georgian national ballet tour 2022

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The National Ballet of Georgia "Suhishvili" - tour 2022

8th of June 2022, 20:00hrs  Summer theatre Varna

The two-hours programme of The Georgian National Ballet is a continuous story of the adventure of the Georgian spirit, the change, which took place in its existence.

Tickets : 30, 40, 50, 60 and 80 BGN Box office : Palace of Culture and Sports, Summer theatre Online : EVENTIM  

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About Sukhishvili Georgian National Ballet

Genres: Ballet , Folklore

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Royal National Ballet with the “Fire of Georgia” Program

March 3, 2022 @ 8:00 pm.

Parking is limited, so please plan ahead.

Royal National Ballet with the “Fire of Georgia” Program

Direct from Tbilisi, Georgia, World Touring brings the Royal National Ballet with a new program “Fire of Georgia”.

From the first days of its creation, the Dance Company has been the pride of its country. The Company has enjoyed wide popularity, due to its original, distinguished choreography and high professionalism of dancers. The dancers were charmingly wholehearted and skillful. The staging was sleek and the costumes were gorgeous…” After their recent tour to Poland, the Polish newspaper wrote: “The performance of the Royal National Ballet was an unexpected revelation for us.”

The Dance Company’s popularity is growing every year; and with every year grows the number of people around the world that admires their vital talent. The Dance Company of Georgia is very anxious to come to the United States for the first time, where their performances will be met with great anticipation, appreciation and love.

For all ticket sales and inquiries please click the Buy Tickets button below only.  Please note: The Wilshire Ebell Theatre does not sell tickets for this event.

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The Royal National Ballet presents a world premiere of the newly staged performances. This is a first precedent of this kind of performance  in the history of Georgian and world Choreography as it does not have an analogue. The performance called a "Georgian Fire" and combines Georgian folk dances, with wonderful decorations and technical support.

From 30 to 50 Professional dancers will present the show, supported with the new musical entourage and supplementary slide show decorations.

New performance of the “Royal National Ballet” is different from any other performances in the world. It is complete innovation and novelty at the show market.

The show unites world’s, folklore and classical dance elements, acrobatics, stunts, temperament, grace, synchrony, artistic, athletic boys and appealing girls. We would like also to inform you that the show includes 8 small dancers from "Academy Of Dance", who add beauty to the performance and makes it more commercial!

"Fire Of Georgia"

The Georgian Traditional Dance Program

1. Kolkhuri Suita

4. Mountainous Variations

5. Khevsuruli Suita

6. Swordplay

7. Old Tbilisi Bohemia

8. Dance by the spring water

9. Lezginka

10. Mkhedruli (Competition)

Chief Balletmaster

Gela Potskhishvili

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Rescheduling Friday's two performances of Four Divo to Saturday afternoon and evening. Questions, email info@thesharon .

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The Royal National Ballet: Fire of Georgia Direct from Tbilisi, democratic state of Georgia

Friday, March 24th, 2023 / 7:00 pm

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Where to See Georgian Folk Dance, Polyphony & Live Music in Tbilisi

Given Georgia’s rich heritage of folk song and dance, it’s little wonder that seeing a live show tops many first time visitors’ list of things to do when visiting Tbilisi .

The capital is not necessarily the best place to see such performances – Svaneti, Kakheti and Guria regions are both famed for their polyphony traditions, and Folk House in Zugdidi is one of my favourite places to listen to traditional music in Western Georgia.

This being Tbilisi though, there is a huge range of entertainment on offer, from live folk and jazz music to Georgian national dance. There are even a few opportunities to hear Georgia’s UNESCO-listed polyphonic chanting in the capital.

There’s always something on in Tbilisi, whether it’s a show at the Opera and Ballet Theatre on Rustaveli Avenue – or at the opposite end of the spectrum, an impromptu performance in a bar or restaurant in the old town (or sometimes at your guesthouse !).

This guide shows you where to find Georgian folk dance and live music in Tbilisi any night of the week. If you know of any other venues I should add to the list, please share the details in the comments below.

Please note: This post contains affiliate links, meaning I may earn a commission if you make a purchase by clicking a link (at no extra cost to you). Learn more.

Where to see Georgian folk dance in Tbilisi

Sukhishvili.

If you’re lucky enough to be in Tbilisi when Sukhishvili is performing at the New Theatre in Marjanishvili or at their purpose-built outdoor stage, Takara, do not pass up the chance to see them! Georgia’s world-famous dance troupe blends traditional Kartuli and contemporary dance for a one-of-a-kind experience.

I’ve been lucky enough to see them twice now, once during a special rehearsal and again in spring 2022 when they did a special program to raise money for Ukraine.

Sukhishvili only performs in Tbilisi when they’re not touring. Usually they do a run of shows in winter/early spring plus a few dates in summer . Dates are announced on their Facebook Page and tickets are sold online via TKT.GE . Be warned that it always sells out fast – you have to get in early as soon as tickets are released.

An accessible way to see a Georgian traditional dance show in Tbilisi at any time of year (usually paired with folk singing and polyphony) is with a ‘dinner and a show’ . I’ve been to more than a couple over the years – they’re touristy and a bit naff, but always good fun.

Here is a list of restaurants that currently offer dinner and a show in Tbilisi. Always call ahead to confirm times and prices. Reservations are essential.

Mravaljamieri

This restaurant takes its name from a popular folk song whose title translates to ‘may you live a long life’. Georgian folk dance and song is staged every evening in the main dining hall from 7pm until 10pm. They also have a large outdoor seating area and private gazebos.

The dinner and show ‘package’ requires a minimum order of 50 GEL’s worth of food and drink per person. The food gets mixed reviews, but they do serve Qvevri wine .

Find it on the riverside in Didube, 10 minutes by taxi from Freedom Square.

Contact via Facebook to confirm times and prices, and to make a booking.

• Location: Baratashvili Left Bank (Chugureti-Didube)

Ethno Tsiskvili

Located in Dighomi (15 minutes by taxi from Freedom Square), Ethno Tsiskvili is by far the most popular restaurant in Tbilisi to see national dance. I personally think the food is overrated – I much prefer eating next door at one of my favourite restaurants in Tbilisi , Kakhelebi.

If you’re here for the show though, you won’t be disappointed. They go all-out at Tsiskvili, opulent costumes, fire-tipped swords and all . Performances commence every night at 7.30pm, starting with polyphonic singing and folk music from the house ensemble, Ethno Art Band Gordela. Dancers come on in between sets, lighting up the floor. For something more laid back, the massive complex has another dining area with live jazz .

Food prices are a little higher than what you might be used to in Tbilisi, and they also enforce a strict dress code (‘business casual’) – so don’t come in your day clothes (long pants for men and a collared shirt are preferable; women can get away with a skirt or dress).

Reservations (first by email, then by follow-up phone call) are absolutely essential. Tables often book out weeks in advance during high season. Book via the website .

Their second restaurant, Tsiskvili in Vake , also has live music and cover bands (but no dancing).

• Location: Beliashvili Street (Dighomi)

Restaurant Ethnographer

Also located on Beliashvili Street, close to Tsiskvili, restaurant Ethnographer has live music and dance nightly. It’s a bit more relaxed and dare I say, less touristy.

There is no dress code or minimum spend that I’m aware of. Bookings are recommended, though – contact them through Facebook or by phone on +995 577362828.

In The Shadow of Metekhi

Located on the opposite side of the river in Avlabari (5 minutes by taxi or 30 minutes by foot from Freedom Square), this restaurant is convenient to the Old Town. The huge outdoor terrace overlooking the river makes it a favourite.

Normally they host live music and traditional dance every night of the week . Currently there are vocal performances by quartet Ranina and folk dance on Friday nights only from 7-9.30pm.

• Location: Ketevan Dedofali Avenue (Avlabari)

Old City Wall

Located close to Freedom Square, Old City Wall is a typical Georgian restaurant that hosts music and dance during high season starting from 8pm.

They require a minimum number of bookings before they will call in the troupe – so be sure to message them on Facebook at least a few days in advance. Staff are very responsive.

• Location: Shota Nishnianidze Street (close to Freedom Square)

Located on Abano Street in Old Tbilisi amongst the Abanotubani Sulfur Baths , this restaurant gets mixed reviews for both the food and service – but the location can’t be beat if you want to see national dance in the centre of the city. The dance and song program takes place every evening from 8pm onwards.

• Location: Abano Street (Kala)

Restaurant Tabla

Located in Vake (10 minutes by taxi from Freedom Square), Tabla is a beautiful restaurant with a large upstairs dining hall, a stage and a dance floor. This place gets better reviews for its food and service – but portions are a little small in my opinion. One major pet peeve is that you can only order mini khinkali dumplings in the main restaurant – big khinkali are only served downstairs!

Tabla hosts Georgian music and dance every Friday and Saturday night from 8pm-10pm. In summer high season, they sometimes put on extra shows on weekdays plus vocal recitals by a resident quartet.

• Location: Chavchavadze Avenue (Vake)

Khinkali House

There are several branches of this popular restaurant chain in Tbilisi, including one in Avlabari district. There is no set schedule for live music and folk dance at present – performances happen sporadically, mostly on weekends.

Contact via Facebook to check if any performances are scheduled during your visit.

• Location: Avlabari, Rustaveli Avenue & Isani

This restaurant is located further out in Okrokana, a 15-minute drive west from the centre of the city, but boasts wonderful views from its hilltop location. As well as classic Georgian fare they offer live music and dance courtesy of a resident ensemble.

• Location: Tbilisi-Kojori Street

Where to hear Georgian Polyphony in Tbilisi

Interested in learning more about Georgia’s folk music traditions? Voices of the Ancestors is a podcast ‘about Georgian polyphonic songs and the women who sing them’. Listen here .

Didgori Folk Ensemble

What Sukhishvili is to Georgian folk dance, Didgori is to polyphonic singing. This highly regarded ensemble is usually off touring Europe and playing festivals over the summer, but you can often catch them live in Tbilisi in autumn or winter. In previous years they have performed at Orbeliani Square and the Opera Theatre, as well as appearing at special events and folk festivals around Georgia.

The all-male ensemble is named after Didgori mountain where the Georgians defeated the Seljuks in an epic battle. The traditional chants and three-voice polyphony pieces they perform are intimately tied to this legend and beginnings of Christianity in Georgia.

Check their Facebook Page for event listings and updates.

Aristaeus Ethno Wine Bar (temporarily closed)

Run by the same folks behind Georgian Kalata, my favourite place in Tbilisi to buy edible Georgian souvenirs , Aristaeus is a delightful little wine bar with a great atmosphere.

Georgian Polyphony, courtesy of a local quartet, is normally performed every Thursday, Friday and Saturday night from 8-10pm. It pairs perfectly with a bottle of Georgian wine and an artisanal sharing plate

More info on their Facebook Page . Aristaeus is currently closed – I will return to update this info when things change.

Churches in Tbilisi with polyphonic chanting

Liturgical chanting is a traditional part of Orthodox mass, thus there are plenty of opportunities to hear Georgian polyphony in various churches around the city . Traditionally, mass includes three-voiced polyphony without any instrumental accompaniment. I’ve witnessed groups of up to six men chanting.

Sunday morning mass starts at 9am with the chanting at around 10am approximately. My two favourite churches to hear polyphony are Anchiskhati Basilica in Old Tbilisi, home of the world-famous Anchiskhati Choir (it’s a squeeze to get inside the tiny building, but the acoustics are amazing); and the Holy Lord’s Transfiguration Convent (Queen Darejan’s Palace) in Avlabari. The latter has speakers in the churchyard so you can sit on a bench in the fresh air and listen to the nuns singing inside.

People don’t seem to mind visitors dropping in – but please be discrete. I usually stand close to the door and keep my head down. Remember to dress appropriately and refrain from taking photos inside.

Bars & restaurants with live music in Tbilisi

If you’re looking for something more low-key with a local vibe, there are dozens more bars and restaurants in Tbilisi with live jazz, blues, rocks, acoustic and folk music. This list brings together a handful of the most popular.

Side note: For the best wine, cocktail and craft beer bars in Tbilisi, see this list .

I’m not covering techno music or clubs here – that’s a whole other article. Rather, these are places where you can sit (or stand) with a drink, listen to local artists and meet people. All are casual spaces that welcome walk-ins, though some now require a booking so it’s best to check ahead.

Shin-Aura (local favourite for live music in Tbilisi)

My favourite place for lunch in Tbilisi (their set menu is terrific), Shin-Aura transforms into a bar and event space after dark. The leafy outdoor area here is gorgeous, and the range of local wines spectacular.

Performances range from piano-vocal duos to jazz and folk, poetry recitals and literary events. During the season (from around April onwards) Shin-Aura hosts ‘Folk Fridays’.

The Saburtalo location is a bit of a trek if you’re staying in Old Tbilisi, but the payoff is that Shin-Aura has a truly local feel.

See event listings and announcements on their Facebook Page .

I can’t guarantee you’ll hear live music at this beautiful restaurant on Pavle Ingorokva Street – but regardless, this is an excellent place in central Tbilisi for a meal. On weekends, owned Luarsab Togonidze often pulls out his guitar and belts out a duet or two with his wife or daughter. We were lucky enough to catch an impromptu performance on our most recent visit.

More information and bookings on their Facebook Page .

Jazz Cafe Konka

At the opposite end of the spectrum, Jazz Cafe Konka (AKA Cafe Bar “Konka Station”) is located right in the middle of Old Town near the Cotton Rows. Live jazz, blues and rock from the likes of the Reso Kiknadze quintet takes place every Thursday, Friday and Saturday night in winter and every night of the week from late April, normally starting at 9pm. Accompanied by local wine and Georgian fare.

This Georgian-European restaurant off Agmashenebeli Avenue hosts live performances every night from 9pm. Friday nights are dedicated to jazz. The program is usually finalised a month in advance and includes both established local musicians and up-and-comers.

This low-key jazz bar off Kote Afkhazi Street pairs 80s New York jazz performances with cocktails and soul food. You can catch a live band any night of the week from 9pm onwards.

Backstage76

This cult-classic bar recently moved to a new location in Vake Park. As well as regular music performances (including quirky acts such as live accordion), they also host Latin nights, stand-up comedy nights and magic shows. In short: There’s always something fun happening at Backstage76.

Part of IOTA Hotel in Sololaki, Ghumeli is a restaurant first and foremost that specialises in oven-baked dishes and steaks. Live music performances (normally a duo or quintet) take place on Friday and Saturday evenings from 8.30pm.

Movement Theatre

Founded in 2001, Movement Theatre is an indie performance space that hosts ballet and theatrical performances as well as contemporary art exhibitions and music. Jam sessions are normally held on Fridays from 8pm. Find it in Mushtaidi Garden, near Dinamo Stadium.

See event listings and announcements on their website or Facebook Page .

Ikalto Micro Art Residence

This is a laid-back indie gallery in a ninth-floor apartment building in Saburtalo. Entrance costs 5 tetri – the cost of riding the elevator! Piano and acoustic performances are often staged alongside exhibitions and residencies by young artists.

Woland’s Speakeasy

This popular cocktail lounge near Liberty Square hosts a rotating program of karaoke, quiz nights and live music. This is a great place to meet other travellers.

Drunk Owl Bar

This popular bar in the Old Town (not far from the Meidan Bazaar) has live music most Friday, Saturday and Sunday nights from 8 or 9pm. As the name suggests, this is a place to get tanked up on cheap booze. The atmosphere is great and staff are friendly, but the majority of the clientele are tourists.

MacLaren’s Irish Pub

Popular among expats, this pub on Shardeni screens sports events and hosts regular live music, ranging from subdued acoustic performances by Georgian artists in the early evenings, to late-night extravaganzas with rock-and-roll cover bands.

Where is your favourite place for folk dance or live music in Tbilisi? Share your recommendations below and I’ll add them to the list.

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Hi Emily!! The Old City Wall restaurant often has traditional singing, which is truly superb in its cavern setting. They are very responsive to questions about live performances via messenger in English. The food and wine are reasonably priced for such a lovely atmosphere. I highly recommend it!!

Terrific suggestion Robin, thank you for that! I will add them to the list and hopefully check it out myself next time I’m in town. Cheers!

Thanks for all the info! Will be in Georgia in March and making plans based on your recommendations!

Wonderful, glad to hear that! Let me know if there’s anything else I can help with.

Before the pandemic, the branch of Khinkali House behind Avlabari Metro (and others? not sure) had a surprisingly good music and dance performance. You could get a table in the balcony where it was calmer and had a good view. We went with my cousin’s kids, and they were especially nice to them and always made sure the kids were in the front row when people stood up to watch. I don’t think their khinkali are as special as lots of people seem to, but they’re certainly standard.

Fantastic tip, Em! I didn’t realise they had live music. I will add this to the list. Thank you for sharing!

Irina KOUZNETSOVA ✿

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• v.26(4); 2022 Jul

Language: English | Russian

Investigation of genetic polymorphism of Russian rape and turnip rape varieties using SSR and SRAP markers

Изучение генетического полиморфизма российских сортов рапса и сурепицы с использованием ssr- и srap-маркеров, i.a. klimenko.

Federal Williams Research Center of Forage Production and Agroecology, Lobnya, Moscow region, Russia

V.T. Volovik

A.a. antonov, v.a. dushkin, a.o. shamustakimova, yu.m. yu.m. mavlyutov.

Rapeseed (Brassica napus L.) and turnip rape (B. rapa L. subsp. campestris (L.)) are important agricultural plants widely used for food, fodder and technical purposes and as green manure. Over the past decades, a large number of perspective varieties that are being currently cultivated in every region of Russia have been developed. To increase the breeding eff iciency and facilitate the seed production, modern molecular-genetic techniques should be introduced as means to estimate species and varietal diversity. The objective of the presented research study was to investigate DNA polymorphism of the rapeseed and turnip rape varieties developed at Federal Williams Research Center of Forage Production and Agroecology and detect informative markers for varietal identif ication and genetic certif ication. To genotype 18 gDNA samples, 42 and 25 combinations of respective SSR and SRAP primers were used. The results obtained demonstrate that SRAP markers were more effective for polymorphism analysis: 36 % of the tested markers revealed genetic polymorphism compared with only 16.7 % of microsatellite loci. Molecular markers to detect differences at interspecif ic and intervarietal levels have also been found. For the investigated set, such microsatellite loci as Na12A02, Ni2C12, Ni02-D08a, Ra02-E01, Ni03H07а and SRAP-marker combinations as F13-R9, Me4- R7, F11-Em2, F10-R7, F9-Em2 and F9-R8 proved to be informative. Application of the two marker techniques made it possible to detect a higher level of DNA polymorphism in plants of different types (spring and winter varieties) if compared against the intervarietal differences within a species or a group. According to Nei’s genetic diversity index, in the cluster of winter rapeseed, VIK 2 and Gorizont varieties had the longest genetic distance, and in the spring cluster, these were Novosel and Veles. A high level of similarity was found between Vikros and Bizon winter rapeseed varieties. The results obtained have a high practical value for varietal specif ication of seed material and genetic certif ication of rapeseed and turnip rape varieties.

Рапс (Brassica napus L.) и сурепица (B. rapa L. subsp. campestris (L.)) – важные сельскохозяйственные культуры, широко используются для продовольственных, кормовых и технических целей, а также в качестве сидератов. За последние десятилетия создано большое количество перспективных сортов, культивируемых практически во всех регионах России. Для повышения эффективности селекционного процесса и успешного развития семеноводства необходимо внедрять современные молекулярно-генетические методы оценки видового и сортового разнообразия. Цель настоящей работы заключалась в изучении ДНК-полиморфизма сортов рапса и сурепицы селекции Федерального научного центра кормопроизводства и агроэкологии им. В.Р. Вильямса и выявлении информативных маркеров для сортовой идентификации и генетической паспортизации. Для генотипирования 18 образцов геномной ДНК использовали 42 и 25 комбинаций SSR- и SRAP-праймеров соответственно. Результаты показали, что маркеры SRAP более эффективны для анализа полиморфизма изучаемого материала: 36 % от общего числа испытанных маркеров демонстрировали генетический полиморфизм, тогда как для микросателлитных локусов этот показатель равнялся 16.7 %. Определены молекулярные маркеры для выявления различий на межвидовом и межсортовом уровнях. Информативными для исследуемой выборки сортов оказались микросателлитные локусы Na12A02, Ni2C12, Ni02-D08a, Ra02-E01, Ni03H07а и комбинации SRAP-маркеров F13-R9, Me4-R7, F11-Em2, F10-R7, F9-Em2 и F9-R8. Анализ сортового материала по двум системам маркирования показал более высокий уровень ДНК-полиморфизма у образцов растений разного типа развития (яровой/озимый) в сравнении с различиями между сортами в пределах вида или группы. Согласно индексам генетического разнообразия Нея, в кластере сортов озимого рапса наибольшей генетической удаленностью выделялись ВИК 2 и Горизонт, среди яровых – Новосёл и Велес. Высокий уровень сходства обнаружен между яровыми сортами рапса Викрос и Бизон. Полученная информация имеет практическое значение для контроля сортовой принадлежности и генетической паспортизации семенного материала сортов рапса и сурепицы.

Introduction

Cabbage oilseed crops such as rapeseed (Brassica napus L.) and turnip rape (B. rapa L. subsp. campestris (L.)) are cultivated in almost every region of Russia, and, for the foreseeable future, are regarded as the main reserve for increasing the production of vegetable oil and fodder protein. These plants are widely used in food, fodder, technical purposes and as green manure that increases soil fertility thanks to the plants’ root remains containing up to 6 tons of organic maters, 80 kg of nitrogen, 60 kg of phosphorus and 90 kg of potassium per hectare. As for their food and fodder properties, rapeseed and turnip rape exceed many other cultivated crops since their seeds are 40–48 % fat and 21–33 % protein and contain a high amount of essential amino acids (Volovik, 2015). Rapeseed can provide livestock with green forage from early spring to late fall thanks to their cold hardiness and fast regrowth after mowing. They are also an excellent silage material, and their seeds and seed by-pass products are processed to produce seed cake and coarse meal. In the recent years the varieties of rapeseed and turnip rape with low or no erucic-acid content became available and seed production has increased more than 7 times to reach the world’s third place after soybeans and cotton. Russia’s short-term plans are to increase rapeseed planting acreage to 2.5 mln he.

As for Russian research institutions working intensely to select cabbage oilseed crops, the leading ones are All-Russian Research Institute of Rapeseed, All-Russian Research Institute of Oilseed Crops and All-Russian Williams Fodder Research Institute. For the two last decades, they have produced the perspective varieties of rapeseed, turnip rape, white mustard and oil radish that have been recommended for oil production, livestock and poultry green forage, combination fodder, seed cake and coarse meal production. In 2021, “State Register” of the Russian Federation included 13 varieties of rapeseed and 3 varieties of turnip rape selected by Federal Williams Research Center of Forage Production and Agroecology (Kosolapov et al., 2019; State Register…, 2021).

For preservation and rational use of newly available varieties, intensification of the selection process and protection of intellectual property, modern and effective methods to estimate species and varietal diversity at a genetic level are to be introduced. One of such techniques that has been successfully applied in the recent years is molecular DNA markers, which, if compared against the traditional morphological indicators, possess a number of advantages. These include a high level of polymorphism; even genome distribution; reliability; a possibility to automate the assay procedure that does not depend on environmental conditions or a plant development phase (Agarwal et al., 2008; Khlestkina, 2011; Chesnokov, 2018). If the most informative and convenient DNA markers are selected, their capabilities to estimate the genetic variability of selection material are regarded as unlimited.

Laboratory for Molecular and Genetic Studies in Federal Williams Research Center of Forage Production and Agroecology has been developing a system for DNA identification and genetic certification of Russian fodder crops. For the time being, the varietal identification techniques have been adapted for perennial legume grasses such as red clover and different species of alfalfa (Klimenko et al., 2020a, b). The assay uses samples of the summary total DNA obtained through a modified method from an arbitrary selected sample of every variety’s germinants. Two types of molecular markers were used: SSR (simple sequence repeats), which detect the variability of microsatellite genome sequences, and SRAP (sequence related amplified polymorphism), which is based on PCR with a pair of primers for amplification of intron/exon regions (open reading frames). The techniques have been tested on different species of fodder crops to optimize the amplification conditions, detection and analysis of results.

A problem of reliable varietal identification is particularly topical for rapeseed due to its limited genetic variability conditioned by the intensive selection aimed at higher content and quality of oil. Currently, a significant number of published studies have been devoted to using different DNA markers for estimation of the genetic diversity of rapeseed varieties and hybrids (Plieske, Struss, 2001; Snowdon, Friedt, 2004; Klyachenko et al., 2018; Mozgova et al., 2019); to genetic mapping (Piquemal et al., 2005; Gao et al., 2007; Geng, 2012) and marking the genes of economically valuable traits (Chen et al., 2010; Ananga et al., 2012). However, only a few such studies have investigated Russian varieties. Four varieties of winter and spring rapeseed (Podmoskovniy, Vikros, VIK 2 and Severyanin) were studied by Byelorussian researchers to identify the gene alleles determining the concentration of oleic and linolic acids in rapeseed oil (Lemesh et al., 2015). The same varieties were investigated to detect the DNA markers of the genes responsible for erucic-acid synthesis (Amosova et al., 2014). Microsatellite markers were used to study the genetic polymorphism of Russian varieties Ratnik and SNK- 198 (Satina, 2010) as well as the genetic homogeneity of spring rapeseed varieties Bulat and Forward (Rogozhina et al., 2015). Such winter varieties as Stolychniy, Laureat, Gorizont, Nord and Severyanin were investigated to detect the quantitative trait loci (QTLs) associated with high winter hardiness (Mozgova et al., 2019).

The objective of the presented study was to investigate DNA polymorphism of rapeseed and turnip rape varieties developed by breeders of Federal Williams Research Center of Forage Production and Agroecology and to identify the informative markers for varietal differentiation and genetic certification.

Materials and methods

Plant material. The study investigated 15 varieties of winter (Severyanin, Stolychniy, VIK 2, Nord, Laureat, Gorizont, Garant) and spring (Vikros, Novik, Novosel, Veles, Grant, Podmoskovniy, Lugovskoy, Bizon) rapeseed and 3 varieties of winter (Zarya) and spring (Nadezhda, Svetlana) turnip rape.

DNA extraction and PCR analysis. The gDNA was extracted from 30 germinants of each abovementioned variety (bulk samples) using the basic SDS method (Kirby, Cook, 1967; Dellaporta et al., 1983) with some modifications (Klimenko et al., 2020b). The quality and concentration of the obtained DNA fractions were verified with agarose gel (1.5 %) electrophoresis and using a Nabi spectrophotometer (MicroDigital, South Korea).

To carry out SSR analysis, 42 markers from the database Brassica info (https://www.brassica.info) and available publications were applied. The efficiency of the primers devised for these markers had been demonstrated in the studies devoted to development of the technology of rapeseed genotyping (Satina, 2010) and selection of the samples with low erucic-acid and glucosinolate content (Hasan et al., 2008). A part of the markers included in the analysis was used for hybridization control and detection of Alternaria blight resistant genotypes in Indian mustard (B. juncea L.) (Chandra et al., 2013; Sharma et al., 2018).

The PCR-mixture of 20 μl contained 3 μl 10 × PCR buffer (Taq Turbo Buffer), 0.5 μl 50 × dNTPs mix, 0.4 μl Taq polymerase (5U), forward and reverse primers (0.1 μl each, 100 μm) and 0.1 μl of DNA sample (20 ng/μl). The amplification was performed in a T-1000 thermal cycler (Bio-Rad, USA) at two different temperature regimes. The first amplification program was an initial 3-min denaturation at 95 °C followed by 30 cycles of 30 s at 94 °C, 30 s at 55–57 °C, 30 s at 72 °C and a final 5-min elongation at 72 °C (Satina, 2010). The second program included an initial 5-min denaturation at 95 °C followed by 39 cycles of 1 min at 94 °C, 2 min at 46–51 °C (depending on the primer pair in use), 2 min at 72 °C and a final 10-min elongation at 72 °C (Chandra et al., 2013). The reproducibility of obtained results was attested in three-fold replication.

SRAP analysis was carried out using 25 primer combinations comprised from 10 single oligonucleotides: F9, F13, Me4, F10, F11, R9, R7, Em2, R14, R8 (Li, Quiros, 2001; Rhouma et al., 2017). The amplification program was an initial 4-min denaturation at 94 °C followed by 10 cycles with changing temperature and duration parameters (1 min at 94 °C, 1 min at 35 °C, 1 min at 72 °C); followed by 30 cycles (1 min at 94 °C, 1 min at 50 °C, 1 min at 72 °C) and a final 5-min elongation step run at 72 °C. The PCR-mixture composition was similar to that used for the microsatellite analysis.

PCR-products were separated using 90-min 50-V agarosegel electrophoresis (4 % MetaPhorR Agarose, Rockland or 1.6 % LE, Lonza, USA). As the reference markers, 20 bp DNA Ruler (Bio-Rad), 100 kb DNA Ladder (Thermo Fisher Scientific, USA) and 100 bp + 1.5 kb (SibEnzyme, Russia) were applied.

Analysis of the obtained results. PCR-product detection and size measurement was performed using a GelDoc XR+ imaging system (Bio-Rad) and the ImageLab software (Bio- Rad Lab., Inc.) for molecular-mass markers. The obtained results were transformed into a binary matrix, and PopGene v. 1.32 (Yeh et al., 2000) was applied to determine such genetic diversity indices as the effective number of alleles per locus; Shannon’s index; expected heterozygosity; Nei’s genetic distance (Nei, Li, 1979). Polymorphism information content (PIC) for every pair of primers was calculated by the formula presented in the study (Chesnokov, Artemyeva, 2015). To build the genetic similarity dendrogram, the unweighted pair group method with arithmetic averages was applied in NTSYSpc v 2.10 (Rohlf, 2000).

To obtain gDNA from the rapeseed and turnip rape germinants, a modified SDS method was used. The applied protocol proved more effective and less costly compared to other known protocols and commercial reagents kits. The results of electrophoresis and spectrophotometry attested to the DNA’s high concentration and purification degree from protein compounds and polysaccharides for all experimental samples (Fig. 1, 2).

Lanes 1–15 (rape varieties): Severyanin, Stolychniy, VIK 2, Nord, Laureat, Gorizont, Garant, Vikros, Novik, Novosel, Veles, Grant, Podmoskovniy, Lugovskoy, Bizon; 16–18 (turnip rape varieties): Zarya, Nadezhda, Svetlana.

SSR-analysis

For genotyping the full variety collection, out of 42 SSR primers, 7 primers providing stable and reproducible amplification were selected (Table 1).

Analysis of the amplification fragments obtained using the listed primers detected 42 alleles. Their number per locus was 6 on average, varying from 3 (Ni2C12 and Bna.M.010) to 10 (Ra02-E01a). The fragment size varied from 110 bps (Ni2C12) to 1200 bps (Ni02-D08a). The maximum allele frequency was registered for Bna.M.010 (0.83), and the minimum – for Ni03H07a (0.27); the mean value was 0.42. The primers developed for Ni03H07a, Ni02-D08a and Ra02-E01a markers made it possible to detect 8–10 alleles per locus and had the highest PIC (0.82).

SRAP-analysis

Based on the results of preliminary testing, the initial 25 combinations of SRAP primers were reduced to 10 pairs, amplifying stable polymorphic DNA fragments (Table 2). In total, 53 PCR fragments of 132–1674 nucleotide pairs in size were obtained. One combination contained from 4 (F9-R9) to 7 (F10-R8, F11-Em2, F10-R7) amplicons. A part of the markers proved to be informative to detect the amplification fragments for differentiating the type of plants (winter/ spring). Using 6 combinations made it possible to obtain the amplicons specific for varieties identification (marked with a star in the Table 2).

Fig. 3 demonstrates the electrophoregram of PCR results with the F9-R8 primer combination. Significant DNA profile differences were found between winter (I) and spring (II) rapeseed varieties (joined in curly brackets). The arrows mark the variety-specific PCR products characteristic for Stolychniy winter rapeseed (508 bps) and Nadezhda spring turnip rape (700 bps) as well as the absence of an amplicon in size of 460 bps in spring rapeseed Podmoskovniy though it was a specific characteristic for other varieties in this group. The performed analysis demonstrated that it is possible to identify rapeseed varieties Grant and Novosel with 3 marker combinations (F11-Em2, F10-R7 and Me4-R7), and Gorizont and Lugovskoy – with 2 (F13-R9 and Me4-R7). Variety VIK 2 was identified with SRAP primers F9-Em2, and spring ones Veles – with F10-R7. Specific DNA spectra for rapeseed varieties Stolychniy, Podmoskovniy and turnip rape Nadezhda were obtained with F9-R8 combination.

Winter rapeseed varieties: Severyanin (1), Stolychniy (2), VIK 2 (3), Nord (4), Laureat (5), Gorizont (6), Garant (7); spring rapeseed varieties: Vikros (8), Novik (9), Novosel (10), Veles (11), Grant (12), Podmoskovniy (13), Lugovskoy (14), Bizon (15). Winter turnip rape: Zarya (16); spring turnip rape: Nadezhda (17), Svetlana (18). H2O control (19). M – molecular weight marker (100 кb DNA Ladder).

The performed analysis demonstrated that it is possible to identify rapeseed varieties Grant and Novosel with 3 marker combinations (F11-Em2, F10-R7 and Me4-R7), and Gorizont and Lugovskoy – with 2 (F13-R9 and Me4-R7). Variety VIK 2 was identified with SRAP primers F9-Em2, and spring ones Veles – with F10-R7. Specific DNA spectra for rapeseed varieties Stolychniy, Podmoskovniy and turnip rape Nadezhda were obtained with F9-R8 combination

The obtained data were transformed into a binary matrix to calculate Nei’s genetic distances (Table 3). The lowest genetic similarity coefficient (0.7069) was found between rapeseed varieties Gorizont, Novosel and Grant, the highest – between spring varieties Vikros and Bizon (1.0) as well as Veles and Bizon (0.9655). A similarly high genetic distance (0.3228) indicated significant differences between pairs: Grant and VIK 2, and Lugovskoy and Stolychniy. Low distance values and high genetic similarity were demonstrated by spring varieties Bizon and Vikros (zero distance) and winter varieties Garant, Severyanin, Stolychniy, Nord, Laureat (0.0174).

Notе. According to the data of 1 (Rhouma et al., 2017); 2 (Сатина, 2010); 3 (Chandra et al., 2013).

The results of PCR analysis for SSR and SRAP markers were used to determine the genetic variability indices and build an UPGMA dendrogram depicting the varieties’ phylogenetic relationships. The variety material had a low degree of genetic heterogeneity, while higher values of expected heterozygosity (He) and the number of effective alleles (ne) were determined with SSR markers: 0.25 on average against 0.14 and 1.47 per locus if compared to 1.24, respectively. However, the SRAP method has enabled obtaining more PCR products applicable for varietal differentiation (Table 4).

Notе. No. 1–15 – rapeseed varieties Severyanin, Stolychniy, VIK 2, Nord, Laureat, Gorizont, Garant, Vikros, Novik, Novosel, Veles, Grant, Podmoskovniy, Lugovskoy, Bizon.

Analysis of the UPGMA dendrogram demonstrated that the winter/spring rapeseed varieties were divided into two distinguishable clusters (Fig. 4). The first one united such winter cultivars as Severyanin, Garant, Stolychniy, Nord, Laureat, Gorizont, VIK 2; the second – all the spring ones. In the winter cluster VIK 2 and Gorizont were the most distant from the other varieties. The distances between Stolychniy, Nord, Laureat as well as between Garant and Severyanin were much shorter, which was confirmed by their high genetic similarity indices being 0.9655 and 0.9828, respectively (see Table 3). The most distant among spring rapeseed were twozero varieties Novosel, Grant and Lugovskoy, which had the longest genetic distances in the cluster (0.3469 and 0.3228). Bizon and Vikros belonged to one subgroup, sharing a common branch of the dendrogram.

The bulk strategy of DNA sampling from 30 germinants per variety has significantly reduced the labor efforts and cost of the research if compared to the traditional method of individual sample genotyping. The method has proved its efficiency for different cultures especially in large-scale studies of vast populations (Liu et al., 2018). However, this approach is only justified if the analyzed set of samples is representative. For cross-pollinating species with a high level of intrapopulation variations, it should include at least 30–50 plants per variety, which significantly increases the likelihood of registering a rare alleles, the occurrence of which in the population does not exceed 10 % (Crossa, 1989; Semerikov et al., 2002). The plants of winter rapeseed are known for their high self-pollination capacity (up to 70 % of flowers) (Shpaar, 2012), many varieties are linear; while in spring rapeseed this capacity reaches 40 % (Osipova, 1998). That’s why in our study we used budk samples that combined 30 seedlings from each variety.

A significant part of SSR primers tested in our study generated monomorphic amplification fragments. They did not allow us to properly estimate the genetic variability and had low reproducibility in replicated experiments. A proportion of the markers proven effective for intervarietal DNA polymorphism detection comprised 16.7 %, being much lower than in other studies (Plieske, Struss, 2001; Hasan et al., 2008; Tian et al., 2017). It was probably due to the composition of the tested collection that had a narrow genetic basis considering the varieties’ pedigree. At the same time, such parameters of genetic variability as the number of allelic variants, singleallele frequency, PIC and He were comparable to those found in published data (Satina, 2010; Klyachenko et al., 2018).

In general, the used markers made it possible to detect DNA polymorphism between rapeseed and turnip rape as well as between the winter and spring varieties within each species. However, Na12A02 marker turned out to be variety-specific for Bizon winter rapeseed and Zarya spring turnip rape, and Ra02-E01а – for VIK 2 winter rapeseed and Svetlana spring turnip rape. The unique alleles of Podmoskovniy and Lugovskoy rapeseed were detected using Ni02-D08a loci. The indicated markers can be used for varietal DNA identification and genetic certification.

SSR primers for the markers of Indian mustard’s Alternaria blight resistance genes (Chandra et al., 2013), such as Ni02- D08a, Ni03H07a and RA02-E01a, proved to be the most effective. Their application enabled us to detect the specific amplification fragments for linear winter rapeseed variety VIK 2. They also proved effective for Gorizont, which had been obtained on the base of VIK 2 by seed freezing followed by their selection at low-temperature stress. These two varieties share high winter hardiness and are resistant to Alternaria blight. Thereby the results of our study can be useful for further selection of perspective breeding material and QTL analysis on disease resistance.

Among the spring rapeseed, Veles variety turned out to be substantially different while Lugovskoy and Garant had many similarities in the studied microsatellite parts of regions of the genome. Veles is a new perspective variety that has been approved for use since 2021 and was selected based on Vikros using the method of chemical mutagenesis, producing a high frequency of nucleotide changes. This is possibly the reason for Veles having unique alleles in three loci: Ni2C12, Ra02- E01a, Na12A02. For Vikros variety, a specific DNA profile was also obtained with Ni2C12 marker.

Rapeseed Grant was selected using the method of interspecies and intervarietal hybridization of early-maturing foreign breeding samples and the high-yielding varieties Lugovskoy and Vikros, developed at Federal Williams Research Center of Forage Production and Agroecology. Their common origin is probably the reason for the genetic similarity found between Grant and Lugovskoy varieties.

In general, SSR analysis failed to achieve optimum effect in identification of the investigated varieties: from the total set, including 42 primers for microsatellite genome loci, only four were attested as variety-specific for rapeseed, and only one (Ni03H07а) – for Nadezhda spring turnip rape.

For further investigation of DNA polymorphism, SRAP analysis was applied. SRAP is the third generation of molecular markers that were initially designed for the genes of B. oleracea L. (Li, Quiros, 2001) and are successfully used these days for genetic variability estimation and genetic mapping in different plants (Aneja et al., 2012; Rhouma et al., 2017; Liu et al., 2018). This is a cheap, effective and highly reproducible technique

2017; Liu et al., 2018). This is a cheap, effective and highly reproducible technique

The final dendrogram of phylogenetic relations made it possible to visually estimate the degrees of genetic similarities and differences of the studied material. For instance, close placing of such rapeseed varieties as Stolychniy, Nord and Laureat was probably determined by the features of their origin: they were selected for winter hardiness from a combination, in which one of the parental forms was Promin’, a well-known winter rapeseed variety

Garant, selected for winter hardiness, and Severyanin, which was obtained by seed freezing in a climatic chamber and the following individual-family selection, turned out to be in the common subgroup and at a short genetic distance (0.0174) from each other. In addition to high winter hardiness, these varieties are resistant to lodging and to damage by pathogenic fungi

A two-zero spring variety Novosel takes a special position in his group (Nei’s distance is 0.3469). Novosel was developed based on the foreign breeding samples and Russian varieties Lugovskoy and Vikros, characterized by early maturing and high yield. Specific properties of the new breeding achievement are shorter maturation period in comparison to standard varieties and high resistance to Alternaria blight.

Spring rapeseed Bizon and Vikros take the common branch of the dendrogram. The varieties were developed using the method of interspecies hybridization but from different parental forms; characterized by high yield productivity, early maturation and low glucosinolate content.

The presented study has proved the efficiency of SSR and SRAP markers for estimation of DNA polymorphism in rapeseed and turnip rape varieties developed in Federal Williams Research Center of Forage Production and Agroecology. During the study, SRAP technique has demonstrated a higher level of informativity: 36 % of the tested markers were polymorphic, while for the microsatellite loci this rate did not exceed 16.7 %.

Both techniques of molecular analysis enabled detecting the DNA markers for identification of 10 out of 15 rapeseed varieties tested and for 2 turnip rape samples. Microsatellite loci Na12A02, Ni2C12, Ra02-E01 and Ni02-D08a allowed obtaining unique PCR products for Bizon, Veles, Vikros, VIK 2, Podmoskovniy and Lugovskoy rapeseed varieties. Marker Ni03H07а proved effective for identifying Nadezhda turnip rape. In the used SRAP test kit, such primers as F13-R9, Me4- R7, F11-Em2, F10-R7, F9-Em2 and F9-R8 proved effective for detecting variety-specific amplicons or obtaining unique DNA profiles for different types of plants (winter/spring) in rapeseed varieties Grant, Novosel, Gorizont, Stolychniy, Lugovskoy, Podmoskovniy and in spring turnip rape Svetlana.

The results of the study can be used for development of the perspective breeding samples and hybrids, for genetic certification and seed material purity control.

Conflict of interest

The authors declare no conflict of interest.

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Acknowledgments

The presented investigation was supported by the means of the federal budget, directed for performing the government assignment (project No. 0442-2019-0001АААА-А19-119122590053-0).

Contributor Information

I.A. Klimenko, Federal Williams Research Center of Forage Production and Agroecology, Lobnya, Moscow region, Russia .

V.T. Volovik, Federal Williams Research Center of Forage Production and Agroecology, Lobnya, Moscow region, Russia .

A.A. Antonov, Federal Williams Research Center of Forage Production and Agroecology, Lobnya, Moscow region, Russia .

V.A. Dushkin, Federal Williams Research Center of Forage Production and Agroecology, Lobnya, Moscow region, Russia .

A.O. Shamustakimova, Federal Williams Research Center of Forage Production and Agroecology, Lobnya, Moscow region, Russia .

Yu.M. Yu.M. Mavlyutov, Federal Williams Research Center of Forage Production and Agroecology, Lobnya, Moscow region, Russia .