Inter-simple Sequence Repeat Analysis Research Articles

Genetic diversity among genotypes of Parkia platycephala (Benth.), a typical tree of northeastern Brazil

Parkia platycephala (Fabaceae) is a useful tree. The leaves are rich in protein, fiber and minerals and are good quality, low-cost fodder for animals, making it a great option for animal feed during the dry season in Northeast Brazil. In addition, wood is used in small buildings and as fuel. With the aim of exploring genetic variation within the species, we have assessed the molecular and morpho-agronomic characteristics of 10 accessions (F1 - F10) of the species originating from the same location and maintained in the Forage Collection at Embrapa Meio Norte. Clustering analyses based on the amplicons generated by 12 inter simple sequence repeat (ISSR) primers and on 36 qualitative/quantitative markers separated the accessions into two groups, the constituents of which were dependent on the characteristics considered. The most divergent genotypes according to ISSR analysis were F3 and F8. Morpho-agronomic analysis identified F2 and F7 as the most divergent, while the traits that contributed most (36.5%) to total diversity were, in order of importance, mature inflorescence length, stem length, immature inflorescence width, seed weight and pedicel length. The results revealed that there is sufficient genetic variability among the studied accessions. These accessions with greater diversity are candidates for actions that promote the conservation, domestication and genetic improvement of the species.

Изучение генетического разнообразия образцов коллекции розы эфиромасличной института сельского хозяйства Крыма с применением ISSR-маркеров

Essential-oil rose is a valuable crop widely used in production. The compliance with international quality standards is relevant for domestic production of essential-oil roses. In this regard an important stage of work in essential-oil roses breeding field is assessment of their genetic polymorphism which is necessary for parental form selection, new samples study. To protect property rights of such cultivars of the Research Institute of Agriculture of Crimea as Zolushka, Lan, Lada, Legrina and Raduga that are included in the "The state register of breeding achievements permitted for cultivation", it is necessary to develop genetic passports. An available method for genetic diversity study is inter-simple sequence repeats analysis also used for crops genetic passportization. In 2022, to analyze RIAC essential-oil rose collection genetic diversity with ISSR-PCR, ten rose cultivars were studied, some of which originated from Bulgaria and the others were bred in Crimea. Eight most effective ISSR primers were selected, their optimal annealing temperature was determined and informative parameters such as PIC, EMR and MI were calculated. All primers showed 100% polymorphism of samples. According to ISSR-PCR data, genetic distances matrix was constructed and on its basis samples clustering by UPGMA method was carried out. Due to a dendrogram analysis, two Crimean cultivars Aura and Zolushka were included in the same group with cultivars from Bulgaria. That may indicate genetic proximity of these cultivars to Bulgarian ones which are the world quality standard. At the same time Raduga cultivar is genetically removed from all varieties. The work result is development of effective methodology for assessing RIAC essential oil rose collection genetic diversity using ISSR markers.

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.

Phenological, morphological and genetic characterization of local grapevine (Vitis labrusca L.) genotypes grown in the Black Sea Region in Northern Turkey

Vitis labrusca has become widely naturalized in the Black Sea region of Northern Turkey. The aim of this work was to evaluate the morphological, phenological, and genetic characteristics of V. labrusca accessions grown in the Black Sea region during the last three growing seasons. Local genotypes were described according to the Organisation Internationale de la Vigne et du Vin (OIV) ampelographic descriptor, including shoot length (cm), mature leaf size, bunch length and width, berry length and width, berry weight, number of berries, Total Soluble Solids (TTS) and titratable acidity. Additionally, phenological data, including bud burst, flowering, veraison and harvest date, were recorded. The accessions were characterized at the molecular level, and genetic relationships were assessed by means of Inter Simple Sequence Repeat (ISSR) markers. Using 6 ISSR primers, seventy-seven bands were obtained, of which 69 were polymorphic with a mean polymorphic rate of 88.68%. These ISSR primers produced polymorphism information content (PIC) values ranging from 0.48 to 0.5. The genetic similarity ranged from 0.08 to 0.83 among the genotypes. According to the dendrogram based on the ISSR analysis, Accessions 8 and 5 were genetically related, with a coefficient of similarity of 0.83, while Accession 3 was the most distantly related genotype, with a coefficient of similarity of 0.08. These results demonstrated that ISSR markers can be used for genetic diversity analysis among V. labrusca genotypes. Our results also described characteristics of new V. labrusca genotypes that could be valuable for future Marker-Assisted Selection (MAS) and grapevine breeding.
 Key words: Diversity, foxy species, Inter-Simple Sequence Repeat markers (ISSRs), Breeding, polymorphism

Genetic stability analysis associated with salt stress and salicylic acid in fenugreek (Trigonella foenum-graecum) plants

Abstract. Alhasnawi AN. 2023. Genetic stability analysis associated with salt stress and salicylic acid in fenugreek (Trigonella foenum-graecum) plants. Biodiversitas 24: 3601-3608. Salt damage to plants has been attributed to a combination of many factors like the accumulation of toxic metal ions or osmotic stress. This study attempted to understand the mitigating effects of the salicylic acid (SA) treatment on the salt-stressed seed germination ability of the fenugreek plants (Trigonella foenum-graecum); SA pre-treatment along with the salt tolerance capacity have also been tested. The effects of salinity and applied SA on biochemical traits and genetic stability of plantshave been estimated in this study. Respective molecular and biochemical changes occurring in plants as well as genetic analysis have been performed. The results indicated that exogenous application of SA(0.4 mmol) improved the biochemical traits in the seedlings. The evaluation of the genetic stability of the plants was also conducted. Whereas DNA marker-based genetic stability estimates are considered a reliable, accurate, and advanced technique.The Inter-Simple Sequence Repeats (ISSR) analysis has been performed with the help of 7 ISSRs primers. The 6 ISSRs primers generated 54 fragments in total, with an average of 4.5 bands for every primer. The ISSR did not reveal any polymorphic bands. Finally, it has been concluded that exogenous SA can be effectively used as a potential growth regulator for improving the biochemical traits and salinity stress tolerance of the fenugreek plants and stimulating its medicinal properties. Furthermore, ISSR markers were seen to be a good tool as potential diagnostic markers for testing the genetic stability of plants.

An Effective and Low-Cost Method for Genomic DNA Extraction from Cyclanthera Pedata Species (A Nutraceutical Plant) without Liquid Nitrogen for Inter Simple Sequence Repeat Analyses

An Effective and Low-Cost Method for Genomic DNA Extraction from Cyclanthera Pedata Species (A Nutraceutical Plant) without Liquid Nitrogen for Inter Simple Sequence Repeat Analyses

ISSR Analysis to Detect Genetic Variation Among Some Lentil Genotypes in Syria

Lentil (Lens culinaris Medik) plays a significant role in human and animal nutrition. It is important to generate baseline information on genetic diversity of local and introduced lentil in Syria. Therefore, this study aimed to assess diversity and relationship among two local varieties and 35 introduced lentil lines from ICARDA using 20 Inter Simple Sequence Repeats (ISSR) primers.18 primers were able to amplify DNA bands with a total of 184 bands. Primers ISSR5, ISSR11, ISSR13, ISSR15 and ISSR16 presented 100% polymorphism percentage. The mean value of polymorphism percentage was 87.31%. ISSR primers could identify 19 unique bands in all genotypes, 12 of them were positive bands (present) and 7 were negative bands (absent). Primer ISSR11 was able to detect 3 positive unique bands in genotype Ln20. The UPGMA (unweighted pair group method with arithmetical averages) clustering based on ISSR data grouped 37 lentil genotypes into 6 clusters. The lowest genetic distance (0.20) was observed between (Ln15 and Ln16, Ln21 and Ln22, Ln27 and Ln28) which proves that they are closely related. While Ln24 presented the highest genetic distance (0.49) with both Ln18 and Idleb3 which refers to their genetic divergence.

Morphological and Genetic Analysis of Momordica cochinchinensis (Lour.) Spreng. (Gac) from Different Accessions in Malaysia

Momordica cochinchinensis or gac fruit is a 'superfruit' that is well-known in Vietnam. Gac is an orange fruit that is ovoid in shape and has a soft spiny texture. In Malaysia, gac fruit is a new and less known plant. This study aimed to characterize gac fruit using morphological analysis involving both vegetative and reproductive parts and to characterize the genetic diversity in gac fruit by using Inter-simple sequence repeat (ISSR) analysis. Four different gac accessions were collected from different areas (Kota Damansara (Selangor), Melaka Tengah (Melaka), Hulu Langat (Selangor) and Kuantan (Pahang)) were cultivated under tropical conditions in Kuantan, Pahang. The gac accessions showed differences in morphological characters. Generally, the gac fruits were reddish-orange in colour, the leaf was dark green on the adaxial part and light green on the abaxial part, and the female and male flower was light yellow and white in color. The fruit weight ranged from 193.72 g (GD) to 334.70 g (GH) with varied shapes and spike density. DNA extraction was following the CTAB method. All 30 primers showed high levels of polymorphism (83%) and the polymorphism information content (PIC) with the mean of 0.48. Nei's genetic distance coefficient ranged between 0.27 and 0.6 with the mean value of 0.41. Dendrogram based on UPGMA analysis grouped the four gac accessions into two main groups. Cluster I consisted of accession GD, GM and GH while cluster II consisted of only GX. Results from both morphological and molecular analysis showed genetic diversities in all four gac fruits studied.

Determination of Genetic variation among Beauveria bassiana and Metarhizium anisopliae Isolates from Palestine and other Geographic regions using ITS and β-tubulin Sequences, RFLP and ISSR Data

In this work we report the genetic variation of ten isolates of entomopathogenic fungi, four of which were isolated from Palestine while the remaining six were from other geographic regions. The variation was determined using molecular-based techniques including; sequence analysis of β-tubulin (BT) gene, the Internal Transcribed Spacers (ITS) 1 & 2 of the rDNA, Restriction Fragment Length Polymorphism (RFLP), and Inter-Simple Sequence Repeat (ISSR) analysis. The sequencing data of ITS regions confirmed the taxonomic identity of six isolates as Metarhizium anisopliae while the identity of the other four as Beauveria bassiana. These data also revealed that the two Palestinian Beauveria b. isolates are probably new strains since theyshowed 99% homology with the best match sequences reported in the gene bank. The restriction digestion of the ITS2 sequences produced a fingerprint for the PAL-M01 isolate. The ITS and BT sequences were able to distinguish the B-Bot genotype from the other Beauveria b. isolates. In contrast, the ITS could discriminate the three Metarhizium isolates: Ma-7, PAL-M01, and PPRC. Moreover, the ISSR analysis detected high levels of polymorphism among the Beauveria b. (80%) and the Metarhizium a. (90%) isolates. The fact that the PPRC and B-Bot isolates areobviously genetically different from their Metarhizium a. and Beauveria b. counterparts could be attributed to their distant geographic origins. The molecular data obtained in this work can be utilized for specific identification of each ofthese isolates which, would facilitate the local use of these fungi as biocontrol agents. To the best of our knowledge, this is the first study, in Palestine, attempting to discriminate between EPF isolates using advanced molecular techniques.

Induction of Axillary Bud Swelling of Hevea brasiliensis to Regenerate Plants through Somatic Embryogenesis and Analysis of Genetic Stability.

To overcome rubber tree (RT) tissue culture explant source limitations, the current study aimed to establish a new Hevea brasiliensis somatic embryogenesis (SE) system, laying the technical foundation for the establishment of an axillary-bud-based seedling regeneration system. In this study, in vitro plantlets of Hevea brasiliensis Chinese Academy of Tropical Agricultural Sciences 917 (CATAS 917) were used as the experimental materials. Firstly, the optimum conditions for axillary bud swelling were studied; then, the effects of phenology, the swelling time of axillary buds (ABs), and medium of embryogenic callus induction were studied. Plantlets were obtained through somatic embryogenesis. Flow cytometry, inter-simple sequence repeat (ISSR molecular marker) and chromosome karyotype analysis were used to study the genetic stability of regenerated plants along with budding seedlings (BSs) and secondary somatic embryo seedlings (SSESs) as the control. The results show that the rubber tree's phenology period was mature, and the axillary bud induction rate was the highest in the 2 mg/L 6-benzyladenine (6-BA) medium (up to 85.83%). Later, 3-day-old swelling axillary buds were used as explants for callogenesis and somatic embryogenesis. The callus induction rate was optimum in MH (Medium in Hevea) + 1.5 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) + 1.5 mg/L 1-naphthalene acetic acid (NAA) + 1.5 mg/L Kinetin (KT) + 70 g/L sucrose (56.55%). The regenerated plants were obtained after the 175-day culture of explants through callus induction, embryogenic callus induction, somatic embryo development, and plant regeneration. Compared with the secondary somatic embryo seedling control, axillary bud regeneration plants (ABRPs) were normal diploid plants at the cellular and molecular level, with a variation rate of 7.74%.

Micropropagation, Bioactive compounds, and Molecular diversity responses of Gardenia jasminoides Ellis using Gamma radiation

Mutation stimulation is a viable and well-established breeding approach for plants enhancement, induction bioactive compounds, and genetic variety generation to create novel plant mutants. The objective of this study was to mutate Gardenia jasminoides Ellis plant by gamma radiation (0, 15, 20, 25, 30, 35, and 40 Gy) as well as to determine the morphological and biochemical parameters of in vitro and in vivo treated plants. Consequently, the study has to detect the genetic diversity of the obtained mutants. The results indicated that gardenia plant irradiated with gamma at dose of 15 Gy showed the greatest growth rate. In addition, the maximum contents of chlorophyll (a, b) and carotenoids were detected in shootlets mutated with 15 Gy of gamma rays. Whereas, the highest total phenolic (TPC) and flavonoid contents (TFC), anthocyanin amounts, and antioxidant capacity were found in shootlets exposed to 40 Gy of gamma rays in vitro and in vivo. Furthermore, the suspected genetic variation in Gardenia jasminoides Ellis after treatment with γ-irradiation was evaluated using inter-simple sequence repeat (ISSR) analysis which produced a total of 98 bands, with the percentage of polymorphic bands ranging from 30 to 60%. According to the similarity matrix, the dendrogram was constructed into three clusters. Consequently, the results concluded that γ-irradiation of Gardenia jasminoides Ellis has given a sufficient number of induced mutations. Subsequently, the ISSR analysis has provided a powerful molecular marker for identifying mutants. Therefore, the in vitro  radiation-induced mutation could be a useful technique for assisting breeding programs for new Gardenia cultivars.

Micropropagation Protocol and Genetic Stability of the Salix myrtilloides Plants Cultivated In Vitro

Salix myrtilloides L. is a relict species, threatened with extinction in many European countries. To prevent the loss of the species, tissue culture was established to produce plant material for reintroduction in natural habitats. Micropropagation was chosen as a method to obtain new plants. S. myrtilloides shoots were disinfected with NaOCl, AgNO3, or with a two-step disinfection with NaOCl, and then placed on MS medium supplemented with BA at 1 mg·dm-3 and IBA at 0.1 mg·dm-3. Regenerated shoots were cultivated in presence of BA, KIN, and 2iP to select the treatment with the highest multiplication rate. The obtained plants were acclimatized to ex vitro conditions. Inter-simple sequence repeat (ISSR) and flow cytometric analyses were conducted on in vitro regenerated plants to check their genetic stability. The best disinfection results were obtained when explants were treated with 1.5% NaOCl for 20 min. The highest multiplication rate and good quality plants were noted in the control media, without growth regualtors and in presence of kinetin at 0.5 mg·dm-3. Flow cytometry and ISSR analyses confirmed genetic stability in plantlets, which indicated the possibility to use the in vitro obtained plants for reintroduction.

Genetic and phenotype recovery of Ananas comosus var. MD2 in response to ionizing radiation

Due to their sessile nature, plants are exposed to various environmental stressors such as exposure to high levels of harmful ultraviolet (UV), ionizing, and non-ionizing radiations. This exposure may result in various damages, ranging from DNA and chromosomal aberrations to phenotypic abnormalities. As an adaptation, plants have evolved efficient DNA repair mechanisms to detect and repair any damage caused by exposure to these harmful stressors to ensure their survival. In this study, the effects of gamma radiation (as a source of ionizing radiation) on clonal Ananas comosus var. MD2 was evaluated. The morphology and physiology of the clonal plantlets before and after exposure to gamma radiation were monitored at specific time intervals. The degree of genetic variation between the samples pre- and post-irradiation was also analyzed by using inter-simple sequence repeat (ISSR) markers. The resulting data revealed that the heights of the irradiated plantlets were significantly reduced (compared to control), but improved with the recovery period. Irradiated samples also exhibited relatively good photosynthetic efficiency that further improved as the plantlets recover. These observations were supported by the ISSR analysis, where the genetic dissimilarities between the irradiated samples and control were reduced by 0.1017, after 4 weeks of recovery. Overall, our findings suggested that the phenotype recovery of the clonal A. comosus var. MD2 plantlets was contributed by their ability to detect and repair the DNA lesions (as exemplified by the reduction in genetic dissimilarity after 4 weeks) and hence allow the plantlets to undergo phenotype reversion to normal plant stature.

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.

Inter simple sequence repeats and morphological traits to identify cultivated cotton varieties (Gossypium barbadense L.) in Egypt

Egyptian cultivated Cotton significantly impacted Egypt's economy, as it is well-known worldwide. This study aims to determine how much genetic and phenotypic variation exists in five different varieties of Egyptian Cotton using Inter Simple Sequence Repeats (ISSR) as a molecular marker and twenty-one quantitative and qualitative morphological traits as a taxonomic source in the development and evolution of this plant. Eleven ISSR primers were used, producing a total of 134 bands with a polymorphism percentage of 67%. Positive and negative significant Pearson correlations were found among the studied morphological traits in line with the phenotypic correlations in some characteristics. The genotypic correlation coefficient was higher in magnitude than that of phenotypic correlation. The five varieties were grouped into two major clusters using the UPGMA method based on morphological and ISSR analysis. The first one included G86 and G89 varieties, while the second cluster included G80 and G95; the G90 was separated from the other four varieties. This genetic relationship may be attributed to their similar ancestors. The information from this study should help with cotton breeding efforts to attain a high level of germplasm diversity and develop new high-yielding types to enhance cotton production and quality.

Assessing the Impact of Geographical Distribution and Genetic Diversity on Metabolic Profiles of a Medicinal Plant, Embelia ribes Burm. f.

The extensive use of Embelia ribes Burm. f. (Embelia) in tribal medicine proclaimed global attention as a promising candidate in complementary and alternative medicine. The knowledge of chemical blends is a prerequisite for the selection of raw materials for herbal medicine formulations; however, the influence of geographical distance and genetic diversity on the metabolome of Embelia fruits is unknown. Therefore, we collected Embelia fruits from four locations across the Western Ghats of India and analyzed the metabolic profile and genotypic diversity of Embelia fruits by liquid chromatography-tandem mass spectrometry (LC-MS/MS) and inter simple sequence repeats (ISSR), respectively. LC-MS/MS analysis yielded 583 compounds; however, the trimmed data resulted in 149 compounds. Further, MS/MS analysis identified 36 compounds, among which we reported 30 compounds for the first time from Embelia. These compounds belong to 11 compound classes that suggest location-specific chemical blends of Embelia fruits. Multivariate analysis showed 94% compound diversity across the accessions. ISSR analysis suggests 95% polymorphism across the accessions. A significant positive correlation (80%) between metabolomics and genotypic data matrices validates the genotype's influence in tuning Embelia's metabolic profiles. We conclude that the chemical profiles of Embelia are location-specific, which can be explored for the selection of herbal trade sustainably.

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.

Genetic, Phenotypic, and Pathogenic Variation Among Athelia rolfsii, the Causal Agent of Peanut Stem Rot in China.

Peanut stem rot caused by Athelia rolfsii is a serious soilborne disease worldwide and is becoming increasingly important in China. A total of 293 A. rolfsii isolates were collected from four representative peanut producing provinces in northern, central, and southern China. These isolates were assigned to 45 mycelial compatibility groups (MCGs) through pairing testing. The MCG diversity among isolates was greater in the southern sampled provinces compared with the northern provinces. A high level of genetic variability was found among the isolates from Guangdong Province in southern China. Variations were found in mycelial growth rate and sclerotial number, size, and dry weight of isolates sampled from places in different latitudes. Size and dry weight of sclerotia were positively correlated with latitude (P < 0.01), but the number of sclerotia was negatively correlated with latitude (P < 0.01). All tester isolates were pathogenic on peanut but varied in disease index. Inter-simple sequence repeat analysis and unweighted pair-group method with arithmetic average clustering resulted in three distinct clusters that were associated with the geographical location of the collection sites and sclerotial traits but were not associated with virulence of these isolates. These findings imply that genetic diversity, morphological traits, and virulence among A. rolfsii isolates varied in diverse geographical regions in China, and genetic diversity and sclerotial traits might be affected by latitude.

Detection of genetic variation among five basil species by using ISSR, SCOT and SRAP markers

Background: Eight inter-simple sequence repeats (ISSR), 14 Start codon-targeted (SCoT), and eleven sequence-related amplified polymorphisms were used to determine genetic diversity among five basil species (Ocimum tenuiflorum, Ocimum kilimandscharicum Guerk, Ocimum basilicum L., Ocimum americanum, and Ocimum basilicum var. minimum). Results: All of the markers' PCR products can be used to discriminate between the different basil species. The ISSR has the highest polymorphism percentage (73.1%), followed by SCoT (57.2%) and SRAP (55.6%). According to the combined data from the three markers, the basil species used were clustered into four groups, each with extremely excellent coordination between the similarity matrix and dendrograms. Species-specific markers have been identified. The SCoT analysis generates the highest number of species-specific marker (17) compared with ISSR and SRAP analysis which generated 14 and 13 markers respectively. Conclusions: These markers can be considered as a useful marker to improve Ocimum characteristics.

Detection of genetic variation among five basil species by using ISSR, SCOT and SRAP markers

Background: Eight inter-simple sequence repeats (ISSR), 14 Start codon-targeted (SCoT), and eleven sequence-related amplified polymorphisms were used to determine genetic diversity among five basil species (Ocimum tenuiflorum, Ocimum kilimandscharicum Guerk, Ocimum basilicum L., Ocimum americanum, and Ocimum basilicum var. minimum). Results: All of the markers' PCR products can be used to discriminate between the different basil species. The ISSR has the highest polymorphism percentage (73.1%), followed by SCoT (57.2%) and SRAP (55.6%). According to the combined data from the three markers, the basil species used were clustered into four groups, each with extremely excellent coordination between the similarity matrix and dendrograms. Species-specific markers have been identified. The SCoT analysis generates the highest number of species-specific marker (17) compared with ISSR and SRAP analysis which generated 14 and 13 markers respectively. Conclusions: These markers can be considered as a useful marker to improve Ocimum characteristics.