Prolamin Research Articles

Application of CRISPR/Cas9 system to knock out GluB gene for developing low glutelin rice mutant

The nutritional quality improvement is among the most integral objective for any rice molecular breeding programs. The seed storage proteins (SSPs) have greater role to determine the nutritional quality of any cereal grains. Rice contains relatively balanced amino acid composition and the SSPs are fractioned into albumins (ALB), globulins (GLO), prolamins (PRO) and glutelins (GLU) according to differences in solubility. GLUs are further divided into subfamilies: GluA, GluB, GluC, and GluD depending on resemblance in amino acid. The GLU protein accounts for 60–80% of total protein contents, encoded by 15 genes located on different chromosomes of rice genome. The clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system was employed to knockout Glu-B (LOC-Os02g15070) gene in non-basmati rice PK386 cultivar. The mutant displayed two base pair and three base pair mutation in the targeted regions. The homozygous mutant plant displayed reduction for both in total protein contents and GLU contents whereas, elevation in GLO, ALB and PRO. Moreover, the mutant plant also displayed reduction in physio-chemical properties e.g., total starch, amylose and gel consistency. The agronomic characteristics of both mutant and wild type displayed non-significant differences along with increase in higher percentage of chalkiness in mutant plants. The results obtained from scanning electron microscopy showed the loosely packed starch granules compared to wild type. The gene expression analysis displayed the lower expression of gene at 5 days after flowering (DAF), 10 DAF, 15 DAF and 20 DAF compared to wild type. GUS sub-cellular localization showed the staining in seed which further validated the results obtained from gene expression. Based on these findings it can be concluded Glu-B gene have significant role in controlling GLU contents and can be utilized in breeding programs to enhance the nutritional quality of rice, and may serve as healthy diet for patient allergic with high GLU contents.

Extraction and characterization of proteins present in concentrates from quality protein maize (QPM)

Quality protein maize (QPM) represents an alternative protein source for food. This work aims to characterize fractions, molecular weights, and thermal stability of the proteins present in QPM protein concentrates obtained by isoelectric precipitation. Flours from two treatments, non-nixtamalized and nixtamalized, as well as three types of maize varieties: Sac Beh (QPM white), Chichén Itzá (QPM yellow), and Blanco Uxmal (control), were obtained. The experiment was conducted using a bifactorial 2x3 design. Four isoelectric precipitation pHs were evaluated, having the highest yield and maximum protein precipitation at pH 2,5 at 4 °C. The relative protein fractions in the maize varieties in both treatments showed more elevated amounts of prolamins (PR) and glutelins (GT) compared to the control. All QPM showed higher GT and lower PR. The denaturing electrophoretic profile (SDS-PAGE) showed molecular weights for the concentrates ranging from 17,6 to 225 kDa (non-nixtamalized), from 12,9 to 132,2 kDa (nixtamalized), and from the fractions with weights from 10,2 to 220,7 kDa. The thermograms showed a change in thermal stability in the concentrates from non-nixtamalized flour; there were no thermal transitions in the nixtamalized ones due to the denaturation of the proteins when obtaining the samples during nixtamalization of all the varieties studied.

Stanovení prolaminů ječmene v pivu a pivovarských materiálech.

Prace se zabýva možnostmi imunochemicke detekce hordeinove frakce bilkovin jecmene a připadných stěpných derivatů, respektive charakterizaci monoklonalnich protilatek připravených za ucelem vývoje rychleho, specifickeho a citliveho stanoveni hordeinových bilkovin v potravinařstvi, konkretně v pivu. Byla vyvinuta metoda nepřime kompetitivni ELISA prolaminů jecmene a stanoveny pracovni koncentrace vsech imunoreagencii. Dale byly testovany křižove interakce jednotlivých monoklonalnich protilatek s proteiny ostatnich cerealii. Detekcni limit metody cinil 2 ppm, přicemž linearni rozsah kalibracni křivky se pohybuje v rozmezi 1-100 ppm.

IBS patients complaining of adverse reaction to flour have unrecognized IgE to prolamins from wheat

IBS Patients Complaining of Adverse Reaction to Flour Have Unrecognized IgE to Prolamins from Wheat. Francesca Polato, Gastroenterological and Surg Dept, Padova Univ, Padova Italy; Barbara Simonato, Andrea Curioni, Gabriella Pasini ~, Oept Di Biotecnologie Agrarie, Univ di Padova, Padova Italy; Carla Gemignani, Allergologic Unit, Univ di Padova, Padova Italy; Mario Plebani, Lab Medicine, Azienda Hosp Padova, Padova Italy; Franca Oe Lazzari, Gastroenterological and Surg Dept, Padova Univ, Padova Italy

5'-flanking region responsible for endosperm-specific expression of rice prolamine chimeric gene in transgenic tobacco.

The 5' upstream region (-680-(+)40) containing the complete signal peptide coding sequence of the rice seed storage prolamine gene was amplified in vitro with polymerase chain reaction from the genome of Chinese super rice cultivar Zhonghua 8. Physical map and DNA sequence analysis showed strong homology with the 5'-flanking region of rice prolamine gene reported by Kim in 1988. No changes in the signal peptide coding sequence and a long leader sequence with several small ORFs were found. Chimeric gene containing 5'-flanking region of the prolamine gene has been transcriptionally fused with the beta-glucuronidase reporter gene. The fusion junction was confirmed by both physical map and DNA sequence analysis. The resultant chimeric gene was used to transform the tobacco explants by Ti binary system of Agrobacterium tumefaciens LBA4404. With dot and Southern blotting hybridization, three transgenic tobacco plants with the copies of chimeric GUS genes as many as 20 were obtained. Histochemical analysis revealed that the GUS activity in the endosperm tissues of tobacco seeds at the developmental stage was about 20 DAP. No GUS activity was found in leaves, stems, roots and flowers of the transgenic tobacco plants. Therefore, we concluded that the 5'-upstream cis-elements from -680 to -18 were enough to confer the endosperm-specific and temporal expression of rice prolamine gene.

The proteins of Bajri (Pennisetum typhoideum)

Bajri contains about two-fifths of its proteins as a prolamin and one-fifth as a globulin. The latter can be fractionated into two proteins, a small part A coagulating at 43°–45° C. and the larger part B coagulating at 83°–85° C.

A preliminary note on a prolamin isolated from a legume seed

A preliminary note on a prolamin isolated from a legume seed