Inter-simple Sequence Repeat Markers Research Articles

Thymus algeriensis Boiss. et Reut., a north African endemic plant species: genetic diversity and population structure as assessed by molecular markers, a pioneer step for conservation implications.

Thymus algeriensis Boiss. et Reut. is one of the most widespread North African species of the genus Thymus L. The species is subshrub growing primarily in subtropical biome of Morocco, Algeria, Tunisia and Libya. In Tunisia, the plant species is under high pressure of anthropogenic activities including over-collecting. The assessment of genetic diversity and population structure of T. algeriensis is a pioneer step to retrace its evolutionary history and to perform appropriate conservation strategies of the plant species. Seven wild populations growing, widely, in different bioclimatic zones were selected and analysed using two molecular markers systems. Fifteen Simple Sequence Repeats (SSRs) and fifteen Inter-Simple Sequence Repeats (ISSRs) were used to characterize genetically 140 different genotypes. The results showed a high molecular variation within populations and among the studied genotypes. The intra-populations genetic diversity revealed by SSRs was higher (P = 80.95%, Na = 2.143 and He = 0.364) than that based on ISSRs (P = 78.12%, Na = 1.632, He = 0.265 and I = 0.398). As demonstrated by inbreeding coefficients, a significant level of differentiation and a low level of gene flow were detected among studied populations (FST = 0.161 for SSRs and ΦST = 0.197 for ISSRs). Furthermore, the results of ISSRs marker suggest land strips as barriers in population genetic structure. While SSRs marker reflects a relatively structured bioclimatic patterns of studied populations. The Bayesian analysis showed a specific adaptation of populations to local environments. The used molecular markers (ISSRs and SSRs) seem to be effective in deciphering genetic polymorphism of Tunisian genotypes of T. algeriensis. Therefore, the genetic structure of the studied genotypes could constitute a starting point for further conservation, characterization and breeding programs.

Genetic diversity and morpho-physiological assessment of drought tolerance in rapeseed (Brassica napus L.) cultivars

Abstract Water deficit is one of the most important abiotic stresses constraining crop production in rapeseed. Understanding the mechanisms of adaptation to this stress is essential for the development and production of drought-tolerant genotypes. For this reason, this research study aims to investigate the importance of genetic diversity in identifying genotypes with a high degree of drought tolerance through assessing effectiveness of inter simple sequence repeat (ISSR) markers on 14 genotypes of rapeseed in a factorial design. Morphological and physiological characteristics were studied after the early stages of growth; in order to evaluate the genetic diversity among genotypes, 18 different ISSR markers were used. A total of 106 clear and scalable loci were amplified, of which 60 bands (56.6%) were polymorphic. The highest polymorphism information content belonged to marker number 9 with the amount of 0.365 (85.7%). Gene variation ranged from 0.081 to 0.365 and the rapeseed genotypes were divided into three groups by cluster analysis (unweighted pair group method with arithmetic mean method). The analysis of molecular variance showed that 70% of the total variation was observed within populations and 30% of this variation occurred among populations. In addition, t-test was used for comparing oil content percentage among different genotypes in control and stress levels. Adriana had the highest amount of seed oil with 36.47%, whereas Karaj 2 had the lowest amount with 27.28 and Cooper had the highest decrease in oil content percentage under stress conditions. Overall, the genotypes Likord, Hyola 401 and Sarigol 32 were identified as the most drought-tolerant.

Unraveling the genetic diversity of Ceiba pubiflora (Malvaceae) in isolated limestone outcrops: Conservation strategies.

Seasonally Dry Tropical Forests (SDTFs) located on limestone outcrops are vulnerable to degradation caused by timber logging and limestone extraction for cement production. Some of these forests represent the last remnants of native vegetation cover, functioning as isolated islands. Ceiba pubiflora (Malvaceae) is a tree frequently found on limestone outcrops in the central region of Brazil. This study aimed to evaluate the genetic diversity and identify suitable populations for the establishment of Management Units (MUs) for conservation. Inter-simple sequence repeat markers were employed to assess the genetic diversity in ten populations sampled from the Caatinga, Cerrado, and Atlantic Forest biomes. The species exhibited substantial genetic diversity (HT = 0.345; PLP = 97.89%). Populations SAH, JAN, and MON demonstrated elevated rates of polymorphic loci (> 84.2%) along with notable genetic diversity (He > 0.325). Additionally, these populations were the primary contributors to gene flow. The analysis of molecular variance (AMOVA) indicated that most genetic variation occurs within populations (91.5%) than between them. In the Bayesian analysis, the ten populations were clustered into five groups, revealing the presence of at least three barriers to gene flow in the landscape: 1) the Central Plateau or Paranã River valley; 2) near the Espinhaço mountains or the São Francisco River valley; and 3) around the Mantiqueira mountain range, Chapada dos Veadeiros plateau, and disturbed areas. A positive and statistically significant correlation was observed between genetic (θB) and geographic distances (r = 0.425, p = 0.008). Based on these findings, we propose the establishment of Management Units in Minas Gerais state, encompassing the (1) southern region (MIN population), (2) central region (SAH population), and (3) north region (MON population), as well as in Goiás state, covering the (4) Central Plateau region. These units can significantly contribute to preserving the genetic diversity of these trees and protecting their habitat against ongoing threats.

Molecular, morphological, palynological and biochemical characterization of six accessions of two Mangifera indica L cultivars (Keitt and Ewais) native to Egypt

AbstractAccurate characterization of crops or fruits leads to successful improvements in breeding programs. Mango (Mangifera indica L.) is the second most common fruit in Egypt. It is a heterozygous plant; consequently, the possibility of incorrect characterization is significant. Therefore, this study used six accessions of two M. indica cultivars, ‘Keitt’ and ‘Ewais’, to produce precise molecular characterization and prove their genetic relatedness using inter-simple sequence repeat (ISSR) and start codon targeted (SCoT) markers. Pollen morphology was studied for the first time for these two Egyptian cultivars to determine their correlation. Morphological traits were quantitatively and qualitatively investigated. In addition, phytochemical analyses were performed to correlate the six mango accessions. One of these accessions (accession 3) was commercially identified as ‘Ewais’ cv., but the obtained result revealed that it does not belong to ‘Ewais’ cv. This finding was proved molecularly using ISSR and SCoT markers and morphologically. Similarly, phytochemical analyses revealed a difference between accession 3 and the other accessions under investigation. Polymorphic information content (PIC) and resolving power (RP) data revealed that the SCoT technique was more informative than ISSR for studying these genotypes of mango. Accordingly, the present findings prove the necessity for molecular, morphological, palynological, and biochemical investigations for accurate characterization of mango genotypes and genetic relatedness for improving breeding programs.

Genetic diversity of Guava varieties (Psidium guajava L.) based on morphology and ISSR molecular markers

Abstract. Hien PTT, Cham LTT, Hang VTT. 2024. Genetic diversity of Guava varieties (Psidium guajava L.) based on morphology and ISSR molecular markers. Biodiversitas 25: 1037-1045. Guava (Psidium guajava L.) is a fleshy-fruited representative belonging to the Myrtaceae family that has attracted attention due to its economic benefits. In this study, seventeen guava varieties in the Mekong Delta and Hanoi were collected and genetically validated by both morphology and DNA markers. The results revealed that fruit shape is a powerful indicator of guava discrimination. There are 12 out of 15 Inter Simple Sequence Repeats (ISSR) markers that show informative band patterns and 93 amplicons are generated with the size in length 200-2000 bp. The average polymorphic band and PIC value are 97.85% and 36%, respectively. Based on the dendrogram constructed by the Unweighted Pair Group Method using arithmetic Averages (UPGMA) method, samples were grouped into two main groups and the similarity level ranged from 55-98%. Group 2 contains Oi Tim (Purple guava), a special variety that showed high differences in terms of morphology and DNA profiles. Such information reflects the diversity of guava germplasm in Viet Nam, which is useful for conservation, and breeding strategies. The result revealed that 11 varieties expressed genetic diversity through both phenotypic traits and DNA profiles. Such information illustrates the genetic diversity among Psidium guajava, contributing to germplasm resources and plant breeding programs.

Empirical phenotyping and genome-wide association study reveal the association of panicle architecture with yield in Chenopodium quinoa.

Chenopodium quinoa manifests adaptability to grow under varying agro-climatic scenarios. Assessing quinoa germplasm's phenotypic and genetic variability is a prerequisite for introducing it as a potential candidate in cropping systems. Adaptability is the basic outcome of ecological genomics of crop plants. Adaptive variation predicted with a genome-wide association study provides a valuable basis for marker-assisted breeding. Hence, a panel of 72 quinoa plants was phenotyped for agro morphological attributes and association-mapping for distinct imperative agronomic traits. Inter simple sequence repeat (ISSR) markers were employed to assess genetic relatedness and population structure. Heatmap analysis showed three genotypes were early maturing, and six genotypes were attributed for highest yield. The SD-121-07 exhibited highest yield per plant possessing green, glomerulate shaped, compact density panicle with less leaves. However, SJrecm-03 yielded less exhibiting pink, intermediate shape, intermediate density panicles with less leaves. The phenotyping revealed strong correlation of panicle architecture with yield in quinoa. A genome-wide association study unraveled the associations between ISSR makers and agro-morphological traits. Mixed linear modes analysis yielded nine markers associated with eight traits at p ≤ 0.01. Moreover, ISSR markers significantly associated with panicle shape and leafiness were also associated with yield per plant. These findings contribute to the provision of authenticity for marker-assisted selection that ultimately would support quinoa breeding programs.

Conservation Strategies for Aquilaria sinensis: Insights from DNA Barcoding and ISSR Markers.

The evergreen tree species Aquilaria sinensis holds significant economic importance due to its specific medicinal values and increasingmarket demand. However, the unrestricted illegal exploitation of its wild population poses a threat to its survival. This study aims to contribute to the conservation efforts of A. sinensis by constructing a library database of DNA barcodes, including two chloroplast genes (psbA-trnH and matK) and two nuclear genes (ITS and ITS2). Additionally, the genetic diversity and structure were estimated using inter-simple sequence repeats (ISSR) markers. Four barcodes of 57 collections gained 194 sequences, and 1371 polymorphic bands (98.63%) were observed using DNA ISSR fingerprinting. The Nei's gene diversity (H) of A. sinensis at the species level is 0.2132, while the Shannon information index (I) is 0.3128. The analysis of molecular variance revealed a large significant proportion of total genetic variations and differentiation among populations (Gst = 0.4219), despite a relatively gene flow (Nm = 0.6853) among populations, which were divided into two groups by cluster analysis. There was a close genetic relationship among populations with distances of 0.0845 to 0.5555. This study provides evidence of the efficacy and dependability of establishing a DNA barcode database and using ISSR markers to assess the extent of genetic diversity A. sinensis. Preserving the genetic resources through the conservation of existing populations offers a valuable proposition. The effective utilization of these resources will be further deliberated in subsequent breeding endeavors, with the potential to breed agarwood commercial lines.

Genotyping of Vanilla planifolia Jacks ex Andrews from the Totonacapan region using Inter-Simple Sequence Repeat markers

Vanilla is the only genus of plants in the Orchidaceae family used in the food industry, being the most widely used flavoring worldwide and the second most valuable economically. Cultivation is of great biocultural importance in the Totonacapan region, in the state of Veracruz, as this region is considered its center of origin and distribution. However, little is known regarding the number of genotypes present in this area, as well as the genetic diversity, which is vital for the improvement of this crop. Therefore, the objective of this study was to perform the genotyping of 60 Vanilla planifolia accessions from the Germplasm Bank of The National Institute of Forestry, Agriculture, and Livestock Research (Instituto Nacional de Investigaciones Forestales, Agrícolas y Pecuarias - INIFAP), Experimental Field Ixtacuaco, which were selected and collected in the Totonacapan region. For this purpose, the number of V. planifolia genotypes was determined based on ISSR (Inter-Simple Sequence Repeat) molecular markers. Three DNA extraction protocols from plants and fungi described in the literature were evaluated, and a fourth protocol was adapted with different procedures from the previous protocols, seeking to obtain a high concentration and good quality and integrity. Once quality DNA was obtained, fourteen ISSR primers were evaluated, of which three primers that showed the highest number of amplified bands were selected. Subsequently, the genotyping of the 60 accessions was carried out, resulting in nine genotypes with primer 808 and six genotypes with primers 812 and 856. These results allowed the genotyping of Vanilla planifolia accessions with ISSR markers, which will help evaluate the genetic diversity of this crop in the Totonacapan region in the future.

Genetic Variation among Aeluropus lagopoides Populations Growing in Different Saline Regions

Aeluropus lagopoides is a halophytic grass growing in different sabkhas of Saudi Arabia. In this study, 14 inter-simple sequence repeat (ISSR) and 15 sequence-related amplified polymorphism (SRAP) molecular markers were selected to investigate the genetic diversity within and among five natural populations of A. Lagopiodes. The genetic diversity varied within and among populations. ISSR markers were slightly more efficient than SRAP markers in evaluating genetic diversity. Average polymorphism information content, effective number of alleles, Nei’s genetic diversity, and Shannon’s information index values of ISSR markers were higher than SRAP. Analysis of molecular variance revealed about 40% genetic variation among the population and 60% within the population. Overall, the genetic diversity was lowest in Jouf (40%), while the Qaseem populations were the highest (60%). Jizan populations were highly dissimilar to other regions. A Mantel test indicated a positive correlation between geographic and genetic distance. The cluster analysis showed three groups; the first group comprises Jouf and Salwa populations, the second group comprises Qareenah and Qaseem, and the third group comprises the Jizan population. This observation matched the geographic distribution of the species. These findings can help in the conservation of a diverse population of A. lagopoides in saline regions as well as rehabilitation of these degraded unique habitats.

Genetic diversity among Vanda celebica, Vanda dearei, and their hybrids based on ISSR markers

A cross between two different orchid species is a way to improve the horticultural performance of the hybrids. Inter-simple sequence repeat (ISSR) is a method of molecular markers that could be used to analyze genetic diversity among parents and their hybrid progenies. The purpose of this study was to examine the genetic diversity of the parent species of Vanda dearei and Vanda celebica, the hybrid progenies of (♀ Vanda dearei x ♂ Vanda celebica)-cross and the hybrid progenies of their reciprocal cross. DNA amplification using 5 ISSR primers, namely UBC 810, UBC 815, UBC 834, UBC 835, and UBC 880, produced 60 DNA bands with a size of 200-2800 bp, with the percentage of polymorphism ranging from 81.8%-100% and the percentage of monomorphism as much as 0%-18.2%. Analysis of genetic diversity showed that the genetic diversity among hybrid progenies of (♀ V. dearei x ♂ V. celebica) was 8%. In comparison, those among hybrid progenies of its reciprocal cross (♀ V. celebica x ♂ V. dearei) were 13%. In conclusion, deploying V. celebica as a female parent and V. dearei as a male parent will most likely produce higher diversity among its progenies, compared to the reciprocal cross.
 Keywords: DNA bands; diversity; marker, orchid; reciprocal cross

The population structure of the Fusarium brachygibbosum species complex in the grassland biome of South Africa

Fusarium brachygibbosum Padwick is a phytopathogen with a widespread distribution, infecting various host plants. In South Africa, there is a limited number of studies on the genetic diversity of fusaria, particularly in undisturbed soils. In the current study, the genetic diversity of F. brachygibbosum was investigated using inter-simple sequence repeat (ISSR) and mating type genes. The F. brachygibbosum isolates were collected from four different geographic regions within the grassland biome of South Africa. A total of ten ISSR primers produced 64 distinct and reproducible amplicons, with 100% polymorphism. The ISSR markers revealed high levels of intraspecific variation, which is depicted by various genetic diversity indexes. In the analysis of mating type genes, the MAT1-1-2 gene was found among the four populations, but the MAT1-2-1 gene was only present in the population from the Groenkloof Nature Reserve. The data from this study will contribute substantially to knowledge of grassland fusaria of South Africa and their population structure in the grassland biome.

Conservation Genetics of the Only Honeysuckle Azalea (Rhododendron luteum) Population Present in Greece

Honeysuckle azalea (Rhododendron luteum) has only a single population occurrence in Greece, on Lesvos Island of the north-eastern Aegean Sea. The genetic diversity of this population was studied in a population of n = 37 individuals randomly sampled in a transect spanning between the highest and lowest natural altitudinal distribution limits in Mt. Ordymnos, SW Lesvos. A modified DNA extraction and isolation protocol was used to overcome problems of DNA quality due to secondary metabolite activity. Genetic variation was investigated based on molecular Inter Simple Sequence Repeat (ISSR) markers. Results showed the presence of a sufficient amount of genetic diversity for the maintenance of adaptive potential. Genetic diversity was lower but comparable to that of other Rhododendron species sampled from the centre of their natural distribution, despite the relatively small population size, negative anthropogenic pressure and population isolation due to the island environment. Some structuring of genetic diversity was indicated based on a PCoA analysis and the genetic distance dendrogram, while spatial autocorrelation was highly significant. Results point towards the need to assign a protection status to the whole area of the species’ natural distribution on Lesvos Island. Moreover, it is proposed that an in situ Gene Conservation Unit (GCU) be established in the core of this population in Lesvos as part of the Network of the European Forest Genetic Resources Programme, while the establishment of ex situ conservation is also advised.

Genetic Diversity Analysis of Bitter Gourd (Momordica charantia L.) Germplasm using Molecular Markers

Bitter gourd [Momordica charantia L.] is an important cucurbitaceous crop of medicinal and nutraceutical value. The present study was conducted to compare the efficacy of random amplified polymorphic DNA (RAPD) and inter simple sequence repeats (ISSR) molecular markers for genetic diversity analysis of bitter gourd. Twenty-one genotypes of bitter gourd, collected from different parts of India, were used for diversity analysis using two different types of markers. RAPD and ISSR markers yielded an average of 7.16 and 5.73 amplicon per primer, respectively. In the present study, RAPD and ISSR showed 89 and 63% polymorphic bands, respectively. Combined data analysis of RAPD and ISSR markers provided 73.66% polymorphism with 5.33 polymorphic amplicons per primer. The better discriminatory power of ISSR markers over RAPD markers may be due to comparatively higher values of average polymorphic information content (0.175) as well as the diverse nature of the genotypes. The information generated in this study would be helpful in designing breeding strategies for bitter gourd improvement.

POLYMORPHISM Inter Simple Sequence Repeats (ISSR) PRIMERS FOR GENETIC DIVERSITY FOR POOTI PLANTS (Hopea gregaria)

Pooti (Hopea gregaria) is a medium size tree that can grow up to 35 m from the Dipterocarpaceae family. Since 1998, the International Union for Conservation of Nature (IUCN) has reported the pooti tree as endangered, and the species has a narrow distribution, found only in Southeast Sulawesi. One of the efforts to overcome the rarity of the Pooti plant is through a tree breeding program. However, this tree breeding program will only be successful with the support of genetic diversity information. The study of genetic diversity can be done using a molecular-based approach using Inter Simple Sequence Repeats (ISSR) markers. Samples of Pooti leaves used in the study were taken from Botanical Forest Park Nipa-Nipa (Tahura) and Protected Forest Nanga-Nanga, Kendari City, Southeast Sulawesi Province. A total of 50 samples were extracted using the CTAB (Cetyl Trimethyl Ammonium Bromide) method. Ten types of primers that have been carried out obtained seven primers that can produce bright polymorphic bands and can be continued for study genetic diversity, namely UBC 810, UBC 813, UBC 814, UBC 822, UBC 823, UBC 827, and UBC 830.

Physical, Organoleptic, and Phytochemical Valuation of the Promising Greek Kiwifruit Genotype Arta Kiwifruit

To evaluate a new kiwifruit genotype named “Arta Kiwifruit”, a comparative analysis of fruit physical characteristics and biochemical composition in comparison to the widely cultivated Hayward cultivar took place, both at harvest and after a storage period for two successive years. The findings revealed that “Arta Kiwifruit” holds a significant advantage, as it features a larger fruit size, is approximately 34% heavier than “Hayward”, and boasts a distinct shape characterized by a bigger length. Both genotypes exhibited similar dry matter content. No significant difference was observed in protein content, or total phenolic compounds between the two genotypes, while “Hayward” exhibited a significantly higher concentration of sugars and higher sweetness index. “Hayward” demonstrated also increased levels of chlorophyll at harvest, along with higher concentrations of carotenoids. “Hayward” during the first year presented higher antioxidant activity both at harvest and after storage. On the other hand, “Arta Kiwifruit” displayed elevated amino acid concentrations compared to “Hayward”, both at harvest and after storage. Phenotypic traits, as well as the genetic analysis using inter-simple sequence repeats (ISSR) markers, further confirmed the distinct genetic profile of “Arta Kiwifruit”, highlighting its potential significance for kiwifruit breeding programs and commercial use.

Genetic variability of wild palms Euterpe precatoria, Euterpe oleracea and Mauritia flexuosa with molecular markers ISSR

Wild palms constitute a group of plants of particular economic importance in the tropics, being a valuable source for obtaining construction materials, fabrics, fuel, food, ornamental and medicinal plants. Despite their economic, social, and environmental importance, the genetic variability existing in wild palms, mainly of the genus Mauritia and Euterpe, is unknown, which limits their potential use and the identification of promising genotypes to be included in recombination programs. The genetic variability of three wild palm species (Euterpe oleracea, Euterpe precatoria, and Mauritia flexuosa) was evaluated using Inter-Simple Sequence Repeat (ISSR) markers. A total of 51 genotypes were analyzed with seven ISSR primers, which generated a total of 153 bands, with a percentage of polymorphic loci of 90.24%. The cluster analysis revealed the formation of six groups at a similarity level of 37%, grouping the palm genotypes by species and geographical origin. Molecular variance analysis revealed that within group variations contributed more to genetic diversity (89%) than between group variations (11%). The average value of Fst was 0.20, demonstrating moderate genetic differentiation. The high genetic variability found in this study indicates the valuable genetic potential present in palm germplasm, which could be used for future improvement programs of the species.

Evaluation of Biological Control of Sorghum Strains Using Bacillus Thuringiensis and Pseudomonas Aeruginosa Under Drought Stress

Background: Sorghum is an economically significant staple food crop for more than half a billion people in developing nations, especially in arid and semi-arid locations where drought stress is a significant limiting factor. Despite usually being regarded as tolerant, sorghum suffers severely from drought stress, which lowers its productivity and nutritional quality throughout its principal cultivation areas. Objective: Improvements in DNA fingerprinting by ISSRs, SSRs, and RAPD markers have also been employed in sorghum genetic modification (GMOs) to enhance the economic characteristics of this crop. Materials and methods: To provide a natural defence against pests, the most tolerant plants among the seven varieties of sorghum bicolour were selected and planted in the second season of 2020–2021 under treatment with two microorganisms, B. thuringiensis and P. aeruginosa. This study considered seven varieties of sorghum bicolour planted under 50% water deficiency in 2019–2020. Genetic variability analysis of sorghum genotypes was performed using seven Inter-Simple Sequence Repeat (ISSR) primers, six Simple Sequence Repeat (SSR) primers, and five Random Amplified Polymorphic DNA (RAPD) primers. Seven Sorghum bicolour accessions were collected from various regions of Egypt and their phylogenetic relationships were evaluated. Additionally, DNA fingerprinting and analyses of the genetic diversity and evolutionary linkages in the sorghum germplasm employed the (ISSR) molecular marker technique. Results and conclusion: The Fisher Least Significant difference test (LSD) at P < 0.05, based on RAPD, ISSR, and SSR markers demonstrated a significant connection.  The findings demonstrated that 51 bands with a size range of 100–1500 bp and polymorphism percentage of 72.5% were created using five RAPD primers. Seven ISSR primers generated 45 bands With a 57.8(%) polymorphism percentage, ranging in size from 100 to 3000 bp. six SSR primers generated 28 bands with (67.86%) polymorphism percentage of 67.86 %, ranging in size from 100 to 1500 bp. Morphological characteristics and ISSR, SSR, and RAPD analyses were used to group the UPGMA Dendrogram into groups. Jaccard's coefficient was used to analyse the genetic similarity matrix. The maximum similarity was observed for ISSR between Hybrid Sh1 and Hybrid Sh306 (0.984%), SSR between Hybrid Sh306 and Sudan grass (0.964%), and RAPD between Giza 15 and Indian Millet (0.706%). The classification of sorghum germplasm, breeding initiatives, and conservation efforts rely heavily on the determination of the genetic diversity among sorghum species. Identification of genetic variants, morphological features, and genetic analysis of ISSR, SSR, and RAPD are useful techniques. These findings demonstrate a large ratio of variation in sorghum. This work could serve as a guide for future research on sorghum and aid in the understanding of species and breeding initiatives.

Detected Genetic Markers for Three Varieties of Rice (Oryza sativa L.) under Nano- Particles

Despite the enormous advancements in breeding, fresh and accessible methods of plant development are preferred. Many research have been done to determine the impact of zinc oxide nanoparticles (ZnO NPs) and magnesium nanoparticles (MgONPs) on plant growth. ZnO NPs and MgONPs have been used extensively in agriculture. Few research have looked into the potential impact of ZnO NPs on grain Zn content or cereal yield formation to date. The goal of the study was to demonstrate how varied concentrations of nanoparticles affected root germination in three different rice types (Giza 177, Sakha super 300, and Sakha 108) and the fungus Fusariummoniliforme, which causes foot rot in rice. Here, we performed a pot experiment to assess the effects that ZnO nanoparticles and MgONPs had on the shoot length, root length, and germination of rice. The experiment involved three dosages of Zn and Mg nanoparticle solution at varying concentrations of 10, 20, and 100 ppm. The outcomes showed that, when compared to the control treatment, ZnO NPs and MgONPs increased the length of the shoot, the length of the roots, and the rate of germination. Using random amplified polymorphic DNA (RAPD) and inter simple sequence repeats (ISSR) markers, the genetic makeup of plants treated with 10, 20, and 100 ppm AgNPs was also examined. In vitro rooted shoots were acclimated in a greenhouse before being tested for biochemical stability and phenotypic stability. In vitro, callogenesis and caulogenesis were both reduced by AgNPs at the greatest dose (100 ppm). In order to maximise genome coverage, three rice cultivars from distinct origins were chosen, and they were tested using three RAPD markers and seven selected ISSR markers. AgNPs-treated plants using ISSR markers produced 87 amplified bands in total, 45 polymorphic allelic variants, and 36 monomorphic allelic variations. The Pearson correlation coefficient was used in the Mantel test to determine whether there was a significant association between the Jaccard's dissimilarity matrices based on ISSR and RAPD markers (r = 0.69; P 0.05). Morphological characteristics, ISSR, and RAPD analysis were used to partition the UPGMA Dendrogram into two groupings. Jaccard's coefficient was used to analyse the genetic similarity matrix. Sakha Super 300 and Giza 177 had the most genetic similarity (95%) while Sakha 108 and Giza 177 had the lowest genetic similarity (90%) according to ISSR and RAPD study. The classification of rice germplasm, breeding initiatives, and conservation efforts all heavily rely on the determination of genetic diversity within the rice species. To find genetic variations, morphological characteristics, ISSR, and RAPD analysis are useful techniques. The findings demonstrated the large ratio of variation in rice. This work successfully established the possibility of using MgONPs and ZnO NPs as high-performing fertilisers to improve rice output and quality.This study could serve as a guide for future research on rice and could support efforts to understand the species and improve breeding stock.

Molecular Variation in Some Taxa of Genus Astragalus L. (Fabaceae) in the Iraqi Kurdistan Region

Astragalus L. is one of the main genera of blossoming plants, and its diversity of species and forms is well known. The Astragalus L. taxa make a significant contribution to the mountainous and steppe ecosystems of the Kurdistan region of Iraq. Although the species within this genus have been the subject of several molecular studies, the evolutionary relationships among these species remain unidentified. Despite extensive research, little is known about the phylogenetic relationships among the various subgenera of Astragalus. This research is intended to investigate the molecular variation of 33 species of the genus Astragalus L. found in the Kurdistan area of Iraq. For molecular validations, three separate techniques (nuclear ribosomal DNA (rDNA), inter simple sequence repeats (ISSR), and conserved DNA-derived polymorphism (CDDP)) were used. With respect to Astragalus L. indentations, universal ITS1 and ITS4 gene sequencing was used, and the discovered sequences were subjected to BLAST searches in the NCBI database. A phylogenetic tree was generated with two main clades. Regarding detecting genetic diversity between the taxa, 24 molecular markers (14 ISSR and 10 CDDP) were used. High values of polymorphic materials and gene diversity were detected. ISSR markers had an average of 22.71 polymorphic bands per primer, while CDDP markers had the highest mean values for polymorphic information content (0.37), Shannon’s information index (0.27), expected heterozygosity (0.19), and unbiased expected heterozygosity (0.23). Cluster and principal coordinate analyses divided the Astragalus L. taxa into four main groups using the two molecular data sets. According to the results of the molecular variance analysis, the highest variation was detected within sections, with values of 92.01 and 89.48% for ISSR and CDDP markers, respectively. These outcomes suggest the effectiveness of molecular markers and the ITS region in determining and identifying genetic correlations between Astragalus species.

The potential of gamma irradiation on antioxidant capacity and genomic alterations in Calendula officinalis

There are lines of evidence that ionizing radiations such as gamma rays can cause different biological effects on plants. Marigold (Calendula officinalis L.) is a member of the family Asteraceae. It possesses profound amounts of active ingredients. The aim of this study was to evaluate the changes imposed upon different dose levels of gamma radiation on some features of Calendula officinalis such as antioxidant activity, total phenolic compounds and flavonoid contents, antibacterial activity and genomic alterations. Calendula officinalis seeds were exposed to different doses of Gamma radiation (0, 10, 15, 20 and 25 GY). Total phenolics, flavonoids, antioxidant activity (measured by DPPH assay) using methanolic extracts of plants and antibacterial activity measured by the disc diffusion assay showed significant differences to the control samples. The samples treated with 10 GY gamma rays showed the highest total phenol and flavonoid contents. Antioxidant activity significantly differed between Gamma rays dose levels and it was the highest at 25 GY. Four bacterial strains including E. coli, Bacillus subtilis and Pseudomonas aeroginosa were used for the antibacterial assay. Extracts from plants treated with 25 GY gamma rays showed the highest antibacterial activity against the 4 bacterial strains. Random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) markers were used to study the genetic variation. The polymorphism information content (PIC) for RAPD primers ranged from 3% to 13% and ranged from 6 to 13% for ISSR primers. Results indicated that ISSR markers were more efficient than RAPD markers, as they detected 25.57% polymorphic DNA bands compared to 21.31% polymorphism for RAPD markers.