Abstract
Our previous studies have found that gene APRR2 is the candidate gene of w controlling the green color of immature fruit in cucumber, and its allele aprr2, which controls the white color, has a 101-amino acid residues deletion at the C-terminus compared with APRR2. Here, we characterized the APRR2 and analyzed its function in cucumber. qPCR analysis and prokaryotic expression of APRR2 and aprr2 clarified the relationship between APRR2 and aprr2 and confirmed that APRR2/aprr2 is a gene controlling cucumber color. Sequence and phylogenetic relationship analysis revealed that gene APRR2 and aprr2 with REC domain and DNA-binding B-motif are placed into subclass I of PRR subfamily. Subcellular localization assay indicated that APRR2 is a nuclear protein, but aprr2 is not. These results suggest APRR2 gene is a transcription factor in cucumber. Additionally, the expression analysis of Class-I KNOTTED1-like homeobox genesTKN4 and TKN2 involved in the chloroplast development showed they were in line with APRR2 expression, therefore, APRR2 together withTKN4, TKN2 likely contribute to the mechanisms related to chloroplast development.
Similar content being viewed by others
Abbreviations
- TCS:
-
Two-component regulatory systems
- RR:
-
Response regulator
- PRR:
-
Pseudo-response regulators
- APRR2 :
-
Arabidopsis pseudo-response regulator 2 gene;
- KNOX :
-
Class-I KNOTTED1-like homeobox
- IPTG:
-
Isopropyl-β-d-thiogalactoside
- SDS-PAGE:
-
Sodium dodecyl sulfate-polyacrylamide gel electrophoresis
- YEB:
-
Agrobacterium rhizogenes culture medium
- LB:
-
Lysogeny broth
- PCR:
-
Polymerase chain reaction
- Rt-PCR:
-
Real-time quantitative PCR
- cDNA:
-
DNA complementary to RNA
- GLK2 :
-
Golden2-like
- REC:
-
Receiver
- RLD:
-
Receiver-like domain
- PRRs:
-
Pseudo-response regulators
- ARRS:
-
Response regulators
- CTK:
-
Cytokinin
- CLM9:
-
Calmodulin9-like
References
Agarwal P, Reddy MP, Chikara J (2011) WRKY: its structure, evolutionary relationship, DNA-binding selectivity, role in stress tolerance and development of plants. Mol Biol Rep 38(6):3883–3896
Argyros RD et al (2008) Type B response regulators of Arabidopsis play key roles in cytokinin signaling and plant development. Plant Cell 20(8):2102–2116
Avivi Y, et al (2000) Clausa, a tomato mutant with a wide range of phenotypic perturbations, displays a cell type-dependent expression of the homeobox gene LeT6/TKn2. Plant Physiol 124(2):541–552
Baranowskij N, Frohberg C, Prat S, Willmitzer L (1994) A novel DNA binding protein with homology to Myb oncoproteins containing only one repeat can function as a transcriptional activator. EMBO J 13(22):5383–5392
Brand A, Borovsky Y, Meir S, Rogachev I, Aharoni A, Paran I (2012) pc8.1, A major QTL for pigment content in pepper fruit, is associated with variation in plastid compartment size. Planta 235(3):579–588
Chen Y-S, Chao Y-C, Tseng T-W, Huang C-K, Lo P-C, Lu C-A (2016) Two MYB-related transcription factors play opposite roles in sugar signaling in Arabidopsis. Plant MolBiol 93:1–13
Cortleven A, Schmulling T (2015) Regulation of chloroplast development and function by cytokinin. J Exp Bot 66(16):4999–5013
Di Giacomo E, Laffont C, Sciarra F, Iannelli MA, Frugier F, Frugis G (2017) KNAT3/4/5-like class 2 KNOX transcription factors are involved in Medicago truncatula symbiotic nodule organ development. New Phytol 213(2):822–837
Dityatkin SY, Lisovskaya KV, Panzhava NN, Iliashenko BN (1972) Frozen-thawed bacteria as recipients of isolated coliphage DNA. Biochimica Et Biophysica Acta 281(3):319–323
Fitter DW, Martin DJ, Copley MJ, Scotland RW, Langdale JA (2002) GLK gene pairsregulate chloroplast development in diverse plant species. Plant J 31(6):713–727
Frugis G, et al. (2001) Overexpression of KNAT1 in lettuce shifts leaf determinate growth to a shoot-like indeterminate growth associated with an accumulation of isopentenyl-type cytokinins. Plant Physiol 126(4):1370–1380
Hall LN, Rossini L, Cribb L, Langdale JA (1998) GOLDEN 2: a novel transcriptionalregulator of cellular differentiation in the maize leaf. Plant Cell 10(6):925–936
Hannig G, Makrides SC (1998) Strategies for optimizing heterologous protein expression in Escherichia coli. Trends Biotechnol 16(2):54–60
Hay A, Tsiantis M (2010) KNOX genes: versatile regulators of plant development and diversity. Development 137(19):3153–3165.
Heyl A, et al (2013) Nomenclature for members of the two-component signaling pathway of plants. Plant Physiol 161(3):1063–1065
Holsters M, de Waele D, Depicker A, Messens E, van Montagu M, Schell J (1978) Transfection and transformation of Agrobacterium tumefaciens. Mol Gen Genet MGG163(2):181–187
Horsch RB, Fraley RT, Rogers SG, Sanders PR, Lloyd A, Hoffmann N (1984) Inheritance of functional foreign genes in plants. Science 223(4635):496
Hwang I, Sheen J (2001) Two-component circuitry in Arabidopsis cytokinin signal transduction. Nature 413(6854):383–389
Hwang I, Chen HC, Sheen J (2002) Two-component signal transduction pathways in Arabidopsis. Plant Physiol 129(2):500–515
Inskeep WP, Bloom PR (1985) Extinction coefficients of chlorophyll a and B in n, n-dimethylformamide and 80% acetone. Plant Physiol 77(2):483–485
Jasinski S et al (2005) KNOX action in Arabidopsis is mediated by coordinate regulation of cytokinin and gibberellin activities. Curr Biol 15(17):1560–1565
Jia G, Li B, Zhang D, Zhang T, Li Z, Dai J, Wang S (2009) Cloning and characterization of a novel R1-MYB transcription factor in maize. Prog Nat Sci 19(9):1089–1096
Jin H, Martin C (1999) Multifunctionality and diversity within the plant MYB-gene family. Plant MolBiol 41(5):577–585
Kusaba S, Kano-Murakami Y, Matsuoka M, Tamaoki M, Sakamoto T, Yamaguchi I, Fukumoto M (1998) Alteration of hormone levels in transgenic tobacco plants overexpressing the rice homeobox gene OSH1. Plant Physiol 116(2):471–476
Le DT, Nishiyama R, Watanabe Y, Mochida K, Yamaguchi-Shinozaki K, Shinozaki K, Tran L-SP (2011) Genome-wide survey and expression analysis of the plant-specific NAC transcription factor family in soybean during development and dehydration stress. DNA Res 18(4):263–276
Li C, Li D, Shao F, Lu S (2015) Molecular cloning and expression analysis of WRKY transcription factor genes in Salvia miltiorrhiza. BMC Genomics 16:200
Liu H, et al (2016) Map-based cloning, identification and characterization of the w gene controlling white immature fruit color in cucumber (Cucumis sativus L.). Theor Appl Genet 129(7):1247–1256
Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2–∆∆CT method. Methods 25(4):402–408
Lohrmann J et al (2001) The response regulator ARR2: a pollen-specific transcription factor involved in the expression of nuclear genes for components of mitochondrial complex I in Arabidopsis. Mol Gen Genom 265(1):2–13
Makino S, et al (2000) Genes encoding pseudo-response regulators: insight into His-to-Asp phosphorelay and circadian rhythm in Arabidopsis thaliana. Plant Cell Physiol 41(6):791
Mierendorf RC, Morris BB, Hammer B, Novy RE (1998) Expression and purification of recombinant proteins using the pET system. Mol Diag Infectious Dis 13:257–292
Mizuno T (2005) Two-component phosphorelay signal transduction systems in plants: from hormone responses to circadian rhythms. Biosci Biotechnol Biochem 69(12):2263–2276
Mothes K (1960) Über das Altern der Blätter und die Möglichkeit ihrer Wiederverjüngung. Naturwissenschaften 47(15):337–351
Nadakuduti SS, Holdsworth WL, Klein CL, Barry CS (2014) KNOX genes influence a gradient of fruit chloroplast development through regulation of GOLDEN2-LIKE expression in tomato. Plant J 78(6):1022–1033
Nagano Y (2000) Several features of the GT-factor trihelix domain resemble those of the Myb DNA-binding domain. Plant Physiol 124(2):491–494
Ori N, Juarez MT, Jackson D, Yamaguchi J, Banowetz GM, Hake S (1999) Leaf senescence is delayed in tobacco plants expressing the maize homeobox gene knotted1 under the control of a senescence-activated promoter. Plant Cell 11(6):1073–1080
Pan Y, et al (2013) Network inference analysis identifies an APRR2-like gene linked to pigment accumulation in tomato and pepper fruits. Plant Physiol 161(3):1476–1485
Parnis A, Cohen O, Gutfinger T, Hareven D, Zamir D, Lifschitz E (1997) The dominant developmental mutants of tomato, Mouse-ear and Curl, are associated with distinct modes of abnormal transcriptional regulation of a knotted gene. Plant Cell 9(12):2143–2158
Richards DE, Peng J, Harberd NP (2000) Plant GRAS and metazoan STATs: one family? Bioessays 22(6):573–577
Richmond AE, Lang A (1957) Effect of kinetin on protein content and survival of detached Xanthium leaves. Science 125(3249):650–651
Riechmann JL et al (2000) Arabidopsis transcription factors: genome-wide comparative analysis among eukaryotes. Science 290(5499):2105–2110
Rose A, Meier I, Wienand U (1999) The tomato I-box binding factor LeMYBI is a member of a novel class of Myb-like proteins. Plant J 20(6):641–652
Sakai H, Honma T, Aoyama T, Sato S, Kato T, Tabata S, Oka A (2001) ARR1, a transcription factor for genes immediately responsive to cytokinins. Science 294(5546):1519–1521
Satbhai SB et al (2011) Pseudo-response regulator (PRR) homologues of the moss Physcomitrella patens: insights into the evolution of the PRR family in land Plants. DNA Res 18(1):39–52
Scofield S, Murray JA (2006) KNOX gene function in plant stem cell niches. Plant Mol Biol 60(6):929–946
Seo PJ, Kim S-G, Park C-M (2008) Membrane-bound transcription factors in plants. Trends Plant Sci 13(10):550–556
Smita S, Katiyar A, Chinnusamy V, Pandey DM, Bansal KC (2015) Transcriptional regulatory network analysis of MYB transcription factor family genes in Rice. Front Plant Sci. 10.3389/fpls.2015.01157doi:10.3389/fpls.2015.01157
Stock AM, Robinson VL, Goudreau PN (2000) Two-component signal transduction. Annu Rev Biochem 69(1):183–215
Sun W, Cao Z, Li Y, Zhao Y, Zhang H (2007) A simple and effective method for protein subcellular localization using Agrobacterium-mediated transformation of onion epidermal cells. Biologia 62(5):529–532
Tanaka R, Kobayashi K, Masuda T (2011). Tetrapyrrole metabolism in Arabidopsis thaliana. The Arabidopsis Book: e0145
Tsuda K, Ito Y, Sato Y, Kurata N (2011) Positive autoregulation of a KNOX Gene Is essential for shoot apical meristem maintenance in rice. Plant Cell 23(12):4368–4381
Wan H, Zhao Z, Qian C, Sui Y, Malik AA, Chen J (2010) Selection of appropriate reference genes for gene expression studies by quantitative real-time polymerase chain reaction in cucumber. Anal Biochem 399(2):257–261
Waters MT, Moylan EC, Langdale JA (2008) GLK transcription factors regulate chloroplastdevelopment in a cell-autonomous manner. Plant J 56(3):432–444
Wu Y, Wang Y, Mi XF, Shan JX, Li XM, Xu JL, Lin HX (2016) The QTL GNP1 Encodes GA20ox1, which increases grain number and Yield by increasing cytokinin activity in rice panicle meristems. PLoS Genet 12(10):e1006386
Yan JH, Wang B, Zhong YP, Yao LM, Cheng LJ, Wu TL (2015) The soybean R2R3 MYB transcription factor GmMYB100 negatively regulates plant flavonoid biosynthesis. Plant MolBiol 89(1–2):35–48
Yanai O et al (2005) Arabidopsis KNOXI proteins activate cytokinin biosynthesis. Curr Biol 15(17):1566–1571
Acknowledgements
This research was supported by grants from the National Natural Science Foundation of China (Project#31372074), National Key Research and Development Plan (2016YFD0101705)of China, Xi’an Agricultural Science and Technology Innovation Project of China (NC1501(1)) and Shaanxi Province Agricultural Science and Technology Innovation and Research Project of China (2016NY-048). We acknowledge the assistance provided by Dr. Fang Cheng (Northwest A&F University of Shaanxi).
Author information
Authors and Affiliations
Contributions
JL and HL conceived and designed the study. JL and HL constructed the prokaryotic expression, qPCR analysis, sequence and phylogenetic relationship analysis, and wrote the paper. JJ performed the subcellular localization. MC and JX. performed some of the field work. HM, SC and YL provided valuable research ideas. ZC and HL led and supervised the study.
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare that there are no competing interests.
Additional information
Jianqing Jiao, Hanqiang Liu and Jia Liu have contributed equally to this work.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Jiao, J., Liu, H., Liu, J. et al. Identification and functional characterization of APRR2 controlling green immature fruit color in cucumber (Cucumis sativus L.). Plant Growth Regul 83, 233–243 (2017). https://doi.org/10.1007/s10725-017-0304-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10725-017-0304-1