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Study of genetic modifications of flower development and methylation status in phytoplasma infected Brassica (Brassica rapa L.)

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A Correction to this article was published on 15 November 2022

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Abstract

Background

The plants of B. rapa (syn. B. campestris) are the most important food crop of Pakistan for the production of cooking oil. Brassica plants infected by phytoplasma exhibit floral abnormalities including phyllody, virescence, hypertrophied sepal and aborted reproductive organs and affected flower developmental genes which reduces the yield manifold.

Methods and results

The expression level of flower developmental genes in healthy and phytoplasma infected brassica were compared by using semi-quantitative reverse transcription polymerase chain reaction and DNA hybridization. In infected brassica, LEAFY (LFY) gene, controlling the development and maintenance of floral organ, and directly involved in controlling the homeotic gene expression was affected, while APETALA2, regulate the production of sepals and petals, were not altered. Whereas the genes WUSCHEL, APETALA3 and AGAMOUS, were significantly down-regulated, that were responsible for the identity of shoot and central meristem, petals and stamens production, and stamens and carpels development, respectively. The GLUB gene, controlling the production of β-1,3-glucanases enzyme, was highly up-regulated. According to DNA hybridization results, AGAMOUS and APETALA3 were restricted to floral organs territories in healthy and phytoplasma infected brassica, indicating that their expression was tissue-specific. These outcomes indicated that flower abnormalities of phytoplasma infected B. rapa are linked with DNA methylation in the expression of homeotic genes regulating flower development.

Conclusions

Azacitidine act as a DNA demethylating reagent. By applying the foliar spray of azacitidine during the flower development, cells of Phytoplasma infected plants exhibits demethylation of DNA when treated with azacitidine chemical that incorporated as analogue of cytosine during the cell division stage. B. rapa showed the up-regulation of gene expression level significantly that restore the normal production of flowers, ultimately increase the oil production throughout the world.

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References

  1. Scarth R, Tang T (2006) Modification of brassica oil using conventional and transgenic approaches. Health Sci 41:67–71

    Google Scholar 

  2. Liefting LW, Shaw M, Kirkpatrick BC (2004) Sequence analysis of two plasmids from the phytoplasma beet leafhopper transmitted virescence agent. Microbiology 150:1809–1817

    Article  CAS  PubMed  Google Scholar 

  3. Hogenhout, (2011) Diverse targets of phytoplasma effectors: from plant development to defense against insects. Annu Rev Phytopathol 49:175–195

    Article  PubMed  Google Scholar 

  4. Calari A, Paltrinieri S, Contaldo N, Sakalieva D, Mori N, Duduk B, Bertaccini A (2011) Molecular evidence of phytoplasmas in winter oilseed rape, tomato and corn seedlings. Bull Insectology 64:157–158

    Google Scholar 

  5. Davis RE, Lee IM (2000) Phytoplasma. In: Lederberg J (ed) Encyclopedia of microbiology, 2nd edn. Academic Press, Cambridge, pp 640–646

    Google Scholar 

  6. Ahmad JN, Sharif MZ, Ahmad SJN, Tahir M, Bertaccini A (2019) Molecular identification and characterization of phytoplasmas in insect vectors of chickpea phyllody disease in Punjab Pakistan. Phytopathogenic Mollicutes 9(1):105–106

    Article  Google Scholar 

  7. Sharif MZ, Ahmad SJN, Tahir M, Ziaf K, Zhang SH, Ahmad JN (2019) Molecular identification and characterization of phytoplasmas associated with carrot, cabbage and onion crops and their insect vectors in Punjab Pakistan. Pak J Agric Sci 56(2):1–8

    Google Scholar 

  8. Jung HY, Sawayanagi T, Wongkaew P, Miyata S, Ugaki M, Hibi T, Namba S (2003) ‘Candidatus phytoplasma oryzae’, a novel phytoplasma taxon associated with rice yellow dwarf disease. Int J Syst Evol Microbiol 53:1925–1929

    Article  CAS  PubMed  Google Scholar 

  9. Arocha Y, Lopez M, Pinol B, Fernandez M, Picornell B, Almeida R, Palenzuela WMR, Jones P (2005) ‘Candidatus phytoplasma graminis’ and ‘Candidatus phytoplasma caricae’, two novel phytoplasmas associated with diseases of sugarcane, weeds and papaya in Cuba. Int J Syst Evol Microbiol 55:2451–2463

    Article  CAS  PubMed  Google Scholar 

  10. Seemuller E, Schneider B (2004) ‘Candidatus phytoplasma mali’, ‘Candidatus phytoplasma pyri’ and ‘Candidatus phytoplasma prunorum’, the causal agents of apple proliferation, pear decline and European stone fruit yellows, respectively. Int J Syst Evol Microbiol 4:1217–1226

    Article  Google Scholar 

  11. Al-Saady NA, Akhtar JK, Alberto C, Ali MS, Assunta B (2008) ‘Candidatus phytoplasma omanense’, associated with witches’-broom of Cassia italica (Mill.) Spreng. in Oman. Int J Syst Evol Microbiol 58:461–466

    Article  CAS  PubMed  Google Scholar 

  12. Valiunas D, Staniulis J, Davis RE (2006) ‘Candidatus Phytoplasma fragariae’, a novel phytoplasma taxon discovered in yellows diseased strawberry, Fragaria x ananassa. Annu Rev Phytopathol 56:277–281

    CAS  Google Scholar 

  13. Bertaccini A (2007) Phytoplasmas: diversity, taxonomy and epidemiology. Front Biol 12:673–689

    CAS  Google Scholar 

  14. Schneider B, Torres E, Martin MP, Schroder M, Behnke HD, Seemuller E (2005) ‘Candidatus phytoplasma pini’, a novel taxon from Pinus silvestris and Pinus halpensis. Int J Syst Evol Microbiol 55:303–307

    Article  CAS  PubMed  Google Scholar 

  15. Curkovic-Perica M, Lepedus H, Seruga-Music M (2007) Effect of indole-3-butyric acid on phytoplasmas in infected Cathan anthusroseus shoots grown in vitro. FEMS Microbiol Lett 268:171–177

    Article  PubMed  Google Scholar 

  16. Olivier CY, Galka B (2008) Consequences of phytoplasma infection on canola crop production in the Canada prairies. In: Endure International Conference, Diversifying crop protection, La grande-Motte, France, Book of Abstracts, pp 1–4

  17. Verdin E, Pascal S, Jean-Luc D, Elia C, Fouad J, Souheir EZ, Brigitte G, Joseph MB, Monique G (2003) ‘Candidatus phytoplasma phoenicium’ sp. nov., a novel phytoplasma associated with an emerging lethal disease of almond trees in Lebanon and Iran. Annu Rev Phytopathol 53:833–838

    CAS  Google Scholar 

  18. Adwas A, Elsayed A, Azab A (2019) Oxidative stress and antioxidant mechanisms in human body. J Appl Biotechnol Bioeng 6(1):43–47

    Google Scholar 

  19. Firrao G, Gibb K, Streten C (2005) Short taxonomic guide to the genus ‘Candidatus Phytoplasma’. Plant Pathol 87:249–263

    Google Scholar 

  20. Himeno M, Neriya Y, Minato N, Miura C, Sugawara K, Ishii Y, Amaji Y, Kakizawa S, Oshima K, Namba S (2011) Unique morphological changes in plant pathogenic phytoplasma-infected petunia flowers are related to transcriptional regulation of floral homeotic genes in an organ-specific manner. Plant J 67:971–979

    Article  CAS  PubMed  Google Scholar 

  21. Kitamura Y, Hosokawa M, Uemachi U, Yazawa S (2009) Selection of ABC genes for candidate genes of morphological changes in hydrange a floral organs induced by phytoplasma infection. Sci Hortic 122:603–609

    Article  CAS  Google Scholar 

  22. Pracros P, Renaudin J, Eveillard S, Mouras A, Hernould M (2006) Tomato flower abnormalities induced by stolbur phytoplasma infection are associated with changes of expression of floral development genes. Mol Plant Microbe Interact 19:62–68

    Article  CAS  PubMed  Google Scholar 

  23. Reeves PH, Coupland G (2001) Analysis of flowering time control in arabidopsis by comparison of double and triple mutants. Plant Physiol 126:1085–1091

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Sugio A, Maclean AM, Kingdom HN, Grieve VM, Manimekalai R, Hogenhout SA (2011) Diverse targets of phytoplasma effectors: from plant development to defense against insects. Annu Rev Phytopathol 49:175–195

    Article  CAS  PubMed  Google Scholar 

  25. Smaczniak C, Immink RG, Angenent GC, Kaufmann K (2012) Developmental and evolutionary diversity of plant MADS-domain factors: insights from recent studies. Development 139(17):3081–3098

    Article  CAS  PubMed  Google Scholar 

  26. Ahmad JN, Garcion C, Teyssier T, Hernould M, Gallusci P, Pracros P, Renaudin J, Evellerd S (2013) Effects of stolbur phytoplasma infection on DNA methylation processes in tomato plants. Plant Pathol 62:205–216

    Article  CAS  Google Scholar 

  27. Prunet N, Morel P, Negrutiu I, Trehin C (2009) Time to stop: flower meristem termination. Plant Physiol 150:1764–1772

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Su YT, Chen J-C, Lin C-P (2011) Phytoplasma-induced floral abnormalities in catharanthus roseus are associated with phytoplasma accumulation and transcript repression of floral organ identity genes. Mol Plant Microbe Interact 24:1502–1512

    Article  CAS  PubMed  Google Scholar 

  29. Zheng X, Chen L, Li M, Lou Q, Xia H, Wang P, Li T, Liu H, Luo L (2013) Transgenerational variations in DNA methylation induced by drought stress in two rice varieties with distinguished difference to drought resistance. PLoS ONE 8:e80253

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Mason G, Noris E, Lanteri S, Acquadro A, Accotto PG, Portis E (2008) Potentiality of methylation-sensitive amplification polymorphism (MSAP) in identifying genes involved in tomato response to tomato yellow leaf curl sardinia virus. Plant Mol Biol Rep 26:156–173

    Article  CAS  Google Scholar 

  31. Cao X, Fan G, Deng M, Zhao Z, Dong Y (2014) Identification of genes related to paulownia witches’ broom by AFLP and MSAP. Int J Mol Sci 15:14669–14683

    Article  PubMed  PubMed Central  Google Scholar 

  32. Jacobsen SE, Sakai H, Finnegan EJ, Cao X, Meyerowitz EM (2000) Ectopic hyper-methylation of flower-specific genes in Arabidopsis. Curr Biol 10:179–186

    Article  CAS  PubMed  Google Scholar 

  33. Yia H, Lihong L (2013) DNA methylation changes in response to sulfur dioxide stress in Arabidopsis plants. Procedia Environ Sci 18:37–42

    Article  Google Scholar 

  34. Manning K, Tor M, Poole M et al (2006) A naturally occurring epigenetic mutation in a gene encoding an SBP-box transcription factor inhibits tomato fruit ripening. Nat Genet 38:948–952

    Article  CAS  PubMed  Google Scholar 

  35. Takeno K (2010) Epigenetic regulation of photoperiodic flowering. Plant Signal Behav 5:788–791

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Ahrens U, Seemuller E (1992) Detection of DNA of plant pathogenic mycoplasma-like organisms by a polymerase chain reaction that amplifies a sequence of the 16S rRNA gene. Phytopathology 82:828

    Article  CAS  Google Scholar 

  37. Foerster AM, Scheid OM (2010) Analysis of DNA methylation in plants by bisulfite sequencing. In: Kovalchuk I, Zemp FJ (eds) Plant Epigenetics. Humana Press, Totowa NJ USA, pp 1–11

    Google Scholar 

  38. Teyssier E, Bernacchia G, Maury S et al (2008) Tissue dependent variations of DNA methylation and endoreduplication levels during tomato fruit development and ripening. Planta 228:391–399

    Article  CAS  PubMed  Google Scholar 

  39. Mazzucato A, Olimpieri I, Siligato F, Picarella ME, Soressi GP (2008) Characterization of genes controlling stamen identity and development in a parthenocarpic tomato mutant indicates a role for the DEFICIENS ortholog in the control of fruit set. Physiol Plant 132:526–537

    Article  CAS  PubMed  Google Scholar 

  40. Lohmann JU, Weigel D (2002) Building beauty: the genetic control of floral patterning. Dev Cell 2:135–142

    Article  CAS  PubMed  Google Scholar 

  41. Ahmad JN, Renaudin J, Eveillard S (2014) Expression of defence genes in stolbur phytoplasma infected tomatoes, and effect of defense stimulators on disease development. Eur J Plant Pathol 139:39–51

    Article  CAS  Google Scholar 

  42. MacLean AM, Sugio A, Makarova OV et al (2011) Phytoplasma effector SAP54 induces indeterminate leaf-like flower development in Arabidopsis plants. Plant Physiol 157:831–841

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Bird A (2002) DNA methylation patterns and epigenetic memory. Genes Dev 16:6–21

    Article  CAS  PubMed  Google Scholar 

  44. Zilberman D, Gehring M, Tran RK, Ballinger T, Henikoff S (2007) Genome-wide analysis of Arabidopsis thaliana DNA methylation uncovers an interdependence between methylation and transcription. Nat Genet 39(61):9–629

    Google Scholar 

  45. Weigel D, Alvarez J, Smyth DR, Yanofsky MF, Meyerowitz EM (1992) LEAFY controls floral meristem identity in Arabidopsis. Cell 69:843–859

    Article  CAS  PubMed  Google Scholar 

  46. Lenhard M, Bohnert A, Jurgens G, Laux T (2001) Termination of stem cell maintenance in Arabidopsis floral meristems by interactions between WUSCHEL and AGAMOUS. Cell 105:805–814

    Article  CAS  PubMed  Google Scholar 

  47. Yanofsky MF, Ma H, Bowman JL, Drews GN, Feldmann KA, Meyerowitz EM (1990) The protein encoded by the Arabidopsis homeotic gene agamous resembles transcription factors. Nature 34:35–39

    Article  Google Scholar 

  48. Kaufmann K, Melzer R, Theissen G (2005) MIKC-type MADS domain proteins: structural modularity, protein interactions and network evolution in land plants. Gene 347:183–198

    Article  CAS  PubMed  Google Scholar 

  49. Robles P, Pelaz S (2005) Flower and fruit development in Arabidopsis thaliana. Int J Dev Biol 49:633–643

    Article  CAS  PubMed  Google Scholar 

  50. Cettul E, Firrao G (2010) Effects of phytoplasma infection on Arabidopsis thaliana development. In: Brown DR, Bertaccini A (eds) 18th Congress of the international organization for mycoplasmology chiancianoterme. Italy

    Google Scholar 

  51. Sharma V (2013) Pathogenesis related defence functions of plant chitinases and β-1,3-glucanases. Vegetos 26:215–218

    Article  Google Scholar 

  52. Cordero MJ, Raventos D, San Segundo B (1994) Differential expression and induction of chitinases and β-1,3-glucanases in response to fungal infection during germination of maize seeds. Mol Plant Microbe Interact 7:23–31

    Article  CAS  Google Scholar 

  53. Khan MA, Jafri S, Rana IA, Ullah I (1992) Improvement of wheat protein by supplementation with potato flour. Pak J Agric Sci 13:101–106

    Google Scholar 

  54. Ward ER, Payne GB, Moyer MB, Williams SC, Dincher SS, Sharkey KC, Beck JJ, Taylor HT, Ahl-Goy P, Meins FJ, Ryals JA (1991) Differential regulation of b-1,3-glucanase messenger RNAs inresponse to pathogen infection. Plant Physiol 96:390–397

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  55. Villanea F, Bolnick D, Monroe C, Worl R, Cambra R, Leventhal A (2013) Brief communication: evolution of a specific O allele (O1v (G542A)) supports unique ancestry of Native Americans. Am J Phys Anthropol 151:649–657

    Article  PubMed  Google Scholar 

  56. Llamas B, Holland ML, Chen K, Cropley JE, Cooper A, Suter CM (2012) High-resolution analysis of cytosine methylation in ancient DNA. PLoS ONE 7:1–6

    Article  Google Scholar 

  57. Jofuku KD, Den-Boer BGW, Van-Montagu M, Okamuro JK (1994) Control of Arabidopsis flower and seed development by the homeotic gene APETALA2. Plant Cell 6:1211–1225

    CAS  PubMed  PubMed Central  Google Scholar 

  58. Zhang M, Kimatu JN, Xu K, Liu B (2010) DNA cytosine methylation in plant development. J Genet Genom 37:1–12

    Article  Google Scholar 

  59. Buoso S, Pagliari L, Musetti R, Martini M, Marroni F, Schmidt W, Santi S (2019) ‘Candidatus Phytoplasma solani’interferes with the distribution and uptake of iron in tomato. BMC Genomics 20(1):703

    Article  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

The FICP international research grant run by Principal Investigator Dr. Jam Nazeer Ahmad and Co- principal investigator Dr. Samina Jam Nazeer Ahmad for research funding. All the equipment and chemical I used for my research work were funded by this project. The authors extend their appreciation to the researchers supporting project no (RSP-2021/293) at King Saud University, Riyadh, Saudi Arabia.

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Authors and Affiliations

  1. Institute of Botany, University of the Punjab Lahore, Punjab, Pakistan

    Mohammad Aijaz Ahmad & Shakil Ahmed

  2. Department of Botany, University of Agriculture Faisalabad, Punjab, Pakistan

    Samina Jam Nazeer Ahmad

  3. Department of Agricultural Engineering, Khwaja Fareed University of Engineering and Information Technology, Rahim Yar Khan, Punjab, 64200, Pakistan

    Adnan Noor Shah

  4. Department of Entomology, Faculty of Agriculture, University of Agriculture, Faisalabad, Pakistan

    Jam Nazeer Ahmad

  5. Department of Food Sciences & Nutrition, College of Food & Agriculture Sciences, King Saud University, Riyadh, 11451, Saudi Arabia

    Wahidah H. Al-Qahtani

  6. Integrated Molecular Plant Physiology Research (IMPRES), Department of Biology, University of Antwerp, Antwerp, Belgium

    Hamada AbdElgawad

  7. Department of Botany, Division of Science and Technology, University of Education, Lahore, Pakistan

    Anis Ali Shah

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  1. Mohammad Aijaz Ahmad
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Ahmad, M.A., Ahmad, S.J.N., Shah, A.N. et al. Study of genetic modifications of flower development and methylation status in phytoplasma infected Brassica (Brassica rapa L.). Mol Biol Rep 49, 11359–11369 (2022). https://doi.org/10.1007/s11033-022-07743-0

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