pp 1–14 | Cite as

Role of a cotton endoreduplication-related gene, GaTOP6B, in response to drought stress

  • Yanfei Tian
  • Huihui Gu
  • Zhuxuan Fan
  • Gongyao Shi
  • Jiachen Yuan
  • Fang WeiEmail author
  • Yan Yang
  • Baoming TianEmail author
  • Gangqiang Cao
  • Jinyong HuangEmail author
Original Article


Main conclusion

Cotton GaTOP6B is involved in cellular endoreduplication and a positive response to drought stress via promoting plant leaf and root growth.

Drought is deemed as one of adverse conditions that could cause substantial reductions in crop yields worldwide. Since cotton exhibits a moderate-tolerant phenotype under water-deficit conditions, the plant could therefore be used to characterize potential new genes regulating drought tolerance in crop plants. In this work, GaTOP6B, encoding DNA topoisomerase VI subunit B, was identified in Asian cotton (Gossypium arboreum). Virus-induced gene silencing (VIGS) and overexpression (OE) were used to investigate the biological function of GaTOP6B in G. arboreum and Arabidopsis thaliana under drought stress. The GaTOP6B-silencing plants showed a reduced ploidy level, and displayed a compromised tolerance phenotype including lowered relative water content (RWC), decreased proline content and antioxidative enzyme activity, and an increased malondialdehyde (MDA) content under drought stress. GaTOP6B-overexpressing Arabidopsis lines, however, had increased ploidy levels, and were more tolerant to drought treatment, associated with improved RWC maintenance, higher proline accumulation, and reduced stomatal aperture under drought stress. Transcriptome analysis showed that genes involved in the processes like cell cycle, transcription and signal transduction, were substantially up-regulated in GaTOP6B-overexpressing Arabidopsis, promoting plant growth and development. More specifically, under drought stress, the genes involved in the biosynthesis of secondary metabolites such as phenylpropanoid, starch and sucrose were selectively enhanced to improve tolerance in plants. Taken together, the results demonstrated that GaTOP6B could coordinately regulate plant leaf and root growth via cellular endoreduplication, and positively respond to drought stress. Thus, GaTOP6B could be a competent candidate gene for improvement of drought tolerance in crop species.


Arabidopsis Cotton Drought tolerance Endoreduplication Gossypium arboreum Topoisomerase VIB Transcriptome 



Differentially expressed genes


Open reading frame


Relative water content


Topoisomerase VIB


Virus-induced gene silencing



This work was financially supported by the Program for Science and Technology Innovation Talents in Universities of Henan Province (No. 19HASTIT014), and the National Natural Science Foundation of China (No. 31600995) and the fund of the Science and Technology Cooperation Projects of Henan Province of China (No. 152106000055).

Compliance with ethical standards

Conflict of interest

The authors declared that they have no conflicts of interest to this work.

Ethical standards

The experiments in this study comply with the current laws of China.

Supplementary material

425_2018_3067_MOESM1_ESM.jpg (150 kb)
Supplementary material 1 Bioinformatic analysis of GaTOP6B (JPEG 149 kb)
425_2018_3067_MOESM2_ESM.jpg (17 kb)
Supplementary material 2 Subcellular localization of GaTOP6B (JPEG 16 kb)
425_2018_3067_MOESM3_ESM.jpg (36 kb)
Supplementary material 3 Characterization of CLA-silencing plantlets and GaTOP6B-overexpressing Arabidopsis (JPEG 36 kb)
425_2018_3067_MOESM4_ESM.jpg (57 kb)
Supplementary material 4 Survival rate and DAB staining and chlorophyll content in GaTOP6B-silencing cotton under drought stress (JPEG 57 kb)
425_2018_3067_MOESM5_ESM.jpg (209 kb)
Supplementary material 5 Gene expression analysis (JPEG 209 kb)
425_2018_3067_MOESM6_ESM.jpg (58 kb)
Supplementary material 6 COG classification of DEGs in transgenic Arabidopsis compared to WT under 200 mM mannitol treatment (JPEG 57 kb)
425_2018_3067_MOESM7_ESM.jpg (63 kb)
Supplementary material 7 (JPEG 62 kb)
425_2018_3067_MOESM8_ESM.doc (42 kb)
Supplementary material 8 (DOC 42 kb)


  1. An XJ, Deng ZY, Wang T (2011) OsSpo11-4, a rice homologue of the archaeal TopVIA protein, mediates double-strand DNA cleavage and interacts with OsTopVIB. PLoS One 6:e20327CrossRefGoogle Scholar
  2. Anders S, Huber W (2010) Differential expression analysis for sequence count data. Genome Biol 11:R106CrossRefGoogle Scholar
  3. Bergerat A, de Massy B, Gadelle D, Varoutas PC, Nicolas A, Forterre P (1997) An atypical topoisomerase II from Archaea with implications for meiotic recombination. Nature 386:414–417CrossRefGoogle Scholar
  4. Blattner FR (2015) TOPO6: a nuclear single-copy gene for plant phylogenetic inference. Plant Syst Evol 302:239–244CrossRefGoogle Scholar
  5. Breuer C, Stacey NJ, West CE, Zhao Y, Chory J, Tsukaya H, Azumi Y, Maxwell A, Roberts K, Sugimoto-Shirasu K (2007) BIN4, a novel component of the plant DNA topoisomerase VI complex, is required for endoreduplication in Arabidopsis. Plant Cell 19:3655–3668CrossRefGoogle Scholar
  6. Breuer C, Braidwood L, Sugimoto K (2014) Endocycling in the path of plant development. Curr Opin Plant Biol 17:78–85CrossRefGoogle Scholar
  7. Butt HI, Yang Z, Chen E, Zhao G, Gong Q, Yang Z, Zhang X, Li F (2017) Functional characterization of cotton GaMYB62L, a novel R2R3 TF in transgenic Arabidopsis. PLoS One 12:e0170578CrossRefGoogle Scholar
  8. Ceccarelli M, Santantonio E, Marmottini F, Amzallag GN, Cionini PG (2006) Chromosome endoreduplication as a factor of salt adaptation in Sorghum bicolor. Protoplasma 227:113–118CrossRefGoogle Scholar
  9. Chen T, Li W, Hu X, Guo J, Liu A, Zhang B (2015) A cotton MYB transcription factor, GbMYB5, is positively involved in plant adaptive response to drought stress. Plant Cell Physiol 56:917–929CrossRefGoogle Scholar
  10. Cheng XQ, Zhu XF, Tian WG, Cheng WH, Hakim Sun J, Jin SX, Zhu HG (2017) Genome-wide identification and expression analysis of polyamine oxidase genes in upland cotton (Gossypium hirsutum L.). Plant Cell Tiss Organ Cult 129:237–249CrossRefGoogle Scholar
  11. Clough SJ, Bent AF (1998) Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J 16:735–743CrossRefGoogle Scholar
  12. Corbett KD, Berger JM (2003) Structure of the topoisomerase VI-B subunit: implications for type II topoisomerase mechanism and evolution. EMBO J 22:151–163CrossRefGoogle Scholar
  13. De Veylder L, Larkin JC, Schnittger A (2011) Molecular control and function of endoreplication in development and physiology. Trends Plant Sci 16:624–634CrossRefGoogle Scholar
  14. Eady C, Weld R, Lister C (2000) Agrobacterium tumefaciens-mediated transformation and transgenic-plant regeneration of onion (Allium cepa L.). Plant Cell Rep 19:376–381CrossRefGoogle Scholar
  15. Elmaghrabi AM, Rogers HJ, Francis D, Ochatt S (2017) PEG induces high expression of the cell cycle checkpoint gene WEE1 in embryogenic callus of Medicago truncatula: potential link between cell cycle checkpoint regulation and drought. Front Plant Sci 8:1479CrossRefGoogle Scholar
  16. Gao X, Shan L (2013) Functional genomic analysis of cotton genes with Agrobacterium-mediated virus-induced gene silencing. Methods Mol Biol 975:157–165CrossRefGoogle Scholar
  17. Gegas VC, Wargent JJ, Pesquet E, Granqvist E, Paul ND, Doonan JH (2014) Endopolyploidy as a potential alternative adaptive strategy for Arabidopsis leaf size variation in response to UV-B. J Exp Bot 65:2757–2766CrossRefGoogle Scholar
  18. Hartung F, Puchta H (2001) Molecular characterization of homologues of both subunits A (SPO11) and B of the archaebacterial topoisomerase 6 in plants. Gene 271:81–86CrossRefGoogle Scholar
  19. Hartung F, Angelis KJ, Meister A, Schubert I, Melzer M, Puchta H (2002) An archaebacterial topoisomerase homolog not present in other eukaryotes is indispensable for cell proliferation of plants. Curr Biol 12:1787–1791CrossRefGoogle Scholar
  20. Hase Y, Trung KH, Matsunaga T, Tanaka A (2006) A mutation in the uvi4 gene promotes progression of endo-reduplication and confers increased tolerance towards ultraviolet B light. Plant J 46:317–326CrossRefGoogle Scholar
  21. Jain M, Tyagi AK, Khurana JP (2006) Overexpression of putative topoisomerase 6 genes from rice confers stress tolerance in transgenic Arabidopsis plants. FEBS J 273:5245–5260CrossRefGoogle Scholar
  22. Jain M, Tyagi AK, Khurana JP (2008) Constitutive expression of a meiotic recombination protein gene homolog, OsTOP6A1, from rice confers abiotic stress tolerance in transgenic Arabidopsis plants. Plant Cell Rep 27:767–778CrossRefGoogle Scholar
  23. Kirik V, Schrader A, Uhrig JF, Hülskamp M (2007) MIDGET unravels functions of the Arabidopsis topoisomerase VI complex in DNA endoreduplication, chromatin condensation, and transcriptional silencing. Plant Cell 19:3100–3110CrossRefGoogle Scholar
  24. Leclercq J, Martin F, Sanier C, Clement-Vidal A, Fabre D, Oliver G, Lardet L, Ayar A, Peyramard M, Montoro P (2012) Over-expression of a cytosolic isoform of the HbCuZnSOD gene in Hevea brasiliensis changes its response to a water deficit. Plant Mol Biol 80:255–272CrossRefGoogle Scholar
  25. Lefebvre V, North H, Frey A, Sotta B, Seo M, Okamoto M, Nambara E, Marion-Poll A (2006) Functional analysis of Arabidopsis NCED6 and NCED9 genes indicates that ABA synthesized in the endosperm is involved in the induction of seed dormancy. Plant J 45:309–319CrossRefGoogle Scholar
  26. Lopienska-Biernat E, Molcan T, Paukszto L, Jastrzebski JP, Myszczynski K (2018) Modelling studies determining the mode of action of anthelmintics inhibiting in vitro trehalose-6-phosphate phosphatase (TPP) of Anisakis simplex s.l. Exp Parasitol 184:46–56CrossRefGoogle Scholar
  27. Lubbers EL, Chee PW, Saranga Y, Paterson AH (2007) Recent advances and future prospective in molecular breeding of cotton for drought and salinity stress tolerance. In: Jenks MA, Hasegawa PM, Jain SM (eds) Advances in molecular breeding towards drought and salt tolerant crops. Springer, Dordrecht, pp 775–796CrossRefGoogle Scholar
  28. Ma LF, Li Y, Chen Y, Li XB (2016) Improved drought and salt tolerance of Arabidopsis thaliana by ectopic expression of a cotton (Gossypium hirsutum) CBF gene. Plant Cell Tiss Organ Cult 124:583–598CrossRefGoogle Scholar
  29. Miao BH, Han XG, Zhang WH (2010) The ameliorative effect of silicon on soybean seedlings grown in potassium-deficient medium. Ann Bot 105:967–973CrossRefGoogle Scholar
  30. Mustafa R, Shafiq M, Mansoor S, Briddon RW, Scheffler BE, Scheffler J, Amin I (2016) Virus-induced gene silencing in cultivated cotton (Gossypium spp.) using tobacco rattle virus. Mol Biotechnol 58:65–72CrossRefGoogle Scholar
  31. Nir I, Shohat H, Panizel I, Olszewski N, Aharoni A, Weiss D (2017) The tomato DELLA protein PROCERA acts in guard cells to promote stomatal closure. Plant Cell 29:3186–3197CrossRefGoogle Scholar
  32. Pirrello J, Deluche C, Frangne N, Gevaudant F, Maza E, Djari A, Bourge M, Renaudin JP, Brown S, Bowler C, Zouine M, Chevalier C, Gonzalez N (2018) Transcriptome profiling of sorted endoreduplicated nuclei from tomato fruits: how the global shift in expression ascribed to DNA ploidy influences RNA-Seq data normalization and interpretation. Plant J 93:387–398CrossRefGoogle Scholar
  33. Radziejwoski A, Vlieghe K, Lammens T, Berckmans B, Maes S, Jansen MA, Knappe C, Albert A, Seidlitz HK, Bahnweg G, Inze D, De Veylder L (2011) Atypical E2F activity coordinates PHR1 photolyase gene transcription with endoreduplication onset. EMBO J 30:355–363CrossRefGoogle Scholar
  34. Saleem MF, Sammar Raza MA, Ahmad S, Khan IH, Shahid AM (2016) Understanding and mitigating the impacts of drought stress in cotton—a review. Pak J Agric Sci 53:609–623Google Scholar
  35. Scholes DR, Paige KN (2015) Plasticity in ploidy: a generalized response to stress. Trends Plant Sci 20:165–175CrossRefGoogle Scholar
  36. Sekmen AH, Ozgur R, Uzilday B, Turkan I (2014) Reactive oxygen species scavenging capacities of cotton (Gossypium hirsutum) cultivars under combined drought and heat induced oxidative stress. Environ Exp Bot 99:141–149CrossRefGoogle Scholar
  37. Shinozaki K, Yamaguchi-Shinozaki K (2007) Gene networks involved in drought stress response and tolerance. J Exp Bot 58:221–227CrossRefGoogle Scholar
  38. Sugimoto-Shirasu K, Stacey NJ, Corsar J, Roberts K, McCann MC (2002) DNA topoisomerase VI is essential for endoreduplication in Arabidopsis. Curr Biol 12:1782–1786CrossRefGoogle Scholar
  39. Villeneuve AM, Hillers KJ (2001) Whence meiosis. Cell 106:647–650CrossRefGoogle Scholar
  40. Wang NN, Zhao LL, Lu R, Li Y, Li XB (2015) Cotton mitogen-activated protein kinase4 (GhMPK4) confers the transgenic Arabidopsis hypersensitivity to salt and osmotic stresses. Plant Cell Tiss Organ Cult 123:619–632CrossRefGoogle Scholar
  41. Woolfenden HC, Bourdais G, Kopischke M, Miedes E, Molina A, Robatzek S, Morris RJ (2017) A computational approach for inferring the cell wall properties that govern guard cell dynamics. Plant J 92:5–18CrossRefGoogle Scholar
  42. Yin Y, Cheong H, Friedrichsen D, Zhao Y, Hu J, Mora-Garcia S, Chory J (2002) A crucial role for the putative Arabidopsis topoisomerase VI in plant growth and development. Proc Natl Acad Sci USA 99:10191–10196CrossRefGoogle Scholar
  43. Zhao F, Ma J, Li L, Fan S, Guo Y, Song M, Wei H, Pang C, Yu S (2016) GhNAC12, a neutral candidate gene, leads to early aging in cotton (Gossypium hirsutum L.). Gene 576:268–274CrossRefGoogle Scholar
  44. Zhou L, Wang NN, Kong L, Gong SY, Li Y, Li XB (2014) Molecular characterization of 26 cotton WRKY genes that are expressed differentially in tissues and are induced in seedlings under high salinity and osmotic stress. Plant Cell Tiss Organ Cult 119:141–153CrossRefGoogle Scholar
  45. Zhou L, Wang NN, Gong SY, Lu R, Li Y, Li XB (2015) Overexpression of a cotton (Gossypium hirsutum) WRKY gene, GhWRKY34, in Arabidopsis enhances salt-tolerance of the transgenic plants. Plant Physiol Biochem 96:311–320CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Zhengzhou Research Base, State Key Laboratory of Cotton BiologyZhengzhou UniversityZhengzhouPeople’s Republic of China
  2. 2.School of Agricultural SciencesZhengzhou UniversityZhengzhouPeople’s Republic of China
  3. 3.School of Chemical Engineering and EnergyZhengzhou UniversityZhengzhouPeople’s Republic of China

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