, VvGA2ox3, and VvGA2ox4 were up-regulated at 12 DBF, full bloom, and 5 DAF, respectively. Conversely, most GA biosynthetic genes, VvGA20oxs and VvGA3oxs, were down-regulated at near full bloom, and the timing of their peak expression was changed. These results suggest that GA application at pre-bloom changes the GA biosynthesis into GA catabolic pathway at near full bloom by altering the transcription level and timing of GA oxidase genes during grapevine inflorescence development.

Fruit quality, anthocyanin and total phenolic contents, and antioxidant activities of 45 blueberry cultivars grown in Suwon, Korea.

Blueberry fruits from 45 commercial cultivars (39 northern highbush and 6 half highbush blueberry) grown in Suwon, Korea were analyzed for fruit size, soluble solids content, titratable acidity, total anthocyanin content, total phenolic content, and antioxidant activity. Fruit characteristics varied widely among the 45 blueberry cultivars. Fruit weight ranged from 0.9 to 3.6 g, soluble solids content from 8.3 to 14.3 °Brix, and titratable acidity from 0.8% to 3.6%. Antioxidant activity ranged from 0.7 to 2.1 mg of quercetin equivalents per gram of fresh berries in different blueberry cultivars. Among the 45 blueberry cultivars, high amounts of anthocyanins and polyphenols, and high antioxidant activity were observed in 'Elliott', 'Rubel', 'Rancocas', and 'Friendship'.

Transcriptional profiling of ESTs responsive to Rhizobium vitis from 'Tamnara' grapevines (Vitis sp.).

Genes related with defense responses were screened from the cDNA library constructed with Rhizobium vitis-inoculated or salicylic acid (SA)-treated 'Tamnara' grapevine (Vitis sp.) leaves. Among 13,728 expressed sequence tags (ESTs) from 'Tamnara' grapevine upon R. vitis inoculation and SA treatment, 6776 unigenes containing 1915 contigs and 4860 singletons were obtained. In gene ontology analysis, there were about 3200 clones related with biological process, 3555 with molecular function, and 3354 with cellular component genes. Proteins of secretory organ (35%), plasma membrane (30%), endoplasmic reticulum (20%), and vacuole (11%) were predicted. Photosynthesis-related genes and defense-related genes were most abundant. Among ESTs, 199 resistance-related ones were mapped to the genome of Vitis vinifera L. with three markers, GLP1-12, MHD98, and MHD145, which are known to be linked to resistance against powdery mildew. Approximately, 120 simple sequence repeats (SSRs) detected in cDNAs could be used as EST-derived SSR markers in disease resistant grape breeding.

Crystallization and preliminary X-ray diffraction studies of alpha-cyclodextrin glucanotransferase isolated from Bacillus macerans.

Single crystals of alpha-cyclodextrin glucanotransferase isolated from Bacillus macerans have been grown with polyethylene glycol 6000 as a precipitating agent by sitting-drop vapour diffusion at room temperature. The crystals were suitable for X-ray analysis and diffracted to at least 2.0 A (space group P2(1)2(1)2(1)), with unit-cell parameters a = 66.79 (2), b = 79.66 (1), c = 141.16 (1) A. Assuming the asymmetric cell to be occupied by a monomer of 74 kDa, the unit cell contains 42.6% solvent with a crystal Volume per protein mass, V(M), of 2.53 A(3) Da(-1).