Annona crassiflora is a fruit-bearing tree species native to the Cerrado that has ecological and economic potential, mainly due to the production of attractive and useful fruits for a number of species, including humans. To provide a basis for its conservation and breeding, the objective was to assess the diversity and genetic structure of natural populations of the species using inter-simple sequence repeat (ISSR) molecular markers. Eight populations were analysed in the state of Minas Gerais, Brazil, with 24 individuals randomly sampled in each population, reaching a total of 192 trees. Ten ISSR primers were used, resulting in high genetic diversity for the combined data (H* = 0.35 and I* = 0.52). However, a wide range of representative values was observed in the individual assessment of populations, with JAN, GM and MC standing out for their low genetic diversity, resulting in H* of 0.19, 0.23 and 0.24 and I* of 0.28, 0.34 and 0.35, respectively. An analysis of molecular variance showed greater variation within populations, indicating gene flow (Nm), but genetic differentiation between populations was moderate. Bayesian analysis, although resulting in four genetic groups, revealed the presence of a majority group for the GM and JAN populations. We propose measures to maintain these populations, such as raising awareness of local extractivism and planting genetically divergent seedlings. Furthermore, we recommend including of all populations in conservation and breeding programmes, aiming to cover the maximum genetic variation for the species.

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.

The genus Lilium contains a number of ornamental crop species, which are commercially important in many countries. As they are vegetatively propagated, maintaining genetic stability is essential for their efficient conservation. In this study, we investigated the genetic stability of regenerated plants of three cultivars (L. bolanderi ‘Lenora’, L. bolanderi ‘Mount Duckling’ and L. bolanderi ‘Mount Dragon’) and one variety (L. callosum var. flavum) after cryopreservation, compared with fresh (donor) and non-cryopreserved plants using morphological traits and ISSR markers. No differences in morphological parameters including flower, stigma and pollen colour, floral spots, floral direction or polymorphic bands were observed between control (fresh and non-cryopreserved) and cryopreserved plantlets. In addition, based on the resulting UPGMA dendrogram, the four taxa were divided into different clusters. All cryopreserved, non-cryopreserved and fresh plants in each group could be grouped together in a single cluster with more than 97 or 100% similarity. The results suggest a very low level or the absence of genetic variation in terms of morphological and genetic stability among the plants regenerated after cryopreservation.

Bread wheat (Triticum aestivum) is one of the most important food crops in the world. Its physiological and morphological traits are closely related to yield. Therefore, it is generally important to discover the genomic region associated with these traits. In this research, associations between 21 simple sequence repeat (SSR) markers and 10 inter-simple sequence repeat (ISSR) markers with some traits related to root structure at the embryonic and seedling stages and also some agromorphological traits at the whole plant stage were evaluated on a set of 102 wheat genotypes. A highly significant coefficient of variation among different genotypes was observed in all measured traits. A high level of polymorphism with SSR and ISSR markers was obtained. Genetic structure analysis revealed two distinct subpopulations. Significant correlations were found between genomic markers and evaluated traits. A total of nine markers, including four SSR markers in 1A, 3A, 5A and 2B chromosomal regions and five different ISSR markers were related to the studied traits. Several molecular markers were significantly associated with more than one phenotypic trait, suggesting the possible presence of pleiotropic or indirect effects. The phenotypic variation justified by these alleles ranged from 4 to 15%. Obtained genetic information can be targeted for further validation and genetic analysis in the relevant populations or other breeding sets.

Phenotypic plasticity and rapid evolution are two important strategies by which invasive species adapt to a wide range of environments and consequently are closely associated with plant invasion. To test their importance in invasion success of Crofton weed, we examined the phenotypic response and genetic variation of the weed by conducting a field investigation, common garden experiments, and intersimple sequence repeat (ISSR) marker analysis on 16 populations in China. Molecular markers revealed low genetic variation among and within the sampled populations. There were significant differences in leaf area (LA), specific leaf area (SLA), and seed number (SN) among field populations, and plasticity index (PIv) for LA, SLA, and SN were 0.62, 0.46 and 0.85, respectively. Regression analyses revealed a significant quadratic effect of latitude of population origin on LA, SLA, and SN based on field data but not on traits in the common garden experiments (greenhouse and open air). Plants from different populations showed similar reaction norms across the two common gardens for functional traits. LA, SLA, aboveground biomass, plant height at harvest, first flowering day, and life span were higher in the greenhouse than in the open-air garden, whereas SN was lower. Growth conditions (greenhouse vs. open air) and the interactions between growth condition and population origin significantly affect plant traits. The combined evidence suggests high phenotypic plasticity but low genetically based variation for functional traits of Crofton weed in the invaded range. Therefore, we suggest that phenotypic plasticity is the primary strategy for Crofton weed as an aggressive invader that can adapt to diverse environments in China.

DNA fragment analysis, based on amplification of intersimple sequence repeats by the polymerase chain reaction (ISSR-PCR), was used to assess genetic relationships of 217 U.S. accessions of common cocklebur. Twenty-four polymorphic markers were generated from six primers. Analysis of genetic similarity by clustering procedures resulted in separation of the accessions into two main clusters. Accessions within these two clusters were designated as either northern or southern genotypes. Forty-four of 48 accessions analyzed from Washington, Michigan, Iowa, and Ohio were northern genotypes, whereas 67 of 68 accessions analyzed from Mississippi, Arkansas, South Carolina, and North Carolina were southern genotypes. Accessions from Kansas, Missouri, and Illinois included 40 and 56 northern and southern genotypes, respectively, indicating a transition zone. In Illinois, accessions collected from northern and southern counties tended to be northern and southern genotypes, respectively. We conclude that much of the genetic variation among U.S. common cocklebur accessions is distributed along a latitudinal gradient.

Lanceleaved waterplantain is an exotic weed of rice that was introduced into Australia in the 1930s. Since its introduction, it has spread throughout much of the rice-growing region in southeastern Australia. The variability of lanceleaved waterplantain in these regions was studied using polymerase chain reaction-based, inter simple sequence repeat (ISSR) analysis. Deoxyribonucleic acid fingerprints from samples of the weed from southeastern Australia were compared between locations and with two samples of common waterplantain, a closely related species. The analysis indicated that there were two distinct groups of lanceleaved waterplantain that correlated with location. From the results of multidimensional scaling analysis, it is hypothesized that the Griffith group did not arise from hybridization between lanceleaved waterplantain and common waterplantain, and that it is more likely that the group arose from a separate introduction into the area. It is also suggested that there is seed movement between areas in the Murray Valley and Colleambally Irrigation areas. The implications of this variation for biological control of the weed are discussed.

Intraspecific genetic variation may contribute significantly to invasiveness and control problems, but has been characterized to date in relatively few invasive weed species. We examined 56 intersimple sequence repeat (ISSR) loci in 220 individuals from 11 invading populations of yellow toadflax sampled across five western states. All populations showed high levels of genetic diversity. Estimated values for Shannon's diversity measure ranged from 0.217 to 0.388, and for expected heterozygosity from 0.178 to 0.260. Nei's total gene diversity index (HT), on the basis of all individuals across all populations, was 0.267. Partitioning of genetic variance using analysis of molecular variance revealed 1.7% of genetic variation among regional population groups, 29.1% among populations within groups, and 69.2% within populations, consistent with expectations for an outcrossing species but suggesting little geographic differentiation. Pairs of adjacent individuals identical at all ISSR loci that appeared to be ramets of a single clone were detected in only one population. This indicates that patch expansion in yellow toadflax is driven more by sexual reproduction via seed than by rhizomatous clonal spread, at least at the spatial scale of sampling for this study. Eight populations had significant values for Mantel's R at P = 0.05, suggesting some fine-scale positive genetic structuring, possibly from restricted gene flow. Population clustering on the basis of Nei's genetic distance between populations and unweighted pair group method with arithmetic mean did not reflect geographic location. It is likely that multiple introductions of this species have occurred across the Intermountain West, followed by extensive genetic recombination. High levels of genetic diversity within yellow toadflax populations pose management challenges, as already seen in reports of variable response to herbicide application and limited impacts of biocontrol agent releases.

Recent advances in molecular methods and statistical analyses provide weed scientists with powerful tools for examining the genetic structure of weedy plant populations. The value of these studies depends on effective sampling protocols; however, there is little consensus on how to sample plant populations for genetic diversity analyses. In this review, we draw on published literature that incorporates sampling theory and spatial statistics in population genetic analyses to identify key factors to consider when designing a sampling strategy. We discuss how sampling design is affected by research objectives, biology of the study species, population structure, marker choice, and the genetic parameters to be investigated, and we offer suggestions on defining sampling units and developing sampling protocols.

Although there is evidence that interspecific hybridization can initiate invasion by nonnative plants, there are few documented examples of novel hybridization events between introduced plant species already exhibiting invasive behavior. We conducted morphometric and molecular analyses of toadflax plants with intermediate morphology found at two sites in Montana, which were co-invaded by yellow toadflax and Dalmatian toadflax. Field-collected putative hybrid plants had intermediate morphometric scores (mean 0.47, on a scale of 0.0 = indistinguishable from Dalmatian toadflax to 1.0 = indistinguishable from yellow toadflax) for a suite of phenotypic traits that differentiate the parent species (leaf length : width ratio, growth form, seed morphology, inflorescence type, and ventral petal shape). Inter-simple sequence repeat (ISSR) analysis of a subset of these putative hybrids revealed combinations of species-diagnostic bands, confirming the presence of DNA from both parent species. Controlled interspecific hand-pollinations generated viable first generation (F1) hybrid plants that also had intermediate morphometric scores (mean 0.46) and a mix of species-diagnostic ISSR bands from both parents. The hand-generated F1 hybrids crossed readily with both parent species to produce viable first generation backcrossed (BC1) plants. Our results confirm that hybridization is occurring between invasive populations of yellow toadflax and Dalmatian toadflax, and that the hybrid progeny are viable and fertile. This example of hybridization between alien congeners is of concern as the parent taxa are already known to be highly invasive. Further research is needed to assess the invasive potential of hybrid toadflax populations, and the likelihood of introgressive trait transfer between the parent species.

Mexican sunflower is a native species of North and Central America that was introduced into China early last century, but it has widely naturalized and become a harmful invasive plant in tropical and subtropical regions in South China. Inter-simple sequence repeat (ISSR) markers were employed to assess genetic diversity and variation in Mexican sunflower populations from China and neighboring regions. The karyotypes of populations were also studied. Our research showed high levels of genetic diversity in all populations. The lowest genetic diversity estimates were represented in two populations in Laos, suggesting prevention of new introductions into Laos is critical. Partitioning of genetic variance revealed that genetic variation was mostly found within populations, and unweighted pair group method with arithmetic means (UPGMA) analysis showed that the introductions into China and Laos were independent. There were no obvious correlations between genetic relationships and geographic distance of populations in China, consistent with the human associated dispersal history of Mexican sunflower. Previous cytological data and our chromosome count (2n = 34) and karyotype analysis showed chromosome stability among populations. The high levels of genetic diversity within invasive Mexican sunflower populations could be challenging for its management in China, and further expansion and potential negative effects on ecological systems of this plant should be monitored.

Molecular variability of vanilla (Vanilla planifolia) and three wild species was assessed to explore the possible sources of variation that can be used for crop improvement. A total of 154 ISSR loci were analysed by the UPGMA, assignment tests of individuals (STRUCTURE) and indices of genetic diversity. The assignment tests were done at two levels: first considering the four species and then only the accessions of V. planifolia. The molecular analysis indicated 99.3% polymorphism among all species and 70.45% within V. planifolia. The UPGMA showed the separation of these four species into three groups and grouped V. planifolia accessions into three subgroups. The more genetically differentiated accessions were of the Rayada morphotype and a wild accession was from Oaxaca, followed by a wild accession from Quintana Roo; all the commercial accessions of V. planifolia (Mansa morphotype) were grouped together. The STRUCTURE analysis differentiated between V. planifolia and the three wild species, and among the accessions of the Mansa and Rayada morphotypes and the wild accessions. The STRUCTURE analysis also indicated the presence of mixed individuals. These results are of great importance since the accessions of V. planifolia that are genetically more differentiated are the most threatened due to the scarcity of these individuals, the destruction of habitat and replacement by the commercial morphotype. These individuals should be salvaged and used to expand the genetic background of vanilla.

In Uruguay, onion (Allium cepa L.) germplasm is mainly derived from the genetic material introduced by several waves of European immigrants and subsequently multiplied by household farmers, resulting in a wealth of locally adapted populations. This study examined the genetic diversity in a collection of 27 local onion populations and two cultivars derived from them. A total of 843 onion plants were fingerprinted, and 83 inter-simple sequence repeat polymorphic bands were generated. Analysis of molecular variance showed high diversity within the populations (66% of the total variation). Some short-day populations from different geographical origins were grouped together by the unweighted pair-group method using arithmetic averages, principal coordinate analysis and cluster analysis, while the more extensively sampled long- and intermediate-day populations showed a widespread distribution, with no significant subgrouping among them. This weakly structured gene pool is probably the consequence of seed and bulb exchange between farmers and natural inter-pollination. Nevertheless, a Bayesian analysis allowed the distinction of four genetic backgrounds of alleles in the whole collection, and populations were predominately assigned to each genetic background. In addition, mitochondrial variants determining normal (N) pollen fertility or the sterile S or T types were analysed for the same set of plants using specific primers. Most accessions showed a proportion of male-sterile individuals. Cytoplasm type T was the most represented (26 out of 29 accessions), and cytoplasm type S was found in a low proportion of individuals in seven populations. Uruguayan onion germplasm maintains a low degree of genetic differentiation despite the small cultivated area and intense seed exchange, probably due in part to different market purposes based on the growing cycle.

Blackberry is a fruiting berry species with very high nutrient contents. With the recent increasing consumer demand for blackberries, new sources of germplasm and breeding techniques are required to improve blackberry production. This study was carried out to evaluate the genetic diversity (GD) and relationship among 55 blackberry (Rubus fruticosus) mutants derived from γ-ray treatment (52 lines) and N-methyl-N′-nitrosourea (MNU) treatment (three lines) using an inter-simple sequence repeat marker. A total of 18 bands were amplified with an average of 3.6 bands per primer. Among them, eight bands were identified to be polymorphic with a rate of 44.4%. In addition, the GD information content values were highest in the 60 Gy treatment population and the GD values were higher in the γ-ray treatment populations than in the MNU treatment population. According to a cluster analysis, all the mutant lines can be classified into five categories, and the genetic distance was greatest between the 80 Gy-irradiated population and other populations. These results indicate that mutant lines have high GD and can be effectively utilized for improving blackberry breeding.

Inter simple sequence repeats (ISSR) and random amplified DNA polymorphism (RAPD) markers were used to estimate the genetic diversity among 14 tropical silkworm races (Bombyx mori L.) for identifying potential parents to be used for hybrid preparation for commercial exploitation. High polymorphism (70.91 and 74.70%) was revealed by ISSR and RAPD markers. The dendrograms generated, using unweighted pair group method using arithmetic average from these markers, grouped the 14 silkworm races into two major groups, which corroborates the differences in cocoon characteristics. Discriminant function analysis of ISSR and RAPD markers identified three functions for cocoon weight and two functions for shell weights, respectively. Step-wise multiple regression analysis identified six ISSR markers (834500, 8841700, 8841850, 8271500, 8401500 and 7891250) and seven RAPD markers (834500, 885900, 8101400, 884900, 8361500, 7891250 and 7621700) significantly associated with cocoon and shell weights. The genetically divergent parents, identified through this study, can be used for the preparation of hybrids for commercial utilization. Similarly, the DNA markers identified can be utilized for marker-assisted selection along with those identified through quantitative trait locus mapping by others. Thus the information generated in this study is useful in silkworm breeding programmes in the tropical sericulture industry.

Inter simple sequence repeat markers were used to assess the genetic diversity and population genetic structure in 12 populations of Nothapodytes nimmoniana from Western Ghats of India. A total of 16 selected primers produced 103 discernible bands, with 76 (73.7%) being polymorphic. The Nei's gene diversity (h) ranged from 0.1166 to 0.2124, with an average of 0.1518 at the population level and 0.2965 at the species level indicating high genetic diversity. The Shannon's index (I) was estimated to be 0.2189 within populations (range 0.1703–0.2947) and 0.4352 at the species level. The analysis of molecular variance showed that the genetic variation was found mainly within populations (73%), but variance among populations was only 27% and its value, ΦPT = 0.271, P < 0.001, implied that high genetic differentiation among populations. In addition, Nei's differentiation coefficient (GST) was found to be high (0.4882) and the gene flow (Nm) was low (0.5242), confirming the high population genetic differentiation. The unweighted pair-group method using arithmetic average clustering elicited similar results. Based on this, we propose conservation strategy for this plant species.

The pollination ecology, breeding system and population genetic structure of three climbing Bauhinia species B. championii (4 populations, 23 individuals), B. corymbosa (2 populations, 25 individuals) and B. glauca (8 populations, 76 individuals) were studied in Hong Kong, southern China. We hypothesize that the climbing Bauhinia species will attract targeted pollinators to achieve out-cross success and high levels of self-incompatibility will be expected to maintain diversity, with local population expansion relying on vegetative propagation. All three species have inflorescences consisting of numerous small, pale, fragrant flowers, which show diurnal anthesis. Field observations revealed that all three species are predominantly pollinated by bees (particularly Apis mellifera) and butterflies (Graphium and Papilio species), although B. championii is also pollinated by wasps and flies. Bauhinia corymbosa and B. glauca have sucrose-dominant nectar, whereas B. championii has hexose-dominant nectar. In controlled-pollination experiments fruit and seed set were generally highest following artificial out-crossing. The index of self-incompatibility of B. championii is 1.07, indicating self-compatibility; B. corymbosa and B. glauca were obligately self-incompatible. The population genetic structure and variation of the Bauhinia species was investigated using ISSR markers. Generally the three species have moderate within-population (mean HS = 0.206) and high among-population genetic variation (mean GST = 0.284). No correlation exists between the geographical and genetic distance, possibly due to the small local population size. All three species showed high levels of heterozygosity as expected for predominantly out-crossing long-lived K-selected species.

When cultivating genetically modified varieties, the spontaneous gene flow between crop and wild relatives could be of concern. We analyzed spontaneous hybridization between a transgenic male-sterile line of oilseed rape (Brassica napus, 2n=38, AACC) and, as pollen donors, three European populations of wild radish (Raphanus raphanistrum, 2n=18, Rr,Rr) and a variety of cultivated radish (Raphanus sativus, 2n=18, RR). Seeds showed size and shape dimorphism that correlated to the frequency of hybrids. The offspring were scored morphologically and analyzed using DNA markers (inter-simple sequence repeats) to quantify hybrid frequencies. Seed set ranged from 0.4–1.2 seeds per pod, and 0.02–0.6 seeds per pod were confirmed as hybrids. The frequency of confirmed hybrids differed significantly among populations of R. raphanistrum. In the cross with a French population, all offspring were hybrids; in the cross with a Swiss population, 53% of the offspring were hybrids; and in the cross with a Danish population, only 2% of the offspring were found to be hybrids. The remaining offspring apparently belonged to two groups: the majority was B. napus-like plants, possibly of matromorphic origin, and a minority from the Danish cross seemed to carry fragments of the Raphanus genome. In the cross with a cultivated R. sativus, all offspring were found to be hybrids. This is the first report on spontaneous hybridization between B. napus and R. sativus. Hybrids from all cross-combinations had low pollen fertility (0–15%). If R. raphanistrum occurs where male-sterile B. napus is cultivated, large regional differences in hybridization frequencies between the species could complicate environmental risk assessment of transgenic oilseed rape.