Cloning and Expression Analysis of a Pollen Development Gene MF21 in Broccoli

doi:10.13560/j.cnki.biotech.bull.1985.2015.04.014

Abstract

Abstract: In this study, the MF21 gene were cloned from a broccoli maintenance line ‘WN12-95B’. Bioinformatic analysis showed that the full length of MF21 gene was 468 bp and encoded 151 amino acid peptides. The relative molecular weight and pI of MF21 was 16.70 kD and 8.56, respectively. Meanwhile, the MF21 protein was speculated as a hydrophobic protein. The MF21 protein include a 22 amino acid length signal peptide, and also contain two CK2phosphorylation sites, four N-myristoylaton sites and one tyrosine kinase phosphorylation site. Random coils and β-strands were main components of the two-dimension structure. Based on the molecular evolution, we found that the MF21 gene had approximate evolution relationship between broccoli and Brassica carinata. Theexpression analysis of MF21 gene by qRT-PCR indicated that this gene was mainly expressed in bud of broccoli and had tissue specificity. Theexpressive abundance of bud in different development stage had greatdifferences between the Ogu CMS line and its maintenance line.

Key words: broccoli, pollen development, MF21, gene clone, expression analysis

Pei Xuli, Jing Zan’ge, Tang Zheng, Wang Yan, Wang Zhen, Chen Zhongwen, Luo Tiankuan, Zhang Xiaoling. Cloning and Expression Analysis of a Pollen Development Gene MF21 in Broccoli[J]. Biotechnology Bulletin, 2015, 31(3): 102-107.

References

[1] Yang X, Makaroff CA, Ma H. The Arabidopsis MALE MEIOCYTE DEATH1 gene encodes a PHD-finger protein that is required for male meiosis[J]. The Plant Cell, 2003, 15(6)：1281-1295.
[2] Zhou S, Wang Y, Li W, et al. Pollen Semi-Sterility1 encodes a kinesin-1-like protein important for male meiosis, anther dehiscence, and fertility in rice[J]. The Plant Cell, 2011, 23(1)：111-129.
[3] Jiang J, Yu Y, Dong H, et al. BcMF21 is important for pollen development and germination in Brassica campestris ssp. Chinensis[J]. Molecular Biology Reports, 2014, 41(1)：537-544.
[4] Jiang J, Yu X, Miao Y, et al. Sequence characterization and expression pattern of BcMF21, a novel gene related to pollen development in Brassica campestris ssp. chinensis[J]. Molecular Biology Reports, 2012, 39(7)：7319-7326.
[5] 张强, 黄鹂, 曹家树. 白菜多聚半乳糖醛酸酶基因 BcMF6 的克隆, 序列分析及其表达[J]. 园艺学报, 2007, 34(1)：117-124.
[6] 宋江华, 朱熠鹏, 余小林, 等. BcA9启动子调控的反义 BcMF12 对白菜基因沉默的效果[J]. 中国农业科学, 2008, 41(5)：1410-1415.
[7] 李焰焰, 聂传朋, 曹家树. 十字花科蔬菜 BcMF13 同源基因的克隆及进化分析[J]. 应用与环境生物学报, 2011, 17(2)：207-210.
[8] 韩莹琰, 张爱红, 范双喜, 曹家树. 十字花科植物C2H2型锌指蛋白新基因 BcMF20 同源序列克隆与进化分析[J]. 核农学报, 2011, 25(5)：916-921.
[9] Tingley WG, Ehlers MD, Kameyama K, et al. Characterization of protein kinase A and protein kinase C phosphorylation of the N-methyl-D-aspartate receptor NR1 subunit using phosphorylation site-specific antibodies[J]. Journal of Biological Chemistry, 1997, 272(8)：5157-5166.
[10] Kaneko K, Tabuchi M, Sueyoshi N, et al. Cellular localization of CoPK12, a Ca2+/calmodulin-dependent protein kinase in mushroom Coprinopsis cinerea, is regulated by N-myristoylation[J]. Journal of Biochemistry, 2014, 156(1)：51-61.
[11] Chakraborty A, Werner JK Jr, Koldobskiy MA, et al. Casein kinase-2 mediates cell survival through phosphorylation and degradation of inositol hexakisphosphate kinase-2[J]. Proceedings of the National Academy of Sciences, 2011, 108(6)：2205-2209.
[12] 蒋晶晶. 白菜花粉发育和授粉受精相关基因的表达分析与功能验证[D]. 杭州：浙江大学, 2012.
[13] Nagaharu U. Genome analysis in Brassica with special reference to the experimental formation of B. napus and peculiar mode of fertilization[J]. Journal of Japanese Botany, 1935, 7：389-452.
[14] Udall JA, Quijada PA, Osborn TC. Detection of chromosomal rearrangements derived from homeologous recombination in four mapping populations of Brassica napus L. [J]. Genetics, 2005, 169(2)：967-979.
[15] Mascarenhas JP. Gene activity during pollen development[J]. Annual Review of Plant Biology, 1990, 41(1)：317-338.