Plant Cell Reports

, Volume 32, Issue 10, pp 1531–1542 | Cite as

Transcriptome analysis of cytoplasmic male sterility and restoration in CMS-D8 cotton

  • Hideaki Suzuki
  • Laura Rodriguez-Uribe
  • Jiannong Xu
  • Jinfa Zhang
Original Paper


Key message

A global view of differential expression of genes in CMS-D8 of cotton was presented in this study which will facilitate the understanding of cytoplasmic male sterility in cotton.


Cytoplasmic male sterility (CMS) is a maternally inherited trait in higher plants which is incapable of producing functional pollen. However, the male fertility can be restored by one or more nuclear-encoded restorer genes. A genome-wide transcriptome analysis of CMS and restoration in cotton is currently lacking. In this study, Affymetrix GeneChips© Cotton Genome Array containing 24,132 transcripts was used to compare differentially expressed (DE) genes of flower buds at the meiosis stage between CMS and its restorer cotton plants conditioned by the D8 cytoplasm. A total of 458 (1.9 %) of DE genes including 127 up-regulated and 331 down-regulated ones were identified in the CMS-D8 line. Quantitative RT-PCR was used to validate 10 DE genes selected from seven functional categories. The most frequent DE gene group was found to encode putative proteins involved in cell wall expansion, such as pectinesterase, pectate lyase, pectin methylesterase, glyoxal oxidase, polygalacturonase, indole-3-acetic acid-amino synthetase, and xyloglucan endo-transglycosylase. Genes in cytoskeleton category including actin, which plays a key role in cell wall expansion, cell elongation and cell division, were also highly differentially expressed between the fertile and CMS plants. This work represents the first study in utilizing microarray to identify CMS-related genes by comparing overall DE genes between fertile and CMS plants in cotton. The results provide evidence that many CMS-associated genes are mainly involved in cell wall expansion. Further analysis will be required to elucidate the molecular mechanisms of male sterility which will facilitate the development of new hybrid cultivars in cotton.


Cotton Cytoplasmic male sterility (CMS) CMS-D8 Microarray Differentially expressed genes 


  1. Ahmed FE, Hall AE, DeMason DA (1992) Heat injury during floral development in cowpea (Vigna unguiculata, Fabaceae). Am J Bot 79:784–791CrossRefGoogle Scholar
  2. Aitken A (2006) 14–3-3 proteins: a historic overview. Semin Cancer Biol 16:162–172PubMedCrossRefGoogle Scholar
  3. Amagai M, Ariizumi T, Endo M, Hatakeyama K, Kuwata C, Shibata D, Toriyama K, Watanabe M (2003) Identification of anther-specific genes in a cruciferous model plant, Arabidopsis thaliana, by using a combination of Arabidopsis macroarray and mRNA derived from Brassica oleracea. Sex Plant Reprod 15:213–220Google Scholar
  4. Baniwal SK, Bharti K, Chan KY, Fauth M, Ganguli A, Kotak S, Mishra SK, Nover L, Port M, Scharf KD et al (2004) Heat stress response in plants: a complex game with chaperones and more than twenty heat stress transcription factors. J Biosci 29:471–487PubMedCrossRefGoogle Scholar
  5. Becker JD, Boavida LC, Carneiro J, Haury M, Feijo JA (2003) Transcriptional profiling of Arabidopsis tissues reveals the unique characteristics of the pollen transcriptome. Plant Physiol 133:713–725PubMedCrossRefGoogle Scholar
  6. Blackmore S, Wortley AH, Skvarla JJ, Rowley JR (2007) Pollen wall development in flowering plants. New Phytol 174:483–498PubMedCrossRefGoogle Scholar
  7. Blanchoin L, Boujemaa-Paterski R, Henty JL, Khurana P, Staiger CJ (2010) Actin dynamics in plant cells: a team effort from multiple proteins orchestrates this very fast-paced game. Curr Opin Plant Biol 13:714–723PubMedCrossRefGoogle Scholar
  8. Bourquin V, Nishikubo N, Abe H, Brumer H, Denman S, Eklund M, Christiernin M, Teeri TT, Sundberg B, Mellerowicz EJ (2002) Xyloglucan endotransglycosylases have a function during the formation of secondary cell walls of vascular tissues. Plant Cell 2002:3073–3088Google Scholar
  9. Bunney TD, van Walraven HS, de Boer AH (2001) 14–3-3 protein is a regulator of the mitochondrial and chloroplast ATP synthase. Proc Natl Acad Sci USA 98:4249–4254PubMedCrossRefGoogle Scholar
  10. Campbell P, Braam J (1999) Xyloglucan endotransglycosylases: diversity of genes, enzymes and potential wall-modifying functions. Trends Plant Sci 4:361–366PubMedCrossRefGoogle Scholar
  11. Carlsson J, Lagercrantz U, Sundstro 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 49:452–462PubMedCrossRefGoogle Scholar
  12. Carpita NC, Gibeaut DM (1993) Structural models of primary cell walls in flowering plants: consistency of molecular structure with the physical properties of the walls during growth. Plant J 3:1–30PubMedCrossRefGoogle Scholar
  13. Cecchetti V, Altamura M, Falasca G, Costantino P, Cardarelli M (2008) Auxin regulates Arabidopsis anther dehiscence, pollen maturation, and filament elongation. Plant Cell 20:1760–1774PubMedCrossRefGoogle Scholar
  14. Cheng H, Qin L, Lee S, Fu X, Richards DE, Cao D, Luo D, Harberd NP, Peng J (2004) Gibberellin regulates Arabidopsis floral development via suppression of DELLA protein function. Development 131:1055–1064PubMedCrossRefGoogle Scholar
  15. Cheng Y, Dai X, Zhao Y (2006) Auxin biosynthesis by the YUCCA flavin monooxygenases controls the formation of floral organs and vascular tissues in Arabidopsis. Genes Dev 20:1790–1799PubMedCrossRefGoogle Scholar
  16. De Rienzo F, Gabdoulline RR, Menziani MC, Wade RC (2000) Blue copper proteins: a comparative analysis of their molecular interaction properties. Protein Sci 9:1439–1454PubMedCrossRefGoogle Scholar
  17. Dharmasiri N, Dharmasiri S, Weijers D, Lechner E, Yamada M, Hobbie L, Ehrismann JS, Jürgens G, Estelle M (2005) Plant development is regulated by a family of auxin receptor F box proteins. Dev Cell 9:109–119PubMedCrossRefGoogle Scholar
  18. Dieterich JH, Braun HP, Schmitz UK (2003) Alloplasmic male sterility in Brassica napus (CMS ‘Tournefortii-Stiewe’) is associated with a special gene arrangement around a novel atp9 gene. Mol Genet Genomics 269:723–731PubMedCrossRefGoogle Scholar
  19. Dong X, Kim WK, Lim YP, Kim YK, Hur Y (2013) Ogura-CMS in Chinese cabbage (Brassica rapa ssp. pekinensis) causes delayed expression of many nuclear genes. Plant Sci 199–200:7–17PubMedCrossRefGoogle Scholar
  20. Dzeja P, Terzic A (2009) Adenylate kinase and AMP signaling networks: metabolic monitoring, signal communication and body energy sensing. Int J Mol Sci 10:1729–1772PubMedCrossRefGoogle Scholar
  21. Endo M, Matsubara H, Kokubuna T, Masuko H, Takahata Y, Tsuchiya T, Fukuda H, Demura T, Watanabe M (2002) The advantages of cDNA microarray as an effective tool for identification of reproductive organ-specific genes in a model legume, Lotus japonicas. FEBS Lett 514:229–237PubMedCrossRefGoogle Scholar
  22. Feng C, Wilson HL, Hurley JK et al (2003) Essential role of conserved arginine 160 in intramolecular electron transfer in human sulfite oxidase. Biochemistry 42:12235–12242PubMedCrossRefGoogle Scholar
  23. Fujii S, Toriyama K (2008) DCW11, down-regulated gene 11 in CW-type cytoplasmic male sterile rice, encoding mitochondrial protein phosphatase 2c is related to cytoplasmic male sterility. Plant Cell Physiol 49:633–640PubMedCrossRefGoogle Scholar
  24. Fujii S, Komatsu S, Toriyama K (2007) Retrograde regulation of nuclear gene expression in CW-CMS of rice. Plant Mol Biol 63:405–417PubMedCrossRefGoogle Scholar
  25. Fujii S, Yamada M, Toriyama K (2009) Cytoplasmic male sterility-related protein kinase, OsNek3, is regulated downstream of mitochondrial protein phosphatase 2C, DCW11. Plant Cell Physiol 50:828–837PubMedCrossRefGoogle Scholar
  26. Goldberg RB, Beals TP, Sanders PM (1993) Anther development: basic principles and practical applications. Plant Cell 5:1217–1229PubMedGoogle Scholar
  27. Hadfield KA, Bennett AB (1998) Polygalacturonases: many genes in search of a function. Plant Physiol 117:337–343PubMedCrossRefGoogle Scholar
  28. Hanson MR, Bentolila S (2004) Interactions of mitochondrial and nuclear genes that affect male gametophyte development. Plant Cell 16:S154–S169PubMedCrossRefGoogle Scholar
  29. Honys D, Twell D (2003) Comparative analysis of the Arabidopsis pollen transcriptome. Plant Physiol 132:640–652PubMedCrossRefGoogle Scholar
  30. Honys D, Twell D (2004) Transcriptome analysis of haploid male gametophyte development in Arabidopsis. Genome Biol 5:R85PubMedCrossRefGoogle Scholar
  31. Huang J, Lee SH, Lin C, Medici R, Hack E, Myers AM (1990) Expression in yeast of the T-urf13 protein from Texas male-sterile maize mitochondria confers sensitivity to methomyl and to Texascytoplasm-specific fungal toxins. EMBO J 9:339–347PubMedGoogle Scholar
  32. Huang S, Cerny RE, Qi Y, Bhat D, Aydt CM, Hanson DD, Malloy KP, Ness LA (2003) Transgenic studies on the involvement of cytokinin and gibberellin in male development. Plant Physiol 131:1270–1282PubMedCrossRefGoogle Scholar
  33. Jiang CZ, Lu F, Imsabai W (2008) Silencing polygalacturonase expression inhibits tomato petiole abscission. J Exp Bot 59:973–979PubMedCrossRefGoogle Scholar
  34. Kieber J, Rothenberg M, Roman G, Feldmann K, Ecker J (1993) CTR1, a negative regulator of the ethylene response pathway in Arabidopsis, encodes a member of the raf family of protein kinases. Cell 72:427–441PubMedCrossRefGoogle Scholar
  35. Klinman JP (1996) Mechanisms whereby mononuclear copper proteins functionalize organic substrates. Chemical Rev 96:2541–2562CrossRefGoogle Scholar
  36. Knox RB (1984) The pollen grain. In: Johri BM (ed) Embryology of angiosperms. Springer-Verlag, BerlinGoogle Scholar
  37. Lee HF, Mak BS, Chi CS et al (2002) A novel mutation in neonatal isolated sulphite oxidase deficiency. Neuropediatrics 33:174–179PubMedCrossRefGoogle Scholar
  38. Ma H (2005) Molecular genetic analyses of microsporogenesis and microgametogenesis in flowering plants. Annu Rev Plant Biol 56:393–434PubMedCrossRefGoogle Scholar
  39. Mayfield JA, Fiebig A, Johnstone SE, Preuss D (2001) Gene families from the Arabidopsis thaliana pollen coat proteome. Science 292:2482–2485PubMedCrossRefGoogle Scholar
  40. Murray JAH, Crockett N (1992) In: Murray JAH (ed) Antisense RNA and DNA. Wiley-Liss, New York, pp 1–49Google Scholar
  41. Pacini E (1990) Tapetum and microspore function. In: Blackmore S, Knox RB (eds) Microspores: evolution and ontogeny. Academic, London, pp 213–237Google Scholar
  42. Pang M, Stewart JMcD, Zhang J (2011) A mini-scale hot borate method for the isolation of total RNA from a large number of cotton tissue samples. Afr J Biotechnol 10:15430–15437CrossRefGoogle Scholar
  43. Park JH, Halitschke R, Kim HB, Baldwin IT, Feldmann KA, Feyereisen R (2002) A knock-out mutation in allene oxide synthase results in male sterility and defective wound signal transduction in Arabidopsis due to a block in jasmonic acid biosynthesis. Plant J 31:1–12PubMedCrossRefGoogle Scholar
  44. Pauly M, Qin Q, Greene H, Albersheim P, Darvill A, York WS (2001) Changes in the structure of xyloglucan during cell elongation. Planta 212:842–850PubMedCrossRefGoogle Scholar
  45. Pelletier G, Budar F (2007) The molecular biology of cytoplasmically inherited male sterility and prospects for its engineering. Curr Opin Biotechnol 18:121–125PubMedCrossRefGoogle Scholar
  46. Persson S, Paredez A, Carroll A, Palsdottir H, Doblin M, Poindexter P, Khitrov N, Auer M, Somerville CR (2007) Genetic evidence for three unique components in primary cell-wall cellulose synthase complexes in Arabidopsis. Proc Natl Acad Sci USA 104:15566–15571PubMedCrossRefGoogle Scholar
  47. Piffanelli P, Ross JHE, Murphy DJ (1998) Biogenesis and function of the lipidic structures of pollen grains. Sex Plant Reprod 11:65–80CrossRefGoogle Scholar
  48. Pina C, Pinto F, Feijo JA, Becker JD (2005) Gene family analysis of the Arabidopsis pollen transcriptome reveals biological implications for cell growth, division control and gene expression regulation. Plant Physiol 138:744–756PubMedCrossRefGoogle Scholar
  49. Queitsch C, Hong SW, Vierling E, Lindquist S (2000) Heat shock protein 101 plays a crucial role in thermotolerance in Arabidopsis. Plant Cell 12:479–492PubMedGoogle Scholar
  50. Queitsch C, Sangster TA, Lindquist S (2002) Hsp90 as a capacitor of phenotypic variation. Nature 417:618–624PubMedCrossRefGoogle Scholar
  51. Ramachandran S, Christensen HE, Ishimaru Y, Dong CH, Chao-Ming W, Cleary AL, Chua NH (2000) Profilin plays a role in cell elongation, cell shape maintenance, and flowering in Arabidopsis. Plant Physiol 124:1637–1647PubMedCrossRefGoogle Scholar
  52. Sanders P, Bui A, Weterings K, McIntire K, Hsu YC, Lee P, Truong M, Beals T, Goldberg R (1999) Anther developmental defects in Arabidopsis thaliana male-sterile mutants. Sex Plant Reprod 11:297–322CrossRefGoogle Scholar
  53. Sangster TA, Bahrami A, Wilczek A, Watanabe E, Schellenberg K, McLellan C, Kelley A, Kong SW, Queitsch C, Lindquist S (2007) Phenotypic diversity and altered environmental plasticity in Arabidopsis thaliana with reduced Hsp90 levels. PLoS ONE 2:e648PubMedCrossRefGoogle Scholar
  54. Schmid M, Davison TS, Henz SR, Pape UJ, Demar M, Vingron M, Scholkopf B, Weigel D, Lohmann JU (2005) A gene expression map of Arabidopsis thaliana development. Nat Genet 37:501–506PubMedCrossRefGoogle Scholar
  55. Scott R, Hodge R, Paul W, Draper J (1991) The molecular biology of anther differentiation. Plant Sci 80:167–191CrossRefGoogle Scholar
  56. Scott RJ, Spielman M, Dickinson HG (2004) Stamen structure and function. Plant Cell 16:S46–S60PubMedCrossRefGoogle Scholar
  57. Singh D, Jermakow A, Swain S (2002) Gibberellins are required for seed development and pollen tube growth in Arabidopsis. Plant Cell 14:3133–3147PubMedCrossRefGoogle Scholar
  58. Taylor PE, Glover JA, Lavithis M, Craig S, Singh MB, Knox RB, Dennis ES, Chaudhury AM (1998) Genetic control of male fertility in Arabidopsis thaliana: structural analyses of postmeiotic developmental mutants. Planta 205:492–505PubMedCrossRefGoogle Scholar
  59. Theerakulpisut P, Xu H, Singh MB, Pettitt JM, Knox RB (1991) Isolation and developmental expression of Bcpl, an anther-specific cDNA clone in Brassica campestris. Plant Cell 3:1037–1084Google Scholar
  60. Tissier AF, Marillonnet S, Klimyuk V, Patel K, Torres MA, Murphy G, Jones JDG (1999) Multiple independent defective Suppressor-mutator transposon insertions in arabidopsis: a tool for functional genomics. Plant Cell 11:1841–1852PubMedGoogle Scholar
  61. Vizcay-Barrena G, Wilson ZA (2006) Altered tapetal PCD and pollen wall development in the Arabidopsis ms1 mutant. J Exp Bot 57:2709–2717PubMedCrossRefGoogle Scholar
  62. Wan CY, Wilkins TA (1994) A modified hot borate method significantly enhances the yield of high-quality RNA from cotton (Gossypium hirsutum L.). Anal Biochem 223:7–12PubMedCrossRefGoogle Scholar
  63. Wang Z, Zou Y, Li X, Zhang Q, Chen L, Wu H, Su D, Chen Y, Guo J, Luo D, Long Y, Zhong Y, Liu YG (2006) Cytoplasmic male sterility of rice with Boro II cytoplasm is caused by a cytotoxic peptide and is restored by two related PPR motif genes via distinct modes of mRNA silencing. Plant Cell 18:676–687PubMedCrossRefGoogle Scholar
  64. Wilkins TA, Smart LB (1996) Isolation of RNA from plant tissue. In: Krieg PA (ed) A laboratory guide to RNA: isolation, analysis, and synthesis. Wiley-Liss, Inc., New York, pp 21–42Google Scholar
  65. Willats WG, McCartney L, Mackie W, Knox JP (2001) Pectin: cell biology and prospects for functional analysis. Plant Mol Biol 47:9–27PubMedCrossRefGoogle Scholar
  66. Wise RP, Gobelman-Werner K, Pei D, Dill CL, Schnable PS (1999) Mitochondrial transcript processing and restoration of male fertility in T-cytoplasm maize. J Hered 90:380–385PubMedCrossRefGoogle Scholar
  67. Xu H, Knox RB, Taylor PE, Singh MB (1995) Bcpl, a gene required for male fertility in Arabidopsis. Proc Natl Acad Sci USA 92:2106–2110PubMedCrossRefGoogle Scholar
  68. Zenoni S, Ferrarini A, Giacomelli E, Xumerle L, Fasoli M, Malerba G, Bellin D, Pezzotti M, Delledonne M (2010) Characterization of transcriptional complexity during Berry development in Vitis vinifera using RNA-seq. Plant Physiol 152:1787–1795PubMedCrossRefGoogle Scholar
  69. Zhang JF, Stewart JMcD (2001) Inheritance and genetic relationships of the D8 and D2–2 restorer genes for cotton cytoplasmical male sterility. Crop Sci 41:289–294CrossRefGoogle Scholar
  70. Zhang W, Sun Y, Timofejeva L, Chen C, Grossniklaus U, Ma H (2006) Regulation of Arabidopsis tapetum development and function by DYSFUNCTIONAL TAPETUM1 (DYT1) encoding a putative bHLH transcription factor. Development 133:3085–3095PubMedCrossRefGoogle Scholar
  71. Zhang JF, Turley RB, Stewart JMcD (2008) Comparative analysis of gene expression between CMS-D8 restored plants and normal non-restoring fertile plants in cotton by differential display. Plant Cell Rep 27:553–561PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2013

Authors and Affiliations

  • Hideaki Suzuki
    • 1
    • 2
  • Laura Rodriguez-Uribe
    • 1
  • Jiannong Xu
    • 3
  • Jinfa Zhang
    • 1
  1. 1.Department of Plant and Environmental SciencesNew Mexico State UniversityLas CrucesUSA
  2. 2.Molecular Genetics and MicrobiologyUniversity of New Mexico Health Sciences CenterAlbuquerqueUSA
  3. 3.Department of BiologyNew Mexico State UniversityLas CrucesUSA

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