• Mohsen Ebrahimi

      Articles written in Journal of Genetics

    • Chalcone synthase genes from milk thistle (Silybum marianum): isolation and expression analysis

      Sepideh Sanjari Zahra Sadat Shobbar Mohsen Ebrahimi Tahereh Hasanloo Seyed-Ahmad Sadat-Noor Soodeh Tirnaz

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      Silymarin is a flavonoid compound derived from milk thistle (Silybum marianum) seeds which has several pharmacological applications. Chalcone synthase (CHS) is a key enzyme in the biosynthesis of flavonoids; thereby, the identification of 𝐶𝐻𝑆 encoding genes in milk thistle plant can be of great importance. In the current research, fragments of 𝐶𝐻𝑆 genes were amplified using degenerate primers based on the conserved parts of Asteraceae 𝐶𝐻𝑆 genes, and then cloned and sequenced. Analysis of the resultant nucleotide and deduced amino acid sequences led to the identification of two different members of 𝐶𝐻𝑆 gene family, 𝑆𝑚𝐶𝐻𝑆1 and 𝑆𝑚𝐶𝐻𝑆2. Third member, full-length cDNA (𝑆𝑚𝐶𝐻𝑆3) was isolated by rapid amplification of cDNA ends (RACE), whose open reading frame contained 1239 bp including exon 1 (190 bp) and exon 2 (1049 bp), encoding 63 and 349 amino acids, respectively. In silico analysis of SmCHS3 sequence contains all the conserved CHS sites and shares high homology with CHS proteins from other plants. Real-time PCR analysis indicated that 𝑆𝑚𝐶𝐻𝑆1 and 𝑆𝑚𝐶𝐻𝑆3 had the highest transcript level in petals in the early flowering stage and in the stem of five upper leaves, followed by five upper leaves in the mid-flowering stage which are most probably involved in anthocyanin and silymarin biosynthesis.

    • Characterization of specific DNA markers at VRN-H1 and VRN-H2 loci for growth habit of barley genotypes


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      Requirement of vernalization is an important factor which plays a crucial role in cereals to transit from vegetative to reproductive phase. There are three types of growth habit in barley: winter, spring and facultative types; in which spring type does not require vernalization but winter and facultative genotypes require full and partial vernalization, respectively. Combination of two loci,Vrn-h1 and Vrn-h2, regulates vernalization in barley genotypes. Specific DNA markers have been identified for growth habit regulator genes in barley. In this study, we examined 24 barley genotypes using specific primers for detecting Vrn-h1 and Vrn-h2 loci. Results showed that among all differently suggested primer combinations, a few markers were precisely correlated with seasonal growth habit in barley. The specific markers of 600, 600 and 200 bps were verified for ZCCT-Ha, ZCCT-Hb and ZCCT-Hc loci, respectively. Our field growth habit test showed that cultivar Bahman as a winter growth habit, where all the others genotypes exhibited spring growth habit. By using specific primers for Vrn-h1, only Bahman cultivar produced 616 bp and 830 bp fragments and spring genotypes showed 574 bp or 616 bp alleles without any amplification for 830 bp fragments. Therefore, presence of 616 bp and 830 bp alleles together in each genotype can be considered as an informative marker for winter growth habit in barley. These informative markers can be used easily in barley breeding programmes for detection of growth habit types in the seedling stage.

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