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| Analysis of Transactivation Activity and Expression Characteristics of PoMYB1 in Paeonia ostii 'Feng Dan Bai' |
| LIU Peng, LI Wei, XU Zhuang-Zhuang, LIU Qing-Hua, WANG Kui-Ling*, HAO Qing* |
| College of Landscape Architecture and Forestry, Qingdao Agricultural University, Qingdao 266109, China |
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Abstract Pollination and fertilization are key reproductive processes that affects the seed setting of plants. MYB (v-myb avian myeloblastosis viral oncogene homolog) transcription factor is one of the important transcription factor families involved in the regulation of these processes. An unigene sequence sharing high homology with MYB family gene was obtained from gynoecium transcriptome database of previously constructed tree peony (Paeonia ostii 'Feng Dan Bai') and a complete coding region gene named as PoMYB1 (GenBank No. MT360920) was cloned with the expressed sequence tag and rapid amplification of cDNA ends (RACE) methods. PoMYB1 contained a 864 bp long ORF, indicating that it could encode 267 amino acids residues with 2 typical MYB domains. Bioinformatics analysis showed that the molecular formula of PoMYB1 protein was C1434H2235N415O441S12, the relative molecular weight was 32.73 kD and the isoelectric point was 7.01, the fat coefficient was 68.99, and the instability coefficient was 48.18. These characteristics showed that it was an unstable protein. Amino acids alignment revealed that PoMYB1 was composed of highly conserved and incompletely duplicated R2R3-MYB domains. PoMYB1 had 3 characteristic conserved motifs. Phylogenic analysis showed that the PoMYB1 shared the highest similarity with that of PqMYB, followed by MrMYB1, which were obtained from P. qiui and Morella rubra, both belonged to R2R3-MYB transcription factor family of subgroup 6. The expression characteristics of PoMYB1 gene were analyzed by qRT-PCR. The results indicated that the highest in sepal and followed by leaf and petal, which showed that it played a role in the development of flowers and leaves. The highest expressed increment appeared in 24 h after fertilization, and the highest abundance in 48 h after fertilization was observed, which speculated its putative function in double fertilization. Besides, construction of yeast (Saccharomyces cerevisiae) recombinant vectors containing 3 different missing fragments of PoMYB1 and introduced into yeast cells and the transactivation activation activity of PoMYB1 was also verified by yeast one hybrid. The results indicated that PoMYB1 was a transcriptional activator, and the transcriptional regulatory regions in the C-terminus. The study provides theoretical basis for exploring the molecular mechanisms underlying double fertilization in P. ostii and establishes a foundation for further research transcriptional regulation mechanism of PoMYB1.
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Received: 22 March 2020
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Corresponding Authors:
* wkl6310@163.com; haoqing5608@163.com
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[1] 丁爱琴, 李绍翠, 刘庆超, 等. 2019. 月季RhMYB96转录激活与表达特性分析[J]. 农业生物技术学报, 27(3): 411-420.
(Ding A Q, Li S C, Liu Q C, et al.Analysis of transactivation activity and expression characteristics of RhMYB96 in Rosa hybrida[J]. Journal of Agricultural Biotechnology, 27(3): 411-420.)
[2] 董兆磊. 2010. '凤丹'(Paeonia ostii 'Feng Dan')生殖生物学的初步研究[D]. 硕士学位论文, 北京林业大学, 导师: 胡永红, pp. 32-35.
(Dong Z L.2010. Studies on the reproductive biology of Paeonia ostii 'Feng Dan'[D]. Thesis for M.S., Beijing Forest University, Supervisor: Hu Y H, pp. 32-35.)
[3] 李国君, 赵娟. 2017. 玉米ZmWRKY41基因克隆及转录激活验证[J]. 山西农业大学学报(自然科学版), 37(6): 381-389.
(Li G J, Zhao J.Cloning and transcriptional activation of ZmWRKY41 transcription factor from maize[J]. Journal Shanxi Agricultural University (Natural Science Edition), 37(6): 381-389.)
[4] 李真, 郝青, 李伟, 等. 2019. '凤丹白'牡丹中参与受精过程的PobHLH基因的克隆与表达[J]. 植物生理学报, 55(3): 281-289.
(Li Z, Hao Q, Li W, et al.Cloning and Expression of PobHLH involved in fertilization in Paeonia ostii 'Fengdanbai'[J]. Plant Physiology Journal, 55(3): 281-289.)
[5] Borg M, Brownfield L, Khatab H, et al.2011. The R2R3 MYB transcription factor DUO1 activates a male germline-specific regulon essential for sperm cell differentiation in Arabidopsis[J]. Plant Cell, 23(2): 534-549.
[6] Chai G H, Wang Z G, Tang X F, et al.2014. R2R3-MYB gene pairs in Populus: Evolution and contribution to secondary wall formation and flowering time[J]. Journal of Experimental Botany, 65(15): 4255-4269.
[7] Cominelli E, Gusmaroli G, Allegra D, et al.2008. Expression analysis of anthocyanin regulatory genes in response to different light qualities in Arabidopsis thaliana[J]. Journal of Plant Physiology, 165(8): 886-894.
[8] Dai X Y, Wang Y Y, Yang A, et al.2012. OsMYB2P-1, a R2R3 MYB transcription factor, is involved in the regulation of phosphate-starvation responses and root architecture in rice[J]. Plant Physiology, 159(1): 169-183.
[9] Dubos C, Stracke R, Grotewold E, et al.2010. MYB transcription factors in Arabidopsis[J]. Trends in Plant Science, 15(10): 573-581.
[10] Feller A, Machemer K, Braun E L, et al.2011. Evolutionary and comparative analysis of MYB and bHLH plant transcription factors[J]. Plant Journal for Cell and Molecular Biology, 66(1): 94-116.
[11] Gao L X, Yang H X, Liu H F, et al.2016. Extensive transcriptome changes underlying the flower color intensity variation in Paeonia ostii[J]. Frontiers in Plant Science, 6(80): 1205-1220.
[12] Gu M, Zhang J, Li H, et al.2017. Maintenance of phosphate homeostasis and root development are coordinately regulated by MYB1, a R2R3-type MYB transcription factor in rice[J]. Journal of Experimental Botany, 68(13): 3603-3615.
[13] Gu Z Y, Zhu J, Hao Q, et al.2019. A novel R2R3-MYB transcription factor contributes to petal blotch formation by regulating organ-specific expression of PsCHS in Tree Peony (Paeonia suffruticosa)[J]. Plant and Cell Physiology, 60(3): 599-611.
[14] Higashiyama T, Kuroiwa H, Kuroiwa T.2003. Pollen-tube guidance: Beacons from the female gametophyte[J]. Current Opinion in Plant Biology, 6(1):36-41.
[15] Hoeren F U, Dolferus R, Wu Y R, et al.1998. Evidence for a role for AtMYB2 in the induction of the Arabidopsis alcohol dehydrogenase gene (ADH1) by low oxygen[J]. Genetics, 149(2): 479-490.
[16] Johnson M A, Harper J F, Palanivelu R. A.2019. Fruitful Journey: Pollen tube navigation from germination to fertilization[J]. Annual Review of Plant Biology, 70(1): 809-837.
[17] Jiao B, Zhao X, Lu W X, et al.2019. The R2R3 MYB transcription factor MYB189 negatively regulates secondary cell wall biosynthesis in Populus[J]. Tree Physiology, 39(7): 1187-1200.
[18] Leydon A, Beale K, Woroniecka K, et al.2013. Three MYB transcription factors control pollen tube differentiation required for sperm release[J]. Current Biology, 23(13): 1209-1214.
[19] Liang Y, Tan Z M, Zhu L, et al.2013. MYB97, MYB101 and MYB120 function as male factors that control pollen tube-Synergid interaction in Arabidopsis thaliana fertilization[J]. PLOS Genetics, 9(11): e1003933.
[20] Makkena S, Lee E, Sack F D, et al.2012. The R2R3 MYB transcription factors FOUR LIPS and MYB88 regulate female reproductive development[J]. Journal of Experimental Botany, 63(15): 5545-5558.
[21] Md A R, Francesca R, Francesca D V, et al.2019. Effects on plant growth and reproduction of a peach R2R3-MYB transcription factor overexpressed in tobacco[J]. Frontiers in Plant Science, 10: 1143-1159.
[22] Rabiger D S, Drews G N.2013. MYB64 and MYB119 are required for cellularization and differentiation during female gametogenesis in Arabidopsis thaliana[J]. PLOS Genetics, 9(9): e1003783.
[23] Suo J, Liang X, Pu L, et al., 2003. Identification of GhMYB109 encoding a R2R3 MYB transcription factor that expresssed specifically in fiber initials and elongating fibers of cotton (Gossypium hirsutum L.)[J]. Biochimica et Biophysica Acta, 1630(1): 25-34.
[24] Wilkins O, Nahal H, Foong Jet al.2009. Expansion and diversification of the Populus R2R3-MYB family of transcription factors[J]. Plant Physiology, 149(2): 981-993.
[25] Wu P, Peng M, Li Z, et al.2019. DRMY1, a Myb-Like protein, regulates cell expansion and seed production in Arabidopsis thaliana[J]. Plant and Cell Physiology, 60(2): 285-302.
[26] Xu Y, Lacuone S, Li S F, et al.2014. MYB80 homologues in Arabidopsis, cotton and Brassica: Regulation and functional conservation in tapetal and pollen development[J]. BMC Plant Biology, 14(1): 278-292.
[27] Yang C J, Song J, Ferguson A, et al.2017. Transcription factor MYB26 is key to spatial specificity in anther secondary thickening formation[J]. Plant Physiology, 175(1): 333-350.
[28] Yao L F, Yang B, Xian B S, et al.2020. The R2R3-MYB transcription factor BnaMYB111L from rapeseed modulates reactive oxygen species accumulation and hypersensitive-like cell death[J]. Plant Physiology and Biochemistry, 147: 280-288.
[29] Zhang Y J, Liang W Q, Shi J X, et al.2013. MYB56 encoding a R2R3 MYB transcription factor regulates seed size in Arabidopsis thaliana[J]. Journal of Integrative Plant Biology, 55(11): 1166-1178.
[30] Zhang X P, Zhao L Y, Xu Z D, et al.2018. Transcriptome sequencing of Paeonia suffruticosa 'Shima Nishiki' to identify differentially expressed genes mediating double-color formation[J]. Plant Physiology and Biochemistry, 123: 114-124. |
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