spfpХранение и переработка сельхозсырьяStorage and Processing of Farm Products2072-96692658-767XРОСБИОТЕХ10.36107/spfp.2024.1.458spfp-458Research ArticleСтатьиArticlesГенетический анализ разнообразия линий нута методом случайной амплификации полиморфной днк (RAPD)Investigation of genetic diversity of some chickpea lines using molecular marker (RAPD)https://orcid.org/0000-0001-6189-5563МасумианМаджидMasomianMajidmasoumian200@yahoo.comhttps://orcid.org/0000-0002-4318-7077ГаравириМахмудGharaviriMahmoodgharaviri@hotmail.comhttps://orcid.org/0009-0001-7237-4314ГоджавандЗахраGhojavandZahrazghojavand89@gmail.comhttps://orcid.org/0000-0003-4796-2066АхангаранМахбубехAhangaranMahboobehahangaran@hotmail.comhttps://orcid.org/0000-0002-9287-0585МашенцеваНаталья ГеннадьевнаMashentsevaNatalyanatali-mng@yandex.ruhttps://orcid.org/0000-0002-3131-0904СинакиДжафар МасудSinakiJafar Masoudjmsinaki2020@gmail.comИранская научно-исследовательская организация науки и технологийИранIranian Research Organization for Science and Technology (IROST)Islamic Republic of IranРоссийский биотехнологический университетРоссияRussian Biotechnological UniversityRussian FederationИсламский университет АзадИранIslamic Azad UniversityIslamic Republic of IranРоссийский биотехнологический университетIslamic Republic of IranRussian Biotechnological University,Islamic Republic of Iran20240205202432195107Copyright © Масумиан М., Гаравири М., Годжаванд З., Ахангаран М., Машенцева Н.Г., Синаки Д., 20242024Масумиан М., Гаравири М., Годжаванд З., Ахангаран М., Машенцева Н.Г., Синаки Д.Masomian M., Gharaviri M., Ghojavand Z., Ahangaran M., Mashentseva N., Sinaki J.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://www.spfp-mgupp.ru/jour/article/view/458Введение

Введение: Для селекции новых сортов нута используют разнообразные генетические ресурсы растений. Например, с помощью RAPD-праймеров и ПЦР-реакции можно амплифицировать случайно повторяющиеся последовательности на геномной ДНК нута для оценки его генетического разнообразия.Настоящее исследование направлено на изучение генетического разнообразия новых сортов нута линии FLIP.

Цель

Цель: анализ генетического разнообразия новых линий нута типа кабули селекции FLIP, полученных в  Международном центре сельскохозяйственных исследований засушливых регионов (ICARDA, ИКАРДА), с помощью RAPD-маркеров и выбора линий с широкой генетической вариацией для дальнейшей селекции уникальных и перспективных генотипов. 

Материалы и методы

Материалы и методы: молекулярные RAPD-маркеры, полученные в результате ПЦР-амплификации случайных участков геномной ДНК с одним праймером произвольной нуклеотидной последовательности, были использованы для изучения генетического разнообразия 18 линий нута кабули, произрастающего на территории Ирана. Выделение ДНК из сортов нута осуществляли методом Деллапорта, амплификацию участков генома проводили с использованием 8 праймеров. После электрофореза агарозный гель окрашивали бромистым этидием и анализировали амплифицированные фрагменты генома. 

Результаты

Результаты: Линии нута были ранжированы на основе присутствия и отсутствия фрагментов ДНК, полученных с помощью RAPD-маркеров. Результаты показывают, что наибольшее количество ПЦР-продуктов нарабатывается с праймера UB763 (90 фрагментов ДНК), а наименьшее количество фрагментов ДНК ПЦР-продуктов нарабатывается с праймера T9 (10 фрагментов ДНК). При использовании праймера T9 у большинства линий амплифицируется продукт размером 700 п.н. При анализе полученных данных процент полиморфизма составил 75%, а коэффициент подобия Жаккара – 96%. 

Выводы

Выводы: Исследование подтвердило, что RAPD-маркеры являются эффективными методами обнаружения генетической изменчивости нута. Установлено, что линии FLIP03-26C/3/98TH70/4/(FLIP93-210C/ FLIP87-8C)//S96086 и FLIP06-93C/4/2002TH 119/5/[(FLIP98-64C/FLIP98-47C)//Sel99ter85488] /3/ FLIP98-022C имеют наибольшую генетическую удаленность друг от друга, и их можно использовать в качестве родительских линий для получения новых сортов нута с высоким генетическим разнообразием. Полученные результаты могут быть применены для разработки эффективных планов долгосрочного управления генетическими ресурсами урожая нута.

Introduction

Introduction: A variety of plant genetic resources are used to select new chickpea varieties. For example, using RAPD primers and PCR reactions, randomly repeated sequences on chickpea genomic DNA can be amplified to assess its genetic diversity. The present study is aimed at studying the genetic diversity of new chickpea varieties of the FLIP line.

Purpose

Purpose: analysis of the genetic diversity of new lines of chickpea of the Kabuli type of FLIP selection, obtained at the International Center for Agricultural Research of the Arid Regions (ICARDA), using RAPD markers and selection of lines with wide genetic variation for further selection of unique and promising genotypes.

Materials and methods

Materials and methods: molecular RAPD markers, obtained as a result of PCR amplification of random sections of genomic DNA with a single primer of an arbitrary nucleotide sequence, were used to study the genetic diversity of 18 lines of Kabuli chickpea growing in Iran. DNA extraction from chickpea varieties was carried out using the Dellaporta method; amplification of genome regions was carried out using 8 primers. After electrophoresis, the agarose gel was stained with ethidium bromide and the amplified genome fragments were analyzed.

Results

Results: Chickpea lines were ranked based on the presence and absence of DNA fragments obtained using RAPD markers. The results show that the largest number of PCR products is generated from the UB763 primer (90 DNA fragments), and the smallest number of DNA PCR product fragments is generated from the T9 primer (10 DNA fragments). When using the T9 primer, a 700 bp product is amplified in most lines. When analyzing the data obtained, the percentage of polymorphism was 75%, and the Jaccard similarity coefficient was 96%.

Conclusions

Conclusions: The study confirmed that RAPD markers are effective methods for detecting genetic variation in chickpea. It was established that lines FLIP03-26C/3/98TH70/4/(FLIP93-210C/ FLIP87-8C)//S96086 and FLIP06-93C/4/2002TH 119/5/[(FLIP98-64C/FLIP98-47C)// Sel99ter85488] /3/ FLIP98-022C have the greatest genetic distance from each other, and they can be used as parent lines to develop new chickpea varieties with high genetic diversity. The findings can be applied to develop effective long-term genetic resource management plans for the chickpea crop.

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