Scientia Agricultura Sinica ›› 2012, Vol. 45 ›› Issue (8): 1568-1575.doi: 10.3864/j.issn.0578-1752.2012.08.012

• HORTICULTURE • Previous Articles     Next Articles

Isolation and Expression of Pathogenesis-Related Protein Gene MhPR8 from Malus hupehensis 

 ZHANG  Ji-Yu, QU  Shen-Chun, XUE  Hua-Bai, GAO  Zhi-Hong, GUO  Zhong-Ren, ZHANG  Zhen   

  1. 1.南京农业大学园艺学院， 南京 210095
    2.江苏省中国科学院植物研究所，南京 210014
    3.中国农业科学院郑州果树研究所，郑州 450009
  • Received:2011-09-13 Online:2012-04-15 Published:2012-01-17

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Abstract: 【Objective】This study aimed to clone a full length cDNA and genomic DNA sequences of MhPR8 from Malus hupehensis and analyze the expression pattern of MhPR8 after treatment with the chemical reagents salicylic acid (SA), methyl jasmonate (MeJA), 1-aminocyclopropane-1-carboxylic acid (ACC), abscisic acid (ABA), abiotic stress NaCl and low temperature (4℃), and biotic factors apple ring spot pathogen (Botryosphaeria berengeriana) and apple aphids (Aphis citricota).【Method】The full length cDNA and genomic DNA sequences of MhPR8 were isolated from the M. hupehensis complete cDNA library treated by SA and genomic DNA using the method of PCR amplification, respectively. Real-time quantitative RT-PCR (qRT-PCR) was performed to determine the expression pattern of MhPR8 in M. hupehensis.【Result】The open reading frame of this gene was 897 bp, encoding 298 amino acids with a molecular weight of 31.67 kD and isoelectric point of 4.77. There was no intron in ORF of MhPR8. MhPR8 contains conserved construction domain GH18_hevamine_XipI_class_III, belongs to Class III chitinase. And this protein has six conserved cysteines, and aspartic acid and glutamic acid play a role of catalysis in plants class III chitinase, which are located in 151th and 153th amino acids, respectively. qRT-PCR results show that during the first 48 hours, MhPR8 was obviously induced by SA, MeJA, and ACC, but weakly induced by ABA. And this gene was clearly induced by low temperature, but weakly induced by NaCl. Infected with B. berengeriana resulted in obvious accumulation of MhPR8 transcripts during the first 96 h. The expression of the MhPR8 was clearly induced by the apple aphid (A. citricota) in leaves and stems of M. hupehensis. 【Conclusion】MhPR8 could be involved with the SA- signal pathway and the jasmonic acid (JA)/ ethylene (ET)-signal pathway in M. hupehensis. MhPR8 might be play a key role in resistance to biotic and abiotic stresses.

Key words: Malus hupehensis, MhPR8, expression, chemical reagents, biotic stress, abiotic stress

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