Sequence-related Amplified Polymorphism Markers Research Articles

Induction of genetic variability with gamma radiation and detection of DNA polymorphisms among radiomutants using sequence-related amplified polymorphism markers in Gaillardia pulchella Foug. plants

Induction of genetic variability with gamma radiation and detection of DNA polymorphisms among radiomutants using sequence-related amplified polymorphism markers in Gaillardia pulchella Foug. plants

Assessing genetic diversity and population structure of Iranian melons (Cucumis melo) collection using primer pair markers in association with resistance to Fusarium wilt.

We evaluated genetic diversity and population structure of Iranian melons (Cucumis melo L.) using combinations of 35 primer pairs: 15 Simple-Sequence-Repeats (SSR); 10 Inter-Simple-Sequence-Repeats (ISSR); and 10 Sequence-related amplified polymorphism (SRAP) markers in association with resistance to melon Fusarium wilt, caused by Fusarium oxysporum f. sp. melonis (FOM). Genetic similarity was determined by simple matching coefficient (SSM) and dendrogram by clustering-analysis with unweighted pair groups using arithmetic averages (UPGMA). By combining ISSR-SSR-SRAP markers, a high degree of variation among the melons was detected. The mean polymorphism information content (PIC), marker index (MI), effective-number of alleles (I), expected heterozygosity (H), and Nei's gene diversity parameters were 0.392, 0.979, 1.350, 0.551 and 0.225, respectively. According to MI, PIC, I, H, and Nei indices evaluation, ISSR6, ISSR9, SRAP3, SRAP5, SSR3 and SSR6 had the best performance in genetic diversity of the related melons population. The 35 primers yielded a total of 264 bands, of which 142 showed polymorphism. Clustering of genotypes based on resistance to Fusarium wilt, and comparison with grouping on SSR, SRAP and ISSR marker revealed a significant compliance between disease severity and molecular marker dendrograms. Thus, increasing the number of molecular markers for genetic diversity provides a powerful tool for future agricultural and conservation tasks.

STUDY OF GENETIC DIVERSITY IN PLUM GENOTYPES USING SRAP MARKERS

The hexaploid European plums (Prunus domestica L. and Prunus domestica subsp. insititia (L.) C.K.Schneid.) are main traditional fruit tree culture in Romania and represent an economically important fruit species with limited information on its genetic structure. Our objective was to fingerprint 33 cultivars using four Sequence-Related Amplified Polymorphism (SRAP) primer pairs to estimate the genetic relationships among local and international cultivars. The primer pairs amplified a total of 70 alleles ranging from 14 to 21 alleles per marker. A wide range of fragment length was detected among the accessions, from 65 to 2000 bp. The mean number of alleles per primer combination was 17.5, with the most alleles obtained with SRAP80 (21 alleles), range between 65-1800 bp and the fewest alleles (14) were obtained with SRAP95, range between 150-2000 bp. The neighbor-joining dendrogram, based on Rogers genetic distance, of the plum germplasm studied, consisted of two main clusters of different sizes: 23 entries were grouped into cluster 1 and the remaining 10 entries were grouped into cluster 2. It is interesting that cultivars representing plum species with 6x and 2x ploidy levels were clustered together. Another interesting aspect observed refers to plum genotypes clustered closely on the dendrogram according to their pedigree, such as 'Dani' (P14) - 'Tita' (P15), 'Roman' (P7) - 'Early Rivers' (P9), 'Zamfira' (P6) - 'Pescăruș' (P33). Accessions within the Romanian landrace/old autochthonous cultivars group were not clustered together. Regarding the PCA, the first two principal axes accounted for 12.91% (CP1) and 10.46% (CP2) of the total variation, respectively, together explaining 23.37% of the total variability. Our results showed that SRAP markers represent valuable tools for genetic diversity study on Prunus domestica. To our knowledge, this is the first study using SRAP markers for characterization of P. domestica germplasm. In the future, this molecular genetic information can be combined with phenological and biochemical data to identify genes, quantitative trait loci (QTL) and molecular markers that can be used to improve the plum crop breeding program.

Assessing the Suitability of Selection Approaches and Genetic Diversity Analysis for Early Detection of Salt Tolerance of Barley Genotypes

Assessment of the salt tolerance of a large genotype collection at the early growth stages may assist in the fast-tracking improvement of salt-tolerant barley genotypes in breeding programs. This study aimed to investigate the ability of traits related to seed germination ability and seedling growth performance with helping of nine sequence-related amplified polymorphism (SRAP) markers to detect the salt tolerance of 70 barley genotypes during the early growth stages. The different genotypes were exposed to three salt concentrations (0, 100, and 200 mM NaCl) and evaluated for salt tolerance by looking at germination percentage, germination index, and mean germination time during eight days as well as the lengths and weights of seedling shoot and root after 21 days from sowing. The results showed that genotypic variations in germination ability and seedling growth performance obviously appeared under 200 and 100 mM NaCl, respectively. The germination traits exhibited a strong correlation among themselves, whereas they had a poor correlation with seedling traits. A strong and positive correlation was only observed for shoot fresh weight with shoot length and root fresh weight under salinity conditions. Principal component analysis revealed that the first two components, which explained 53% of the total variability, succeeded to identify the genotypes with high salt tolerance during only one stage (germination or seedling stage) and both stages. Cluster analysis based on the stress tolerance index of germination and seedling traits grouped 70 genotypes into four key clusters, with genotypes grouped in cluster 1 and cluster 2 being salt tolerant during the germination stage and moderately tolerant during the seedling stage; the opposite was found with the genotypes grouped in cluster 4. According to Ward’s method, the salt tolerance of genotypes that ranked as most salt-tolerant (T) or salt-sensitive (S) remained almost unchanged during germination and seedling stages. In contrast, a change in salt tolerance with both stages was found for the genotypes that ranked as moderately salt-tolerant (MT) and salt-sensitive (MS) genotypes. The nine SRAP markers divided the tested genotypes into two distinct clusters, with clusters B had the most T and MT genotypes. Finally, using appropriate statistical methods presented in this study with SRAP markers will be useful for assessing the salt tolerance of a large number of barley genotypes and selecting the genotypes tolerant of and sensitive to salinity at the early growth stage.

Development of a novel SRAP-SCAR marker for rapid identification of lager and ale types in brewer's yeast.

Beer is a globally consumed and universally popular beverage. According to the fermentation conditions of brewer's yeast, ale yeast and lager yeast are the two major varieties. Normal phenotypic and genotypic approaches are insufficient and time-consuming for identifying these two forms of yeast. Therefore, a method for the rapid and cost-effective identification of lager and ale-type brewer's yeasts is necessary. In this study, we analysed the genetic diversity of 23 industrial brewer's yeasts from around the world using sequence-related amplified polymorphism (SRAP) markers and produced stable sequence characteristic amplification region (SCAR) markers. The specific DNA fragments identified by the SRAP marker were sequenced and primers were constructed; the resultant SCAR marker (757bp) was then confirmed against the indicated brewer's yeast type. The development of SRAP-SCAR marker is more economical, simple, and fast compared to morphological markers.

The efficiency of SCoT, ISSR, and SRAP markers for detecting genetic polymorphism among Egyptian barley genotypes

The results of the present study support the idea that, the tested Egyptian barley cultivars could make a useful case study for genetic diversity research. When those cultivars were examined using three distinct markers (codon targeted (SCoT), sequence-related amplified polymorphism (SRAP) and inter-simple sequence repeats (ISSR)). According to SRAP, ISSR, and SCoT analyses, respectively 85, 76, and 66 polymorphism percentages among the seven tested barley cultivars were detected. The genotype specific markers for each genotype used were determined. The data revealed that Giza127 had the highest number of unique markers, showing six specific SCoT markers, three specific SRAP markers, and one specific ISSR marker. The genetic relationships between the seven Egyptian barley cultivars are explained by the phylogenetic tree built using information gathered from various molecular markers. These gene-based molecular markers demonstrated the reliability of SCoT, ISSR, and SRAP molecular markers for identifying DNA polymorphism levels and genetic relationships in barley.

Molecular Characterization of Olive (Olea europaea L.) Cultivars in Duhok Governorate

The olive tree (Olea europaea L.) was considered one of the most cultivated crops in the Mediterranean region, and it had an economically important value due to it began consumed as a table olive and oil production. In this study, AFLP and SRAP techniques were used to identify 15 cultivars of olive plants that were collected from 12 different locations in the Duhok governorate, eleven primer combinations of sequence-related amplified polymorphism (SRAP) and ten primer combinations of amplified fragment length polymorphism (AFLP) were used to fingerprinting these cultivars. The results show that the SRAP marker had the least genetic distance 0.241between (Shtla falay olive and Khodeiri olive) and the highest value 0.946 was found between (Zawita olive and Sorani olive). While AFLP marker show the lowest value of 0.107 between (Nzarky olive and Dger olive) and the highest value of 0.916 between (Amedi olive and Shtla falay olive). Cluster analysis based on AFLP and SRAP combined data clearly differentiated the genotype and it showed it was a powerful tool to evaluate the genetic distance and fingerprinting of these tested cultivars. Establishing the genetic relationships between cultivars and species, and the construction of genetic maps, had the main aims for the researchers in plant breeding programs and to understand the plant evolution lingues, and as a conclusion, this study demonstrates that AFLP and SRAP markers both are powerful tools in plants tree genetics.

Genetic Diversity Assessment of Calendula officinalis Wilding Plants Locally Grown in Bulgaria, Using SRAP Markers

The pot marigold (Calendula officinalis L.) is a well-known garden plant cultivated also for medicinal purposes. In Bulgaria, C. officinalis is an annual plant, often self-seeding and generating garden escapees. The present study describes assessment of the genetic diversity of wilding C. officinalis plants originating from the region of the Rose Valley, Bulgaria, using SRAP (sequence-related amplified polymorphism) markers. Analysis of the data using the Structure software did not support the existence of a clear genetic structure within this set of analyzed wilding plants. Further comparative SRAP analysis of small sets of C. officinalis plants originating from seeds of wilding plants and seeds derived from various European seed suppliers was carried out. The results suggest that the genetic diversity of the tested wilding plants largely resembles the range of genetic diversity of the plant sets derived from commercially available C. officinalis seeds. The possibility of using wilding C. officinalis plants for selection of breeding lines and varieties, well adapted to the local conditions and suitable for organic growing and production of bioactive extracts and compounds is discussed.

Analysis of genetic diversity among Chinese Cyclocybe chaxingu strains using ISSR and SRAP markers.

BackgroundCyclocybe chaxingu is an edible and medicinal fungal species commonly cultivated in China. The major problems currently facing by growers of C. chaxingu is the random labeling of strains and strains aging and degeneration. Therefore, an evaluation of genetic diversity is essential for the conservation and reproducing programs of this species.MethodsIn the present study, 24 widely cultivated strains were collected from the main producing areas of China, and the genetic diversity analysis was performed. DNA polymorphism among these Chinese C. chaxingu strains was analyzed using inter-simple sequence repeat (ISSR) and sequence-related amplified polymorphism (SRAP) markers.ResultsEight ISSR primers amplified a total of 75 DNA fragments of which 61 (81.33%) were polymorphic. Fifteen SRAP primer combinations amplified 166 fragments of which 132 (79.52%) were polymorphic. Cluster analysis showed that the C. chaxnigu strains fall into five groups with a genetic distance values ranging from 0.06 to 0.60 by ISSR analysis, while the SRAP analysis divided the test strains into four groups within the range of genetic distance from 0.03 to 0.57. The results of the present study reveal a high level of genetic diversity among the widely cultivated C. chaxingu strains.

Morphological and Molecular Characterizations of Musa (ABB) 'Mali-Ong' in Thailand.

Simple SummaryThe banana cultivar Musa (ABB) ‘Mali-Ong’ is widely used in the processed food industry in Thailand, where it determines the quality of the products. However, the sub-cultivars of ‘Nam Wa Mali-Ong’ are almost indistinguishable, with few morphological differences and minimal genetic variation. This study used 77 morphological characteristics and two types of molecular markers to distinguish Nam Wa Mali-Ong from other cultivars. The study also assessed the genetic variation of nine ‘Nam Wa Mali-Ong’ clones and compared them with 10 other samples of bananas with different genomes or chromosome sets. The molecular markers grouped the ‘Nam Wa Mali-Ong’ samples. Four clones (A, B, D, and I) were superior and had higher bunch weights. This study will be useful for germplasm evaluation and future ‘Nam Wa Mali-Ong’ improvements.Musa (ABB) ‘Mali-Ong’ is an economically important banana cultivar in Thailand. We morphologically and molecularly characterized ‘Nam Wa Mali-Ong’. Leaf blade width was the only statistically different morphological character among the clones. To determine genetic variation, nine ‘Nam Wa Mali-Ong’ clones were compared with 10 samples of Musa ABB, AA, and BB cultivars by fingerprinting using seven pairs of sequence-related amplified polymorphism (SRAP) and eight inter simple sequence repeat (ISSR) markers. The SRAP and ISSR primers generated 65 and 76 amplicons, respectively, of which 57 (87.7%) and 62 (81.6%) amplicons, respectively, were polymorphic; the polymorphic information content was 0.28–0.49. The SRAP data revealed two distinct groups: Group I, comprising two subgroups (one including all ABB samples and the other containing the BB genome accessions), and Group II, comprising the AA genome accessions. The ISSR data revealed two groups: Group I, which incorporated the AA (Hom Champa) genome, and Group II, consisting of two subgroups: Subgroup A, comprising only the AA (Hom Chan) accessions, and subgroup B, comprising all the ABB accessions and wild banana M. balbisiana (BB genome). The ‘Nam Wa Mali-Ong’ samples clustered together, regardless of the markers used. SRAP and ISSR markers will be useful for germplasm evaluation and future Musa (ABB) improvements.

Morphometric analysis and genetic diversity in Hypericum L. using sequence related amplified polymorphism

There are about 484 species of Hypericum in the Guttiferae family, which includes Hypericoideae. In Iran, species of this genus are mainly found in the north, northwest, and center of the country, and they are key contributors to the floral elements of the Hyrcanian mountains, Irano-Turanian, and Mediterranean regions (such as the Zagros). Medicinal, commercial, and horticultural values are associated with these plants.The genetic diversity was assessed through Sequence-related amplified polymorphism. To uncover genetic diversity and species characteristics in Hypericum species, were studied through a combination of morphological and molecular data. Eighty-five individuals related to 7 Hypericum were collected in 6 provinces. A total of 76 (Number of total loci) (NTL) DNA bands were produced through polymerase chain reaction amplifications (PCR) amplification of seven Hypericum species. These bands were produced with the combinations of 5 selective primers. The total number of amplified fragments ranged from 10 to 20. According to the SRAP (Sequence-related amplified polymorphism) markers analysis, H. perforatum and H. asperulum had the lowest similarity. This study also detected a significant signature of isolation by distance (Mantel test results). Present results showed that sequence-related amplified polymorphism have the potential to identify and decipher genetic affinity in Hypericum species. Current results have implications in biodiversity and conservation programs. Besides this, present results could pave the way for selecting suitable ecotypes for forage and pasture purposes in Iran. Genus Hypericum (Guttiferae, Hypericoideae) is perennial, belonging to the Hypericaceae family, having 484 species in forms of trees, shrubs, and herbs, distributed in 36 taxonomic sections. The genetic diversity was assessed through Sequence-related amplified polymorphism. To uncover genetic diversity and species characteristics in Hypericum species, were studied through a combination of morphological and molecular data. Eighty-five individuals related to 7 Hypericum were collected in 6 provinces. A total of 76 (Number of total loci) (NTL) DNA bands were produced through polymerase chain reaction amplifications (PCR) amplification of seven Hypericum species. These bands were produced with the combinations of 5 selective primers. The total number of amplified fragments ranged from 10 to 20. According to the SRAP (Sequence-related amplified polymorphism) markers analysis, H. perforatum and H. asperulum had the lowest similarity. This study also detected a significant signature of isolation by distance (Mantel test results). Present results showed that sequence-related amplified polymorphism have the potential to identify and decipher genetic affinity in Hypericum species. Current results have implications in biodiversity and conservation programs. Besides this, present results could pave the way for selecting suitable ecotypes for forage and pasture purposes in Iran.

Determination of Genetic Diversity in Banana and Bell Pepper Lines Using Molecular Markers

Successful hybrid cultivar breeding is depend on the high genetic diversity of the plant sources, as well as the homozygous and genetically distant lines requiring hybridization. The aim of this study is to determine the genetic distance between the inbred lines of pepper (Capsicum annuum L.) in order to increase efficacy of the breeding program. In this study, the genetic distances between the inbred lines of 44 bell peppers and 35 banana peppers were investigated using the SRAP (Sequence Related Amplified Polymorphism) markers. Based on pattern scores, dendrograms were produced by the UPGMA (unweighted pair-group method of mathematical averages method). Out of the 71 primer combinations tested, 50 combinations revealed polymorphisms among the banana pepper lines, and a total of 123 polymorphic bands were obtained. In the bell pepper lines, 24 SRAP primer combinations were tested and 15 combinations had 25 polymorphic bands. Based on the UPGMA cluster analysis, the pepper lines divided into groups as bell peppers and banana peppers. While the genetic similarity among the banana pepper lines varied between 0.62 and 0.98, the genetic similarity among the bell pepper lines varied between 0.54 and 1.00. As a result, it can be stated that the SRAP markers can be used successfully for determining the genetic distances of the pepper inbred lines thus will help the breeding programme.

Eco-biochemical responses, phytoremediation potential and molecular genetic analysis of Alhagi maurorum grown in metal-contaminated soils

BackgroundAlhagi maurorum Medik. (camelthorn) is a dominant desert plant indigenous in various habitats, including the Western Desert of Egypt. The plant is especially prevalent in and around economic iron ore deposits. Nutrient and heavy metal levels in A. maurorum tissues and soil samples were assessed to identify associations between heavy metal levels in plants and soil. The objective was to evaluate this species as an indicator of heavy metal pollution. Photosynthetic pigments, protein, proline, alkaloids, flavonoids, 2,2-diphenyl-1-picrylhydrazylscavenging, reduced glutathione, malondialdehyde, antioxidant enzymes, and stress-related gene expression were assessed to determine their functional roles in metal stress adaptation in ultra- and molecular structure. Additionally, the molecular genetic variation in A. maurorum samples was assessed using co-dominant sequence-related amplified polymorphism (SRAP) and inter simple sequence repeats (ISSR).ResultsA substantial difference in enzymatic and non-enzymatic antioxidants of A. maurorum was observed in samples collected from three sites. A. maurorum is suited to the climate in mineralized regions. Morphologically, the stem shows spines, narrow leaves, and a reduced shoot system. Anatomically, modifications included a cuticle coating on leaves and stems, sunken stomata, a compact epidermis, and a thick cortex. Significant anatomical-physiological differences were observed with varying heavy metal soil content, antioxidative enzyme activities increased as a tolerance strategy, and glutathione levels decreased in response to heavy metal toxicity. Heavy metal accumulation also affected the expression of stress-related genes. The highest levels of expression of GST, G6PDH, 6PGD, nitrate reductase 1, and sulfate transporter genes were found in plants collected from site A1. However, auxin-induced protein exhibited its highest expression in plants collected from A2. Six SRAP combinations yielded 25 scoreable markers with a polymorphism rate of 64%, and 5 ISSR markers produced 11 bands with a polymorphism rate of 36.36% for three A. maurorum genotypes. The ME1xEM7 primer combinations provided the most polymorphic information content and resolving power, making it the most useful primer for differentiating A. maurorum genotypes. SRAP markers exhibited a higher diversity index (0.24) than ISSR markers (0.16).ConclusionsA. maurorum displayed adaptive characteristics for heavy metal sequestration from mining site soils and is proposed as a strong candidate for phytoremediation.

Genetic diversity and verbascoside content in natural populations of Pyrostegia venusta (Ker Gawl.) Miers.

Pyrostegia venusta (Ker Gawl.) Miers occurs in threatened biodiversity hotspots of Cerrado and Atlantic forest biomes in Brazil and is used in traditional medicine to treat various respiratory and skin diseases. This study (i) examined the genetic diversity and structure of six natural populations of P. venusta from different Brazilian regions using sequence-related amplified polymorphism (SRAP) markers; and (ii) compared the intra- and inter-populational levels of the bioactive component verbascoside using high-performance liquid chromatography. The population from Nova Mutum, Mato Grosso, presented the highest genetic variability (Nei index H = 0.2759; Shannon index I = 0.4170; 85.14% polymorphic loci), whereas the population from Araxá, Minas Gerais, presented the lowest genetic variability (H = 0.1811; I = 0.2820; 70.27% polymorphic loci). The intra-populational variability (79%) was significantly higher (p = 0.001) than the inter-populational variability (21%). The populations were clustered into two groups but their genetic differentiation was not associated with geographical origin (Mantel test, r = 0.328; p > 0.05). The verbascoside content significantly differed (p > 0.05) among the six populations and between the individuals from each population. The highest verbascoside levels (> 200µg/mg extract) were detected in populations from Araxá and Serrana, while the lowest verbacoside levels were detected in populations from Paranaíta and Sinop. This is the first report on the use of SRAP markers to analyze genetic variability in the family Bignoniaceae. Our findings shall help to better understand the genetic and chemical diversity of P. venusta populations, as well as provide useful information to select the most appropriate individuals to prepare phytomedicines.

Morphometric analysis and genetic diversity in Glaucium (Papaveraceae) using sequence related amplified polymorphism

Glaucium belongs to the Papaveraceae family. Glaucium is a genus of annual, biennial, and perennial herbaceous plants that thrive on salty soils and near the sea.Glaucium is represented by a total of 10 taxa in Iran. Sequence-related amplified polymorphism was used to estimate genetic diversity. A combination of morphological and genomic data was used to identify genetic diversity and species features in Glaucium species. In eight provinces, 65 people connected to five Glaucium were gathered. Through polymerase chain reaction (PCR) amplification of five Glaucium species, a total of 144 (Number of total loci) (NTL) DNA bands were obtained. These bands were created by combining 10 different selective primers. The total number of amplified fragments varied from seven to twenty-six. The expected unbiased heterozygozity (H) ranged from 0.19 (G. grandiflorum subsp. grandiflorum var. grandiflorum) to 0.33 (G. grandiflorum subsp. grandiflorum var. grandiflorum) (G. oxylobum var. oxylobum). The genetic similarities between five species range from 0.63 to 0.88. The findings of clustering revealed two large groupings. The SRAP (Sequence-related amplified polymorphism) markers study revealed that G. grandiflorum and G. oxylobum var. oxylobum had the least similarity. This investigation also discovered a substantial indication of distance isolation (Mantel test results). The current findings indicate that sequence-related amplified polymorphism can discover and understand genetic affinity in Glaucium species. The current findings have consequences for biodiversity and conservation efforts. Aside from that, the current findings may pave the way for identifying acceptable ecotypes for grazing and pasture uses in Iran.

Genetic Diversity and Relationships within Citrus Species Based On Sequence-Related Amplified Polymorphism Markers (Sraps)

Genetic Diversity and Relationships within Citrus Species Based On Sequence-Related Amplified Polymorphism Markers (Sraps)

Genetic diversity and inferred ancestry of Asian lotus (Nelumbo nucifera) germplasms in Thailand and Vietnam

Tropical lotus (Nelumbo) is an important and unique ecological type of lotus germplasm. Understanding the genetic relationship and diversity of the tropical lotus is necessary for its sustainable conservation and utilization. Using 42 EST-SSR (expressed sequence tag-simple sequence repeats) and 30 SRAP (sequence-related amplified polymorphism) markers, we assessed the genetic diversity and inferred the ancestry of representative tropical lotus from Thailand and Vietnam. In total, 164 and 41 polymorphic bands were detected in 69 accessions by 36 EST-SSR and seven SRAP makers, respectively. Higher genetic diversity was revealed in Thai lotus than in Vietnamese lotus. A Neighbor-Joining tree of five main clusters was constructed using combined EST-SSR and SRAP markers. Cluster I included 17 accessions of Thai lotus; cluster II contained three Thai accessions and 11 accessions from southern Vietnam; and cluster III was constituted by 13 accessions of seed lotus. Consistent with the results from the Neighbor-Joining tree, the genetic structure analysis showed that the genetic background of most Thai and Vietnamese lotus was pure, as artificial breeding has been rare in both countries. Furthermore, these analyses indicate that Thai and Vietnamese lotus germplasms belong to two different gene pools or populations. Most lotus accessions are genetically related to geographical distribution patterns in Thailand or Vietnam. Our findings showed that the origin or genetic relationships of some unidentified lotus sources can be evaluated by comparing morphological characteristics and the data of molecular markers. In addition, these findings provide reliable information for the targeted conservation of tropical lotus and parent selection in breeding novel cultivars of lotus.

Phenotypic variability, race profiling and molecular diversity analysis of Indian populations of Fusarium oxysporum f. sp. lentis causing lentil wilt.

Wilt (Fusarium oxysporum f. sp. lentis; Fol) is one of the major diseases of lentil worldwide. Two hundred and thirty-five isolates of the pathogen collected from 8 states of India showed substantial variations in morphological characters such as colony texture and pattern, pigmentation and growth rate. The isolates were grouped as slow (47 isolates), medium (118 isolates) and fast (70 isolates) growing. The macroconidia and microconidia (3.0-77.5 × 1.3-8.8 µm for macroconidia and 1.8-22.5 × 0.8-8.0 µm for microconidia for length × width) were variable in size and considering the morphological features, the populations were grouped into 12 categories. Seventy representative isolates based on their morphological variability and place of origin were selected for further study. A set of 10 differential genotypes was identified for virulence analysis and based on virulence patterns on these 10 genotypes, 70 Fol isolates were grouped into 7 races. Random amplified polymorphic DNA (RAPD), universal rice primers (URPs), inter simple sequence repeats (ISSR) and sequence-related amplified polymorphism (SRAP) were used for genetic diversity analysis. URPs, ISSR and SRAP markers gave 100% polymorphism while RAPD gave 98.9% polymorphism. The isolates were grouped into seven clusters at genetic similarities ranging from 21 to 80% using unweighted paired group method with arithmetic average analysis. The major clusters include the populations from northern and central regions of India in distinct groups. All these three markers proved suitable for diversity analysis, but their combined use was better to resolve the area specific grouping of the isolates. The sequences of rDNA ITS and TEF-1α genes of the representative isolates were analysed. Phylogenetic analysis of ITS region grouped the isolates into two major clades representing various races. In TEF-1α analysis, the isolates were grouped into two major clades with 28 isolates into one clade and 4 remaining isolates in another clade. The molecular groups partially correspond to the lentil growing regions of the isolates and races of the pathogen.

Genetic diversity and relationship analyses of mango (Mangifera indica L.) germplasm resources with ISSR, SRAP, CBDP and CEAP markers

Forty-eight mango accessions were compared for genetic diversity and relationships based on four molecular marker assays, namely, inter-simple sequence repeat (ISSR), sequence-related amplified polymorphism (SRAP), CAAT-box derived polymorphism (CBDP), and cis-element amplified polymorphism (CEAP). Comparative analyses of the marker assays based on Nei's genetic diversity (H), Shannon's information index (I), polymorphism information content (PIC), the marker index (MI), and resolving power (Rp) revealed that (i) the CEAP markers provided the most information according to the PIC, MI and Rp values; (ii) the CBDP and ISSR markers exhibited valuable information according to the H and I values. The correlation values (r) of the Mantel test ranged from 0.334 (ISSR and CEAP) to 0.819 (CEAP and the combination of the four marker assays). The unweighted pair group method with arithmetic mean (UPGMA) cluster results showed that a single-marker assay can accurately reflect the genetic relationships of some mango samples but cannot accurately reflect the genetic relationships of all mango samples and that when multiple markers are used to evaluate the genetic relationships of mango germplasm resources, the results are more accurate than those of a single-marker assay. The combined cluster analysis of the molecular marker types shows higher discrimination according to genetic relationships. Nine mango accessions were suggested to be likely parental or sister lines based on the combination of the ISSR, SRAP, CBDP, and CEAP marker assays.

MOLECULAR EVALUATION AND PHENOL APPLICATION EFFECTS ON BARLEY INFESTATION BY Rhyzopertha dominica (F.)

Food safety has been a challenge for many years owing to pre and post-harvest losses of agricultural products in the word. In this study, twelve barley cultivars were used to evaluate grain yield, quality parameters and insect pest (Rhyzopertha dominica (F.) infestation using Sequence-Related Amplified Polymorphism (SRAP) marker analysis to study the genetic diversity among the 12 cultivars and Phenol as a chemical application such as modern methods of stored grains protection. SRAP markers confirm that there were highly genetic information differences among all cultivars and their response to insect pest (R. dominica (F.). High polymorphism (83%) and high Polymorphic information content (PIC) (0.84%) was found by primer (me6+em6). The dendrogram of SRAP markers had clustered all the cultivars into two major groups based on their genetic information which could use them in cultivar identification breeding for insect pest control. All the hulless barley had high protein and low phenol content and was more susceptible to R. dominica. Application of the phenol concentration (0.4 g ai/kg) in all the test hulless grain showed high mortality percentages after five days of exposure also caused complete reduction of adult progeny of insect Giza 135 and least damage of grains as lowest Lethal Concentration 50% (LC50%) was 0.33 g ai/kg grain. Moreover, there is no change were found in technological grain properties, thus using phenol will e a positive feature integrated pest management (IPM) programs.