Unravelling rutin content of tartary buckwheat of north western Himalayas and insights into nucleotide polymorphisms in PAL gene to infer the associations with rutin biosynthesis

Aatif Mateen Tak; Ammarah Hami; Basharat Bhat; Sajad Ahmad Bhat; Khalid Z Masoodi; M Ashraf Bhat; M D Shah; Mohd Kamran Khan; Sajad Majeed Zargar

doi:10.1007/s13205-022-03218-y

Unravelling rutin content of tartary buckwheat of north western Himalayas and insights into nucleotide polymorphisms in PAL gene to infer the associations with rutin biosynthesis

3 Biotech. 2022 Aug;12(8):156. doi: 10.1007/s13205-022-03218-y. Epub 2022 Jul 2.

Authors

Aatif Mateen Tak¹, Ammarah Hami¹, Basharat Bhat², Sajad Ahmad Bhat³, Khalid Z Masoodi⁴, M Ashraf Bhat⁴, M D Shah⁵, Mohd Kamran Khan⁶, Sajad Majeed Zargar¹

Affiliations

¹ Proteomics Laboratory, Division of Plant Biotechnology, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, Shalimar, Jammu and Kashmir 190025 Srinagar, India.
² NAIP, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, Shalimar, Jammu and Kashmir 190025 Srinagar, India.
³ Division of Basic Sciences, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, Shalimar, Jammu and Kashmir 190025 Srinagar, India.
⁴ Division of Plant Biotechnology, Sher-e-Kashmir University of Agricultural Sciences & Technology of Kashmir, Shalimar, Srinagar, India.
⁵ Division of Plant Pathology, Sher-e-Kashmir University of Agricultural Sciences and Technology of Kashmir, Shalimar, Jammu and Kashmir 190025 Srinagar, India.
⁶ Department of Soil Science and Plant Nutrition, Faculty of Agriculture, Selcuk University, Konya, 42079 Turkey.

Abstract

Buckwheat (Fagopyrum spp.) has immense nutritional and nutraceutical potential. All the plant parts of buckwheat possess various metabolites, such as rutin, quercetin, vitexin etc. The high content of rutin in this pseudo cereal crop strongly adapts it to grow under adverse environments. In the present study 50 germplasm lines of Fagopyrum tataricum were used for estimation of seed endosperm rutin content through HPLC. Furthermore, molecular analysis of PAL gene (Phenylalanine Ammonia Lyase), an upstream gene in rutin biosynthesis pathway was targeted for detection of SNPs to understand the variations in the concentrations of seed endosperm rutin content, among tartary buckwheat genotypes with highest and lowest seed endosperm rutin content. Three primer pairs were employed for amplification of PAL gene for F. tartaricum (covering whole gene) followed by sequencing. Rutin concentration in seed endosperm of F. tartaricum ranged from 194.86 to 1403.22 ppm with an average of 617.06 ppm. Highest rutin concentration was found in genotype BWZ90 and lowest in BWZ16. Significant variations were observed in the seed endosperm rutin content among the genotypes of tartary buckwheat. Furthermore, alignment of PAL gene sequences of genotypes with high seed endosperm rutin content and low seed endosperm rutin content revealed variations at 21 polymorphic sites. The amino acid sequences obtained from the nucleotide sequences were also aligned and the variations were detected at 19 positions. The putative protein structure showed conformational changes among predicted proteins from two contrasting genotypes for endosperm rutin content. We here established an inventory of seed endosperm rutin content of tartary buckwheat. This study also provided insights about role of these SNPs in rutin biosynthesis. Furthermore, this information can be used for breeding buckwheat for high metabolite contents.

Supplementary information: The online version contains supplementary material available at 10.1007/s13205-022-03218-y.

Keywords: Buckwheat; Nutraceutical; PAL; Protein structure; Rutin.