Scientia Agricultura Sinica ›› 2014, Vol. 47 ›› Issue (10): 1985-1994.doi: 10.3864/j.issn.0578-1752.2014.10.012

• HORTICULTURE • Previous Articles     Next Articles

Genome-wide Identification and Expression Analysis of Auxin Response Factor (ARF) Family in Cucumber

 SHENG  Hui-1, 2 , QIN  Zhi-Wei-1, LI  Wen-Bin-3, ZHOU  Xiu-Yan-1, WU  Tao-1, XIN  Ming-1   

  1. 1、College of Horticulture, Northeast Agricultural University/Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in Northeast China, Ministry of Agriculture, Harbin 150030;
    2、Harbin Academy of Agricultural Sciences,  Harbin 150029;
    3、College of Agriculture, Northeast Agricultural University, Harbin 150030
  • Received:2013-10-16 Online:2014-05-20 Published:2014-03-15

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Abstract: 【Objective】 The objective of this study is to identify the cucumber auxin response factor (ARF), forecast the small RNAs and verify the relationship between ARF with small RNAs and auxin. The expression pattern of ARF during seed germination was analyzed and the effect of ARF on the process of parthenocarpy and seed germination was deduced. 【Method】 The Arabidopsis and rice ARF proteins were used to search the cucumber genome database. Then, the retrieved cucumber ARF family structure was analyzed and the small RNAs were predicted. The predicted gma-MIR160o precursur was built into the pCAMBIA2301. By Agrobacterium-mediated method, it was imported into parthenocarpy cucumber varieties. Transgenic plants were validated by RT-PCR. Using real-time RT-PCR method, the expression patterns of ARF were analyzed in auxin treatment, flowing and seed germination. 【Result】 By comparison with the Arabidopsis and rice ARF protein sequences, there are 18 cucumber ARF protein sequences. ARF proteins were divided into four classes. The number of exons varied from 2 to 18, and the structure was similar among the same class. Phylogenetic tree showed that the similarity of 18 genes was not high. Small RNAs corresponds to ARF genes in cucumber were found. Csa010564, Csa011935, Csa015176, Csa020560 and Csa022361 were miR160 targets. Transgenic test further exhibited mRNA abundance of Csa010564, Csa011935 and Csa015176 decreased, while Csa020560 and Csa022361 rose slightly, indicating that Csa010564, Csa011935 and Csa015176 were miR160 targets. The results of real-time RT-PCR exhibited that mRNA abundance of Csa007296, Csa011935 and Csa015176 in roots, stems and leaves were higher than wild type, indicating the expressions of these genes were positively regulated by auxin. Meanwhile, the expressions in leaves and female flowers were decreased on the second flowering day, while the expression of the ovary was the opposite, especially Csa011935 and Csa015176, indicating that ARF genes play a crucial role in the development of the ovary. In the experiment of the auxin treatment, the expressions in different tissues were up-regulated. The expression of ARF genes in the seed germination was analyzed. The results showed that the expression peak of most genes were 12 h and 48 h in imbibition. 【Conclusion】 The ARF genes were regulated by auxin and the corresponding small RNAs. ARF genes may play a key role in the parthenocarpy and seed germination of cucumber .

Key words: cucumber , ARF , small RNAs , auxin , expression patterns

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