Identification of wheat-barley translocations by sequential GISH and two-colour FISH in combination with the use of genetically mapped barley SSR markers

Genome ◽  
2002 ◽  
Vol 45 (6) ◽  
pp. 1238-1247 ◽  
Author(s):  
E D Nagy ◽  
M Molnár-Láng ◽  
G Linc ◽  
L Láng

Five wheat–barley translocations in a wheat background were characterized through the combination of cytogenetic and molecular genetic approaches. The wheat chromosome segments involved in the translocations were identified using sequential GISH and two-colour FISH with the probes pSc119.2 and pAs1. The barley chromatin in these lines was identified using SSR markers. A total of 45 markers distributed over the total barley genome were selected from a recently published linkage map of barley and tested on the translocation lines. The following translocations were identified: 2DS.2DL–1HS, 3HS.3BL, 6BS.6BL–4HL, 4D–5HS, and 7DL.7DS–5HS. Wheat–barley disomic and ditelosomic addition lines for the chromosomes 3HS, 4H, 4HL, 5H, 5HL, and 6HS were used to determine the correct location of 21 markers and the position of the centromere. An intragenomic translocation breakpoint was detected on the short arm of the barley chromosome 5H with the help of SSR marker analysis. Physical mapping of the SSR markers on chromosomes 1H and 5H was carried out using the intragenomic and the interspecific translocation breakpoints, as well as the centromere, as physical landmarks.Key words: wheat-barley translocations, sequential GISH and FISH, SSR markers, physical mapping.

2002 ◽  
Vol 50 (3) ◽  
pp. 303-311 ◽  
Author(s):  
M. Molnár-Láng ◽  
G Linc ◽  
E. D. Nagy ◽  

New wheat × barley, wheat × Aegilops biuncialis and wheat × rye hybrids were produced with the aim of alien gene transfer from these species into wheat. Amphiploids were produced with the help of colchicine treatment from the last two combinations. The new wheat × barley hybrids were multiplied in tissue culture because of the high degree of sterility and then pollinated with wheat to obtain backcross progenies. Wheat-barley chromosome pairing was detected using genomic in situ hybridization (GISH) in two combinations (Mv9 kr1 × Igri, Asakazekomugi × Manas). In vitro conditions caused an increase in chromosome arm association frequency in both combinations and in fertility in some regenerants. Five wheat-barley translocations were produced in a wheat background and characterized through the combination of cytogenetic and molecular genetic approaches (GISH, FISH and SSR markers). The following translocations were identified: 2DS.2DL-1HS, 3HS.3BL, 6BS.6BL-4HL, 4D-5HS and 7DL.7DS-5HS. Physical mapping of the SSR markers on chromosomes 1H and 5H was carried out using the intragenomic and interspecific translocation breakpoints and the centromere as physical landmarks.  Disomic wheat-Aegilops biuncialis additions were produced after backcrossing the wheat-Ae. biuncialis amphiploids. Fluorescence in situ hybridization (FISH) was carried out using two repetitive DNA clones (pSc119.2 and pAs1) on Ae. biuncialis and its two diploid progenitor species to detect chromosome polymorphism. The 7M and 3M disomic chromosome additions were selected and five more lines still need to be characterized.  The octoploid triticale (Mv9 kr1 × Lovászpatonai) produced in Martonvásár was crossed with a 1RS.1BL wheat cultivar Matador. GISH analysis detected pairing between the 1RS arm of the translocation chromosome and that of Lovászpatonai rye in 32 % of the pollen mother cells, making it possible to select recombinants from this combination. The new recombinants between the 1RS of Petkus and the 1RS of Lovászpatonai rye cultivars are being analysed with the help of microsatellite markers.


Genome ◽  
2008 ◽  
Vol 51 (11) ◽  
pp. 922-927 ◽  
Author(s):  
P. G. Luo ◽  
X. Y. Hu ◽  
Z. L. Ren ◽  
H. Y. Zhang ◽  
K. Shu ◽  
...  

Stripe rust, caused by Puccinia striiormis Westend f. sp. tritici, is one of the most important foliar diseases of wheat ( Triticum aestivum L.) worldwide. Stripe rust resistance genes Yr27, Yr31, YrSp, YrV23, and YrCN19 on chromosome 2BS confer resistance to some or all Chinese P. striiormis f. sp. tritici races CYR31, CYR32, SY11-4, and SY11-14 in the greenhouse. To screen microsatellite (SSR) markers linked with YrCN19, F1, F2, and F3 populations derived from cross Ch377/CN19 were screened with race CYR32 and 35 SSR primer pairs. Linkage analysis indicated that the single dominant gene YrCN19 in cultivar CN19 was linked with SSR markers Xgwm410, Xgwm374, Xwmc477, and Xgwm382 on chromosome 2BS with genetic distances of 0.3, 7.9, 12.3, and 21.2 cM, respectively. Crosses of CN19 with wheat lines carrying other genes on chromosome 2B showed that all were located at different loci. YrCN19 is thus different from the other reported Yr genes in chromosomal location and resistance response and was therefore named Yr41. Prospects and strategies of using Yr41 and other Yr genes in wheat improvement for stripe rust resistance are discussed.


2021 ◽  
pp. 125-133
Author(s):  
Г.Ю. Спотарь ◽  
С.А. Блинова ◽  
А.А. Шварцев ◽  
Я.И. Алексеев ◽  
С.М. Гориславец

С помощью молекулярно-генетических и ампелографических методов проведена идентификация сортов винограда, относящихся к наиболее распространенным в мире техническим сортам западно-европейского происхождения. Генотипирование образцов проводилось с использованием 9-ядерных и 3-хлоропластных микросателлитных маркеров. На основании полученных профилей, по данным базы VIVC было установлено, что образец № 2 является сортом Каберне-Совиньон, образец № 4 - сортом Рисланер. Профиль образца № 1 совпадает с профилем сорта Мерло, за исключением разницы в двух парах нуклеотидов (п.н.) в одном аллеле локуса VVMD27, что можно объяснить редким случаем мутации в микросателлитной последовательности и не является достаточным основанием утверждать, что образец № 1 и Мерло являются разными сортами. Генетические профили образцов № 3 и № 6 соответствовали сортам сортогруппы Темпранильо, № 5 - сортам сортогруппы Рислинг рейнский, №7 - сортам сортогруппы Пино черный. Сорта в сортогруппах, полученные в результате соматических мутаций (связанных в основном с окраской ягод), имели одинаковый профиль. Принадлежность образцов к указанным сортам в сортогруппах была подтверждена ампелографическим методом. Использование для идентификации сортов в сортогруппах 6-9-ти SSR-маркеров в сочетании с ампелографическими методами позволяет получить достоверные результаты без удорожания работ. Однако дифференциация клонов и сортов, полученных в результате соматических мутаций, только SSR-маркерами потребует значительного увеличения их количества на 1-2 порядка либо использования высоковариабельных SSR-маркеров, таких как VRG ( Vitis riparia Götzhof). Таким образом, целесообразен более целенаправленный поиск полиморфизмов непосредственно в генах, отвечающих за определенные хозяйственно ценные признаки. В случае возникновения отличия в окраске ягод для дифференциации возможно использовать полиморфизм локуса гена VvMybA1, при изменении во вкусе и аромате ягод - в локусе гена VviDXS, при изменении лигнификации семян - в локусе гена VviAGL11, при повышении устойчивости к заболеваниям - в локусах соответствующих генов резистентности. The identification of grapes related to the most widespread wine varieties of West-European origin was carried out using molecular-genetic and ampelographic methods. Genotyping of samples was provided using 9- nuclear and 3-chloroplast microsatellite markers. Basing on the profiles obtained according to the VIVC database, it was established that Sample No. 2 is a ‘Cabernet-Sauvignon’ variety, and Sample No. 4 is a ‘Rieslaner’ variety. The profile of Sample No. 1 coincides with the ‘Merlot’ profile, except for the difference in 2 base pairs (bp) in one allele of the VVMD27 locus, which can be explained by a rare case of mutation in microsatellite sequence, and is not a sufficient reason to insist that Sample No. 1 and the ‘Merlot’ are different varieties. The genetic profiles of Samples No. 3 and No. 6 corresponded to the varieties of ‘Tempranillo’ group, No. 5 - to the varieties of ‘Rhein Riesling’ group, and No. 7 - to the varieties of ‘Pinot Noir’ group. The varieties of the groups, obtained as a result of somatic mutations (mainly associated with color of berries), had the same profile. The ampelographic method confirmed the origin of samples in the mentioned groups of varieties. Using of 6-9-SSR-markers in combination with ampelographic methods to identify the varieties of groups allows obtaining reliable results without increasing the cost of work. However, differentiation of clones and varieties in groups with only SSR-markers will require a significant increase in their number by 1-2 orders, or using of highly variable SSR-markers, such as VRG ( Vitis riparia Götzhof). Thus, a more targeted search for polymorphisms directly in genes responsible for certain economically valuable traits is advisable. In case of occurrence a difference in the color of berries, it is possible to use for differentiation the polymorphism of VvMybA1 gene locus, when flavor and aroma of berries change - to use the VviDXS gene locus, when seed lignification changes - the VviAGL11 gene locus, when disease resistance increases - the loci of the corresponding resistance genes.


2020 ◽  
Vol 181 (3) ◽  
pp. 81-90
Author(s):  
A. K. Zatybekov ◽  
Y. T. Turuspekov ◽  
B. N. Doszhanova ◽  
S. I. Abugalieva

Background. Soybean (Glycine max (L.) Merr.) gradually becomes one of the leading legume crops in Kazakhstan. The area under soybeans in the country has been increasing annually and requires the development of adapted cultivars with a higher yield, improved quality characters, and resistance to emerging fungal diseases. The enlargement of the crop’s gene pool also suggests the need to study and document local soybean accessions to meet the standards of the available world soybean collection by using reliable and informative types of DNA markers.Materials and methods. In this study, the soybean collection consisting of 288 accessions from different countries, including 36 cultivars and promising lines from Kazakhstan, was studied. The molecular genetic analysis was performed using nine polymorphic SSR (simple sequence repeats) markers, seven of which (Satt244, Satt565, Satt038, Satt309, Satt371, Satt570 and Sat_308) were associated with resistance to three main fungal diseases of soybean – frogeye leaf spot, fusarium root rot, and purple seed stain.Results. The average PIC (polymorphism information content) value of the analyzed SSR markers constituted 0.66 ± 0.07, confirming their highlevel polymorphism. The principal coordinate analysis suggested that the local accessions were genetically most close to the accessions from East Asia. As the collection showed a robust resistance to three studied fungal diseases in Almaty Region during 2018–2019, the distribution of the studied SSR markers in the population was not significantly associated with resistance to the analyzed diseases under field conditions.Conclusion. SSR genotyping of the soybean collection helped to identify accessions that potentially possess resistance-associated alleles of fungal disease resistance genes. The data obtained can be further used for the development of DNA documentation and the breeding the promising cultivars and lines of soybean. 


2020 ◽  
Vol 17 (4) ◽  
pp. 156
Author(s):  
Surti Kurniasih ◽  
Rubiyo Rubiyo ◽  
Asep Setiawan ◽  
Agus Purwantara ◽  
Sudarsono Sudarsono

<p>Microsatellite or simple sequence repeat (SSR) markers have proven to be an excellent tool for cultivar identification, pedigree analysis, and genetic distance evaluations among organisms. The objectives of this research were to characterize cacao collection of Indonesian Coffee and Cacao Research Institute (ICCRI) and to analyze their genetic diversity using SSR markers. In this research, 39 SSR primer pairs were used to amplify genomic DNA of 29 cacao clones. Amplified SSR fragments for each primer pair were scored as individual band and used to determine genetic distance among evaluated cacao clones. Results of the experiment indicated that all SSR primer pairs evaluated were able to produce SSR markers for 29 cacao clones. The results also indicated that 34 out of 39 microsatellite loci evaluated were polymorphic, while 5 others were monomorphic. The total number of observed alleles among 29 clones was 132. Number of alleles per locus ranged from 4-8, with an average of 5.5 alelles per locus. Results of data analysis indicated that the PIC value was 0.665, the observed heterozigosity (Ho) was 0.651, and the gene diversity (He) was 0.720. The PIC, Ho, and He values were considered high. Genetic distances were evaluated using NTSys version 2.1 and dendrogram was constructed. Results of analysis indicated that 12 cacao clones evaluated were clustered in the first group with diversity coefficient of &lt; 3.75. Nine cacao clones were in the second group but with the same value of diversity coefficient (&lt;7.50). The rest of the cacao clones were in the third group with diversity coefficient of&gt;7.50. Based on those finding, all SSR primer pairs evaluated could be used to analyze cacao genome and be useful for genetic diversity analysis of cacao germplasm. The SSR marker analysis in ICCRI cacao collections resulted in high PIC, high observed heterozygosity, and high genetic diversity.</p><p>Key words: Theobroma cacao L, microsatelite, molecular marker, genetic diversity, heterozygosity</p><p> </p><p><strong>Abstrak</strong></p><p>Marka mikrosatelit atau sekuens sederhana berulang (simple sequence repeat = SSR) terbukti merupakan alat yang bagus untuk identifikasi kultivar, analisis pedigree, dan evaluasi jarak genetik berbagai organisme. Penelitian ini bertujuan untuk:1) karakterisasi kakao koleksi Pusat penelitian Kopi dan Kakao Indonesia menggunakan marka SSR dan 2) analisis keragaman genetik klon-klon kakao koleksi dengan menggunakan marka SSR. Dalam penelitian ini, 39 pasangan primer SSR telah digunakan untuk amplifikasi DNA genomik dari 29 klon kakao. Skoring pita SSR hasil amplifikasi menggunakan masing-masing pasangan primer dilakukan secara terpisah dan digunakan untuk menentukan jarak genetik di antara klon kakao yang dievaluasi. Hasil percobaan menunjukkan bahwa semua pasangan primer SSR yang digunakan mampu menghasilkan pita DNA hasil amplifikasi (marka SSR) untuk 29 klon kakao yang diuji. Hasil penelitian juga menunjukkan bahwa 34 dari 39 lokus SSR yang dianalisis bersifat polimorfik sedangkan lima primer yang lain bersifat monomorfik. Dari 29 klon kakao yang dievaluasi, telah berhasil diamplifikasi sebanyak 132 alel, dengan kisaran antara 4-8 alel/lokus. Rataan jumlah alel per lokus sebanyak 5,50. Hasil analisis data yang dilakukan juga menunjukkan nilai PIC untuk marka SSR yang digunakan sebesar 0,665. Untuk populasi klon kakao yang dievaluasi, diperoleh nilai rataan heterosigositas pengamatan (Ho) sebesar 0,651 dan rataan diversitas gen (He) sebesar 0,720. Nilai PIC Ho dan He yang didapat tergolong tinggi. Berdasarkan analisis keragaman dengan menggunakan program NTSys, diperoleh hasil 12 klon kakao berada dalam grup pertama (koefisien keragaman&lt;3,75) dan9 klon berada dalam grup kedua, dengan koefisien keragaman &lt; 7,50. Sedangkan klon-klon lainnya mempunyai koefisien keragaman &gt; 7,50. Berdasarkan hasil penelitian dan analisis data disimpulkan bahwa marka SSR dapat digunakan untuk menganalisis keragaman genetik plasma nutfah kakao. Tingkat polimorfisme yang dihasilkan marka SSR relatif tinggi. Tingkat heterosigositas plasma nutfah kakao koleksi Puslit Kopi dan Kakao Indonesiarelatif tinggi, dan keragaman genetiknyacukup tinggi.</p><p>Kata kunci : Theobroma cacao L, mikrosatelit, marka molekuler, keragaman genetik, heterosigositas</p>


Genetica ◽  
2010 ◽  
Vol 138 (11-12) ◽  
pp. 1277-1296 ◽  
Author(s):  
Tong Geon Lee ◽  
Yong Jin Lee ◽  
Dae Yeon Kim ◽  
Yong Weon Seo

2010 ◽  
Vol 58 (2) ◽  
pp. 151-158 ◽  
Author(s):  
A. Schneider ◽  
I. Molnár ◽  
M. Molnár-Láng

One way of incorporating useful traits from Aegilops biuncialis (2n=4x=28, U b U b M b M b ) into wheat ( Triticum aestivum L. 2n=6x=42, AABBDD) is to develop first addition then translocation lines. The 2M b , 3M b , 7M b , 3U b , 5U b and 5U b /6U b wheat- Ae. biuncialis addition lines were produced in Martonvásár. To facilitate the exact identification of the addition lines, it was necessary to analyse the fluorescence in situ hybridisation patterns of the parental wheat genotype, Ae. biuncialis and its diploid progenitors ( Ae. umbellulata 2n=2x=14, UU and Ae. comosa 2n=2x=14, MM). The great genetic variability of the Aegilops species causes polymorphism in the fluorescence in situ hybridisation (FISH) patterns of the individual chromosomes. Due to the high level of FISH polymorphism, it is advisable to confirm the identification of the Ae. biuncialis chromosomes with the help of molecular (microsatellite, SSR) markers, so 119 wheat SSR markers were tested on Aegilops biuncialis , on Ae. geniculata (2n=4x=28, U g U g M g M g ), on five wheat- Ae. biuncialis addition lines (2M b , 3M b , 7M b , 3U b , 5U b ) and on an addition series of wheat- Ae. geniculata in order to select SSR markers specific to the U and M genomes of Ae. biuncialis and Ae. geniculata .


PLoS ONE ◽  
2013 ◽  
Vol 8 (4) ◽  
pp. e59542 ◽  
Author(s):  
Stuart J. Lucas ◽  
Bala Anı Akpınar ◽  
Melda Kantar ◽  
Zohar Weinstein ◽  
Fatma Aydınoğlu ◽  
...  

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