Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (7): 1408-1418.doi: 10.3864/j.issn.0578-1752.2013.07.011

• HORTICULTURE • Previous Articles     Next Articles

Expression and Functional Analysis of VpMYBR1 Gene Resistant to Uncinula necator from Vitis pseudoreticulata

 HOU  Hong-Min, WANG  Hao, YIN  Xiang-Jing, YAN  Qin, WANG  Yue-Jin, WANG  Xi-Ping   

  1. College of Horticulture, Northwest A&F University/State Key Laboratory of Crop Stress Biology in Arid Areas/Key Laboratory of Horticultural Crop Biology and Germplasm Development in Northwest China, Ministry of Agriculture, Yangling 712100, Shaanxi
  • Received:2012-06-27 Online:2013-04-01 Published:2013-01-21

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Abstract: 【Objective】 The VpMYBR1 gene was cloned from Vitis pseudoreticulata resistance to powdery mildew clone ‘Baihe-35-1’ leaves infected by Uncinula necator, and its expression model and function were analyzed in this study. This study will provide a theoretical basis for revealing the grape powdery mildew resistance mechanism.【Method】VpMYBR1 was isolated by RT-PCR and rapid amplification of cDNA ends (RACE) approaches, and its expression patterns under different treatments and in different tissues were analyzed by using semi-quantitative RT-PCR and real-time quantitative PCR. The plant expression vector of this gene was constructed, and then transformed into Arabidopsis thaliana by Floral Dip method. The disease resistance identification of transgenic plants was analyzed by infecting powdery mildew, trypan blue staining and analyzing the expression of Arabidopsis defense marker genes.【Result】The assembled VpMYBR1 (GenBank ACC no. HQ284197) cDNA sequence was 539 bp in full length including a 228 bp ORF which encodes a polypeptide of 75 amino acids containing a single Myb-like DNA-binding domain. The VpMYBR1 was expressed in leaves, stems, inflorescence, tendril, and fruit of ‘Baihe-35-1’. The expression of VpMYBR1 was strong in leaves and weak in inflorescence and fruit. The VpMYBR1 was induced by U. necator during 6 to 12 hpi in three grapevine genotypes (V. pseudoreticulata clone ‘Baihe-35-1’, ‘Hunan-1’ and V. quinquangularis clone ‘Shang-24’), the maximum induction of VpMYBR1 was the greatest in powdery mildew resistant genotypes ‘Baihe-35-1’, reaching more than 28-fold. In addition, the VpMYBR1 expression of ‘Baihe-35-1’ induced by defence signal molecular (SA and MeJA) was significantly higher than that of ‘Hunan-1’ and ‘Shang-24’. Then the VpMYBR1 gene was transformed into A. thaliana by Floral Dip method to make further study for verifying its function. The results showed that the expression of VpMYBR1 improved the resistance against the powdery mildew by hypersensitive reaction.【Conclusion】 The results of the study indicate that VpMYBR1 is a founding member of a small family of proteins containing a novel Sant/myb domain which is likely to participate in the regulation of the resistance to U. necator. This research has provided a new candidate gene for improving the disease resistance of V. vinifera by transgenic technology.

Key words: Vitis pseudoreticulata , powdery mildew , VpMYBR1 transcription factor , expression , functional analysis

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