Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China

Yangling, China

Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China

Yangling, China
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He J.,Northwest Agriculture and Forestry University | He J.,Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China | Wang D.,Northwest Agriculture and Forestry University | Wang D.,Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China | And 4 more authors.
Agri Gene | Year: 2016

Stilbene synthase plays important roles in the secondary metabolic pathways of many plants as polyketide synthase. Increased consumption of its metabolite, resveratrol, is highly beneficial to our health. The stilbene synthase 6 gene (VqSTS6) has been obtained from the Chinese wild Vitis quinquangularis accession ‘Danfeng-2’. Our previous studies have shown that this stilbene synthase gene exhibits higher levels of expression than other VqSTSs in the all six grape organs. To characterize gene expression, we cloned the promoter of VqSTS6 by homologous cloning. Alignment between the VqSTS6 gene sequence and Vitis vinifera VvSTS48 showed 99.24% identity. The promoter of VqSTS6 fused with the β-glucuronidase (GUS) gene was transformed into tomato (Solanum lycopersicum L. ‘Micro-Tom’) and the activity expression in four organs (roots, stems, leaves and fruits) was analyzed. This revealed that expression was much higher in the fruit than in the other three organs. We also used promoter deletion to show that the –518 bp to –411 bp region contained skn-1_motifs are the core functional promoter region in fruit development. These results evidence the special role of VqSTS6 promoter in regulating its expression in a high-resveratrol grapevine. © 2016 Elsevier Inc.


Xie X.,Northwest Agriculture and Forestry University | Xie X.,Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China | Aguero C.B.,University of California at Davis | Wang Y.,Northwest Agriculture and Forestry University | And 2 more authors.
Plant Cell, Tissue and Organ Culture | Year: 2016

A protocol was standardized to regenerate six grape cultivars through meristematic bulk (MB) induction, which was used for genetic transformation. Meristematic bulk induction worked best with Vitis vinifera ‘Thompson Seedless’ (98.4 %), followed by ‘Chardonnay’ (97.6 %), ‘Redglobe’ (90.2 %) and ‘Cabernet Sauvignon’ (86.2 %), and was less successful with Vitis rupestris ‘St. George’ (85.4 %) and ‘101-14 Millardet et de Grasset (Vitis riparia × V. rupestris)’ (79.6 %). Benzylaminopurine and naphthaleneacetic acid was the most effective combination of cytokinin and auxin for MB formation. 100 µg/ml kanamycin was a better antibiotic selection agent than 2.0 µg/ml hygromycin during transformation. The expression of green fluorescent protein was evaluated with in vitro leaves and roots. Transformation efficiency using meristematic slices was a function of the genotype. Transformation efficiency was greatest in Chardonnay (51.7 %), followed by Thompson Seedless (42.3 %), St. George (41.6 %), Redglobe (40 %), Cabernet Sauvignon (35.6 %) and 101-14 Mgt (29.9 %). This study found that MB induction was a fast and simple alternative for genetic transformation of grape cultivars. © 2016 Springer Science+Business Media Dordrecht


Ji W.,Northwest Agriculture and Forestry University | Ji W.,Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China | Wang Y.,Northwest Agriculture and Forestry University | Wang Y.,Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China
Journal of the Science of Food and Agriculture | Year: 2013

BACKGROUND: Since 1982, the embryo rescue technique has been widely applied to embryo germination of stenospermic grapes in cross-breeding programmes. This project aimed to: (1) use embryos to breed new seedless cultivars of Vitis vinifera as the female parents utilising wild Chinese Vitis spp. as the male parents; and (2) develop an efficient method for in vitro embryo rescue and plant development. RESULTS: Among the different genotypes, the productions of hybrid plants were significantly different, ranged from 21.1% ('Ruby Seedless' × 'Beichun') to only 1.1% ('Pink Seedless' × 'Beichun'), except for the combinations from which no surviving seedlings were obtained. We collected hybridisation fruits from 28 June to 3 August, and obtained their best sampling times described within days after flowering. The highest rates of embryo formation (24.3%) and plant development (91.4%) were found when ovules of 'Ruby Seedless' were cultured in MM4 + 500 mg L-1 mashed banana. CONCLUSION: Seven new hybrids of V. vinifera with wild Chinese Vitis spp. were obtained. As a result of early nuclear-free character identification, 17 seedless grape lines were obtained. An efficient system of seedless grape breeding through embryo rescue was also established. © 2013 Society of Chemical Industry.


Yu Y.,Northwest University, China | Yu Y.,Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China | Xu W.,Northwest University, China | Xu W.,Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China | And 10 more authors.
Journal of Experimental Botany | Year: 2011

RING finger proteins comprise a large family and play important roles in regulation of growth and development, hormone signalling, and responses to biotic and abiotic stresses in plants. In this study, the identification and functional characterization of a C4C4-type RING finger protein gene from the Chinese wild grapevine Vitis pseudoreticulata (designated VpRFP1) are reported. VpRFP1 was initially identified as an expressed sequence tag (EST) from a cDNA library constructed from leaves of V. pseudoreticulata inoculated with the grapevine powdery mildew Uncinula necator. Sequence analysis of the deduced VpRFP1 protein based on the full-length cDNA revealed an N-terminal nuclear localization signal (NLS) and a C-terminal C4C4-type RING finger motif with the consensus sequence Cys-X 2-Cys-X 13-Cys-X 1-Cys-X 4-Cys-X 2-Cys-X 10-Cys-X 2-Cys. Upon inoculation with U. necator, expression of VpRFP1 was rapidly induced to higher levels in mildew-resistant V. pseudoreticulata plants. In contrast, expression of VpRFP1 was down-regulated in mildew-susceptible V. vinifera plants. Western blotting using an antibody raised against VpRFP1 showed that VpRFP1 was also induced to higher levels in V. pseudoreticulata plants at 12-48 hours post-inoculation (hpi). However, there was only slight increase in VpRFP in V. vinifera plants in the same time frame, even though a more significant increase was observed at 96-144hpi in these plants. Results from transactivation assays in yeast showed that the RING finger motif of VpRFP1 exhibited some activity of transcriptional activation; however, no activity was seen with the full-length VpRFP1. Overexpression of VpRFP1 in Arabidopsis plants was found to enhance resistance to Arabidopsis powdery mildew Golovinomyces cichoracearum, which seemed to be correlated with increased transcript levels of AtPR1 and AtPR2 in the pathogen-infected tissues. In addition, the Arabidopsis transgenic lines showed enhanced resistance to a virulent bacterial pathogen Pseudomonas syringae pv. tomato DC3000. Taken together, the results suggested that VpRFP1 may be a transcriptional activator of defence-related genes in grapevines. © 2011 The Author.


Jiao Y.,Northwest University, China | Jiao Y.,Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China | Xu W.,Northwest University, China | Xu W.,Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China | And 4 more authors.
Journal of Experimental Botany | Year: 2016

Stilbenes are central phytoalexins in Vitis, and induction of the key enzyme stilbene synthase (STS) is pivotal for disease resistance. Here, we address the potential for breeding resistance using an STS allele isolated from Chinese wild grapevine Vitis pseudoreticulata (VpSTS) by comparison with its homologue from Vitis vinifera cv. 'Carigane' (VvSTS). Although the coding regions of both alleles are very similar (>99% identity on the amino acid level), the promoter regions are significantly different. By expression in Arabidopsis as a heterologous system, we show that the allele from the wild Chinese grapevine can confer accumulation of stilbenes and resistance against the powdery mildew Golovinomyces cichoracearum, whereas the allele from the vinifera cultivar cannot. To dissect the upstream signalling driving the activation of this promoter, we used a dual-luciferase reporter system in a grapevine cell culture. We show elevated responsiveness of the promoter from the wild grape to salicylic acid (SA) and to the pathogen-associated molecular pattern (PAMP) flg22, equal induction of both alleles by jasmonic acid (JA), and a lack of response to the cell death-inducing elicitor Harpin. This elevated SA response of the VpSTS promoter depends on calcium influx, oxidative burst by RboH, mitogen-activated protein kinase (MAPK) signalling, and JA synthesis. We integrate the data in the context of a model where the resistance of V. pseudoreticulata is linked to a more efficient recruitment of SA signalling for phytoalexin synthesis. © 2016 The Author 2016. Published by Oxford University Press on behalf of the Society for Experimental Biology.


Xu L.-F.,Northwest University, China | Xu L.-F.,Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China | Zhou P.,Northwest University, China | Zhou P.,Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China | And 5 more authors.
Journal of Integrative Agriculture | Year: 2013

The soil organic matter and nutrients are fundamental for the sustainability of pear production, but little is known about the spatial distribution of soil organic matter and nutrients in a pear orchard. With the soil of the pear (cv. Dangshansu on P. betulifolia Bunge. rootstock) orchard under clean and sod cultivation models as test materials, the experiment was conducted to evaluate spatial variability of soil organic matter (SOM), total nitrogen (STN), total phosphorus (STP), total potassium (STK), available nitrogen (SAN), and available potassium (SAK) in and between rows at different soil depths (0-60 cm). The SOM, STN, STP, STK, SAN and SAK of the different soil layers under the two tillage models were different in the vertical direction. The SOM, STN, STP and SAN in the 0-20 cm soil layer were higher than those in the 20-40 and 40-60 cm soil layers. The STK of 40-60 cm soil layer was higher than that in the 0-20 and 20-40 cm soil layers. The STK increased with the depth of soil in the vertical direction in the clean cultivated pear orchard. Variability of the SOM, STN, STP, STK, SAN and SAK of sample sites in between rows of the same soil layer was found in the pear orchard soil in the horizontal direction under clean and sod cultivation management systems, except that STK of all sites did not show the difference in identical soil layers in the pear orchard under clean cultivation. The sod cultivation model improved the SOM, STN, and STK in the 0-20 cm soil layer in the pear orchard, and the three components increased by 12.8, 12.7 and 7.3% compared to clean cultivation, respectively. The results can be applicable to plan collection of orchard soil samples, assess orchard soil quality, and improve orchard soil management practices. © 2013 Chinese Academy of Agricultural Sciences.


Hu M.,Northwest University, China | Hu M.,Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China | Qiu Z.,Northwest University, China | Qiu Z.,Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China | And 6 more authors.
Proteome Science | Year: 2012

Background: Breeding for strong red skin color is an important objective of the pear breeding program. There are few reports of proteome research in green skin pear and its red skin bud mutation. The manuscript at hand is one of the first studies dealing with 2D-PAGE-based analysis of pear fruits and leaves, establishing a suitable sample preparation and testing different 2D-PAGE protocols. Therefore, it may grant a basis for further studies on the pear proteome being the studies main goal. A proteomic analysis was conducted on leaves and fruits of 'Zaosu' pear (Pyrus bretschneideri Rehd.) and its red skin bud mutation in order to reveal their genetic differences in the protein level.Results: In the present study, the optimized two-dimensional (2-D) gel electrophoresis system of pear leaf and fruit was set up, and applied to analyze the leaves and fruit protein. The interesting peptide fragments were determined using 4800 Plus MALDI TOF/TOFTM Analyzer mass spectrometer, and the sequence obtained was blasted in NCBInr to identify the differentially-expressed protein. In the 1.5-fold differently-expressed proteins between 'Zaosu' pear and its mutant, 10 out of 35 proteins in fruit and 12 out of 24 ones in leaves were identified successfully. Among the 22 identified proteins, 7 protein spots were related to photosynthesis and energy metabolism; 4 were associated with environmental stress; 4 with disease defense; 2 with amino acid metabolism; 2 with cytoskeleton; 1 with antioxidant function; 1 with calcium metabolism; and 1 with unknown function. Moreover, related physiological index, such as chlorophyll content, Rubisco content and polyphone oxidase activity, were different between 'Zaosu' pear and its mutant.Conclusion: A 2-D gel electrophoresis system of pear leaves and fruits was established, which was suitable for the analysis of proteome comparison. To the best of our knowledge, we have performed the first analysis of the proteomic changes in leaves and fruits of 'Zaosu' pear and its red skin bud mutation. Our study provides important information on the use of proteomic methods for studying protein regulation of 'Zaosu' pear and its red skin bud mutation. © 2012 Hu et al; licensee BioMed Central Ltd.


Wen Y.,Northwest University, China | Wen Y.,Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China | Wang X.,Northwest University, China | Wang X.,Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China | And 3 more authors.
Planta | Year: 2012

Aldehyde dehydrogenases (ALDHs) catalyze the irreversible oxidation of a broad spectrum of reactive aldehydes to their corresponding carboxylic acids. Although the proteins have been studied from various organisms and at different growth stages in plants, their potential roles in pathogen infection have not been examined. Here we isolated and functionally characterized a pathogen-inducible ALDH gene (VpALDH2B4) from Chinese wild grapevine Vitis pseudoreticulata accession Baihe-35-1. When transiently expressed in Arabidopsis leaves, VpALDH2B4 was found to be localized in mitochondria. Escherichia coli expressed GST-VpALDH2B4 exhibited ALDH activity in vitro and was capable of utilizing malondialdehyde (MDA), acetaldehyde and glyceraldehydes as its substrate. Over-expression of VpALDH2B4 in Arabidopsis resulted in hypersensitive response-like cell death, enhanced resistance to downy mildew and powdery mildew presumably via the SA-signaling pathway. The same Arabidopsis transgenic plants also showed enhanced tolerance to salt stress, which is accompanied by less MDA accumulation and upregulation of the stress-responsive superoxide dismutase activity. Taken together, our results suggest that VpALDH2B4 and perhaps its orthologous genes may be involved in responses of plants to stresses imposed by both biotrophic pathogens and high salinity conditions. © 2012 Springer-Verlag.


Xu T.-F.,Northwest University, China | Xu T.-F.,Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China | Zhao X.-C.,Shandong Seed Group Co. | Jiao Y.-T.,Northwest University, China | And 7 more authors.
PLoS ONE | Year: 2014

Previously, VpPR-10.1 was isolated and characterized from a cDNA library of a fungus-resistant accession of Chinese wild grape (Vitis pseudoreticulata). We found that expression of VpPR-10.1 is affected by the fungal pathogen Erysiphe necator. To investigate the biochemical basis of the nuclease activity of VpPR-10.1 and its role in antifungal resistance, we generated recombinant VpPR-10.1 as well as site-directed mutations targeting three conserved amino acid residues among plant PR-10 s: Lys55, Glu149, and Tyr151. We showed that wild-type recombinant VpPR-10.1 exhibits both RNase and DNase activities. Mutant VpPR10.1-Y151H essentially retained all these activities. In contrast, VpPR10.1-K55N, where Lys55 in the P-loop region is mutated to Asn, and VpPR10.1-E149G, where Glu149 is mutated to Gly, lost their nuclease activity, indicating that both residues play a critical role in catalyzing RNA and DNA degradation. Furthermore, VpPR10.1 and VpPR10.1-Y151H inhibited the growth of the cultured fungal pathogen Alternaria alternate. Through transient expression in grapevine, we also demonstrated that VpPR10.1-K55N and VpPR10.1-E149G compromised resistance to E. necator . Finally, we further found that VpPR-10.1 can lead to programmed cell death and DNA degradation when incubated with tobacco BY-2 suspension cells. We show here that Lys55 and Glu149, but not Tyr151, are required for the RNase, DNase and antifungal activities of VpPR-10.1. The strong correlation between the level of VpPR-10.1 nuclease activity and its antifungal property indicates that the former is the biochemical basis for the latter. Taken together, our experiments revealed that VpPR-10.1 is critical in mediating fungal resistance in grape, potentially playing a dual role by degrading pathogen RNA and inducing programmed death of host cells. © 2014 Xu et al.


Zhao H.,Northwest University, China | Zhao H.,Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China | Guan X.,Northwest University, China | Guan X.,Key Laboratory of Horticultural Plant Biology and Germplasm Innovation in Northwest China | And 4 more authors.
Protoplasma | Year: 2013

Glyoxal oxidase (GLOX), an extracellular H2O2-producing enzyme, has been reported in Phanerochaete chrysosporium and Ustilago maydis. We previously isolated a grapevine GLOX gene from the highly resistant to Erysiphe necator Chinese wild Vitis pseudoreticulata accession Baihe-35-1 and designated it as VpGLOX (GenBank accession no. DQ201181). Transient expression of VpGLOX can suppress Powdery Mildew in susceptible genotype were studied. To further investigate the function of the VpGLOX gene, real-time PCR and Western blot analysis were performed to examine expression patterns at transcriptional and translational levels, respectively. The results showed that VpGLOX expression at the transcriptional level increased significantly in the disease-resistant accession Baihe-35-1 after Erysiphe necator inoculation, but no significant changes in the susceptible accession, V. pseudoreticulata accession Guangxi-2 could be observed. As evident from a Western blot analysis, VpGLOX protein increased slightly in Baihe-35-1 after E. necator inoculation, but not statistical significant difference changes in Guangxi-2. The immunolocalization via immunogold electron microscopy showed that VpGLOX was mainly located in the adaxial epidermal cell wall of E. necator-inoculated leaves of both Baihe-35-1 and Guangxi-2. Agrobacterium-mediated transient expression assays revealed that VpGLOX expression could produce H2O2, which may directly play a role in defense mechanism during plant-pathogen interactions. Our results could provide further insight into the biological role of VpGLOX in the defense response against E. necator in V. pseudoreticulata. © 2012 Springer-Verlag Wien.

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