Frequency Of Outcrossing Research Articles

Embryological observations on seed abortion in Hibiscus syriacus L. and physiological studies on nutrients, enzyme activity and endogenous hormones

Under natural conditions, most Hibiscus syriacus L. individuals form very few mature seeds or the mature seeds that do form are of poor quality. As a result, seed yield is poor and seeds have low natural germinability. These phenomena strongly hinder utilization of the excellent germplasm resources of H. syriacus. The study has shown that pollen activity and stigma receptivity were high on the day of anthesis, and the pistils and stamens were fertile. Pollen release and stigma receptivity were synchronous. But in styles following self and cross-pollination, pollen tube abnormalities (distortion and twisting of the pollen tubes) and callose deposition were observed. Cross-pollinated pollen tubes elongated faster and fewer pollen tube abnormalities were observed compared with self-pollinated pollen tubes. And during embryo development, abnormalities during the heart-shaped embryo stage led to embryo abortion. Imbalance in antioxidant enzyme activities and low contents of auxin and cytokinin during early stages of embryo development may affect embryo development. Therefore, a low frequency of outcrossing and mid-development embryo abortion may be important developmental causes of H. syriacus seed abortion. Nutrient deficiencies, imbalance in antioxidant enzyme activities, and a high content of abscisic acid at advanced stages of seed development may be physiological causes of seed abortion.

Smooth pigweed (Amaranthus hybridus) and unresolved Amaranthus spp. from Brazil resistant to glyphosate exhibit the EPSPS TAP-IVS substitution

AbstractThe presence of glyphosate-resistant smooth pigweed (Amaranthus hybridus L.) biotypes has increased in southern Brazil in recent years, presenting the triple amino acid substitution TAP-IVS in 5-enol-pyruvylshikimate-3-phosphate synthase (EPSPS) previously found in Argentina. Some of these biotypes have morphological characteristics of A. hybridus and redroot amaranth (Amaranthus retroflexus L.). The present study aimed to identify, through molecular markers, the herbicide-resistant species of Amaranthus from Brazil that have the TAP-IVS substitution and to analyze the occurrence of pollen-mediated gene flow (PMGF) as the source of the TAP-IVS substitution in these biotypes. Six biotypes were evaluated using internal transcribed spacer (ITS) sequences, of which two (AMACHY-S and CAMAQ-R) were molecularly classified as A. hybridus and four (AMACRET-S, AMACVI-S, ARRGR-R, and SAOJER-R) were unclassified. Interestingly, all the glyphosate-resistant biotypes (ARRGR-R, SAOJER-R, and CAMAQ-R) had the TAP-IVS substitution and an increase in EPSPS relative copy number; however, only CAMAQ-R was confirmed as A. hybridus. Although the biotypes ARRGR-R and SAOJER-R are closely related to A. hybridus and green pigweed (Amaranthus powellii S. Watson), their species identity could not be resolved. The biotype SAOJER-R also was resistant to acetolactate synthase (ALS)-inhibiting herbicides due to a tryptophan to leucine substitution at position 574 in ALS. The evaluation of 119,746 seedlings in an intraspecific hybridization study of A. hybridus indicated an outcrossing frequency of 0.09%. In contrast, an absence of interspecific hybridization (A. hybridus × unclassified biotype, AMACVI-S) was found after screening 111,429 offspring. Unclassified biotypes might be derived from one or more ancient hybridization events and subsequently evolved the triple mutation independently. Alternatively, such biotypes could have evolved from recent hybridization events, which occur at a frequency below the level of detection in our study.

Frequency of Outcrossing and Isolation Distance in Faba Beans (Vicia faba L.)

Faba beans (Vicia faba L.) constitute a partially outcrossing species requiring an isolation distance to maintain genetic purity when more than one variety is grown in field conditions. This information is crucial for seed growers and faba bean breeders. A study was conducted at the University of Sydney’s Plant Breeding Institute, Narrabri, over two years to examine the extent of natural outcrossing using a creamy white flower characteristic as a morphological marker, which is controlled by a single recessive gene. The white-flowered genotype (IX225c) was grown in paired rows of 150 m length in four directions from a central 480 m2 plot of the normal flowered genotype PBA Warda. A beehive was placed in the central plot at the flowering time and natural pollination was allowed. At maturity, seed samples were taken from the white-flowered genotype at designated intervals along each axis and 100 seeds from each sample were grown in the glasshouse/birdcage to the 4–5 leaf stage and the proportion of plants displaying a stipule spot pigmentation (normal flower color and spotted stipule are linked) was used to determine the percentage of outcrossing. Maximum outcrossing of 2.28% occurred where both genotypes were grown side by side (0 m) and the degree of outcrossing decreased as the distance along each axis from the central plot increased. At a 6 m distance, the outcrossing was less than 1%; however, on occasion, it increased to 1% beyond a distance of 100 m, indicating the volatile and unpredictable nature of bee flights. Distance had a major effect on outcrossing but the direction and its interaction had no effect. The results suggest that to limit outcrossing to below 0.5%, a distance of more than 150 m between plots of different faba beans cultivars would be required. It also indicated that Australian faba bean genotypes are mostly self-fertile and a relatively narrow isolation distance will ensure self-fertilization in seed production and breeding programs.

Transgene Bioconfinement: Don’t Flow There

The adoption of genetically engineered (GE) crops has led to economic and environmental benefits. However, there are regulatory and environmental concerns regarding the potential movement of transgenes beyond cultivation. These concerns are greater for GE crops with high outcrossing frequencies to sexually compatible wild relatives and those grown in their native region. Newer GE crops may also confer traits that enhance fitness, and introgression of these traits could negatively impact natural populations. Transgene flow could be lessened or prevented altogether through the addition of a bioconfinement system during transgenic plant production. Several bioconfinement approaches have been designed and tested and a few show promise for transgene flow prevention. However, no system has been widely adopted despite nearly three decades of GE crop cultivation. Nonetheless, it may be necessary to implement a bioconfinement system in new GE crops or in those where the potential of transgene flow is high. Here, we survey such systems that focus on male and seed sterility, transgene excision, delayed flowering, as well as the potential of CRISPR/Cas9 to reduce or eliminate transgene flow. We discuss system utility and efficacy, as well as necessary features for commercial adoption.

The Grassland Fragmentation Experiment in the Swiss Jura Mountains: A Synthesis

We synthesize findings from a 7-year fragmentation experiment in species-rich, nutrient-poor, dry calcareous grasslands in the north-western Jura mountains, Switzerland. We used a standardized approach with 48 fragments (0.25–20.25 m2) and corresponding control plots in three sites. The 5-m-wide isolation area around the fragments was maintained by frequent mowing. Fragments experienced various ecological changes, e.g., plant biomass increased along fragment edges. We examined fragmentation effects on species richness and composition, abundance, genetic diversity, functional diversity, species traits and species interactions (pollination, herbivory, parasitism, disease) in a wide array of invertebrate (gastropods, spiders, woodlice, various insect groups) and plant taxa. Responses to fragmentation differed between taxonomical groups and species. While species richness and individual density were lower in fragments in some groups, the opposite was true for other groups. Fragmentation effects were most pronounced on species interactions; however, some effects only occurred with a delay. For example, fragmentation influenced foraging patterns of bumblebees, affecting pollination, which in turn resulted in a decreased outcrossing frequency and reduced genetic diversity in a focal plant species. We highlight key findings of the experiment and emphasize their implications for grassland conservation.

Food/Feed and Environmental Risk Assessment of Genetically Modified Glufosinate-tolerant Oilseed Rape MS8, RF3 and MS8 x RF3 for Import, Processing and Feed Uses under Directive 2001/18/EC (Notification C/BE/96/01)

Food/Feed and Environmental Risk Assessment of Genetically Modified Glufosinate-tolerant Oilseed Rape MS8, RF3 and MS8 x RF3 for Import, Processing and Feed Uses under Directive 2001/18/EC (Notification C/BE/96/01)

Quantification of Outcrossing Events in Haploid Fungi Using Microsatellite Markers.

Species in genera of the fungal family Ceratocystidaceae are known to have different mating strategies, including heterothallism and homothallism. Of these, species of Ceratocystis, typified by the pathogen Ceratocystis fimbriata all undergo unidirectional mating-type switching. This implies that the pathogens possess the ability to self, but also to undergo sexual outcrossing between isolates of different mating types. In this study, we extended the recently developed microsatellite-based technique to determine the extent to which outcrossing occurs in ascospore masses of haploid fungi to two field collections of Ceratocystis albifundus. In this way, the role of reproductive strategies in shaping population structure and diversity could be better understood. Results showed that a high frequency of outcrossing occurs in isolates of the pathogen from both non-native and native areas. This explains the high level of genetic diversity previously observed in this population despite the fact that this pathogen has the ability to self.

Food, Feed and Environmental Risk Assessment of Glufosinatetolerant Genetically Modified Oilseed Rape T45 for Food and Feed Uses, Import and Processing Under Regulation (EC) No 1829/2003 (Application EFSA/GMO/UK/2005/25)

Food, Feed and Environmental Risk Assessment of Glufosinatetolerant Genetically Modified Oilseed Rape T45 for Food and Feed Uses, Import and Processing Under Regulation (EC) No 1829/2003 (Application EFSA/GMO/UK/2005/25)

Genetic diversity of preserved rice seed samples from the Mikawa area, Japan, stored in the Meiji era.

Preserved rice (Oryza sativa L.) seeds stored for nearly a century as an emergency food stocks from the Mikawa area were investigated for their genetic diversity. Morphologically, the seeds appeared to be typical Japonica. One chloroplast INDEL petN-trnC, two nuclear INDELs Acp1 and Cat1, and three SNP markers in Starch synthase IIa were amplified to characterize the molecular profile. The efficiency of amplification varied among the markers. Most of preserved seeds were classified as Japonica, but some were identified as Indica. The heterozygous genotypes detected suggested a high frequency of outcrossing at that time. On the other hand, 21 SSR markers showed quite a high degree of amplification efficiency. Principal coordinate analysis and STRUCTURE analysis based on the SSR polymorphisms proved that the preserved seeds contained alleles that were not detected among current landraces and breeding varieties, and there were the expected three subpopulations among 96 preserved seeds. These results indicated that these preserved seeds from Mikawa area in Meiji era had high genetic diversity and consisted of some subpopulations including Indica landraces with typical Japonica seed shape. These lines were considered to have been lost from current genetic resources.

Preliminary Environmental Risk Assessment of Genetically Modified Oilseed Rape MON 88302 for Food and Feed Uses, Import and Processing under Regulation (EC) No 1829/2003 (Application EFSA/GMO/BE/2011/101)

Preliminary Environmental Risk Assessment of Genetically Modified Oilseed Rape MON 88302 for Food and Feed Uses, Import and Processing under Regulation (EC) No 1829/2003 (Application EFSA/GMO/BE/2011/101)

Environmental Risk Assessment of Glufosinate-Tolerant Genetically Modified Oilseed Rape T45 for Food and Feed Uses, Import and Processing under Regulation (EC) No 1829/2003 (Application EFSA/GMO/UK/2005/25)

Environmental Risk Assessment of Glufosinate-Tolerant Genetically Modified Oilseed Rape T45 for Food and Feed Uses, Import and Processing under Regulation (EC) No 1829/2003 (Application EFSA/GMO/UK/2005/25)

Environmental Risk Assessment of Glufosinate-Tolerant Genetically Modified Oilseed Rape MS8, RF3 and MS8 x RF3 for Import, Processing and Feed Uses under Directive 2001/18/EC (Notification C/BE/96/01)

Environmental Risk Assessment of Glufosinate-Tolerant Genetically Modified Oilseed Rape MS8, RF3 and MS8 x RF3 for Import, Processing and Feed Uses under Directive 2001/18/EC (Notification C/BE/96/01)

Microsatellite and SNAP markers used for evaluating pollen dispersal on Pati tall coconuts and Xenia effect on the production of ‘Kopyor’ fruits

Kopyor coconut is one of the many unique coconut types existed in Indonesia. To overcome the problem of low kopyor fruit yield, it is necessary to study xenia effect on the fruit yield of this coconut. The combination of kopyor coconut and normal coconut population selected at infarmers’ coconut plantations consisted of 33 normal coconut trees, and 9 kopyor coconut trees. All adult trees surrounding the 9 kopyor heterozygous (Kk) palms were evaluated as potential male candidate parents (pollen donors). All samples genotypes were determined using four SNAP markers and six microsatellite marker loci, parentage analysis using CERVUS software version 2.0. Results of the analysis indicate that xenia effect reduced kopyor fruit yields. Kopyor heterozygous (Kk) female parents produced low number of kopyor fruits when they were surrounded by many normal homozygous (KK) pollen donors. Out of 99 harvested progeny arrays from the kopyor heterozygous (Kk) female parents, none exhibited kopyor phenotype. The results also indicate that the pollen dispersal from normal homozygous (KK) donor palms range from 0 m (self pollination) to 54 m (outcrossing). The occurence of outcrossing frequency was at least 95% and the selfing frequency is 5%.

Genetic Subdivision and Variation in Selfing Rates Among Central American Populations of the Mangrove Rivulus, Kryptolebias marmoratus.

We used 32 polymorphic microsatellite loci to investigate how a mixed-mating system affects population genetic structure in Central American populations (N = 243 individuals) of the killifish Kryptolebias marmoratus (mangrove rivulus), 1 of 2 of the world's only known self-fertilizing vertebrates. Results were also compared with previous microsatellite surveys of Floridian populations of this species. For several populations in Belize and Honduras, population structure and genetic differentiation were pronounced and higher than in Florida, even though the opposite trend was expected because populations in the latter region were presumably smaller and highly selfing. The deduced frequency of selfing (s) ranged from s = 0.39-0.99 across geographic locales in Central America. This heterogeneity in selfing rates was in stark contrast to Florida, where s > 0.9. The frequency of outcrossing in a population (t = 1 - s) was tenuously correlated with local frequencies of males, suggesting that males are one of many factors influencing outcrossing. Observed distributions of individual heterozygosity showed good agreement with expected distributions under an equilibrium mixed-mating model, indicating that rates of selfing remained relatively constant over many generations. Overall, our results demonstrate the profound consequences of a mixed-mating system for the genetic architecture of a hermaphroditic vertebrate.

Inferring outcrossing in the homothallic fungus Sclerotinia sclerotiorum using linkage disequilibrium decay.

The occurrence and frequency of outcrossing in homothallic fungal species in nature is an unresolved question. Here we report detection of frequent outcrossing in the homothallic fungus Sclerotinia sclerotiorum. In using multilocus linkage disequilibrium (LD) to infer recombination among microsatellite alleles, high mutation rates confound the estimates of recombination. To distinguish high mutation rates from recombination to infer outcrossing, 8 population samples comprising 268 S. sclerotiorum isolates from widely distributed agricultural fields were genotyped for 12 microsatellite markers, resulting in multiple polymorphic markers on three chromosomes. Each isolate was homokaryotic for the 12 loci. Pairwise LD was estimated using three methods: Fisher's exact test, index of association (IA) and Hedrick's D'. For most of the populations, pairwise LD decayed with increasing physical distance between loci in two of the three chromosomes. Therefore, the observed recombination of alleles cannot be simply attributed to mutation alone. Different recombination rates in various DNA regions (recombination hot/cold spots) and different evolutionary histories of the populations could explain the observed differences in rates of LD decay among the chromosomes and among populations. The majority of the isolates exhibited mycelial incompatibility, minimizing the possibility of heterokaryon formation and mitotic recombination. Thus, the observed high intrachromosomal recombination is due to meiotic recombination, suggesting frequent outcrossing in these populations, supporting the view that homothallism favors universal compatibility of gametes instead of traditionally believed haploid selfing in S. sclerotiorum. Frequent outcrossing facilitates emergence and spread of new traits such as fungicide resistance, increasing difficulties in managing Sclerotinia diseases.

Changes in pollinator assemblages following hurricanes affect the mating system ofLaguncularia racemosa(Combretaceae) in Florida, USA

Abstract:Hurricanes are major disturbance events in Neotropical mangrove communities, potentially affecting the reproductive success of mangrove species. This was the first investigation of changes in mangrove pollinator assemblages following hurricanes, and the effect of these changes on the mating system ofLaguncularia racemosa. Insect pollinator assemblages were investigated in three Florida mangrove communities in 2001–2003, 2005 and 2009; two hurricanes affected the area in 2004. Visitation rates were estimated from 1445 insects observed during 272 10-min intervals; the number of flowers visited by each insect was also recorded. Pollinator diversity was estimated with the Shannon Index. Following the hurricanes, species richness was reduced by 43–65% and diversity declined by 36–70%. Significant declines in insect visitation toL. racemosaresulted in reduced outcrossing frequencies in 2005.Laguncularia racemosaflowers autogamously self-pollinate without insect visitors, so fruit set still occurred. Visitation rates returned to pre-hurricane levels by 2009, but foraging behaviours differed from pre-hurricane patterns; outcrossing was further favoured by reduced frequencies of long foraging bouts and increased frequencies of short foraging bouts. The mixed mating system ofL. racemosaprovides reproductive assurance following hurricane disturbances, when pollinator abundance is low.

The Ghost of Outcrossing Past in Downy Brome, an Inbreeding Annual Grass

We investigated the frequency of outcrossing in downy brome (Bromus tectorum L.), a cleistogamous weedy annual grass, in both common garden and wild populations, using microsatellite and single nucleotide polymorphic (SNP) markers. In the common garden study, 25 lines with strongly contrasting genotypes were planted in close proximity. We fingerprinted 10 seed progeny from 8 individuals of each line and detected 15 first-generation heterozygotes for a t-value (corrected for cryptic crosses) of 0.0082. Different genotypes were significantly overrepresented as maternal versus paternal parents of heterozygotes, suggesting gender-function-dependent genetic control of outcrossing rates. In 4 wild populations (>300 individuals each), expected heterozygosity ranged from 0.149 to 0.336, whereas t-values ranged from 0.0027 to 0.0133, indicating high levels of both genetic diversity and inbreeding. Up to a third of the individuals in each population belonged to groups with identical or nearly identical SNP genotypes, whereas many of the remaining individuals were members of loose clusters of apparently related plants that probably represent descendants from past outcrossing events. Strict inbreeding in some lineages within a population with occasional outcrossing in others may be related to positive selection on adaptive syndromes associated with specific inbreeding lineages, or possibly to among-lineage differences in genetic regulation of outcrossing.

Pollinator-mediated competition between two co-flowering Neotropical mangrove species, Avicennia germinans (Avicenniaceae) and Laguncularia racemosa (Combretaceae)

Three ecological relationships are possible between co-flowering plant species; they may have no effect on one another, compete for pollination services, or facilitate one another by attracting more pollinators to the area. In this study, the pollinator-mediated relationship between two mangrove species with overlapping flowering phenologies was investigated in one south Florida community. Pollinator observations were recorded between 0900 h and 1700 h during June and July, 2008-2010. Insect visitation rates to Avicennia germinans and Laguncularia racemosa were estimated from 522 observation intervals of 10 min during three phenological time periods, when each species flowered alone and when they co-flowered. The number of timed intervals varied between years due to differences in flowering phenology, from four to 42 for A. germinans and from nine to 94 for L. racemosa. Avicennia germinans began flowering first in all years, and insect visitation rates were significantly greater to A. germinans than to L. racemosa (P<0·001). Flowers of both species received visits from bees, wasps, flies and butterflies; Apis mellifera was the most common floral visitor to both species. Visitation rates to L. racemosa increased significantly when A. germinans stopped flowering (P<0·001). However, there was no significant change in visitation rates to A. germinans after L. racemosa began flowering (P=0·628). When they co-flowered, A. germinans outcompeted L. racemosa for pollinators. Laguncularia racemosa hermaphrodites self-pollinate autogamously when not visited by insects, so reduced visitation to L. racemosa flowers reduced the frequency of outcrossing and increased the frequency of selfing. Reduced outcrossing limits male reproductive success in this androdioecious species, which could lead to changes in the breeding system. The degree of overlap in flowering phenologies varied between years, so the effect on the mating and breeding system may differ between years.

Outcrossing frequencies from multiple high erucic acid oilseed rape fields to a central receptor field

Genetically modified oilseed rape is currently grown on about 23 % of the global oilseed rape acreage. In order to separate transgenic and non-transgenic oilseed rape production and to ensure co-existence of different agricultural cultivation schemes, as is specified by the European legislation, confinement measures have to be defined. Pollen-mediated gene flow is the most important means by which transgenes are dispersed between fields. In contrast to the majority of the previous investigations the objective of this study was to assess the extent of gene flow in the case of multiple pollen-donor fields. A high erucic acid rape genotype was used as biochemical marker for the quantification of outcrossing into a low erucic acid oilseed rape variety. Outcrossing data were obtained from two experimental locations. As expected, multiple pollen sources in a fragmented landscape can result in high gene transfer frequencies, thus requiring larger isolation distances than a field design with a single pollen donor source. The results of the study are transferable to homozygous transgenic oilseed rape varieties.

Insect visitation rates and foraging patterns differ in androdioecious and hermaphrodite-only populations of Laguncularia racemosa (Combretaceae) in Florida

Abstract:Insect-pollinated Laguncularia racemosa has a variable breeding system; some populations are androdioecious, with male and hermaphroditic plants, while others lack male plants. We observed the foraging behaviours of insects in three androdioecious and three hermaphrodite-only populations of L. racemosa in Florida. In each population, insect visitation rates were estimated from 30–108 timed intervals. We recorded the number of flowers visited by 144–224 insects during foraging bouts made to 15–40 male and hermaphroditic plants. Male plants in androdioecious populations had significantly more visitors than hermaphroditic plants, increasing the number of vectors carrying pollen from male plants. Further, many insects visited few flowers during foraging bouts, which should increase outcrossing frequency. According to mathematical models, male plants benefit from these combined factors. Plants in hermaphrodite-only populations had significantly more visitors than hermaphroditic plants in androdioecious populations. Proportionately more insects visited many flowers during foraging bouts in hermaphrodite-only versus androdioecious populations. The increased likelihood of geitonogamous self-pollination could help explain the lack of male plants in hermaphrodite-only populations. Differences in pollinator assemblages and the relative abundances of several species were responsible for differences in foraging behaviours: Apis mellifera, Bombus sp., Melissodes sp., Xylocopa sp., Euodynerus sp. and a calliphorid species.