Preview

Vavilov Journal of Genetics and Breeding

Advanced search

A high coverage reference transcriptome assembly of pea (Pisum sativum L.) mycorrhizal roots

https://doi.org/10.18699/VJ20.625

Abstract

Arbuscular mycorrhiza (AM) is an ancient mutualistic symbiosis formed by 80–90 % of land plant species with the obligatorily biotrophic fungi that belong to the phylum Glomeromycota. This symbiosis is mutually beneficial, as AM fungi feed on plant photosynthesis products, in turn improving the efficiency of nutrient uptake from the environment. The garden pea (Pisum sativum L.), a widely cultivated crop and an important model for genetics, is capable of forming triple symbiotic systems consisting of the plant, AM fungi and nodule bacteria. As transcriptomic and proteomic approaches are being implemented for studying the mutualistic symbioses of pea, a need for a reference transcriptome of genes expressed under these specific conditions for increasing the resolution and the accuracy of other methods arose. Numerous transcriptome assemblies constructed for pea did not include mycorrhizal roots, hence the aim of the study to construct a reference transcriptome assembly of pea mycorrhizal roots. The combined transcriptome of mycorrhizal roots of Pisum sativum cv. Frisson inoculated with Rhizophagus irregularis BEG144 was investigated, and for both the organisms independent transcriptomes were assembled (coverage 177x for pea and 45x for fungus). Genes specific to mycorrhizal roots were found in the assembly, their expression patterns were examined with qPCR on two pea cultivars, Frisson and Finale. The gene expression depended on the inoculation stage and on the pea cultivar. The investigated genes may serve as markers for early stages of inoculation in genetically diverse pea cultivars.

About the Authors

A. M. Afonin
All-Russia Research Institute for Agricultural Microbiology
Russian Federation
Pushkin, St. Petersburg


I. V. Leppyanen
All-Russia Research Institute for Agricultural Microbiology
Russian Federation
Pushkin, St. Petersburg


O. A. Kulaeva
All-Russia Research Institute for Agricultural Microbiology
Russian Federation
Pushkin, St. Petersburg


O. Y. Shtark
All-Russia Research Institute for Agricultural Microbiology
Russian Federation
Pushkin, St. Petersburg


I. A. Tikhonovich
All-Russia Research Institute for Agricultural Microbiology; Faculty of Biology, St. Petersburg State University
Russian Federation
Pushkin
St. Petersburg


E. A. Dolgikh
All-Russia Research Institute for Agricultural Microbiology
Russian Federation
Pushkin, St. Petersburg


V. A. Zhukov
All-Russia Research Institute for Agricultural Microbiology
Russian Federation
Pushkin, St. Petersburg


References

1. Alizadeh O. Mycorrhizal symbiosis. Adv. Stud. Biol. 2011;3(6):273281.

2. Alves-Carvalho S., Aubert G., Carrère S., Cruaud C., Brochot A.L., Jacquin F., Klein A., Martin C., Boucherot K., Kreplak J., Da Silva C., Moreau S., Gamas P., Wincker P., Gouzy J., Burstin J. Full-length de novo assembly of RNA-seq data in pea (Pisum sativum L.) provides a gene expression atlas and gives insights into root nodulation in this species. Plant J. 2015;84(1):1-19. DOI 10.1111/tpj.12967.

3. Bankevich A., Nurk S., Antipov D., Gurevich A.A., Dvorkin M., Kulikov A.S., Lesin V.M., Nikolenko S.I., Pham S., Prjibelski A.D., Pyshkin A.V., Sirotkin A.V., Vyahhi N., Tesler G., Alekseyev M.A., Pevzner P.A. SPAdes: a new genome assembly algorithm and its applications to single-cell sequencing. J. Comput. Biol. 2012;19(5): 455-477. DOI 10.1089/cmb.2012.0021.

4. Borisov A., Vasil’chikov A., Voroshilova V., Danilova T., Zhernakov A., Zhukov V., Koroleva T., Kuznetsova E., Madsen L., Mofett M., Naumkina T., Nemankin T., Ovchinnikova E., Pavlova Z., Petrova N., Pinaev A., Radutoiu S., Rozov S., Rychagova T., Solovov I., Stougaard J., Topunov A., Weeden N., Tsyganov V., Shtark O., Tikhonovich I. Regulatory genes of garden pea (Pisum sativum L.) controlling the development of nitrogen-fixing nodules and arbuscular mycorrhiza: a review of basic and applied aspects. Appl. Biochem. Microbiol. 2007;43(3):237-243. DOI 10.1134/S0003683807030027.

5. Borisov A.Y., Danilova T.N., Koroleva T.A., Naumkina T.S., Pavlova Z.B., Pinaev A.G., Shtark O.Y., Tsyganov V.E., Voroshilova V.A., Zhernakov A.I., Zhukov V.A., Tikhonovich I.A. Pea (Pisum sativum L.) regulatory genes controlling development of nitrogenfixing nodule and arbuscular mycorrhiza: fundamentals and application. Biologia (Bratislava). 2004;59(13):137-144.

6. Bushnell B. BBMap. 2014. Available at: sourceforge.net/projects/bbmap/

7. Davidson N.M., Hawkins A.D.K., Oshlack A. SuperTranscripts: a data driven reference for analysis and visualisation of transcriptomes. Genome Biol. 2017;18:148. DOI 10.1186/s13059-017-1284-1.

8. Demchenko K., Winzer T., Stougaard J., Parniske M., Pawlowski K. Distinct roles of Lotus japonicus SYMRK and SYM15 in root colonization and arbuscule formation. New Phytol. 2004;163(2):381-392. DOI 10.1111/j.1469-8137.2004.01123.x.

9. Duc G., Messager A. Mutagenesis of pea (Pisum sativum L.) and the isolation of mutants for nodulation and nitrogen fixation. Plant Sci. 1989;60(2):207-213. DOI 10.1016/0168-9452(89)90168-4.

10. Engvild K.C. Nodulation and nitrogen fixation mutants of pea, Pisum sativum. Theor. Appl. Genet. 1987;74(6):711-713. DOI 10.1007/BF00247546.

11. Franssen S.U., Shrestha R.P., Bräutigam A., Bornberg-Bauer E., Weber A.P. Comprehensive transcriptome analysis of the highly complex Pisum sativum genome using next generation sequencing. BMC Genomics. 2011;12(1):227. DOI 10.1186/1471-2164-12-227.

12. Gobbato E. Recent developments in arbuscular mycorrhizal signaling. Curr. Opin. Plant Biol. 2015;26:1-7. DOI 10.1016/j.pbi.2015.05.006.

13. Goodstein D.M., Shu S., Howson R., Neupane R., Hayes R.D., Fazo J., Mitros T., Dirks W., Hellsten U., Putnam N., Rokhsar D.S. Phytozome: a comparative platform for green plant genomics. Nucleic Acids Res. 2012;40(D1):D1178-D1186. DOI 10.1093/nar/gkr944.

14. Grabherr M.G., Haas B.J., Yassour M., Levin J.Z., Thompson D.A., Amit I., Adiconis X., Fan L., Raychowdhury R., Zeng Q., Chen Z., Mauceli E., Hacohen N., Gnirke A., Rhind N., di Palma F., Birren B.W., Nusbaum C., Lindblad-Toh K., Friedman N., Regev A. Full-length transcriptome assembly from RNA-Seq data without a reference genome. Nat. Biotechnol. 2011;29(7):644-652. DOI 10.1038/nbt.1883.

15. Grunwald U., Nyamsuren O., Tamasloukht M., Lapopin L., Becker A., Mann P., Gianinazzi-Pearson V., Krajinski F., Franken P. Identification of mycorrhiza-regulated genes with arbuscule development-related expression profile. Plant Mol. Biol. 2004;55(4):553-566. DOI 10.1007/s11103-004-1303-y.

16. Gutjahr C., Parniske M. Cell and developmental biology of arbuscular mycorrhiza symbiosis. Annu. Rev. Cell Dev. Biol. 2013;29(1):593-617. DOI 10.1146/annurev-cellbio-101512-122413.

17. Hoagland D.R., Arnon D.I. The water-culture method for growing plants without soil. Circular. California Agric. Exp. Station. 1950; 347(2).

18. Huerta-Cepas J., Szklarczyk D., Forslund K., Cook H., Heller D., Walter M.C., Rattei T., Mende D.R., Sunagawa S., Kuhn M., Jensen L.J., von Mering C., Bork P. eggNOG 4.5: a hierarchical orthology framework with improved functional annotations for eukaryotic, prokaryotic and viral sequences. Nucleic Acids Res. 2016;44(D1):D286D293. DOI 10.1093/nar/gkv1248.

19. Jacobi L.M., Kukalev A.S., Ushakov K.V., Tsyganov V.E., Provorov N.A., Borisov A.Y., Tikhonovich I. Genetic variability of garden pea (Pisum sativum L.) for symbiotic capacities. Pisum Genet. 1999; 31:44-45.

20. Kaschuk G. Sink Stimulation of Leaf Photosynthesis by the Carbon Costs of Rhizobial and Arbuscular Mycorrhizal Fungal Symbioses. Wageningen, Wageningen Univ., S.n., 2009.

21. Kerr S.C., Gaiti F., Beveridge C.A., Tanurdzic M. De novo transcriptome assembly reveals high transcriptional complexity in Pisum sativum axillary buds and shows rapid changes in expression of diurnally regulated genes. BMC Genomics. 2017;18(1):221. DOI 10.1186/s12864-017-3577-x.

22. Kreplak J., Madoui M.A., Cápal P., Novák P., Labadie K., Aubert G., Bayer P.E., Gali K.K., Syme R.A., Main D., Klein A., Bérard A., Vrbová I., Fournier C., D’Agata L., Belser C., Berrabah W., Toegelová H., Milec Z., Vrána J., Lee H.T., Kougbeadjo A., Térézol M., Huneau C., Turo C.J., Mohellibi N., Neumann P., Falque M., Gallardo K., McGee R., Tar’an B., Bendahmane A., Aury J.M., Batley J., Le Paslier M.C., Ellis N., Warkentin T.D., Coyne C.J., Salse J., Edwards D., Lichtenzveig J., Macas J., Doležel J., Wincker P., Burstin J. A reference genome for pea provides insight into legume genome evolution. Nat. Genet. 2019;51(9):1411-1422. DOI 10.1038/s41588-019-0480-1.

23. Leppyanen I.V., Shakhnazarova V.Y., Shtark O.Y., Vishnevskaya N.A., Tikhonovich I.A., Dolgikh E.A. Receptor-like kinase LYK9 in Pisum sativum L. is the CERK1-like receptor that controls both plant immunity and AM symbiosis development. Int. J. Mol. Sci. 2018; 19:8. DOI 10.3390/ijms19010008.

24. Livak K.J., Schmittgen T.D. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCT method. Methods. 2001;25(4):402-408. DOI 10.1006/meth.2001.1262.

25. Lu G. Vector NTI, a balanced all-in-one sequence analysis suite. Brief. Bioinformatics. 2004;5(4):378-388. DOI 10.1093/bib/5.4.378.

26. Mamontova T., Afonin A.M., Ihling C., Soboleva A., Lukasheva E., Sulima A.S., Shtark O.Y., Akhtemova G.A., Povydysh M.N., Sinz A., Frolov A., Zhukov V.A., Tikhonovich I.A. Profiling of seed proteome in pea (Pisum sativum L.) lines characterized with high and low responsivity to combined inoculation with nodule bacteria and arbuscular mycorrhizal fungi. Molecules. 2019;24(8):1603. DOI 10.3390/molecules24081603.

27. Manck-Götzenberger J., Requena N. Arbuscular mycorrhiza symbiosis induces a major transcriptional reprogramming of the potato SWEET sugar transporter family. Front. Plant Sci. 2016;7:487. DOI 10.3389/fpls.2016.00487.

28. Morandi D., Sagan M., Prado-Vivant E., Duc G. Influence of genes determining supernodulation on root colonization by the mycorrhizal fungus Glomus mosseae in Pisum sativum and Medicago truncatula mutants. Mycorrhiza. 2000;10(1):37-42. DOI 10.1007/s005720050285.

29. Murray J.D., Muni R.R.D., Torres-Jerez I., Tang Y., Allen S., Andriankaja M., Li G., Laxmi A., Cheng X., Wen J., Vaughan D., Schultze M., Sun J., Charpentier M., Oldroyd G., Tadege M., Ratet P., Mysore K.S., Chen R., Udvardi M.K. Vapyrin, a gene essential for intracellular progression of arbuscular mycorrhizal symbiosis, is also essential for infection by rhizobia in the nodule symbiosis of Medicago truncatula. Plant J. 2011;65(2):244-252. DOI 10.1111/j.1365-313X.2010.04415.x.

30. Park H.J., Floss D.S., Levesque-Tremblay V., Bravo A., Harrison M.J. Hyphal branching during arbuscule development requires Reduced Arbuscular Mycorrhiza1. Plant Physiol. 2015;169(4):2774-2788. DOI 10.1104/pp.15.01155.

31. Pimprikar P., Gutjahr C. Transcriptional regulation of arbuscular mycorrhiza development. Plant Cell Physiol. 2018;59(4):876. DOI 10.1093/pcp/pcy024.

32. Pumplin N., Mondo S.J., Topp S., Starker C.G., Gantt J.S., Harrison M.J. Medicago truncatula Vapyrin is a novel protein required for arbuscular mycorrhizal symbiosis. Plant J. 2010;61(3):482-494. DOI 10.1111/j.1365-313X.2009.04072.x.

33. Shtark O.Y., Danilova T.N., Naumkina T.S., Vasilchikov A.G., Chebotar V.K., Kazakov A.E., Zhernakov A.I., Nemankin T.A., Prilepskaya N.A., Borisov A.U., Tikhonovich I.A. Analysis of pea (Pisum sativum L.) source material for breeding of cultivars with high symbiotic potential and choice of criteria for its evaluation. Ecol. Genet. 2006;4(2):22-28. DOI 10.17816/ecogen4222-28.

34. Shtark O.Y., Sulima A.S., Zhernakov A.I., Kliukova M.S., Fedorina J.V., Pinaev A.G., Kryukov A.A., Akhtemova G.A., Tikhonovich I.A., Zhukov V.A. Arbuscular mycorrhiza development in pea (Pisum sativum L.) mutants impaired in five early nodulation genes including putative orthologs of NSP1 and NSP2. Symbiosis. 2016; 68(1-3):129-144. DOI 10.1007/s13199-016-0382-2.

35. Siddiqui Z.A., Akhtar M.S., Futai K. (Eds.). Mycorrhizae: Sustainable Agriculture and Forestry. Springer, 2008. DOI 10.1007/978-1-4020-8770-7.

36. Simão F.A., Waterhouse R.M., Ioannidis P., Kriventseva E.V., Zdobnov E.M. BUSCO: assessing genome assembly and annotation completeness with single-copy orthologs. Bioinformatics. 2015;31(19): 3210-3212. DOI 10.1093/bioinformatics/btv351.

37. Solaiman Z., Abbott L.K., Varma A. (Eds.). Mycorrhizal Fungi: Use in Sustainable Agriculture and Land Restoration. Springer, Berlin, Heidelberg, 2014. DOI 10.1007/978-3-662-45370-4.

38. Sudheesh S., Sawbridge T.I., Cogan N.O., Kennedy P., Forster J.W., Kaur S. De novo assembly and characterisation of the field pea transcriptome using RNA-Seq. BMC Genomics. 2015;16(1):611. DOI 10.1186/s12864-015-1815-7.

39. Tikhonovich I.A., Andronov E.E., Borisov A.Y., Dolgikh E.A., Zhernakov A.I., Zhukov V.A., Provorov N.A., Roumiantseva M.L., Simarov B.V. The principle of genome complementarity in the enhancement of plant adaptive capacities. Russ. J. Genet. 2015;51(9): 831-846. DOI 10.1134/S1022795415090124.

40. Tisserant E., Kohler A., Dozolme-Seddas P., Balestrini R., Benabdellah K., Colard A., Croll D., … Shachar-Hill Y., Tuskan G., Young J.P.W., Gianinazzi-Pearson V., Martin F. The transcriptome of the arbuscular mycorrhizal fungus Glomus intraradices (DAOM 197198) reveals functional tradeoffs in an obligate symbiont. New Phytol. 2012;193(3):755-769. DOI 10.1111/j.1469-8137.2011.03948.x.

41. Tisserant E., Malbreil M., Kuo A., Kohler A., Symeonidi A., Balestrini R., Charron P., … Rensing S.A., Grigoriev I.V., Corradi N., Roux C., Martin F. Genome of an arbuscular mycorrhizal fungus provides insight into the oldest plant symbiosis. Proc. Natl. Acad. Sci. USA. 2013;110(50):20117-20122. DOI 10.1073/pnas.1313452110.

42. Trapnell C., Williams B.A., Pertea G., Mortazavi A., Kwan G., Van Baren M.J., Salzberg S.L., Wold B.J., Pachter L. Transcript assembly and quantification by RNA-Seq reveals unannotated transcripts and isoform switching during cell differentiation. Nat. Biotechnol. 2010;28(5):511-515. DOI 10.1038/nbt.1621.

43. Trouvelot A., Kough J.L., Gianinazzi-Pearson V. Mesure du taux de mycorhization VA d’un système radiculaire. Recherche de méthodes d’estimation ayant une signification fonctionnelle. Mycorhizae: Physiologie et Génétique. 1986;217-221.

44. Young N.D., Debellé F., Oldroyd G.E.D., Geurts R., Cannon S.B., Udvardi M.K., Benedito V.A., … Schwartz D.C., Rogers J., Quétier F., Town C.D., Roe B.A. The Medicago genome provides insight into the evolution of rhizobial symbioses. Nature. 2011;480:520-524. DOI 10.1038/nature10625.

45. Zhernakov A.I., Shtark O.Y., Kulaeva O.A., Fedorina J.V., Afonin A.M., Kitaeva A.B., Tsyganov V.E., Afonso-Grunz F., Hoffmeier K., Rotter B., Winter P., Tikhonovich I.A., Zhukov V.A. Mapping-by-sequencing using NGS-based 3′-MACE-Seq reveals a new mutant allele of the essential nodulation gene Sym33 (IPD3) in pea (Pisum sativum L.). PeerJ. 2019;7. DOI 10.7717/peerj.6662.

46. Zhukov V.A., Akhtemova G.A., Zhernakov A.I., Sulima A.S., Shtark O.Yu., Tikhonovich I.A. Evaluation of the symbiotic effectiveness of pea (Pisum sativum L.) genotypes in pot experiment. Sel’skokhozyaistvennaya Biologiya = Agricultural Biology. 2017; 52(3):607-614. DOI 10.15389/agrobiology.2017.3.607eng.

47. Zhukov V.A., Shtark O.Y., Nemankin T.A., Kryukov A.A., Borisov A.Y., Tikhonovich I.A. Genetic mapping of pea (Pisum sativum L.) genes involved in symbiosis. Sel’skokhozyaistvennaya Biologiya = Agricultural Biology. 2016;51(5):593-601. DOI 10.15389/agrobiology.2016.5.593eng.

48. Zhukov V.A., Shtark O.Y., Puzanskiy R.K., Avdeeva G.S., Yurkov A.P., Smolikova G.N., Yemelyanov V.V., Kliukova M.S., Shavarda A.L., Kirpichnikova A.A., Zhernakov A.I., Afonin A.M., Tikhonovich I.A., Shishova M.F. Metabolic alterations in pea leaves during arbuscular mycorrhiza development. PeerJ. 2019; DOI 10.7717/peerj.7495.

49. Zhukov V.A., Zhernakov A.I., Kulaeva O.A., Ershov N.I., Borisov A.Y., Tikhonovich I.A. De novo assembly of the pea (Pisum sativum L.) nodule transcriptome. Int. J. Genomics. 2015:695947. DOI 10.1155/2015/695947.

50. Zorin E.A., Kliukova M.S., Kulaeva O.A., Afonin A.M., Tikhonovich I.A., Zhukov V.A. Identification of sequences encoding for NCR-peptides and defensins in the “meta-assembly” of transcriptome of pea (Pisum sativum L.) nitrogen-fixing nodules. Ecol. Genet. 2019;17(3):39-46. DOI 10.17816/ecogen17339-17346.


Review

Views: 1295


Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 License.


ISSN 2500-3259 (Online)