Cloning and sequence analysis of two ARF genes and two Aux/IAA genes in Agapanthus praecox ssp. orientalis

doi:10.3969/j.issn.1004-1524.2019.01.12

›› 2019, Vol. 31 ›› Issue (1): 86-97.DOI: 10.3969/j.issn.1004-1524.2019.01.12

• Horticultural Science • Previous Articles Next Articles

Cloning and sequence analysis of two ARF genes and two Aux/IAA genes in Agapanthus praecox ssp. orientalis

YANG Zhou, LYU Ke, LYU Shan, WANG Junjie, ZHANG Di^*

Department of Landscape Science and Engineering, School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai 200240, China

Received:2018-03-24 Online:2019-01-25 Published:2019-04-09

Abstract

Abstract: Using the Illumina sequencing platform, transcriptome sequencing of Agapanthus praecox ssp. orientalis callus and embryonic callus treated with different concentrations of exogenous auxin picloram (PIC) were performed. Two ARF genes and two Aux/IAA genes that expressed differentially during the auxin signal transduction pathway were obtained and named as ApARF1, ApARF2, ApAux/IAA2 and ApAux/IAA3. To further understand their function in auxin signal transduction during somatic embryogenesis, their full-length cDNA sequences were cloned by RACE technique. The full-length cDNA sequences of ApARF1, ApARF2, ApAux/IAA2 and ApAux/IAA3 were 2 343, 2 888, 1 034 and 821 bp, each contained an open reading frame of 1 770, 2 349, 480 and 549 bp, respectively, which encoded 589, 782, 159 and 182 amino acids, respectively. Through bioinformatic analysis including protein domain analysis, phylogenetic tree analysis, and protein homology analysis, we concluded that ApARF1 and ApARF2 proteins had the typical construction of ARFs, and might be related to plant aging; ApAux/IAA2 lacked a completed domain Ⅳ, ApAux/IAA3 lacked a completed domain Ⅱ; however, they may still have the typical function of Aux/IAA family. We constructed gene over-expression vectors with pHB vector and transformed them into wild-type Arabidopsis thaliana. The phenotypic results showed that compared with the wide-type, the over-expression of ApARF1 and ApARF2 could promote the growth and flowering of transgenic plants, while the over-expression of ApAux/IAA2 and Aux/IAA3 showed a depressing influence on the growth of transgenic plants. The transgenic results combined with the sequence analysis proved that ApARFs and ApAux/IAAs had a regulatory role in auxin signal transduction pathway and plant growth.

Key words: Agapanthus praecox, auxin signal, ARF (auxin response factors), Aux/IAA (auxin/indoleacetic acid proteins), gene cloning, embryonic callus

CLC Number:

YANG Zhou, LYU Ke, LYU Shan, WANG Junjie, ZHANG Di. Cloning and sequence analysis of two ARF genes and two Aux/IAA genes in Agapanthus praecox ssp. orientalis[J]. , 2019, 31(1): 86-97.

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Cloning and sequence analysis of two ARF genes and two Aux/IAA genes in Agapanthus praecox ssp. orientalis

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