Scientia Agricultura Sinica ›› 2014, Vol. 47 ›› Issue (7): 1387-1396.doi: 10.3864/j.issn.0578-1752.2014.07.016

• HORTICULTURE • Previous Articles     Next Articles

Cloning of a Plasma Membrane Aquaporin Gene EjPIP1 in Eriobotrya japonica Leaves and Its Expression Analysis After AM Fungi Inoculation

 ZHANG  Yan-1, 2 , LI  Juan-3, YAO  Qing-1, CHEN  Jie-Zhong-1, HU  You-Li-1   

  1. 1、College of Horticulture, South China Agricultural University, Guangzhou 510642；
    2、Agricultural College，Guizhou University/Guizhou Engineering Research Center for Fruit Crops，Guiyang 550025；
    3、Zhongkai University of Agriculture and Engineering，Guangzhou 510225
  • Received:2013-07-10 Online:2014-04-01 Published:2013-12-09

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Abstract: 【Objective】 The aim of this study was to clone a plasma membrane aquaporin gene EjPIP1 from Eriobotrya japonica variety ‘Zaozhong 6’, and to analyze its sequence characteristics, then to study the effects of AM fungi on the PIP expression pattern and water use efficiency(WUE). 【Method】 A pot experiment with seedlings of loquat cultivar (‘Zaozhong 6’), two treatments (AM and NM) and five replicates were conducted. The leaf water use efficiency was measured by using LI-6400 photosynthesis determination system. The full-length cDNA of EjPIP1 was obtained by the technology of reverse transcription PCR (RT-PCR) and RACE. The cDNA sequence and the deduced amino acid sequence were analyzed by bioinformatics method. The expression patterns of EjPIP1 in leaves and roots by AM fungi inoculation were determined by real-time RT-PCR.【Result】In the effective photosynthetic time(7:00—17:00), the water use efficiency of loquat leaves were significantly increased by AM fungi inoculation. Loquat plasma membrane aquaporin gene EjPIP1 (GenBank Accessin No. JX041627) was cloned. The full-length cDNA of EjPIP1 gene consists of 1 142 bp and contains a 861 bp open reading frame (ORF) encoding 286 amino acid proteins. Two highly conserved NPA (Asn-Pro-Ala, asparagine-proline-alanine) motifs of aquaporin were identified in EjPIP1. The homologue analysis revealed that the amino acids sequence of loquat PIP1 was highly homologous with other species, especially apple (Malus domestica), peach (Prunus persica) and strawberry (Fragaria chiloensis), which was up to 97%, 92% and 90%, respectively. Real-time PCR analysis revealed that EjPIP1 could be expressed in different loquat tissues including root and leaf, and the relative expression in leaf was higher than that in root. Furthermore, under well-watered condition, the expression of EjPIP1 in loquat leaf was significantly down-regulated by AM fungi inoculation, while the expression of EjPIP1 in root was up-regulated. 【Conclusion】 The water use efficiency of loquat leaves was significantly increased by AM fungi inoculation. A plasma membrane intrinsic proteins (PIPs) gene, designated as EjPIP1, was cloned from leaf of loquat (E. japonica ‘Zaozhong 6’). And the expression of this corresponding gene in loquat leaves and roots were influenced by AM fungi, which helped to improve water use efficiency of loquat seedlings after being inoculated with AM fungi.

Key words: Eriobotrya japonica , arbuscular mycorrhizal , aquaporin , cloning , expression analysis

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