The commercialization of imazethapyr-resistant (Clearfield™, CL) rice in the southern United States has raised serious concerns about gene flow to red rice, producing imazethapyr-resistant red rice populations. Our objectives were to determine the impact of planting date, CL cultivars, and red rice biotypes on outcrossing rate; and to investigate the relative contribution of flowering time of CL rice and red rice biotypes, together with air temperature and relative humidity (RH), on outcrossing rate. Field experiments were conducted at Stuttgart, Rohwer, and Kibler, AR, from 2005 to 2007, at three or four planting times from mid-April to late May. ‘CL161’ (inbred cultivar) and ‘CLXL8’ (hybrid) rice were planted in nine-row plots, with red rice planted in the middle row. Twelve red rice biotypes were used. The flowering of red rice and CL rice, air temperature, and RH were recorded. Red rice seeds were collected at maturity. To estimate outcrossing rate, resistance to imazethapyr was evaluated in subsequent years and confirmed using rice microsatellite markers. CLXL8 rice flowered 2 to 4 d earlier than CL161 rice, and flowering was completed within 1 wk in all plantings. The flowering duration of most red rice biotypes ranged from 4 to 17 d. Flowering synchrony of red rice biotypes and CL rice ranged from 0 to 100% at different plantings. In general, CLXL8 had greater flowering overlap and higher outcrossing rate with red rice than did CL161 rice. The outcrossing rate of red rice biotypes ranged from 0 to 0.21% and 0 to 1.26% with CL161 and CLXL8 rice, respectively. The outcrossing rate differed within each planting date (P < 0.05). Outcrossing was generally lower in mid-May and late May than in mid-April and late April planting times. Flowering synchrony and outcrossing rate were not correlated (r2 < 0.01). Outcrossing with CL161 was primarily influenced by red rice biotype. A minimum air temperature of > 24 C in the evening also favors outcrossing with CL161. With CLXL8 rice, outcrossing was most affected by RH. When RH was < 54%, outcrossing was less (0.12%) than when RH was ≥ 54% (0.38%). With CLXL8 rice, a minimum RH of ≥ 54%, from mid-morning to noon, increased outcrossing with red rice. To fully understand the interaction effects of these factors on outcrossing with red rice, controlled experiments are needed.

A population model was constructed to simulate the development of imazamox-resistant jointed goatgrass (AEGCY) in imazamox-resistant (Clearfield™) wheat. The model computed changes in the surface and in the buried AEGCY seed banks for both resistant and susceptible biotypes. Simulations started with an initial density of 1,000 susceptible and zero resistant seeds/m2 in each seed bank. Simulation of continuous, no-till Clearfield wheat production resulted in rapid development of resistant AEGCY without hybridization with wheat and in extremely rapid resistance development with hybridization. In less than 10 yr, the resistant population was growing exponentially in both simulations. Adding a fallow year with tillage into the simulated rotation did not substantially slow down the appearance of resistance but did delay the rate of resistant population increase by several orders of magnitude over 10 yr. Alternating Clearfield and a nonresistant winter wheat in combination with fallowing prevented the establishment of a significant resistant AEGCY population and prevented the susceptible seed population from increasing exponentially. These projections suggest that imazamox-resistant wheat can be a tool for managing AEGCY populations especially if combined with rotations that include fallow and crops other than Clearfield winter wheat.

Efficient natural dispersal of herbicide-resistance alleles via seed and pollen can markedly accelerate the incidence of herbicide-resistant weed populations across an agroecoregion. Studies were conducted in western Canada in 2014 and 2015 to investigate pollen- and seed-mediated gene flow in kochia. Pollen-mediated gene flow (PMGF) from glyphosate-resistant (GR) to non-GR kochia was quantified in a field trial (hub and spoke design) at Saskatoon, Saskatchewan. Seed-mediated gene flow of acetolactate synthase (ALS) inhibitor-resistant kochia as a function of tumbleweed speed and distance was estimated in cereal stubble fields at Lethbridge, Alberta and Scott, Saskatchewan. Regression analysis indicated that outcrossing from GR to adjacent non-GR kochia ranged from 5.3 to 7.5%, declining exponentially to 0.1 to 0.4% at 96 m distance. However, PMGF was significantly influenced by prevailing wind direction during pollination (maximum of 11 to 17% outcrossing down-wind). Seed dropped by tumbleweeds varied with distance and plant speed, approaching 90% or more (ca. 100,000 seeds or more) at distances of up to 1,000 m and plant speeds of up to 300 cm s–1. This study highlights the efficient proximal (pollen) and distal (seed) gene movement of this important GR weed.

Two separate field experiments were conducted to quantify the degree of plant-to-plant outcrossing and pollen-mediated gene flow (PMGF) in wild oat. The purpose of the study was to determine the extent to which pollen movement could contribute to the spread of herbicide resistance in this species. In both experiments, an acetyl-CoA carboxylase inhibitor–resistant (R) wild oat genotype (UM1) was used as the pollen donor and a susceptible (S) genotype (UM5) was used as the pollen receptor. Hybrid progeny resulting from a cross between UM1 and UM5 were identified using the herbicide resistance trait as a marker. In the plant-to-plant outcrossing experiment, single UM5 plants were closely surrounded by 20 homozygous R UM1 plants in hills. By screening seed from the S parent for resistance, outcrossing was determined to range from 0 to 12.3%, with a mean of 5.2% over 10 hills. In the PMGF experiment, single homozygous R UM1 plants were surrounded by UM5 plants arranged in a hexagonal pattern at low and high densities (total of 19 and 37 wild oat plants m−2), growing within spring wheat and flax crops. In the wheat crop, mean wild oat outcrossing was 0.08 and 0.05% at low and high densities, respectively. In the less competitive flax, corresponding outcrossing values were 0.07 and 0.16% at low and high densities, respectively. Distance from the pollen source was a significant factor only for the high-density planting arrangement in flax. Up to 77 R hybrid seeds were recovered from 6 m2 in the PMGF experiment, indicating that PMGF contributes to the evolution of resistance in wild oat populations. However, the contribution of pollen movement to resistance evolution and the spread of resistance in wild oat populations would be relatively small when compared with R seed production and dispersal from a resistant plant.

EDITOR'S NOTE: This manuscript was reviewed by six colleagues whose recommendations varied widely. Lack of repetition was a major concern. The authors address the problem in the last paragraph of the results section. Factors favoring publication included the worldwide importance of wild oats, the minimal data on gene flow in the species, and the fact that the results are consistent with those of other studies cited in this manuscript. The points raised by reviewers who did not favor publication, especially the role of the environment in pollen production and viability, are acknowledged.

R. L. Zimdahl, Editor

The transfer of the imazethapyr-resistant gene from Clearfield™ (CL) rice to red rice is an ecological risk. Flowering synchronization and genetic compatibility between cultivated rice and red rice could influence gene transfer. We examined the (1) variability in maximum outcrossing rate between 12 red rice biotypes and ‘CL161’ rice during their peak flowering overlap in the field and (2) genetic compatibility of red rice biotypes with CL161 rice. Experiments were conducted at Stuttgart, AR, and Fayetteville, AR, from 2005 to 2007. To evaluate the flowering synchrony of red rice and CL161 rice as well as its impact on outcrossing rate, field experiments were conducted at four planting times from early April to late May. The red rice biotypes were planted in the middle row of nine-row CL161 plots and flowering was monitored. Outcrosses were evaluated in subsequent years by herbicide response and simple-sequence-repeat marker assays. To determine compatibility, manual crosses were performed between 12 red rice biotypes and CL161 rice in the greenhouse. The flowering duration of all red rice types ranged from 5 to 16 d after the onset of flowering in contrast to 6 d in CL161 rice. Ten of the twelve types of red rice had ≥ 70% overlap in flowering time with CL161 rice in at least one planting date. The maximum field outcrossing rate between red rice biotypes and CL161 ranged from 0.03 to 0.25%. The field outcrossing rate between red rice biotypes differed (P < 0.01), but flowering synchronization was not directly related to outcrossing rate. Manual crosses resulted in seed sets of 49 to 94%. The majority of red rice biotypes had similar compatibility with CL161 rice. Thus, other factors must contribute to hybridization rates in the field. Follow-up experiments should investigate other plant factors and environmental influence on hybridization rate.

In areas with less favourable conditions for agriculture, informal seed systems permit gene flow through pollen to play a crucial role in the development of new varieties. An important factor with great impact on cross-pollination is the plant breeding system, but so far this is little studied within the context of low-input farming systems. This research studied the chances of cross-pollination within and between rice fields in The Gambia. Size and time of flowering were measured for 28 rice fields in one village. The level of mixture was measured in 90 seed lots of rice collected from four villages. Based on the results, we suggest that in general cross-pollination between different rice genotypes occurs more often within fields than between fields. No clear relationship was found between the level of within-field mixture and socio-economic status of farmers. Some comparison was made with millet, which allowed the identification of various factors influencing pollen flow between different genotypes. Effective pollen flow (between genotypes) is a function of a number of factors, such as the rate of cross-pollination of a crop, number of off-types within fields, variety distinctiveness, farmer expert knowledge, length and reliability of the rainy season, growth duration of different varieties, availability of fields, pest pressure and number of varieties grown per field or per farmer. We hypothesize that a low cross-pollination rate is more favourable for the development of new varieties in farmer fields than a high cross-pollination rate.

Hybridization and introgression between crops and wild relatives may have important evolutionary and ecological consequences such as gene swamping or increased invasiveness. In the present study, we investigated hybridization under field conditions between crop lettuce (Lactuca sativa) and its wild relative prickly lettuce (L. serriola), two cross-compatible, predominantly autogamous and insect pollinated species. In 2003 and 2004, we estimated the rates of hybridization between L. sativa and L. serriola in close-to-reality field experiments carried out in two locations of Northern Switzerland. Seeds set by the experimental wild plants were collected and sown (44 352 in 2003 and 252 345 in 2004). Progeny was screened morphologically for detecting natural hybrids. Prior to the experiment, specific RAPD markers were used to confirm that morphological characters were reliable for hybrid identification. Hybridization occurred up to the maximal distance tested (40 m), and hybridization rates varied between 0 to 26%, decreasing with distance. More than 80% of the wild plants produced at least one hybrid (incidence of hybridization, IH) at 0 m and 1 m. It equaled 4 to 5% at 40 m. In sympatric crop-wild populations, cross-pollination between cultivated lettuce and its wild relative has to be seen as the rule rather than the exception for short distances.

Concerns have been raised in Europe about the efficiency, sustainability, and environmental impact of the first genetically modified crops. The committees and regulators in charge of approving procedures have encouraged a field trial approach for safety assessment studies under current agronomic conditions. We describe the gene flow from sugar beet (Beta vulgaris L.) in a multi-year and multi-crop monitoring study on farmers' fields at two locations that has been carried out since 1995. We analyzed two sugar beet lines that have been genetically transformed for herbicide resistance. One sugar beet has resistance to glufosinate and the other to glyphosate. Large differences among lines, years and locations were observed. These differences provided a broad range of situations to estimate the risks. Sugar beet bolters produced the majority (86%) of the herbicide-resistant seeds harvested in the field. Direct pollen flow from sugar beet bolters to weed beets that were growing within the same field as well as in a neighboring field that was left fallow accounted for only 0.4% of the resistant seeds released over the years and locations. Descendants of the hybrids between the sugar beet and the weed beet produced the remaining 13.6% of resistant seeds. Herbicide-resistant seeds from the progeny of the weed beet were recorded up to 112 m away from the closest transgenic pollen donor. Indications were observed of non-randomness of the weed beet producing resistant progeny. We also analyzed pollen flow to male-sterile bait plants located within and outside of the sugar beet field. Herbicide-resistant pollen flow was recorded up to 277 m, and fitted with an inverse power regression. Using sugar beet varieties with no, or very low, sensitivity to bolting and destroying bolters are two necessary measures that could delay gene flow.

We have assessed the utility of morphological and microsatellite markers for tracing field hybridization between Lolium multiflorum and Lolium perenne in cereal-enclosed gene flow plots. The presence of awns on the inflorescence of F1 hybrids was found to be a reliable, but underscoring, indicator of L. multiflorum paternity in L. perenne derived seed as determined by inheritance of species-specific alleles at the microsatellite locus ‘H01 H06’ in these progeny. A positive correlation was evident in the experimental treatment between the number of pollen donor plants in a given plot and the frequency of hybrid F1 seed harvested from pollen receptor plants in that plot. These experiments have established the utility of naturally occurring heritable markers for the measurement of gene flow rates in field Ryegrass populations, with particular significance for risk assessment modeling of potential gene flow from transgenic grass cultivars.

This study deals with hybrids between Aegilops geniculata and bread wheat, Triticum aestivum L., detected in two Ae. geniculata populations in the “Meseta Central”, Spain’s central plateau where wheat is a major crop. Morphological traits and pentaploid chromosome numbers were used to identify hybrids in 2004 and 2005. The frequency of hybridization under natural conditions was calculated for one of these populations. Six hybrids were detected from a sample of 3158 seeds from this population. The hybrids were mainly sterile but seed set can be found. This study extends the range of documented hybridization to include central Spain. The potential risks associated with natural hybridization in the context of transgenic wheat cultivation are discussed.

The ongoing discussion on the co-existence between genetically modified (GM) and non-GM crops becomes more important in the European Union (EU). With the recent inscription of 17 GM maize varieties in the common EU catalogue of varieties of agricultural plant species, the acreage of transgenic maize for market purposes is expected to increase in some European countries. In the EU, specific tolerance thresholds have been established for the adventitious and technically unavoidable presence of GM material in non-GM produce, and member states are elaborating legal frames to cope with co-existence. As maize is a cross-pollinated crop relying on wind for the dispersal of its pollen, technical management measures will be imposed to reduce cross-fertilization between transgenic and non-transgenic maize. Various biological, physical and analytical parameters have been identified to play a role in the study of cross-fertilization in maize. This variability may hamper the comparison between research results and may complicate the definition of appropriate isolation distances and/or pollen barriers in order to limit out-crossing. The present review addresses these parameters and proposes containment measures in order to not exceed the legal labeling thresholds in maize.

There is now considerable evidence for disruption of plant–pollinator mutualisms in fragmented landscapes. The causes can be myriad, and vary from lack of nesting sites for key insect pollinators, to a decline in pollinator visits when plant population sizes are reduced by fragmentation. One important and well-documented consequence of disruption in pollination systems is reduced seed production in plant populations in fragments as a result of pollen-limitation. When reduced seed production, in turn, has a negative effect on seedling recruitment, populations may enter a spiral towards extinction. Some pollination systems, on the other hand, are surprisingly resilient to anthropogenic changes, and pollen flow between trees in fragments separated by several kilometres has been reported recently. Understanding how reproductive attributes of plants, such as breeding systems and specificity of pollination systems, affect the outcome of habitat fragmentation remains a key challenge.

We monitored pollen-mediated transgene dissemination from commercial transgenic squash CZW-3 into its wild relative Cucurbita pepo ssp. ovifera var. texana (C. texana). Transgenic squash CZW-3 expresses the neomycin phosphotransferase II (nptII) gene and the coat protein (CP) genes of Cucumber mosaic virus (CMV), Zucchini yellow mosaic virus (ZYMV), and Watermelon mosaic virus (WMV); thereby, it is resistant to these three aphid-borne viruses. The rate of NPT II and CP transgene introgression increased with overlapping flowering patterns and a high ratio of transgenic F1 hybrids (C. texana × CZW-3) to C. texana. Transgene transfer also readily occurred from transgenic F1 hybrids into C. texana over three generations in field settings where test plants grew sympatrically and viruses were not severely limiting the growth, and fruit and seed production of C. texana. In contrast, introgression of the transgenes into C. texana was not sustained under conditions of high viral disease pressure. As expected, C. texana progeny that acquired the CP transgenes exhibited resistance to CMV, ZYMV, and WMV. This is the first report on transgene dissemination from a transgenic crop that exhibits disease resistance and hybridizes with a wild plant species without loss of fertility.