Journal of Integrative Agriculture 2016,15(3):528-536 Available online at www.sciencedirect.com A ScienceDirect ELSEVIER RESEARCH ARTICLE Isolation and characterization of an ERF-B3 gene associated with flower abnormalities in non-heading Chinese cabbage Cross Marl XU Yu-chao',HOU Xi-lin',XU Wei-wei',SHEN Lu-lu2,LU Shan-wu',ZHANG Shi-lin',HU Chun-mei' State Key Laboratory of Crop Genetics and Germplasm Enhancement,Ministry of Science and Technology/College of Horticulture,Nanjing Agricultural University,Nanjing 210095.P.R.China Agriculture Committee of Feixi County,Hefei 230001,P.R.China Abstract BrcERF-B3 gene,a member of ethylene-responsive factor family,was screened from a mutant plant in non-heading Chinese cabbage(Brassica rapa ssp.chinensis)by cDNA-AFLP technology.We got full length cDNA of two BrcERF-B3 genes by homology-based cloning from two materials and found that their nucleotide sequences were the same by sequencing.The BrcERF-B3 protein,belonging to the B3 subgroup of the ERF subfamily,shared a close relationship with B.rapa.RT-PCR result showed that BrcERF-B3 expressed only in mutant stamen rather than maintainer stamen.gRT-PCR results indicated that BrcERF-B3 expressed highly during reproductive growth development and in the early of mutant buds,suggesting BrcERF-B3 might be involved in the formation of abnormal flower in mutant.What's more,the expression of BrcERF-B3 was more significant to ABA,MeJA and cold stresses in mutant than in maintainer and was down-regulated in NaCI treatment in two lines,implying BrcERF-B3 might be different roles in biotic and abiotic stresses. Keywords:non-heading Chinese cabbage,stamen-petalody,ethylene-responsive factor,gene expression Plant flower architecture was controlled by some key players (such as transcription factors),some other reports showed 1.Introduction that homeotic conversion of stamens into petaloid structures in the basal Endicott Eschscholzia californica occurred Controlling the fertility was an important goal in crop hy- due to the knocking down of EScaAG1 and 2(Yellina et al. brid breeding.but it was difficult in some crops including 2010).The third and fourth whorls in androecium and non-heading Chinese cabbage.Most of phenotype of gynoecium had a homeotic transformation because of the male sterile lines(CMS or GMS)was instable,whereas the down-regulation of both PapsAG homologs concurrently stamen-dismissing material which was caused by stamen homologous transformation had a relatively stable sterility. (Hands et al.2011). AP2/ERF transcription factor,one of the largest plant transcription factor families,played a crucial role in plant growing and in response to biotic and abiotic stresses Received 12 January,2015 Accepted 8 October,2015 (Riechmannn et al.2000;Sakuma et a/.2002;Zhuang XU Yu-chao,E-mail:1049205908@qq.com; Correspondence HU Chun-mei,Tel:+86-25-84395756 et al.2008).The AP2/ERF superfamily was defined by the E-mail:jjjhcm@njau.edu.cn AP2/ERF domain,which consists about 60-70 amino acids 2016,CAAS.Published by Elsevier Ltd.This is an open and involved in DNA binding (Nakano et al.2006).Based access article under the CC BY-NC-ND license (http:/ on differences in the binding domain sequences,the AP2/ creativecommons.org/licenses/by-nc-nd/4.0/). doi10.1016/S2095-3119(15)61203-5Journal of Integrative Agriculture 2016, 15(3): 528–536 RESEARCH ARTICLE Available online at www.sciencedirect.com ScienceDirect Isolation and characterization of an ERF-B3 gene associated with flower abnormalities in non-heading Chinese cabbage XU Yu-chao1 , HOU Xi-lin1 , XU Wei-wei1 , SHEN Lu-lu2 , LÜ Shan-wu1 , ZHANG Shi-lin1 , HU Chun-mei1 1 State Key Laboratory of Crop Genetics and Germplasm Enhancement, Ministry of Science and Technology/College of Horticulture, Nanjing Agricultural University, Nanjing 210095, P.R.China 2 Agriculture Committee of Feixi County, Hefei 230001, P.R.China Abstract BrcERF-B3 gene, a member of ethylene-responsive factor family, was screened from a mutant plant in non-heading Chinese cabbage (Brassica rapa ssp. chinensis) by cDNA-AFLP technology. We got full length cDNA of two BrcERF-B3 genes by homology-based cloning from two materials and found that their nucleotide sequences were the same by sequencing. The BrcERF-B3 protein, belonging to the B3 subgroup of the ERF subfamily, shared a close relationship with B. rapa. RT-PCR result showed that BrcERF-B3 expressed only in mutant stamen rather than maintainer stamen. qRT-PCR results indicated that BrcERF-B3 expressed highly during reproductive growth development and in the early of mutant buds, suggesting BrcERF-B3 might be involved in the formation of abnormal flower in mutant. What’s more, the expression of BrcERF-B3 was more significant to ABA, MeJA and cold stresses in mutant than in maintainer and was down-regulated in NaCl treatment in two lines, implying BrcERF-B3 might be different roles in biotic and abiotic stresses. Keywords: non-heading Chinese cabbage, stamen-petalody, ethylene-responsive factor, gene expression Plant flower architecture was controlled by some key players (such as transcription factors), some other reports showed that homeotic conversion of stamens into petaloid structures in the basal Endicott Eschscholzia californica occurred due to the knocking down of EScaAG1 and 2 (Yellina et al. 2010). The third and fourth whorls in androecium and gynoecium had a homeotic transformation because of the down-regulation of both PapsAG homologs concurrently (Hands et al. 2011). AP2/ERF transcription factor, one of the largest plant transcription factor families, played a crucial role in plant growing and in response to biotic and abiotic stresses (Riechmannn et al. 2000; Sakuma et al. 2002; Zhuang et al. 2008). The AP2/ERF superfamily was defined by the AP2/ERF domain, which consists about 60–70 amino acids and involved in DNA binding (Nakano et al. 2006). Based on differences in the binding domain sequences, the AP2/ Received 12 January, 2015 Accepted 8 October, 2015 XU Yu-chao, E-mail: 1049205908@qq.com; Correspondence HU Chun-mei, Tel: +86-25-84395756, E-mail: jjjhcm@njau.edu.cn doi: 10.1016/S2095-3119(15)61203-5 1. Introduction Controlling the fertility was an important goal in crop hy￾brid breeding, but it was difficult in some crops including non-heading Chinese cabbage. Most of phenotype of male sterile lines (CMS or GMS) was instable, whereas the stamen-dismissing material which was caused by stamen homologous transformation had a relatively stable sterility. © 2016, CAAS. Published by Elsevier Ltd. This is an open access article under the CC BY-NC-ND license (http:// creativecommons.org/licenses/by-nc-nd/4.0/)