基于比较基因组学的茄科蔬菜(番茄、辣椒、茄子)myb因子多态性研究,寻找花青素积累的DNA标记

O. Babak, N. Nekrashevich, T. Nikitinskaya, K. Yatsevich, A. Kilchevsky
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引用次数: 3

摘要

花青素是高价值的植物抗氧化剂;它们也决定了生物和非生物的抗逆性。本研究的目的是研究茄科蔬菜作物C. annuum和S. melongena的花色苷1等位基因多态性。结果显示,在C. annuum中,Myb113-like1 TF和Myb113like2转录因子与S. melongena中的Myb1基因最接近。获得了这些基因的外显子扩增子,并在花青素果实颜色不同的辣椒和茄子样品中进行了测序。开发了鉴定多态性的引物,并对其与果实花青素积累的关系进行了研究。研究发现,单个核苷酸缺失(Myb113-like1)的果实中花青素积累最少或完全不合成之间存在密切的相关性,并且在辣椒样品中,使用CAPS标记myb113 - acci检测到2个SNP (Myb113-like2)。在茄子样品中,使用SCAR标记MybMel和CAPS标记MybmelPst1检测到6 bp和26 bp的缺失。Myb113-like1 TF中的1Indel对花青素合成的干扰是由阅读框的移位和Myb113-like2 TF中的snp导致氨基酸取代:Lys→Arg和Thr→Lys。在茄子中,6 bp的缺失导致该蛋白中ala和arg的丢失;26 bp的缺失会导致mRNA成熟过程中的紊乱。所开发的标记可以识别myb样TF等位基因,导致果实花青素合成紊乱。选择了不同等位基因的玉米和西瓜作为进一步研究和农业新品种。
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Study of the Myb-factor polymorphism based on comparative genomics of vegetable Solanaceae crops (tomato, pepper, eggplant) to search for DNA markers that differentiate samples by the anthocyans accumulation
Anthocyanins are high-value plant antioxidants; they also determine biotic and abiotic stress resistance. The aim of our research was to study the allelic polymorphism of Antocyanin 1 orthologs in the vegetable Solanaceae crops of C. annuum and S. melongena. The search revealed the following closest genes in C. annuum: Myb113-like1 TF and Myb113like2 transcription factors and Myb1 in S. melongena. Exon amplicons of those genes were obtained and then sequenced in the pepper and eggplant samples with contrasting anthocyanin fruit coloration. Primers to the identified polymorphisms were developed and their correlation with the anthocyanin accumulation in fruits was studied. A close correlation was found between a minimum accumulation or the complete absence of anthocyanin synthesis in fruits with a single nucleotide deletion (Myb113-like1), and in the pepper samples, 2 SNP (Myb113-like2) was detected using the CAPS marker Myb 113-AccI. In the eggplant samples, the deletions of 6 and 26 bp were detected using the SCAR marker MybMel and the CAPS marker MybmelPst1. The disturbance of anthocyanin synthesis in pepper forms with 1Indel in Myb113-like1 TF was determined by a shift in the reading frame and SNPs in Myb113-like2 TF lead to amino acid substitutions: Lys → Arg and Thr → Lys. In the eggplant, a deletion of 6 bp leads to the loss of ala and arg in the protein; a deletion of 26 bp causes disorder during the mRNA maturation. The developed markers allow identifying the Myb-like TF alleles under study, resulting in anthocyanin synthesis disturbance in fruits. C. annuum and S. melongena samples with different alleles were selected for a further study and new varieties in agriculture. 
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