Cloning and expression analysis of GhDET3, a vacuolar H+-ATPase subunit C gene, from cotton

Zhongyi Xiao; Kunling Tan; Mingyu Hu; Peng Liao; Kuijun Chen; Ming Luo

doi:10.1016/S1673-8527(08)60044-2

Volume 35 Issue 5

May 2008

Turn off MathJax

Article Contents

Article Navigation > Journal of Genetics and Genomics > 2008 > 35(5): 307-312

PDF( 608 KB)

Cloning and expression analysis of GhDET3, a vacuolar H+-ATPase subunit C gene, from cotton

doi: 10.1016/S1673-8527(08)60044-2

Key Laboratory of Biotechnology and Crop Quality Improvement, Ministry of Agriculture; Biotechnology Research Center, Southwest University, Chongqing 400716, China

More Information

Corresponding author: E-mail address: luomingyuan@swu.edu.cn (Ming Luo)
Received Date: 2008-01-15
Accepted Date: 2008-01-26
Rev Recd Date: 2008-01-25

Available Online: 2008-05-20

Publish Date: 2008-05-20

Abstract

Abstract

Vacuolar H⁺-ATPase was regarded as a key enzyme promoting the fiber cell elongation in cotton (Gossypium hirsuturm L.) through regulating turgor-driven pressure involved in polarity expansion of single cell fiber. The DET3, a V-ATPase subunit C, plays an important role in assembling subunits and regulating the enzyme activity, and is involved in Brassinosteroid-induced cell elongation. To analyze the function of GhDET3 on the elongation of cotton fibers, seven candidates of ESTs were screened and contigged for a 5′-upstream sequence, and the 3′-RACE technique was used to clone the 3′-downstream sequence for the full length of GhDET3 gene. The full length of the target clone was 1,340 bp, including a 10 bp 5′-UTR, an ORF of 1,134 bp, and a 196 bp 3′-UTR. This cDNA sequence encoded a polypepide of 377 amino acid residues with a predicted molecular mass of 43 kDa and a basic isoelectric point of 5.58. Furthermore, a length of 3,410 bp sequence from genomic DNA ofGhDET3 was also cloned by PCR. The deduced amino acid sequence had a high homology with DET3 from Arabidopsis, rice, and maize. Quantitative real-time PCR (qRT-PCR) analysis showed that the GhDET3 expression pattern was ubiquitous in all the tissues and organs detected. The result also revealed that the accumulation of GhDET3 mRNA reached the highest profile at the fiber elongation stage in 12 DPA (days post anthesis) fibers, compared with the lowest level at the fiber initiation stage in 0 DPA ovules (with fibers). The transcript accumulation in fibers and ovules shared the similar variation tendency. In addition, in vitro ovule culture experiment demonstrated that exogenous 24-EBL treatment to 4 DPA ovules (with fibers) was capable of increasing the expression level of GhDET3, and the mRNA accumulation of GhDET3 increased in transgenic FBP7::GhDET2 cotton fibers in vivo. These results indicate that GhDET3 gene plays a crucial role in cotton fiber elongation.
- cotton fiber,
- GhDET3,
- Brassinosteroids

FullText(HTML)

References(26)

References

[1]	Basra, A.S., Malik, C.P. Development of the cotton fiber Int. Rev. Cytol., 89 (1984),pp. 65-113
[2]	Beasley, C.A., Ting, I.P. Am. J. Bot., 60 (1973),pp. 130-139
[3]	Beasley, C.A., Ting, I.P. Am. J. Bot., 61 (1974),pp. 188-194
[4]	Brown, J.W., Feix, G., Frendewey, D. EMBO J., 5 (1986),pp. 2749-2758
[5]	Cabrera, Y., Poch, H., Peto, C. et al. Plant J., 4 (1993),pp. 6671-6682
[6]	Cosgrove, D.J., Durachko, D.M. Automated pressure probe for measurement of water transport properties of higher plant cells Rev. Scient. Instrum., 10 (1986),pp. 2614-2619
[7]	Frohman, M.A., Martin, G.R., Martin, G.R. Rapid production of full-length cDNAs from rare transcripts amplification using a single gene-specific oligonucleotide primer Proc. Natl. Acad. Sci. USA, 85 (1988),pp. 8998-9002
[8]	Ho, M.N., Hill, K.J., Lindorfer, M.A. et al. J. Biol. Chem., 268 (1993),pp. 221-227
[9]	Hofmann, K., Stoffel, W. TM base-A database of membrane spanning protein segments Biol. Chem., 374 (1993),p. 166
[10]	John, B., Louise, C.S., Ross, J. et al. Fiber diffraction of synthetic α-synuclein filaments shows amyloid-like cross-β conformation Proc. Natl. Acad. Sci. USA, 97 (2000),pp. 4897-4902
[11]	Lawrence, B.S., Fakrieh, V., Masayoshi, M. et al. Genes involved in osmoregulation during turgor-driven cell expansion of developing cotton fibers are differentially regulated Plant Physiol., 116 (1998),pp. 1539-1549
[12]	Li, H.M., Altschmieed, L., Chory, J. Genes Dev., 8 (1994),pp. 339-349
[13]	Luo, M., Xiao, Y.H., Li, X.B. et al. GhDET2, a steroid 5α-reductase, plays an important role in cotton fiber cell initiation and elongation Plant J., 51 (2007),pp. 419-430
[14]	Luo, M., Xiao, Y.H., Hou, L. et al. Acta Genet. Sin., 30 (2003),pp. 175-182
[15]	Omri, D., Felix, F., Nathan, N. Crystal structure of yeast V-ATPase subunit C reveals its stator function EMBO Rep., 5 (2004),pp. 1148-1152
[16]	Schubert, A.M., Benedict, C.R., Berlin, J.D. et al. Cotton fiber development-kinetics of cell elongation and secondary wall thickening Crop Sci., 13 (1973),pp. 704-709
[17]	Schumacher, K., Vafeados, D., McCarthy, M. et al. Genes Dev., 13 (1999),pp. 3259-3270
[18]	Shi, Y.H., Zhu, S.W., Mao, X.Z. et al. Transcriptome profiling, molecular biological, and physiological studies reveal a major role for ethylene in cotton fiber cell elongation Plant Cell, 18 (2006),pp. 651-664
[19]	Stevens, T.H., Forgac, M. Annu. Rev. Cell Dev. Biol., 13 (1997),pp. 779-808
[20]	Sun, Y., Allen, R.D. Mol. Genet. Genomics, 274 (2005),pp. 51-59
[21]	Sun, Y., Fokar, M., Asami, T. et al. Characterization of the Brassinosteroid insensitive 1 genes of cotton Plant. Mol. Biol., 54 (2004),pp. 221-232
[22]	Sze, H. Annu. Rev. Plant Physiol., 36 (1985),pp. 175-208
[23]	Sze, H., Schumacher, K., Müller, M.L. et al. Trends Plant Sci., 7 (2002),pp. 157-161
[24]	Tiwari, S.C., Wilkins, T.A. Can. J. Bot., 73 (1995),pp. 746-757
[25]	Ward, J.M., Reinders, A., Hsu, H.T. et al. Plant Physiol., 99 (1992),pp. 161-169
[26]	Xiao, Y.H., Luo, M., Fang, W.G. et al. PCR walking in cotton genome using YADE method Acta Genet. Sin., 29 (2002),pp. 62-66