vavilovВавиловский журнал генетики и селекцииVavilov Journal of Genetics and Breeding2500-3259Institute of Cytology and Genetics of Siberian Branch of the RAS10.18699/VJ20.625vavilov-2641Research ArticleАКТУАЛЬНЫЕ ТЕХНОЛОГИИ ГЕНЕТИКИ РАСТЕНИЙMAINSTREAM TECHNOLOGIES IN PLANT GENETICSРеференсный транскриптом микоризованных корней гороха посевного (Pisum sativum L.) с высоким покрытиемA high coverage reference transcriptome assembly of pea (Pisum sativum L.) mycorrhizal rootsАфонинА. М.AfoninA. M.

г. Пушкин, Санкт-Петербург

Pushkin, St. Petersburg

afoninalexeym@gmail.comЛеппяненИ. В.LeppyanenI. V.

г. Пушкин, Санкт-Петербург

Pushkin, St. Petersburg

КулаеваО. А.KulaevaO. A.

г. Пушкин, Санкт-Петербург

Pushkin, St. Petersburg

ШтаркО. Ю.ShtarkO. Y.

г. Пушкин, Санкт-Петербург

Pushkin, St. Petersburg

ТихоновичИ. А.TikhonovichI. A.

г. ПушкинСанкт-Петербург

PushkinSt. Petersburg

ДолгихЕ. А.DolgikhE. A.

г. Пушкин, Санкт-Петербург

Pushkin, St. Petersburg

ЖуковВ. А.ZhukovV. A.

г. Пушкин, Санкт-Петербург

Pushkin, St. Petersburg

Всероссийский научно-исследовательский институт сельскохозяйственной микробиологииРоссияAll-Russia Research Institute for Agricultural MicrobiologyRussian FederationВсероссийский научно-исследовательский институт сельскохозяйственной микробиологии; Санкт-Петербургский государственный университет, факультет биологииРоссияAll-Russia Research Institute for Agricultural Microbiology; Faculty of Biology, St. Petersburg State UniversityRussian Federation202002072020244331339Copyright © Афонин А.М., Леппянен И.В., Кулаева О.А., Штарк О.Ю., Тихонович И.А., Долгих Е.А., Жуков В.А., 20202020Афонин А.М., Леппянен И.В., Кулаева О.А., Штарк О.Ю., Тихонович И.А., Долгих Е.А., Жуков В.А.Afonin A.M., Leppyanen I.V., Kulaeva O.A., Shtark O.Y., Tikhonovich I.A., Dolgikh E.A., Zhukov V.A.This work is licensed under a Creative Commons Attribution 4.0 License.https://vavilov.elpub.ru/jour/article/view/2641

Арбускулярная микориза (AM) – это древний мутуалистический симбиоз, который образуют 8090 % видов наземных растений с облигатно биотрофными грибами, принадлежащими к филе Glomeromycota. Этот симбиоз является взаимовыгодным, поскольку грибы AM питаются продуктами фотосинтеза растений, в свою очередь повышая эффективность поглощения питательных веществ растением из окружающей среды. Горох (Pisum sativum L.), широко распространенная сельскохозяйственная культура и важный модельный объект генетики, способен образовывать тройные симбиотические системы, состоящие из растения, грибов AM и клубеньковых бактерий. По мере распространения транскриптомных и протеомных подходов в изучении мутуалистических симбиозов гороха, для повышения разрешающей способности и точности других методов возникла необходимость в референсном транскриптоме, т. е. в знании последовательностей генов, экспрессирующихся в различных экспериментальных условиях. Многочисленные транскриптомные сборки, сконструированные для гороха, не включали микоризованные корни, поэтому целью данного исследования было создание референсной транскриптомной сборки микоризованных корней. Было проведено глубокое РНК-секвенирование транскриптома микоризованных корней Pisum sativum сорта Frisson, инокулированных Rhizophagus irregularis BEG144, и для каждого из организмов получены независимые транскриптомные сборки (покрытие 177х для транскриптома гороха и 45x для транскриптома гриба). Качество сборки транскриптома гороха оценено путем сравнения с уже имеющимися сборками транскриптомов других тканей. Для дополнительной оценки качества сборки, у двух сортов гороха (Frisson и Finale) с помощью qPCR проведен анализ экспрессии генов, специфичных для микоризованных корней, последовательности которых были найдены в созданной сборке. Исследованные гены могут служить маркерами ранних стадий развития арбускулярной микоризы у генетически разнообразных сортов гороха.

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.

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The authors declare that there are no conflicts of interest present.