Responses of Overexpressed Arabidopsis VHA-c3 to Dark，ABA and Sugar

doi:10.13560/j.cnki.biotech.bull.1985.2016.06.013

Abstract

Abstract: For studying the role of vacuole H⁺-ATPase subunit c gene(VHA-c3)in plant growth and abiotic stress response，an over-expression vector of VHA-c3 was constructed and introduced into wild-type Arabidopsis thaliana. The expression levels of VHA-c3 in the transgenic homozygote’s plants were detected by semi-quantitative RT-PCR，then the transgenic homozygote were treated by dark，ABA and sugar. The results indicated that 6 lines of homozygous T2 transgenic plants were obtained，and their corresponding transcript levels were increased. The root lengths of 5 lines were reduced among the dark-grown seedlings. Among the ABA-treated seedlings，3 lines were insensitive in primary root growth and the extent of cotyledon unfolding，and 5 lines were insensitive in seed germination. When treated by glucose and sucrose，5 lines and 6 lines reduced the sensitivity in seed germination，respectively. These results indicate that VHA-c3 may affect the cell expansion of root，and be involved in the signal transduction pathways mediated by ABA and sugar.

Key words: Arabidopsis thaliana, VHA-c3, dark, ABA, sugar

SU Jie, GUO Rong-qi, LI Guo-jing, WANG Rui-gang. Responses of Overexpressed Arabidopsis VHA-c3 to Dark，ABA and Sugar[J]. Biotechnology Bulletin, 2016, 32(6): 89-95.

References

[1]Sze H, Li X, Palmgren M. Energization of the plant cell membranes by H⁺-pumping ATPases：biosynthesis and regulation[J]. The Plant Cell, 1999, 11：677-689.
[2]Dietz KJ, Tavakoli N, Kluge C, et al. Significance of the V-type ATPase for the adaptation to stressful growth conditions and its regulation on the molecular and biochemical level[J]. Journal of Experimental Botany, 2001, 52：1969-1980.
[3]Schumacher K, Krebs M. The V-ATPase：small cargo, large effects[J]. Current Opinion in Plant Biology, 2010, 13(6)：724-730.
[4]Arata Y, Baleja JD, Forgac M. Cysteine-directed cross-linking to subunit B suggests that subunit E forms part of the peripheral stalk of the vacuolar H⁺-ATPase[J]. The Journal of Biological Chemistry, 2002, 77：3357-3363.
[5]Hirata T, Iwamoto-kihara A, Sun-wada GH, et al. Subunit rotation of vacuolar-type proton pumping ATPase：relative rotation of the G as to c subunit[J]. The Journal of Biological Chemistry, 2003, 278：23714-23719.
[6]Zhou A, Bu Y, Takano T, et al. Conserved V-ATPase c subunit plays a role in plant growth by influencing V-ATPase-dependent endosomal trafficking[J]. Plant Biotechnology Journal, 2016, 14(1)：271-283.
[7] Tsiantis MS, Bartholomew DM, Smith JAC. Salt regulation of transcript levels for the c subunit of a leaf vacuolar H⁺-ATPase in the halophyte Mesembryanthemum crystallinum[J]. The Plant Journal for Cell and Molecular Biology, 1996, 9：729-736.
[8]Baisakh N, Ramanarao MV, Rajasekaran K, et al. Enhanced salt stress tolerance of rice plants expressing a vacuolar H⁺-ATPase subunit c1(SaVHAc1)gene from the halophyte grass Spartina alterniflora L?isel[J]. Plant Biotechnology Journal, 2012, 10(4)：453-464.
[9]Feng L, Ding H, Wang J, et al. Molecular cloning and expression analysis of RrNHX1 and RrVHA-c genes related to salt tolerance in wild Rosa rugosa[J]. Saudi Journal of Biological Sciences, 2015, 22(4)：417-423.
[10]Sze H, Schumacher K, Muller ML, et al. A simple nomenclature for a complex proton pump：VHA genes encode the vacuolar H⁺-ATPase[J]. Trends in Plant Science, 2002, 7：157-161.
[11]Padmanaban S, Lin X, Perera I, et al. Differential expression of vacuolar H⁺-ATPase subunit c genes in tissues active in membrane trafficking and their roles in plant growth as revealed by RNAi[J]. Plant Physiology, 2004, 134(4)：1514-1526．
[12]徐萍, 李小方, 曾卫军, 等. 拟南芥VHA-c3基因的特异性表达和调节[J]. 华东师范大学学报：自然科学版, 2006, 2：98-104.
[13]郭荣起, 苏杰, 王福慧, 等. 拟南芥VHA-c3启动子的GUS基因融合表达[J]. 中国生物工程杂志, 2008, 28(7)：58-62．
[14]徐萍, 刘瑞香, 曾卫军, 等. 拟南VHA-c基因在非生物胁迫响应中的作用[J]. 华北农学报, 2006, 21(1)：19-22.
[15]邸娜．拟南芥液泡H⁺-ATPase c亚基基因功能的研究[D]．呼和浩特：内蒙古农业大学, 2006．
[16]于秀敏, 武燕, 王瑞刚. 温度和光照对拟南芥VHA-c基因启动子活性的调节[J]. 河南师范大学学报：自然科学版, 2013, 41(4)：120-123.
[17]张春霖, 尚世辉, 王瑞刚．拟南芥VHA-c基因的矿质营养调节[J]. 河南科技大学学报：自然科学版, 2014, 35(5)：79-81.
[18]Murray MG, Thompson WF. Rapid isolation of high molecular weight plant DNA[J]. Nucleic Acids Research, 1980, 8(19)：4321-4325．
[19]Mersereau M, Pazour GJ, Das A. Efficient transformation of Agrobacterium tumefaciens by electroporation[J]. Gene, 1990, 90(1)：149-151．
[20]Clough SJ, Bent AF. Floral dip：a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana[J]. The Plant Journal：for Cell and Molecular Biology, 1998, 16(6)：735-743.
[21]Smart LB, Vojdani F, Maeshima M, et al. Genes involved in osmoregulation during turgor-driven cell expansion of developing cotton fibers are differentially regulated[J]. Plant Physiology, 1998, 116：1539-1549.
[22]Dietz KJ, Rudloff S, Ageorges A, et al. Subunit E of the vacuolar H⁺-ATPase of Hordeum vulgare L.：cDNA cloning, expression and immunological analysis[J]. The Plant Journal：for Cell and Molecular Biology, 1995, 8：521-529.
[23]Schumacher K, Vafeados D, Mccarthy M, et al. The Arabidopsis det3 mutant reveals a central role for the vacuolar H⁺-ATPase in plant growth and development[J]. Genes﹠Development, 1999, 13：3259-3270.
[24]Gogarten JP, Fichmann J, Braun Y, et al. The use of antisense mRNA to inhibit the tonoplast H⁺-ATPase in carrot[J]. Plant Cell, 1992, 4：851-864.
[25]Barkla BJ, Vera-estrella R, Maldonado-gama M, et al. Abscisic acid induction of vacuolar H⁺-ATPase activity in Mesembryanthemum crystallinum is developmentally regulated[J]. Plant Physiology, 1999, 120(3)：811-820.
[26]Chen Y, Ji F, Xie H, et al. The regulator of G-protein signaling proteins involved in sugar and abscisic acid signaling in Arabidopsis seed germination[J]. Plant Physiology, 2006, 140(1)：302-310．
[27]Chen H, Zhang J, Neff MM. Integration of light and abscisic acid signaling during seed germination and early seedling development[J]. Proceedings of the National Academy of Sciences of the United States of America, 2008, 105(11)：4495-4500．