Abstract
CMS-D8 and its restorer were developed by introducing the cytoplasm and nuclear gene Rf 2 from the wild diploid Gossypium trilobum (D8) into the cultivated tetraploid Upland cotton (Gossypium hirsutum). No information is available on how the Rf 2 gene interacts with CMS-associated genes and how CMS-D8 cytoplasm affects nuclear gene expression. The objective of this study was to identify differentially expressed genes in anther tissues between the non-restoring fertile maintainer ARK8518 (rf 2 rf 2) and its isogenic heterozygous D8 restorer line, ARK8518R (Rf 2 rf 2) with D8 cytoplasm, by mRNA differential display (DD). Out of more than 3,000 DDRT-PCR bands amplified by 31 primer combinations from 12 anchor primers and 8 arbitrary decamer primers, approximately 100 bands were identified as being qualitatively differentially displayed. A total of 38 cDNA fragments including 12 preferentially expressed cDNA bands in anther were isolated, cloned and sequenced. Reverse northern blot analysis showed that only 4 genes, including genes encoding a Cys-3-His zinc finger protein and aminopeptidase, were up-regulated, while 22 genes, including genes for phosphoribosylanthranilate transferase (PAT), starch synthase (SS), 4-coumarate-CoA ligase, electron transporter, calnexin, arginine decarboxylase, and polyubiquitin, were down-regulated in the heterozygous restorer ARK8518R. The down-regulation of SS explains the lack of starch accumulation in sterile rf 2 pollen grains in the heterozygous restored plants. The molecular mechanism of CMS and its restoration, specifically the possible roles of SS and PAT genes in relation to restoration of Rf 2 to CMS-D8, are discussed. This investigation represents the first account of such an analysis in cotton.
Similar content being viewed by others
References
Bae KM, Kwon YS, Cho IH, Yi SI (2006) Use of cDNA-AFLP for transcript profiling in narrow genetic pools; for example, cucumber (Cucumis sativus L). Plant Breed 125:488–492
Carlsson J, Lagercrantz U, Sundstrom J, Teixeira R, Wellmer F, Meyerowitz EM, Glimelius K (2007) Microarray analysis reveals altered expression of a large number of nuclear genes in developing cytoplasmic male sterile Brassica napus flowers. Plant J 49:452–462
Chase CD (2006) Cytoplasmic male sterility: a window to the world of plant mitochondrial–nuclear interactions. Trends Genet 23:81–90
Datta R, Chamusco KC, Chourey PS (2002) Starch biosynthesis during pollen maturation is associated with altered patterns of gene expression in maize. Plant Physiol 130:1645–1656
Fujii S, Komatsu S, Toriyama K (2007) Retrograde regulation of nuclear gene expression in CW-CMS of rice. Plant Mol Biol 63:405–417
Gaillard C, Moffatt BA, Blacker M, Laloue M (1998) Male sterility associated with APRT deficiency in Arabidopsis thaliana results from a mutation in the gene APT1. Mol Gen Genet 257:348–353
Geddy R, Mahé L, Brown GG (2005) Cell-specific regulation of a Brassica napus CMS-associated gene by a nuclear restorer with related effects on a floral homeotic gene promoter. Plant J 41:333–345
Hanson M, Bentolila S (2004) Interactions of mitochondrial and nuclear genes that affect male gametophyte development. Plant Cell 16(Suppl):S154–S169
Hong SJ, Dawson VL, Dawson TM (2005) Identification and evaluation of NO-regulated genes by differential analysis of primary cDNA library expression (DAzLE). Methods Enzymol 396:359–368
Jensen MK, Rung JH, Gregersen PL, Gjetting T, Fuglsang AT, Hansen M, Joehnk N, Lyngkjaer MF, Collinge DB (2007) The HvNAC6 transcription factor: a positive regulator of penetration resistance in barley and Arabidopsis. Plant Mol Biol 65:137–150
Kaul MLH (1988) Male sterility in higher plants. Springer, New York
Kempken F, Pring D (1999) Plant breeding: male sterility in higher plants: fundamentals and applications. Prog Bot 60:139–166
Kim DH, Kang JG, Kim BD (2007) Isolation and characterization of the cytoplasmic male sterility-associated orf456 gene of chili pepper (Capsicum annuum L.). Plant Mol Biol 63:519–532
Liang P, Pardee AB (1992) Differential display of eukaryotic messenger RNA by means of the polymerase chain reaction. Science 257:967–971
Liang P, Pardee AB (1997) Differential display methods and protocols. Humana Press
Lieven S, Goormachtig S, Holsters M (2001) A critical evaluation of differential display as a tool to identify genes involved in legume nodulation: looking back and looking forward. Nucleic Acids Res 29:3459–3468
Linke B, Northnagel T, Borner T (2003) Flower development in carrot CMS plants: mitochondria affect the expression of MADSbox genes homologous to BLOBOSA and DEFICIENS. Plant J 34:27–37
Mandaokar A, Kumar VD, Amway M, Browse J (2003) Microarray and differential display identify genes involved in jasmonate-dependent anther development. Plant Mol Biol 52:775–777
Moffatt B, Somerville CR (1988) Positive selection for male-sterile mutants of Arabidopsis thaliana lacking adenine phosophoribosyltransferase activity. Plant Physiol 86:1150–1154
Pasentsis K, Falara V, Pateraki I, Gerasopoulos D, Kanellis AK (2007) Identification and expression profiling of low oxygen regulated genes from Citrus flavedo tissues using RT-PCR differential display. J Exp Bot 58:2203–2216
Regan SM, Moffatt BA (1990) Cytochemical analyses of pollen development in wild-type Arabidopsis and a male-sterile mutant. Plant Cell 2:877–889
Rose AB, Casselman AL, Last RL (1992) A phosphoribosylanthranilate transferase gene is defective in blue fluorescent Arabidopsis thaliana tryptophan mutants. Plant Physiol 100:582–592
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor, NY
Sanabria M, Dubery IA (2006) Differential display profiling of the Nicotiana response to LPS reveals elements of plant basal resistance. Biochem Biophys Res Commun 344:1001–1007
Schnable PS, Wise RP (1998) The nuclear basis of cytoplasmic male sterility and fertility restoration. Trends Plant Sci 3:175–180
Sheoran IS, Saini HS (1996) Drought-induced male sterility in rice: changes in carbohydrate levels and enzyme activities associated with the inhibition of starch accumulation in pollen. Sex Plant Reprod 9:161–169
Smith AM, Denyer K, Martin C (1997) The synthesis of the starch granule. Annu Rev Plant Physiol Plant Mol Biol 48:67–87
Song P, Yamamoto E, Allen RD (1995) Improved procedure for differential display of transcripts from cotton tissues. Plant Mol Biol Rep 13:174–178
Stewart JM (1992) A new cytoplasmic male sterile and restorer for cotton. Proc Beltwide Cotton Conf National Cotton Council, Memphis, p 610
Takeuchi H, Fujiyuki T, Shirai K, Matsuo Y, Kamikouchi A, Fujinawa Y, Kato A, Tsujimoto A, Kubo T (2002) Identification of genes expressed preferentially in the honeybee mushroom bodies by combination of differential display and cDNA microarray. FEBS Lett 513:230–234
Teixeira RT, Knorpp C, Glimelius K (2005) Modified sucrose, starch, and ATP leveles in two alloplasmic male-sterile lines of B. napus. J Exp Bot 56:1245–1253
Wan C, Wilkins TA (1994) A modified hot borate method significantly enhances the yield of high-quality RNA from cotton (Gossypium hirsutum L.). Anal Biochem 223:7–12
Weyman PD, Pan Z, Feng Q, Gilchrist DG, Bostock RM (2006) A circadian rhythm-regulated tomato gene is induced by arachidonic acid and Phythophthora infestans infection. Plant Physiol 140:235–248
Yamamoto MP, Kubo T, Mikami T (2005) The 5′-leader sequence of sugar beet mitochondrial atp6 encodes a novel polypeptide that is characteristic of Owen cytoplasmic male sterility. Mol Genet Genomics 273:342–349
Zhang JF, Stewart JMcD (2001a) CMS-D8 restoration in cotton is conditioned by one dominant gene. Crop Sci 41:283–288
Zhang JF, Stewart JMcD (2001b) Inheritance and genetic relationships of the D8 and D2-2 restorer genes for cotton cytoplasmical male sterility. Crop Sci 41:289–294
Zhang H, Li S, Yi P, Wan C, Chen Z, Zhu Y (2007) A Honglian CMS line of rice displays aberrant F(0) of F (0)F (1)-ATPase. Plant Cell Rep 26:1065–1071
Acknowledgments
The authors thank Dr. A. L. Nepomuceno for his assistance in DD-RTPCR and Dr. L. Z. Xiong for his assistance in performing reverse northern blot analysis. The research reported here was in part funded by the Arkansas Agricultural Experiment Station and in part by the New Mexico Agricultural Experiment Station.
Author information
Authors and Affiliations
Corresponding authors
Additional information
Communicated by C. Quiros.
Rights and permissions
About this article
Cite this article
Zhang, J., Turley, R.B. & Stewart, J.M. Comparative analysis of gene expression between CMS-D8 restored plants and normal non-restoring fertile plants in cotton by differential display. Plant Cell Rep 27, 553–561 (2008). https://doi.org/10.1007/s00299-007-0492-7
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00299-007-0492-7