Isolation, expression, and evolution analysis of the type 2C protein phosphatase gene BcABI1 involved in abiotic and biotic stress in Brassica campestris ssp. chinensis
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Type 2C protein phosphatase (PP2C) is an important component of protein phosphatases involved in various biological processes, such as plant growth and development, cell cycle regulation, signal transduction, and environmental stress response. ABI1 (ABA insensitive 1), a member of the PP2C gene family, is a negative regulator in abscisic acid (ABA) signal transduction possibly related to stress tolerance, but ABI1 in economical plants is poorly understood. In this study, BcABI1 was isolated from Chinese cabbage [Brassica campestris (syn. B. rapa) L. ssp. chinensis (L) Makino] and 23 other cruciferous plants. Phylogenetic analysis was conducted according to the gene sequence of AtABI1 in Arabidopsis. Three introns are present in ABI1 genes encoding 400–437 amino acids. Multiple alignment of the sequences of 24 ABI1 genes showed that the ABI1 homologous genes of cruciferous plants were highly conserved. Meanwhile, the Ka/Ks values were obtained from orthologous gene pairs between the At genome and other crucifer crop genomes, and results showed that the natural selective pressures on these genes were purifying selection. Subsequently, the structure and expression profiles of BcABI1 were examined under abiotic and biotic stresses. BcABI1 presented a constitutive expression pattern, participating in wound stress and NaCl stress responses, and also induced by biotic stress in Chinese cabbage inoculated with Botrytis cinerea and Sclerotinia sclerotiorum. A significant difference between the pathogenic ability of the two bacteria can be observed, and the S. sclerotiorum caused faster pathogenesis. These results could help to elucidate the function of BcABI1 in plant growth and stress response, and gain valuable insights into Chinese cabbage breeding for the improvement of multiple-stress-tolerance.
KeywordsChinese cabbage PP2C ABI1 ABA Abiotic stress Biotic stress
The authors gratefully acknowledge Dr. Gang Lu for his helpful advice and thank Dr. Zhenning Liu, in particular, for the stimulating discussions and critical reading of the manuscript.
XY and SH proposed and designed the research. SH, FW and LK performed the experiment under abiotic and biotic stresses. LM, ZK and LK performed the statistical analyses, interpretation of experimental results. CC and DH performed the qRT-PCR analysis. LK and SH wrote the manuscript. LK, HD and XY revised the manuscript.
This work was partially supported by the National Key Research and Development Program of China (2016YFD0100204-31), the 948 Project of Agricultural Ministry of China (2014-Z28), National Natural Science Foundation of China (31460521), the Breeding Project of the Sci-tech Foundation of Zhejiang Province (2016C02051-6-1), the Project of the Sci-tech Foundation of Ningbo City(2015C110008), and the Project of Application on Public Welfare Technology in Zhejiang Province(LGN18C150003).
Compliance with ethical standards
The authors declare that they have no financial interests.
- Chen YP, Cao JS, Miao Y, Ye WZ (2000) Analysis of genetic polymorphisms in vegetable crops of Brassica campestris by RAPD markers. J Zhejiang Univ (Agri Life Sci) 26(2):131–136Google Scholar
- Cristianini N, Hahn MW (2007) Introduction to computational genomics: a case studies approach. Cambridge, Cambridge University PressGoogle Scholar
- Darvishi E, Kahrizi D, Ghaheri M, Yari K (2017) Study on highly conserved motifs of PP2C-type protein phosphatases in some plant species. Biharean Biologist 11(2):130–132Google Scholar
- Komatsu K, Nishikawa Y, Ohtsuka T, Taji T, Quatrano RS, Tanaka S, Sakata Y (2009) Functional analyses of the ABI1-related protein phosphatase type 2C reveal evolutionarily conserved regulation of abscisic acid signaling between Arabidopsis and the moss Physcomitrella patens. Plant Mol Biol 70:327–340CrossRefPubMedGoogle Scholar
- Lefoulon C, Boeglin M, Moreau B, Very AA, Szponarski W, Dauzat M, Michard E, Gaillard I, Cherel I (2016) The Arabidopsis AtPP2CA protein phosphatase inhibits the GORK K+ efflux channel and exerts a dominant suppressive effect on phosphomimetic-activating mutations. J Biol Chem 291(12):6521–6533CrossRefPubMedPubMedCentralGoogle Scholar
- Ludwikow A, Ciesla A, Kasprowicz-Maluski A, Mitula F, Tajdel M, Galganski L, Ziolkowski PA, Kubiak P, Maleck A, Piechalak A et al (2014) Arabidopsis protein phosphatase 2C ABI1 interacts with type I ACC synthases and is involved in the regulation of Ozone-induced ethylene biosynthesis. Mol Plant 7(6):960–976CrossRefPubMedGoogle Scholar
- Nagaharu U (1935) Genome analysis in Brassica with special reference to the experimental formation of B. napus and peculiar mode of fertilization. Jpn J Bot 7:389–452Google Scholar
- Rodrigues A, Adamo M, Crozet P, Margalha L, Confraria A, Martinho C, Elias A, Rabissi A, Lumbreras V, Gonzalez-Guzman M et al (2013) ABI1 and PP2CA phosphatases are negative regulators of Snf1-related protein kinase1 signaling in Arabidopsis. Plant Cell 25(10):3871–3884CrossRefPubMedPubMedCentralGoogle Scholar
- Tougane K, Komatsu K, Bhyan SB, Sakata Y, Ishizaki K, Yamato KT, Kohchi T, Takezawa D (2010) Evolutionarily conserved regulatory mechanisms of abscisic acid signaling in land plants: characterization of ABSCISIC ACID INSENSITIVE 1-like type 2C protein phosphatase in the liverwort Marchantia polymorpha. Plant Physiol 152:1529–1543CrossRefPubMedPubMedCentralGoogle Scholar
- Yang JH, Liu DY, Wang XW, Ji CM, Cheng F, Liu BN, Hu ZY, Chen S, Pental D, Ju YH, Li XM, Zhang JH, Wang JL, Liu F, Ma WW, Shopan J, Zheng HK, Mackenzie SA, Zhang MF (2016) The genome sequence of allopolyploid Brassica juncea and analysis of differential homoeolog gene expression influencing selection. Nat Genet 48:1225–1232CrossRefPubMedGoogle Scholar
- Yoshida R, Umezawa T, Mizoguchi T, Takahashi S, Takahashi F, Shinozaki K (2006a) The regulatory domain of SRK2E/OST1/SnRK2.6 interacts with ABI1 and integrates abscisic acid (ABA) and osmotic stress signals controlling stomatal closure in Arabidopsis. J Biol Chem 281(8):5310–5318CrossRefPubMedGoogle Scholar
- Yoshida T, Nishimura N, Kitahata N, Kuromori T, Ito T, Asami T, Shinozaki K, Hirayama T (2006b) ABA-Hypersensitive Germination3 encodes a protein phosphatase 2C (AtPP2CA) that strongly regulates abscisic acid signaling during germination among Arabidopsis protein phosphatase 2Cs. Plant Physiol 140:115–126CrossRefPubMedPubMedCentralGoogle Scholar
- Yoshida T, Fujita Y, Sayama H, Kidokoro S, Maruyama K, Mizoi J, Shinozaki K, Yamaguchi-Shinozaki K (2010) AREB1, AREB2, and ABF3 are master transcription factors that cooperatively regulate ABRE-dependent ABA signaling involved in drought stress tolerance and require ABA for full activation. Plant J 61:672–685CrossRefPubMedGoogle Scholar
- Zhang T, Xiang X, Ye WZ, Yu XL, Cao JS (2006) Molecular cloning and bioinformatic analysis of pollen development related gene BcMF2 from Brassica campestris L. ssp. rapifera. J Zhejiang Univ (Agri Life Sci) 32(6):598–605Google Scholar