Phenylalanine Ammonia Lyase (PAL) gene which plays a key role in bio-synthesis of medicinally important compounds, Rutin/quercetin was sequence characterized for its efficient genomics application. These compounds possessing anti-diabetic and anti-cancer properties and are predominantly produced by Fagopyrum spp. In the present study, PAL gene was sequenced from three Fagopyrum spp. (F. tataricum, F. esculentum and F. dibotrys) and showed the presence of three SNPs and four insertion/deletions at intra and inter specific level. Among them, the potential SNP (position 949th bp G>C) with Parsimony Informative Site was selected and successfully utilised to individuate the zygosity/allelic variation of 16 F. tataricum varieties. Insertion mutations were identified in coding region, which resulted the change of a stretch of 39 amino acids on the putative protein. Our Study revealed that autogamous species (F. tataricum) has lower frequency of observed SNPs as compared to allogamous species (F. dibotrys and F. esculentum). The identified SNPs in F. tataricum didn’t result to amino acid change, while in other two species it caused both conservative and non-conservative variations. Consistent pattern of SNPs across the species revealed their phylogenetic importance. We found two groups of F. tataricum and one of them was closely related with F. dibotrys. Sequence characterization information of PAL gene reported in present investigation can be utilized in genetic improvement of buckwheat in reference to its medicinal value.
A drought-related gene belonging to the Dehydration Responsive Element Binding protein (DREB) family has been reported and characterized in durum wheat. Unlike other DREB-homologous genes, it consists of four exons and three introns and produces three transcripts by an alternative splicing mechanism. The gene sequence was analysed in a number of varieties/lines/accessions of durum wheat, triticale and in wheat genome donors, Aegilops speltoides, A. tauschii and Triticum urartu, in order to evaluate the variability and to detect other interesting molecular features. Herewith, some results are presented. In the exon 1, a single sequence repeat codifying for a stretch of alanine residues variable in length (from 3 to 7), was identified. A novel non-autonomous transposon was identified, encompassing the intron1–intron3 region and this was characterized in detail. Part of the exon 4, containing the APetala2 (AP2) domain, responsible for DNA recognition and binding, was isolated and sequenced in a collection of Aegilops species and A. speltoides accessions from the Fertile Crescent, a region characterized by a high wheat biodiversity. Although the 338 bp-long analysed sequences did not reveal substantial differences in the polymorphic patterns, using a geographic subdivision with three clusters (east, centre and west), they completely separated Aegilops from the A. speltoides genus.
Some of the key genes and regulatory mechanisms controlling drought response in durum wheat have been identified. One of the major challenges for breeders is how to use this knowledge for the achievement of drought stress tolerance. In the present study, we report the expression profiles of the TdDRF1 gene, at consecutive plant growth stages, from different durum wheat genotypes evaluated in two different field environments. The expression of a possible target gene (Wdnh13) of the TdDRF1 gene was also investigated and analogies with the transcript profiles were found. The results of the qRT-PCR highlighted differences in molecular patterns, thus suggesting a genotype dependency of the TdDRF1 gene expression in response to the stress induced. Furthermore, a statistical association between the expression of TdDRF1 transcripts and agronomic traits was also performed and significant differences were found among genotypes, suggesting a relationship. One of the genotypes was found to combine molecular and agronomic characteristics.
Diseases adversely affect grain yield of crop plants. Leaf rust is a major disease of wheat. As race-specific resistance breaks down, introduction of newer sources of resistance often from older accessions is necessary. Linkage drag from the donor accessions adversely impacts grain yield. As a result, CIMMYT breeding effort has shifted to using durable resistance, also referred to as race non-specific host resistance, which allows slow rusting but maintains grain yield. One of the key genes for durable resistance is Lr67 and the resistant form of this gene is absent in CIMMYT elite lines. Hence, we have initiated efforts to convert the susceptible copy of Lr67 into its resistant form directly in these elite lines using gene editing. This would eliminate backcrossing and thus save time as well as eliminate linkage drag that would accompany the resistant copy of the gene if it were to be introgressed from an older accession. As first steps, we have isolated and sequenced the genomic copies from each of the A, B, and D genome of the Lr67 gene from three elite lines and an experimental line. Identification of more than 50 single nucleotide polymorphisms (SNPs) in the open reading frames (ORF) among these lines would be useful in designing the guide RNA molecules with precision. Further, we have streamlined genetic transformation of these elite lines, a prerequisite step for gene editing.