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Wang L.,Key Laboratory of Horticultural Plant Biology | Pan Z.-Y.,Key Laboratory of Horticultural Plant Biology | Guo W.-W.,Key Laboratory of Horticultural Plant Biology
Plant Cell, Tissue and Organ Culture | Year: 2010

In order to predict the performance of growth, development and resistance of a citrus diploid cybrid plant between Citrus unshiu Marc. cv. Guoqing No. 1 (G1) and Hirado Buntan pummelo (Citrus grandis (L.) Osbeck) (HBP), the proteomes of leaves from this cybrid and its parents were investigated. First, the diploid cybrid plant (G1 + HBP) created with its nuclear and chloroplast genomes from HBP and mitochondria genome from G1 was further verified by ploidy and molecular analysis (SSR, CAPS and cp-SSR). Then, 2-DE combined with MALDI-TOF/TOF MS were employed to analyze the variation of proteomes in the cybrid and its parental plants. Significant analysis allowed 90 (P < 0.05) differentially expressed protein spots between G1 and the cybrid, 20 between HBP and cybrid, and 116 between G1 and HBP. The comparative proteome patterns well validated the genetic background of the cybrid and its parental plants, suggesting that there is a good correlation between genome constitution and proteome expression in this cybrid. Seventy differentially expressed spots were selected for MALDI-TOF/TOF analysis and 25 proteins were identified. The identified proteins were mainly involved in photosynthesis, stress response, anti-oxidative stress and metabolism. Five proteins involved in photosynthesis such as Rubisco and Rubisco activase were significantly up-regulated in the cybrid, indicating the photosynthesis is enhanced in the cybrid. Expression analysis of proteins involved in stress response and anti-oxidative stress suggested that the resistance might be improved while anti-oxidative system exhibited a complex effect in the cybrid. The nitrogen, sulfur, carbon and energy metabolism might also be affected in the cybrid plant. © 2010 Springer Science+Business Media B.V. Source

Guan X.,Northwest University, China | Guan X.,Key Laboratory of Horticultural Plant Biology | Zhao H.,Northwest University, China | Zhao H.,Key Laboratory of Horticultural Plant Biology | And 4 more authors.
Vitis - Journal of Grapevine Research | Year: 2013

Shoot tips and/or single-bud-internodes, containing shoot apical meristems (SAM) were presented, of Vitis'6-12-2', derived from a Chinese wild V. pseudoreticulata 'Baihe-35-1' × V. vinifera 'Carignane' cross, were used for Agrobacterium-medinttA genetic transformation. In order to achieve Glyoxal oxidase (VpGLOX) overexpressing plants, the propagation and gene transfer system of shoot tips and/or single-bud-internodes undergoing either micropropagated (starting with microshoot-tips) or callus induced (starting with stems with single bud) in vitro plants were optimized. The results show that the most effective way to gain shoot tips and/or single-bud-internodes undergoing micropropagation procedure was to keep micro-shoot-tips in liquid C 2D4B medium at 80 rpm constant orbital shaking with light, then placed on solidified C2D4B medium with 2.9 μM Gibberellic acid 3 (GA3) for elongation. In vitro stems with single buds gave best results for callus formation and adventitious buds induction on half-strength MS medium with 9.0 μM Thidiazuron (TDZ) and 2.9 fiM mg-L-1 a-Naphthaleneacetic acid (NAA). The highest gene transfer frequency was obtained when explants were infected for 10 min with the concentration of. Agrobacterium tumefaciens with an optical density at 600 nm of 0.4, and then co-cultivated for 3 days. Incubation of shoot tips and/or single-bud-internodes in darkness for 3 days is helpful for enhancing gene transfer efficiency. Polymerase chain reaction (PCR) and PCR-Southern blot analyses were utilized to confirm putative transgenic plants. Up to 45 clones have proven to be transformed, and one of them has been planted out. This method opens a door for the gene transfer of recalcitrant Chinese wild V. pseudoreticulata. Source

Wang S.,Northwest University, China | Wang S.,Key Laboratory of Horticultural Plant Biology | Yu Y.,Northwest University, China | Yu Y.,Key Laboratory of Horticultural Plant Biology | And 6 more authors.
Scientia Horticulturae | Year: 2011

EF-hand calcium-binding protein (CBP) gene plays a critical role during embryonic development in animals, but it remains unclear whether this gene possesses similar function in plants. In this study, a novel CBP gene was isolated from Vitis vinifera cv. Thompson Seedless and designated as VvCBP1. Using a combination of sequence structural investigation, expression pattern analysis of different embryo development in grapevine, and silencing assay of VvCBP1 homolog in tomato, the functional role of VvCBP gene in embryo development of seedless grapevine was studied. Structural investigations showed that VvCBP1 is 907. bp in length, and contains two EF-hand motifs. Expression pattern of this gene is different in embryo between seedless and seeded grapevine species. Moreover, the silencing of homology gene led to the seeds number significantly decreased in tomato. These data demonstrate clearly that VvCBP1 plays an important role in embryo development of seedless grapevine. © 2011 Elsevier B.V. Source

Li Z.,Key Laboratory of Horticultural Plant Biology | Liu G.,Key Laboratory of Horticultural Plant Biology | Zhang J.,Key Laboratory of Horticultural Plant Biology | Lu S.,Key Laboratory of Horticultural Plant Biology | And 2 more authors.
Biologia Plantarum | Year: 2012

We isolated PaAP3, a homolog of the class B MADS-box transcription factor gene APETALA3 (AP3), from the monoecious plant London plane tree (Platanus acerifolia Willd.). PaAP3 encodes a protein that shares good levels of identity with class B genes from Arabidopsis thaliana (35 and 51 % identity with PISTILLATA (PI) and AP3, respectively), and also with class B genes of other woody species (59 % identity with PTD from Populus trichocarpa and 66 % with TraAP3 from Trochodendron aralioides). Reverse transcription polymerase chain reaction showed that PaAP3 was expressed in both the female and male flowers of P. acerifolia, but almost no signal was detected in the vegetative tissues or mature embryos. The PaAP3 expression in male flowers showed a relationship with developmental stage. There was a small transient increase during differentiation of the flower primordia in June, but maximal levels occurred during December when flower development appeared arrested. Increased PaAP3 expression was also detected in March of the following year, corresponding to meiotic divisions of the microspore mother cells, but this was lost by April when the pollen was mature. © 2012 Springer Science+Business Media B.V. Source

Yang C.,Huazhong Agricultural University | Yang C.,Key Laboratory of Horticultural Plant Biology | Yang C.,Key Laboratory of Horticultural Crop Biology and Genetic Improvement Central Region | Liu T.,Huazhong Agricultural University | And 17 more authors.
Molecular Genetics and Genomics | Year: 2015

Corky split vein can develop under long-term boron deficient conditions in Citrus sinensis L. Osbeck cv. Newhall. This symptom only occurs in the upper rather than the lower epidermis of old leaves. Our previous study demonstrated that vascular hypertrophy was involved in the symptoms, and the 3rd developmental stage of corky split vein (BD3) was the critical stage for phenotype formation. Here, we performed an intensive study on the BD3 vein and its control sample (CK3 vein). A lignin test demonstrated that the lignin content in BD3 vein was approximately 1.7 times more than the CK3 vein. Anatomical investigation of the corky split vein indicated that the upper epidermis was destroyed by overgrowing vascular cells, and the increased lignin may contribute to vascular cell differentiation and wounding-induced lignification. In a subsequent small RNA sequencing of the BD3 and CK3 veins, 99 known miRNAs and 22 novel miRNAs were identified. Comparative profiling of these miRNAs demonstrated that the 57 known miRNAs and all novel miRNAs exhibited significant expression differences between the two small RNAs libraries of the BD3 and CK3 veins. Associated with our corresponding digital gene expression data, we propose that the decreased expression of two miRNAs, csi-miR156b and csi-miR164, which leads to the up-regulation of their target genes, SPLs (csi-miR156b-targeted) and CUC2 (csi-miR164-targeted), may promote vascular cell division and orderless stage transition in old leaves. © 2015, Springer-Verlag Berlin Heidelberg. Source

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