vavilovВавиловский журнал генетики и селекцииVavilov Journal of Genetics and Breeding2500-3259Institute of Cytology and Genetics of Siberian Branch of the RAS10.18699/VJ16.19-ovavilov-961Research ArticleГЕНЕТИКА И СЕЛЕКЦИЯ РАСТЕНИЙPLANT GENETICS AND BREEDINGАнализ полиморфизма экзона-4 гена VERNALIZATION-A1 у видов полиплоидной пшеницыAnalysis of the VERNALIZATION-A1 exon-4 polymorphism in polyploid wheatМутеркоА. Ф.MuterkoA. F.

Новосибирск

Novosibirsk

muterko@gmail.comСалинаЕ. А.SalinaE. A.

Новосибирск

Novosibirsk

Федеральное государственное бюджетное научное учреждение «Федеральный исследовательский центр Институт цитологии и генетики Сибирского отделения Российской академии наук»РоссияInstitute of Cytology and Genetics SB RASRussian Federation201723052017213323333Copyright © Мутерко А.Ф., Салина Е.А., 20172017Мутерко А.Ф., Салина Е.А.Muterko A.F., Salina E.A.This work is licensed under a Creative Commons Attribution 4.0 License.https://vavilov.elpub.ru/jour/article/view/961

Одними из ключевых генов в селекции пшеницы, влияющих на адаптивность и урожайность культуры, являются гены VRN1. В недавних исследованиях установлена ассоциация полиморфизма нуклеотидной последовательности четвертого экзона гена VRN-A1 с вариабельностью количественных показателей ряда агрономически ценных признаков озимой пшеницы, таких как чувствительность к продолжительности яровизации, морозостойкость, время выхода в трубку и время колошения. Однако эти исследования были выполнены на ограниченном количестве сортов T. aestivum и не охватывали другие виды полиплоидной пшеницы. В ранних исследованиях не предполагалось наличия более чем одной копии гена VRN-A1 на геном. Кроме того, только недавно было показано, что ген VRN-D4 отличается от VRN-A1 всего по нескольким однонуклеотидным мутациям. В настоящей работе проанализировано распределение гаплотипов экзона-4 гена VRN-A1 в 158 образцах 6 видов тетраплоидной и 5 видов гексаплоидной пшеницы, несущих различные аллели этого гена. Для идентификации гаплотипов экзона-4 гена VRN-A1 был разработан кодоминантный маркер, основанный на модуляции кривизны молекул ДНК. Полиморфизм экзона-4 выявлен исключительно в гексаплоидной пшенице и ассоциирован с наличием не менее чем двух копий гена VRN-A1 в геноме. За исключением одного образца, мутантный тип экзона-4 встречался только в комбинации с диким типом. Разработаны аллель-специфические праймеры, благодаря которым, в отличие от существующих методов, можно исключить гаплотипы гена VRN-D4 в ходе анализа гаплотипов VRN-A1. В результате расширения анализируемого участка идентифицированы дополнительные гаплотипы, ассоциированные с полиморфизмом адениновых трактов в интроне-4. При этом получено расщепление по гаплотипу среди образцов, характеризующихся только одним, диким, типом экзона-4 VRN-A1, а также доминантных аллелей этого гена. На основании установленных ассоциаций между гаплотипом и аллельным вариантом гена VRN-A1 в тетраплоидной пшенице вида Triticum dicoccum идентифицирован новый, предположительно доминантный, аллель этого гена (обозначен Vrn-A1k), несущий 42 п.н. инсерцию в области промотора.

One of the key genes that influence the adaptability of wheat to environments and yield are the VRN1 genes. In recent studies, an association of missense mutations within VERNALIZATION-A1 exon-4 with modulation of quantitative values of such agronomically valuable traits as frost tolerance, vernalization requirement duration and flowering time of wheat was shown. However, these investigations were carried out exclusively in T. aestivum varieties and have not covered other species of polyploid wheat and different VRN-A1 alleles. The earlier studies did not consider more than one copy of VRN-A1 per genome. Furthermore, only recently it was shown that only several SNPs distinguish the VRN-D4 and VRN-A1 genes. In the present study, VRN-A1 exon-4 polymorphism was investigated in 158 accessions of 6 tetraploid and 5 hexaploid wheat species carrying the different VRN-A1 alleles. To identify the VRN-A1 exon-4 haplotypes, a co-dominant marker was designed, based on modulation of the curvature of the DNA molecule. Polymorphism of the VRN-A1 exon-4 was revealed only in hexaploid wheat accessions and was associated with the presence of not less than two copies of VRN-A1 per genome. With the exception of one accession, the mutant type of exon-4 was identified only in combination with the wild type. Furthermore, allele-specific primers were designed to identify the VRN-D4 gene or in order to exclude its impact on the results during analysis of the VRN-A1 haplotypes. By expanding the region being analyzed, additional haplotypes, which are associated with polymorphism of adenine tracts within intron-4, were identified. Haplotype segregation was attained among accessions carrying only intact exon-4 of VRN-A1 and among dominant alleles of this gene. Finally, based on the associations revealed between the VRN-A1 alleles and haplotypes, the new putative dominant VRN-A1 allele (designated Vrn-A1k) carrying a 42-bp insertion within the promoter region was identified in tetraploid wheat of Triticum dicoccum.

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