AFLP markers for the assessment of genetic diversity in european and North American potato varieties cultivated in Iran

Information about the genetic diversity of potato germplasm in Iran is important for variety identification andto enhance the classification of germplasm collections and exploit them in breeding programs and for the development andintroduction of new varieties. AFLP fingerprinting was applied to a group of cultivated potato varieties to find if there is anygeographical differentiation in potato diversity from Europe and North America. The high level of polymorphism within potatovarieties and the high number of variety-specific bands suggest that AFLPs are powerful markers for diversity analysis inpotato varieties. No region-specific AFLP markers were found (present in varieties from the same origin and absent inothers). The UPGMA dendrogram revealed four distinct clusters corresponding almost to the geographical origin of thevarieties. However, the bootstrap support for branches was rather weak. No clusters clearly distinguished varieties fromEurope and North America. Varieties from the same geographical origins however tended to group together within eachcluster. The mean similarity and the UPGMA dendrogram both suggest that North American varieties have nearly identicalgenetic diversity to European varieties. The results of AMOVA revealed large within-region variations which accounted for94.5% of the total molecular variance. The between-region variation, although accounting for only 5.5% of the total variation,was statistically significant. AFLP technology was successfully used to evaluate diversity between different geographicalgroups of potatoes and is recommended for potato genetic studies

Protein electrophoretic profiles and physiochemical indicators of salinity tolerance in sorghum (Sorghum bicolor L.)

The present study was conducted to quantify the response of chlorophyll, protein concentration and electrophoretic patterns of leaf and root soluble proteins to increasing salinity in the rooting medium of three varieties (Payam, Kimia and Jambo) of sorghum (Sorghum bicolor L.) and to evaluate the usefulness of these parameters in identifying salt-resistant varieties. This work was carried out with five concentrations of salinity (0, 50, 100, 150 and 200 mM NaCl) and two sampling time points (sampling after 15 and 30 days salt treatment) using a split split plot design with randomized complete block layout. With increasing salinity, the K+ concentration was found to decrease and the amount of Na+ and Na+/K+ increase significantly (P≤0.05) in roots and shoots. Chlorophyll a, b, total chlorophyll concentration and leaf and root soluble protein contents decreased with increasing salinity. The electrophoretic pattern of soluble proteins of cv. Jambo showed that after 15 days NaCl (200 mM) and 30 days NaCl (100 and 150 mM) treatment, a new polypeptide of molecular weight 50 kDa was expressed. The expression of this polypeptide might have been due to the plant adapting to NaCl via expression of a stress-resistant gene. This polypeptide was synthesized under salt stress and is suggested as a marker protein for salt adaptation.Keywords: Chlorophyll, electrophoretic patterns, soluble proteins, salinity, sorghu

Identification of genes involved in steroid alkaloid biosynthesis in Fritillaria imperialis via de novo transcriptomics

Crown imperial (CI) has been used in traditional medicine. Today it is known that such beneficial effects are due to its richness in steroidal alkaloids (SA). Using de novo transcriptomics, orthologues/paralogues finder, phylogenetic analysis and tissue- and developmental stage-specific expression analysis, we identified ten genes and several TFs involved in the biosynthesis of SA in CI. The comparative analysis of ten genes expression profiles revealed the possibility of their co-regulation, which may imply the possibility of their organization in metabolic gene clusters. Having in mind convergent evolution of steroidal biosynthetic pathways in flowering plants and records of convergent evolution of specific proteins, observed expression patterns open a reasonable interest to investigate the possibility of the existence of genes cluster organization in SA pathway in the family Liliaceae or at least in some species of genus Fritillaria. Obtained results support transcriptomics as useful approach in elucidating genes underlying complex biochemical pathways

Evaluation of Salinity Tolerance of Alfalfa Genotypes during Germination Stage Using Multivariate Analysis

ABSTRACT Alfalfa is one of the most important forage plants with a special position among crops. Germination is one of the most sensitive stages to environmental stresses in crops. Therefore, to evaluate and select the most tolerant genotypes to salinity stress during germination stage, 20 genotypes of alfalfa were compared to each other in a completely randomized design. Four salinity levels including 0 (control), 75, 150, and 225 mM NaCl were applied. Analysis of variance showed significant difference for studied characteristics and indices among salinity stress levels and different genotypes. Principal component analysis using all measured characteristics under salinity stress showed that Nikshahri, Bami, Mesa-Sira, Gomi, Sahandava, Hamedani, Kodi, and Siriver were tolerant genotypes to salinity and Defi, Melissa, Kaiseri, Gargologh, and Diablo verdewere sensitive genotypes to salinity. Cluster analysis using the studied characteristics led to the classification of genotypes into two clusters. The first cluster (salinity tolerant) included Nikshahri, Hamedani, Yazdi, Baghdadi, Bami, Gomi, Kodi, Rahnani, Mesa-Sira, kf15, Sahandava, and Siriver genotypes and the second cluster (sensitive to salinity) involved Defi, Melissa, Kaiseri, Gargologh, Ramandi, Dastgerd, Harpinger, and Diablo verde genotypes

Molecular identification of Nosema species in provinces of Fars, Chaharmahal and Bakhtiari and Isfahan (Southwestern Iran)

Objective: To molecularly identify Nosema species in provinces of Isfahan, Fars, Chaharmahal and Bakhtiari. Methods: One hundred and eighty adult honey bees suspected with nosemosis from provinces of Fars (different counties), Isfahan, and Chaharmahal and Bakhtiari were tested. In order to determine the species of Nosema, previously developed PCR and primers based on 16S rRNA gene were used. PCR products were purified and sent to the Korean company of Macrogen for sequencing. Results: Only Nosema ceranae was determined in all samples based on their molecular profile. Sequences of the 16S rRNA gene were sent to GenBank/NCBI (samples accession numbers KP318660–KP318663). Conclusions: This species currently exists in European honeybee apiaries of Apis mellifera in the studied provinces

Molecular and Parasitological Study of Cryptosporidium Isolates From Cattle in Ilam, West of Iran

Background: Toxoplasmosis is a worldwide spread disease. The present study examined the prevalence of Toxoplasma gondii infection among animals of ediblemeat (cattle and sheep) in Chaharmahal va Bakhtiari Province (Southwest of Iran) in 2012. Furthermore, we attempted for the first time to identify this parasite from the meat products in the province. Methods: The tongue, brain, femur muscle and liver of 50 sheep and 70 cattle as well as 50 samples of meat products were selected and collected to perform molecular survey using Nested-PCR method. Results: Of the studied sheep, 38% were infected. The infection rate in the age groups under 1 year, 1-2 years, and more than 2 years was 25%, 35.29% and 52.94%, respectively. The infection rate in femur muscle, brain, liver and tongue was 28%, 32%, 30% and 16%, respectively. Of the studied cattle, 8.57% were infected. The infection rate in the age groups 1-2 years, 2-4 years, and more than 4 years was 3.7%, 9.09% and 14.28%, respectively. Sheep was infected 6 times more than cattle (OR = 6.53 CI = 2.374-18.005).The infection rate among samples of meat products was 12% (6 samples out of 50 samples). Conclusion: Due to the high rate of this parasitic infection among the slaughtered animals as well as meat products in this region, the use of infected material can be one of the main risk factors of transmission of the parasite to humans

Identification of new S-RNase self-incompatibility alleles and characterization of natural mutations in Iranian almond cultivars

The two main objectives of this research were to identify new S-RNase alleles in Iranian almond cultivars and to characterize naturally occurring mutations in these alleles that may cause self-compatibility. We investigated S genotypes of 22 Iranian almond cultivars using stylar RNase electrophoresis, PCR and DNA sequencing. We report six previously unidentified P. dulcis S-RNase alleles (S (45) , S (46) , S (47) , S (48) , S (49) and S (50) ). Four of 12 tested S-RNases were found to be non-functional in vitro: S (49) , S (50) , S (24) /S (na) and S (25) /S (47) . Detected point mutations in the C3 coding region of S (49) - and S (50) -RNase, leading to the replacement of a highly conserved cysteine and histidine residues, are with the highest probability the reason of these S-RNases inactivity. Results also suggested that ten Iranian almond cultivars display unique S genotype. All presented data confirm Iranian cultivars as valuable almond sources which are of interest to almond breeding and conservation programs

Effect of symbiosis with arbuscular mycorrhizal fungi on salt stress tolerance in GF677 (peach x almond) rootstock

Genus Prunus is of great importance for cultivation, mainly because its main species provide fruits and seeds, valuable ornamental qualifies and timber. However, stone fruit trees and almonds, major cultivars of genus Prunus, are sensitive to salt stress. Such sensitivity causes losses in stone fruit and almond production (ca. 50% of regular yield at high salinity grounds). Having in mind that symbiosis with arbuscular mycorrhiza fungi (AMF) may improve plant tolerance to salt stress and that symbiosis effect should be tested in case-by-case approach, we tested salt stress response of AMF inoculated GF677 rootstock (Prunus dulcis x Prunus persica hybrid) in this study. Adding to that, we tested GF677 symbiosis with two AMF species R. intraradices and F. mosseae. Results showed that under salinity stress AMF inoculated GF677 plants displayed improved physiological parameters (chlorophyll, soluble sugars and proline content) and increased antioxidant enzymes activity in comparison to non-inoculated control plants. Comparison of two AMF species beneficial effects on tested parameters revealed that for total chlorophyll content inoculation with F. mosseae has prevailed, while for total soluble sugars and proline content R. intraradices has prevailed. Finally, GF677 in symbiosis with F. mosseae was selected for molecular studies of salinity response. Since many of plants' genes involved in simultaneous response to salt stress and AMF colonization remained unidentified so far, we performed bioinformatics analysis of freely online available data to find differentially expressed genes common to these two responses. Upon GO classification and networking analysis of genes identified as common to both responses, we selected two most prominent ones (UDPGT73C6 and CYP707A3) and tested their expression profile in leaves and roots of F. mosseae inoculated GF677 rootstocks under salt stress. Even though specific roles of UDPGT73C6 and CYP707A3 are un-characterized in Prunus tree species, results suggested their involvement in response to salt stress and AMF inoculation of GF677 plants, which is in concordance with a scarce knowledge on their roles in other plant species. Based on this study finding we may conclude that symbiosis of GF677 rootstocks with AMF increases plants tolerance to salinity stress, which should be considered in other Prunus tree species as well

Application of amplified fragment length polymorphism (AFLPs) analysis to plant breeding and genetics: Procedures, applications and prospects

8 páginas.In the past century, various methods have been developed for the identification and characterization of prokaryotic and eukaryotic organisms at DNA level. One of these promising methods is the amplified fragment length polymorphism (AFLP) analysis. This method combines universal applicability with high powers of discrimination and reproducibility. Total genomic DNA is digested using two restriction enzymes. Double-stranded nucleotide adapters are ligated to the DNA fragments to serve as primer binding sites for PCR amplification. Primers complementary to the adapter and restriction site sequence, with additional nucleotides at the 3´-end, are used as selective agents to amplify a subset of ligated fragments. Polymorphisms are identified by the presence or absence of DNA fragments following analysis on polyacrylamide gels. An increasing number of reports describe the use of AFLP analysis for molecular characterization, phylogenetic studies, disease diagnostics, high-resolution genetic maps and positional cloning of genes of interest. This review describes the principles, advantages and disadvantages of AFLP analysis, summarizing its applications in different plant fields and the novel applications in the molecular analysis of plant DNA.Peer reviewe

Beneficial effect of mycorrhiza on nutritional uptake and oxidative balance in pistachio (Pistacia spp.) rootstocks submitted to drought and salinity stress

40 Pags.- 9 Tabls.- 8 Figs. The definitive version is available at: https://www.sciencedirect.com/science/journal/03044238Environmentally-friendly management techniques such as arbuscular mycorrhizal fungi (AMF) strategies are urgently required to improve plant nutrition and overcome abiotic stress. In this study, we evaluated the influence of symbiotic AMF on four pistachio rootstocks subjected to drought (100 % as the control, and 80 %, 60 %, and 40 % of field capacity as the treatments) and salinity (0.91 as the control, and 7.57, 16.12, and 24.63 dS/m using NaCl as the treatments) stress for 60 days in two separate experiments. Under both drought and salinity stress conditions, the root and shoot dry weights of the pistachio rootstocks markedly increased in the AMF-inoculated plants (+M). The accumulation of minerals in the leaves and roots of (+M) plants was significantly higher than in the non-inoculated plants (-M). AMF improved the nutritional status (N, P, Mg2+, and Ca2+) of the genotypes under both stress conditions. Mycorrhizal association significantly decreased Na+ and Cl− concentrations in the leaves and roots of the pistachio rootstocks under salinity stress. In (+M) plants under drought stress, hydrogen peroxide (H2O2) and methylglyoxal (MG) concentrations in leaf and root tissues and malondialdehyde (MDA) in the roots, and under salinity stress, the H2O2 and MG concentrations in the leaves and MDA in both tissues were lower than those in the (-M) plants. The Sarakhs rootstock under salinity stress accumulated lower minerals (N, P, K+, and Ca2+) and more antioxidants than the other rootstocks, which could be due to its higher sensitivity to this stress. Both stresses reduced the chlorophyll concentration and increased the oxidant content and electrolyte leakage from the roots and leaves of the rootstocks. Sarakhs rootstock under drought stress and Badami-Riz–e-Zarand and UCB1 rootstocks under salinity stress behaved better in terms of mineral absorption. Thus, they were more tolerant of drought and salinity stress, respectively. Based on these findings, the association of mycorrhizal fungi and pistachio rootstocks is a sustainable strategy for increasing biomass, mineral, and Chl contents, and decreasing oxidant content, thereby enhancing the stress tolerance of plants.This work was funded by Iran National Science Foundation (INSF) and Shahrekord University, of Iran. Partial support was obtained from the Spanish State Research Agency (AEI) co-financed with the European Regional Development Fund (FEDER) (project AGL2017-83358-R to Y.G.; AEI/FEDER, UE), and the Aragón Government (Group A09-20R).Peer reviewe