Seed Storage Protein Profiles Research Articles

Variation in Antimicrobial Activity and Seed Storage Proteins in Three Species of the Medicinal Plant Alstonia

The family Apocynaceae comprises three species of the substantially important Alstonia plant, viz., A. scholaris, A. venenata, and A. macrophylla. The investigation of proteins contained within seeds, that has the potential to provide both precise details and a structural basis for characterizing diversity. The utilization of an electrophoretic technique for protein analysis has been observed in recent scholarly investigations. This implies that certain protein bands exhibit variability, with their presence or absence being detected across different seed arrangement levels in the gel. Furthermore, this implies that the protein bands have been segregated into distinct categories. A study was conducted to analyze the seed storage protein profiles of three distinct Alstonia species through the application of sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The resolution of the seed storage protein of the Alstonia species using 15% SDS was found to alter the banding pattern of the polyacrylamide gel. The SDS-PAGE analysis revealed differential up regulation of proteins across distinct bands. Throughout the examination, it was determined that the three aforementioned species exhibited acommon band, in addition to a protein with a molecular weight of 34 kDa. The number of protein bands attached to A. venenata was the highest (ten bands), while the number of protein bands that adhered to A. scholaris was the lowest (five bands). Further, powdered leaves of A. scholaris, A. venenata, and A. macrophylla were investigated for antibacterial and the results, A. macrophylla leaf powder is most effective against Staphylococcus aureus, followed by A. venenata and A. scholaris at 1 μg/ mL. In addition, the findings support the inference that the A. scholaris leaf powder effectively inhibited the growth of Aspergillus niger. A. venenata and A. macrophylla, both at 1μg/mL, are, nonetheless, also effective against A. niger. A. macrophylla has the largest zone of inhibition (11.5 mm) against A. niger.

Assessment of Seed Protein Quality of A Transgenic Chickpea Event Expressing Cry2Aa Protein

Background: Evaluation of the nutritional composition of genetically modified (GM) crops is mandatory for their deregulation. Chickpea is known for its high-quality protein and demonstrating that the seed protein quality of transgenic chickpea remains unaltered is important for its acceptance. Amino acid content, seed storage protein profile and the digestibility of chickpea protein are important determinants of seed protein quality. Thus, in the present study, we assessed the effect of Bt (Cry2Aa) gene expression on the Bt chickpea seed protein quality. Methods: We assessed the amino acid profile, in vitro protein digestibility and factors affecting protein digestibility like trypsin inhibitor, tannins and phytic acid contents of the transgenic Bt chickpea expressing a codon modified Cry2Aa gene and its non-transgenic counterpart. Furthermore, the seed storage proteins were also fractionated and separated on SDS-PAGE followed by mass spectroscopy of the major peptides. Result: Amino acid profile and factors affecting protein digestibility revealed no significant variations between transgenic and non-transgenic chickpeas. Seed storage protein profile confirmed the presence of legumin, vicilin and albumin. No potential change in the digestibility pattern of seed proteins was revealed. Our findings suggest no potential unintended changes in chickpea seed protein quality due to the expression of Cry2Aa gene.

Compositional analysis of transgenic Bt-chickpea resistant to Helicoverpa armigera

ABSTRACT Transgenic chickpeas expressing high levels of a truncated version of the cry1Ac (trcry1Ac) gene conferred complete protection to Helicoverpa armigera in the greenhouse. Homozygous progeny of two lines, Cry1Ac.1 and Cry1Ac.2, had similar growth pattern and other morphological characteristics, including seed yield, compared to the non-transgenic counterpart; therefore, seed compositional analysis was carried out. These selected homozygous chickpea lines were selfed for ten generations along with the non-transgenic parent under contained conditions. A comparative seed composition assessment, seed storage proteins profiling, and in vitro protein digestibility were performed to confirm that these lines do not have significant alterations in seed composition compared to the parent. Our analyses showed no significant difference in primary nutritional composition between transgenic and non-transgenic chickpeas. In addition, the seed storage protein profile also showed no variation between the transgenic chickpea lines. Seed protein digestibility assays using simulated gastric fluid revealed a similar rate of digestion of proteins from the transgenic trcry1Ac lines compared to the non-transgenic line. Thus, our data suggest no unintended changes in the seed composition of transgenic chickpea expressing a trcry1Ac gene.

Anti-stress phytohormones impact on proteome profile of green gram (Vigna radiata) under salt toxicity

Anti-stress phytohormones impact on proteome profile of green gram (Vigna radiata) under salt toxicity

Studies on genetic diversity in some selected medicinal plants of family -Solanaceae using reproductive characters and protein profiling

Reproductive characters and protein profiles of five medicinal plants (Solanum nigrum, Solanum xanthocarphum, Withania somnifera, Capsicum annum and Datura stramonium) of family Solanaceae were estimated through gel electrophoresis. The present investigation revealed that the highest percentage pollen viability was recorded in Datura stramonium (84%) which revealed that this genus has highest reproductive power as well as power of division and lowest percentage pollen viability was obtained in Withania somnifera (67%), which means this genus become scarce. Total seed storage protein profile was examined using sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). A total of 51 bands were scored, out of them 50 were polymorphic with a total of 98.03% polymorphism and only 1 band was monomorphic with 1.96% monomorphism. The electrophoresis of the total seed proteins revealed protein bands in the range of 125 KD to less than 13 KD molecular weight. The similarity index calculated on the basis of presence and absence of bands ranged from 0.029 to 0.333. A dendrogram constructed based on UPGMA (unweighted pair group method using arithmetic averages) showed distinct separation of the collected species into three clusters. This study revealed high genetic variability among the selected members and suggests possibilities for improvement through hybridization programs.

Seed storage protein profiling of sulphur applicated chickpea varieties grown under rainfed condition

Seed storage protein profiling of sulphur applicated chickpea varieties grown under rainfed condition

Genetic Diversity and Population Structure in Upland Rice (Oryza sativa L.) of Mizoram, North East India as Revealed by Morphological, Biochemical and Molecular Markers.

Upland rice landraces from different villages of Mizoram, Northeast India were analyzed for seed morphology, amylose content, aromatic characteristic, seed storage protein profiling and genetic diversity. Results revealed variation in grain length, width, weight and shape. Protein profiling showed polypeptide bands ranging from 7 to 10 with similarity coefficient from 0.556 to 1.000 in the studied populations. Population genetic analysis using simple sequence repeats markers revealed a total of 63 alleles with a high level of gene diversity at 0.6468. High values of Fst and PIC estimates were found at 0.7239 and 0.5984 respectively. The Biruchuk population was found to be the most genetically diverse cultivar and least gene diversity was found in Tuikuk buh. The UPGMA trees based on seed morphology, seed storage protein profiling and simple sequence repeats diversity showed the grouping of rice cultivars into three clusters which were further supported by model-based STRUCTURE analysis. This finding is the first-hand report in upland rice of the state and can be useful for selecting suitable rice lines for prebreeding and germplasm conservation of indigenous hill rice cultivars of Mizoram.

Genetic variation among Saudi tomato (Solanum lycopersicum L.) landraces studied using SDS-PAGE and SRAP markers.

Genetic diversity among seven Saudi tomato landraces collected from different regions of the country was assessed using SDS-PAGE and molecular (sequence-related amplified polymorphism- SRAP) markers. A total of 19 alternative protein bands with different mobility rates were identified within a molecular weight range of 9.584–225 KDa, with 53% polymorphism. Specific protein bands were observed in the “Hail 548” and “Qatif 565” landraces. Genetic similarity based on Jaccard’s coefficient ranged from 0.53 to 1.00, with an average of 0.72. For molecular evaluation, 143 amplicons (fragments) were generated using 27 SRAP primer pair combinations, of which 88 were polymorphic across all the landraces. The PIC values ranged from 0.46 to 0.90, with an average of 0.76. All landraces showed an average of 0.66 similarity coefficient value. The UPGMA dendrogram supported by principal coordinate analysis (PCoA) revealed clusters of the landraces that almost corresponded to their geographical origin. Thus, seed storage protein profiling based on SDS-PAGE and SRAP markers can efficiently be used to assess genetic variability among tomato germplasms. The information obtained in the analysis will be of great interest in the management of ex situ collections for utilization in breeding programs or for direct use in quality markets.

Relationship of Parental Genetic Distance with Heterosis and Specific Combining Ability in Sesame (Sesamum indicum L.) Based on Phenotypic and Molecular Marker Analysis.

The genetic distance analysis for selection of suitable parents has been established and effectively used in many crops; however, there is dearth of conclusive report of relationship of genetic distance analysis with heterosis in sesame. In the present study, an attempt was made to estimate the associations of genetic distances using SSR (GDSSR), seed-storage protein profiling (GDSDS) and agro-morphological traits (GDMOR) with hybrid performance. Seven parents were selected from 60 exotic and Indian genotypes based on genetic distance from clustering pattern based on SSR, seed-storage protein, morphological traits and per se performance. For combining ability analysis, 7 parents and 21 crosses generated from 7×7 half diallel evaluated at two environments in a replicated field trial during pre-kharif season of 2013. Compared with the average parents yield (12.57gplant-1), eight hybrids had a significant (P<0.01) yield advantage across environments, with averages of 26.94 and 29.99% for better-parent heterosis (BPH) and mid-parent heterosis (MPH), respectively, across environments. Highly significant positive correlation was observed between specific combining ability (SCA) and per se performance (0.97), while positive non-significant correlation of BPH with GDSSR (0.048), and non-significant negative correlations with GDMOR (-0.01) and GDSDS (-0.256) were observed. The linear regressions of SCA on MPH, BPH and per se performance of F1s were significant with R2 value of 0.88, 0.84 and 0.95 respectively. The present findings revealed a weak association of GDSSR with F1's performance; however, SCA has appeared as an important factor in the determination of heterosis and per se performance of the hybrids. The present findings also indicated that parental divergence in the intermediate group would likely produce high heterotic crosses in sesame.

Rapid Development and Characterization of Chromosome Specific Translocation Line of Thinopyrum elongatum with Improved Dough Strength.

The protein content and its type are principal factors affecting wheat (Triticum aestivum) end product quality. Among the wheat proteins, glutenin proteins, especially, high molecular weight glutenin subunits (HMW-GS) are major determinants of processing quality. Wheat and its primary gene pool have limited variation in terms of HMW-GS alleles. Wild relatives of wheat are an important source of genetic variation. For improvement of wheat processing quality its wild relative Thinopyrum elongatum with significant potential was utilized. An attempt was made to replace Th. elongatum chromosome long arm (1EL) carrying HMW-GS genes related to high dough strength with chromosome 1AL of wheat with least or negative effect on dough strength while retaining the chromosomes 1DL and 1BL with a positive effect on bread making quality. To create chromosome specific translocation line [1EL(1AS)], double monosomic of chromosomes 1E and 1A were created and further crossed with different cultivars and homoeologous pairing suppressor mutant line PhI. The primary selection was based upon glutenin and gliadin protein profiles, followed by sequential genomic in situ hybridization (GISH) and fluorescent in situ hybridization (FISH). These steps significantly reduced time, efforts, and economic cost in the generation of translocation line. In order to assess the effect of translocation on wheat quality, background recovery was carried out by backcrossing with recurrent parent for several generations and then selfing while selecting in each generation. Good recovery of parent background indicated the development of almost near isogenic line (NIL). Morphologically also translocation line was similar to recipient cultivar N61 that was further confirmed by seed storage protein profiles, RP-HPLC and scanning electron microscopy. The processing quality characteristics of translocation line (BC4F6) indicated significant improvement in the gluten performance index (GPI), dough mixing properties, dough strength, and extensibility. Our work aims to address the challenge of limited genetic diversity especially at chromosome 1A HMW-GS locus. We report successful development of chromosome 1A specific translocation line of Th. elongatum in wheat with improved dough strength.

Assessment of genetic diversity among chickpea (Cicer arietinum L.) genotypes using seed storage protein profiling and rapd markers

Seed storage protein profiling and RAPD were used in present study for the analysis of genetic diversity among 15 chickpea genotypes. SDS-PAGE of seed storage protein differentiated 15 genotypes into three major clusters. RAPD analysis was done using highly polymorphic ten primers which amplified 104 bands with 2.70 average polymorphic bands. RAPD analysis reported 24.4 average per cent polymorphism and highest in OPH-03. OPH-19 reported highly informative primer based on PIC and MI value. Similarity coefficients of RAPD data were ranged from 0.55 to 0.95. Dendrogram generated three major clusters which distinguished Fusarium wilt susceptible and resistant genotypes in cluster I and cluster II, respectively, whereas cluster III included both resistant and susceptible genotypes. RAPD marker was found more informative than seed storage protein profiling for diversity analysis. Further, no specific banding was detected that could discriminate resistant from susceptible genotypes critically.

Characterization of pearl millet hybrids and their parental lines by seed storage protein markers

The variability of seed storage-proteins of 21 pearl millet genotypes were analyzed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Electrophorogram for 21 Pearl millet genotypes (seven hybrids, six female parent, five male parent and three Open Pollinated Variety(OPV)) were scored and Jaccard‘s similarity index (JSI) was calculated. Genetic diversity of Pearl millet genotypes was evaluated by constructing the dendrogram with Unweighted Pair Group Method Arithmetic Average (UPGMA) algorithm using NTSYSPC version 2.01 software. Six mobility bands that were polymorphic (Rm=0.33, 0.39, 0.43, 0.51, 0.69 and 0.75) were used for genotyping. Though it proved an efficient method to distinguish a few genotypes (H 90/4-5, H77/833-2, G 73-107, H 77/29-2, ICMA 95222A, HHB 146, MS 843A and ICMA 89111A) based on banding pattern. Intensity of bands were also some extant helped to differentiate genotypes especially Band with Rm value of 0.43 was light for H 77/29-2, band with Rm value of 0.51 was very light for G73-107 and band with Rm value of 0.75 was medium intensity for H 90/4-5. It is concluded that seed storage protein profiles could be useful markers in the studies of genetic diversity and classification of Pearl millet genotypes.

Comparison of eating quality and seed storage protein among Korean rice landraces

Improving eating quality and nutrition is one of the most important objectives in rice breeding program. Palatability factors and seed protein patterns of 69 Korean rice landraces were compared in this study. Of seven palatability factors, setback viscosity (SBV) had the largest variance, ranging from -144 to 45. Water content (WC) had the lowest variance, ranging from 11.0 to 13.0. In correlation analysis, eating quality was positively correlated with all palatability factors except gelatinization temperature (GT). In principal component analyses (PCA), the first PC with Eigen value of 5.06 explained 42.2% of the total variance. Cool paste viscosity (CPV) was the variable that had the largest positive loading. The second PC with Eigen value of 2.09 explained an additional 17.4% of the total variance. SBV was a variable that had the highest negative loading. Peak viscosity (PKV) had positive variance between PC1 and PC2. The 69 Korean rice landraces were clustered into four groups based on physicochemical properties and palatability factors, with groups I and II showing higher EQ than other groups. SDS-PAGE analyses revealed five seed storage patterns in the 69 Korean rice landraces. Among them, 84% (58 landraces) had pattern I. These results indicate that it is possible to develop high palatability cultivars using Korean rice landraces. In addition, screening of landraces based on variation in seed storage protein profile using SDS-PAGE could be highly effective for the identification of valuable rice genetic resources.

Identification of seed storage protein markers for drought tolerance in mungbean

&lt;p&gt;&lt;strong&gt;A set of 292 mungbean germplasm accessions including 62 popularly adapted local land races and two wild forms (&lt;em&gt;Vigna radiata&lt;/em&gt; var. sublobata), important breeding lines and standard ruling varieties were screened for drought stress tolerance at seedling stage. Eight genotypes e.g., C. No. 35, OUM 14-1, OUM 49-2, Pusa 9072, OM 99-3, Banapur local B, Nipania munga, Kalamunga 1-A) have been identified to possess drought tolerance. Globulin seed storage protein profiling was carried out in 19 selected mungbean genotypes comprising eight drought tolerant, seven drought sensitive, two wild forms of mungbean (TCR 20 and TCR 213) and two standard checks (LGG 460 and T 2-1&lt;/strong&gt;&lt;strong&gt;)&lt;/strong&gt;&lt;strong&gt; to explore differentially expressed polypeptides. Seed protein profiles revealed 15 scorable polypeptide bands with molecular weights ranging from 10.0 to 102.2kD. A specific 12.8kD polypeptide band was present in all above drought tolerant test genotypes including the wild accession TCR 20. Such a polypeptide band may serve as useful biochemical marker for identification of drought tolerant genotypes in mungbean. &lt;/strong&gt;&lt;/p&gt;&lt;p&gt;&lt;strong&gt; Key words: &lt;/strong&gt;Genetic diversity, seed storage protein profile, wild and cultivated &lt;em&gt;Vigna radiata.&lt;/em&gt;&lt;strong&gt; &lt;/strong&gt;&lt;/p&gt;

Revealing contrasting genetic variation and study of genetic diversity in urdbean (Vigna mungo (L.) Hepper) using SDS-PAGE of seed storage proteins

&lt;p&gt;&lt;strong&gt;Total seed storage protein profiles of 20 urdbean genotypes including the popular variety T9 were analysed by Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). 14 genotypes could be clearly identified based on genotype-specific seed protein fingerprints while rest of the test genotypes were categorized into three protein types. Dendrogram based on electrophoretic data clustered the genotypes into seven groups at 78.5% phenon level. TU 95-1 with TU 12-25-4 revealed lowest similarity index value (0.33) followed by TU 95-1 with PU 30 and KU 96-3(SI=0.35). Clustering pattern revealed distinctly divergent group formed by TPU 95-1 and TPU 4. These may serve as a valuable source genotype in recombination breeding. &lt;/strong&gt;&lt;/p&gt;&lt;p&gt;&lt;strong&gt;Key words: &lt;/strong&gt;Seed storage protein profiling, SDS-PAGE, Genetic variation, urdbean.&lt;strong&gt;&lt;/strong&gt;&lt;/p&gt;

Characterization of Five Rice Varieties Using Morphological Traits and Seed Storage Protein Profiling

Rice is one of the most important crops and a major source of nutrition for about 2.5 billion people around the globe that belongs to Poaceae family. Repeated use of selected rice breeding lines in various breeding programs not only limits the genetic basis but also develop susceptibility to the various abiotic and biotic stresses. In these circumstances, genetic variability of the existing rice germplasm should be maintained for several economical traits by conserving land race genotypes. The present study was taken to understand the genetic variability and relatedness among five cultivars of Oryza sativa viz. Jaya and Uma (improved) and Odachen, Chennellu and Vetteri Black (traditional) cultivated in Kerala using seed protein profiling and morphological characteristics. SDS PAGE of grain protein of five edible rice varieties showed about 50% polymorphism. The UPGMA dendrogram of the protein profile generated revealed two main clusters (traditional and improved) at 61% homology. Thus the variation showed by the improved cultivars may be due to the effect of the selection procedures which made them improved and evolved rice variety. Heat map obtained during the analysis showed a clear picture about the morphological values through a color gradient, where the morphological traits apart from number of grains per panicle were mostly at the lower side for the improved varieties than the traditional varieties.Scatter plot obtained through Principle Cordinate Analysis showed morphological cohesiveness among the improved varieties while all the traditional varieties clustered far apart which shows its genetic distinctness.

Revealing genetic diversity in land races and wild accessions of mungbean [&lt;italic&gt;Vigna radiata&lt;/italic&gt; (L.) Wilczek] using SDS-PAGE of seed storage proteins

Total seed storage protein profiles of 74 mungbean land races, three wild accessions and a popular variety ‘Jyoti’ of Odisha were analysed by Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). 32 genotypes could be clearly identified based on genotype-specific seed protein fingerprints while rest of the test genotypes were categorized into eight protein types. Genotypes included in each protein type had 100% homology and some of these could be duplicates. In this pursuit, a few specific polypeptide markers have been detected for identification of the land races/ genotypes. Dendrogram based on electrophoretic data clustered the genotypes into seven groups at 70% phenon level. Paralakhemundi local, Samarjhola local and Phulbani local-D; and three wild accessions (TCR 20, TCR 213 and TCR 243) were comparatively divergent from other genotypes. Besides, Jyoti, Kalahandi local 2A, Sikri local, kodala local A and TCR 20 were identified to be protein rich with high seed yield. TCR 20 being morphologically similar to mungbean, moderately high protein content and high yielding as well as resistant to drought and bruchids; it may serve as a valuable source genotype in recombination breeding

Chromosome number, genome size, seed storage protein profile and competence for direct somatic embryo formation in Algerian annual Medicago species

The genome size of eight species from different wild Algerian populations of diploid and one tetraploid Medicago L., initially analysed for chromosome numbers and seed storage proteins, has been assessed by flow cytometry. Genome size ranged from 2C = 0.94 pg in Medicago orbicularis (L.) Bartal. to 2C = 1.80 pg in Medicago laciniata (L.) Miller. The competence for direct somatic embryo formation in liquid medium was studied for 4 of these species having distinct genome sizes: M. orbicularis (2C = 0.94 pg); M. truncatula Gaertn. (2C = 1.08 pg); M. scutellata (L.) Miller (2C = 1.11 pg); M. arabica (L.) Huds. (2C = 1.22 pg). M. orbicularis, with the smallest genome size, formed somatic embryos most quickly, with a high frequency of reactive explants and with numerous somatic embryos per explant. It was followed by M. truncatula, M. scutellata and M. arabica, which in fact represents the order of increasing genome size.

Evaluation of Genetic Diversity in Vigna radiata (L.) Using Protein Profiling and Molecular Marker (RFLP)

Seed storage protein profiling through SDS-PAGE and DNA Restriction Fragment Length Polymorphism (RFLPs) were surveyed in 5 accessions of mungbean (Vigna radiata L.) sampled from NBPGR Thrissur, Kerala, India, to evaluate genetic diversity. The protein profile showed a low level of polymorphism (26.66%). The similarity index calculated ranged from (42.30-50%). The dendrogram constructed revealed two clusters. Cluster 1st comprises two accessions IC 251432 and IC 251433. Cluster 2nd comprises 3 cultivars viz., IC 251431, IC 251430 and IC 251434. Cluster 2nd also revealed a 100% homology between IC 251431 and IC 251430 accessions. The RFLP analysis was carried out by single restriction enzyme EcoRI. The RFLP profile revealed a high level of polymorphism (90.90%). A total of 11 clear bands were scored. The only band with fragment size 630 kb was monomorphic while, the rest were polymorphic. Similarity index ranged from (42.82-11.11%). The UPGMA analysis divided the accessions into 2 clusters viz., cluster 1st comprising 4 accessions (IC 251431, IC 251432, IC 251433 and IC 251430). Cluster 2nd includes only one accession (IC 251434) occupying a distinct position in constructed dendrogram.

Phylogenetic relationship and germplasm evaluation of different taxa of the genus Cucurbita using seed storage protein profiling

Seed protein profiling of 34 lines of Cucurbita pepo L. from different geographic regions of the world were evaluated for their polypeptide patterns and their phylogenetic relationship with other taxa of the genus Cucurbita. Considerable variations were observed in the polypeptide patterns of various lines on the SDS-polyacrylamide gels under reduced conditions, i.e. seed protein extract with 2-mercaptoethanol. The variations were observed in five different molecular weight regions, i.e. in the range 64–70, 53–60, 30–41, 20–26 and 18–20 kDa. Cluster analysis showed 100% genetic similarity between C. pepo and C. pepo var. pepo whereas it is quite distinct from C. pepo var. texana and C. pepo var. fraterna. On the basis of electrophoretic profiling C. pepo var. texana and C. pepo var. fraterna should be considered as different species and also supports the origin of C. pepo from C. texana.