Introgression from local cultivars is a driver of agricultural adaptation in Argentinian weedy rice.

Weedy rice, a pervasive and troublesome weed found across the globe, has often evolved through fertilization of rice cultivars with little importance of crop-weed gene flow. In Argentina, weedy rice has been reported as an important constraint since the early 1970s, and, in the last few years, strains with herbicide-resistance are suspected to evolve. Despite their importance, the origin and genetic composition of Argentinian weedy rice as well its adaptation to agricultural environments has not been explored so far. To study this, we conducted genotyping-by-sequencing on samples of Argentinian weedy and cultivated rice and compared them with published data from weedy, cultivated and wild rice accessions distributed worldwide. In addition, we conducted a phenotypic characterization for weedy-related traits, a herbicide resistance screening and genotyped accessions for known mutations in the acetolactate synthase (ALS) gene, which confers herbicide resistance. Our results revealed large phenotypic variability in Argentinian weedy rice. Most strains were resistant to ALS-inhibiting herbicides with a high frequency of the ALS mutation (A122T) present in Argentinian rice cultivars. Argentinian cultivars belonged to the three major genetic groups of rice: japonica, indica and aus while weeds were mostly aus or aus-indica admixed, resembling weedy rice strains from the Southern Cone region. Phylogenetic analysis supports a single origin for aus-like South American weeds, likely as seed contaminants from the United States, and then admixture with local indica cultivars. Our findings demonstrate that crop to weed introgression can facilitate rapid adaptation to agriculture environments.

The role of intraspecific crop-weed hybridization in the evolution of weediness and invasiveness: Cultivated and weedy radish (Raphanus sativus) as a case study.

PREMISE: The phenotype of hybrids between a crop and its wild or weed counterpart is usually intermediate and maladapted compared to that of their parents; however, hybridization has sometimes been associated with increased fitness, potentially leading to enhanced weediness and invasiveness. Since the ecological context and maternal genetic effects may affect hybrid fitness, they could influence the evolutionary outcomes of hybridization. Here, we evaluated the performance of first-generation crop-weed hybrids of Raphanus sativus and their parents in two contrasting ecological conditions. METHODS: Using experimental hybridization and outdoor common garden experiments, we assessed differences in time to flowering, survival to maturity, plant biomass, and reproductive components between bidirectional crop-weed hybrids and their parents in agrestal (wheat cultivation, fertilization, weeding) and ruderal (human-disturbed, uncultivated area) conditions over 2 years. RESULTS: Crop, weeds, and bidirectional hybrids overlapped at least partially during the flowering period, indicating a high probability of gene flow. Hybrids survived to maturity at rates at least as successful as their parents and had higher plant biomass and fecundity, which resulted in higher fitness compared to their parents in both environments, without any differences associated with the direction of the hybridization. CONCLUSIONS: Intraspecific crop-weed hybridization, regardless of the cross direction, has the potential to promote weediness in weedy R. sativus in agrestal and ruderal environments, increasing the chances for introgression of crop alleles into weed populations. This is the first report of intraspecific crop-weed hybridization in R. sativus.

Agricultural weeds: the contribution of domesticated species to the origin and evolution of feral weeds.

Agricultural weeds descended from domesticated ancestors, directly from crops (endoferality) and/or from crop-wild hybridization (exoferality), may have evolutionary advantages by rapidly acquiring traits pre-adapted to agricultural habitats. Understanding the role of crops on the origin and evolution of agricultural weeds is essential to develop more effective weed management programs, minimize crop losses due to weeds, and accurately assess the risks of cultivated genes escaping. In this review, we first describe relevant traits of weediness: shattering, seed dormancy, branching, early flowering and rapid growth, and their role in the feralization process. Furthermore, we discuss how the design of "super-crops" can affect weed evolution. We then searched for literature documenting cases of agricultural weeds descended from well-domesticated crops, and describe six case studies of feral weeds evolved from major crops: maize, radish, rapeseed, rice, sorghum, and sunflower. Further studies on the origin and evolution of feral weeds can improve our understanding of the physiological and genetic mechanisms underpinning the adaptation to agricultural habitats and may help to develop more effective weed-control practices and breeding better crops. © 2022 Society of Chemical Industry.

Rapid Evolution of Seed Dormancy During Sunflower De-Domestication.

Hybridization between crops and their wild relatives may promote the evolution of de-domesticated (feral) weeds. Wild sunflower (Helianthus annuus L.) is typically found in ruderal environments, but crop-wild hybridization may facilitate the evolution of weedy populations. Using 1 crop-specific mitochondrial marker (CMS-PET1) and 14 nuclear SSR markers, we studied the origin and genetic diversity of a recently discovered weedy population of sunflower (named BRW). Then, using a resurrection approach, we tested for rapid evolution of weedy traits (seed dormancy, herbicide resistance, and competitive ability) by sampling weedy and wild populations 10 years apart (2007 and 2017). All the weedy plants present the CMS-PET1 cytotype, confirming their feral origin. At the nuclear markers, BRW showed higher genetic diversity than the cultivated lines and low differentiation with one wild population, suggesting that wild hybridization increased their genetic diversity. We found support for rapid evolution towards higher seed dormancy, but not for higher competitive ability or herbicide resistance. Our results highlight the importance of seed dormancy during the earliest stages of adaptation and show that crop-wild hybrids can evolve quickly in agricultural environments.

Molecular characterization of imidazolinone-resistant Brassica rapa × B. napus hybrids.

Herbicide-resistant oilseed rape (Brassica napus) cultivation in our country entails the risk of gene transfer to related wild species. One of these species is the wild turnip (B. rapa), an important weed of winter crops widely distributed in the Pampas region. Despite hybridization risks, Clearfield ® oilseed rape is available in Argentina. In 2008, a B. rapa population, which was sympatric to an imidazolinone-resistant and a conventional oilseed rape cultivar, was located on a farm in the main cropping area of the country. Herbicide-resistant individuals were found in the progeny of this population in a herbicide screening test. Therefore, a molecular characterization using cleaved amplified polymorphic sequence (CAPS) and simple sequence repeat (SSR) markers was conducted on these plants to determine their hybrid nature and to establish the origin of the imidazolinone resistance trait. The results of this study, along with information of field records, confirmed that the resistant plants were first generation interspecific hybrids. Imidazolinone resistance had been effectively transferred from the herbicide-resistant oilseed rape, even in the particular situation of pollen competition. Oilseed rape resistant cultivars are becoming more common in the country. So, considering that seed loss and crop volunteers are common in these species, it is crucial to avoid the dispersion of new resistant weed biotypes as they reduce the effectiveness of chemical control technologies.

Diversity of Naturalized Hairy Vetch (Vicia villosa Roth) Populations in Central Argentina as a Source of Potential Adaptive Traits for Breeding.

Hairy vetch (Vicia villosa ssp. villosa Roth) is native of Europe and Western Asia and it is the second most cultivated vetch worldwide. Hairy vetch is used as forage species in semiarid environments and as a legume cover crop in sub-humid and humid regions. Being an incompletely domesticated species, hairy vetch can form spontaneous populations in a new environment. These populations might contain novel and adaptive traits valuable for breeding. Niche occupancy based on geographic occurrence and environmental data of naturalized populations in central Argentina showed that these populations were distributed mainly on disturbed areas with coarse soil texture and alkaline-type soils. Low rainfall and warm temperatures during pre- and post-seed dispersal explained the potential distribution under sub-humid and semiarid conditions from Pampa and Espinal ecoregions. Conversely, local adaptation along environmental gradients did not drive the divergence among recently established Argentinian (AR) populations. The highest genetic diversity revealed by microsatellite analysis was observed within accessions (72%) while no clear separation was detected between AR and European (EU) genotypes, although naturalized AR populations showed strong differentiation with the wild EU accessions. Common garden experiments were conducted in 2014-16 in order to evaluate populations' germination, flowering, and biomass traits. European cultivars were characterized by low physical seed dormancy (PY), while naturalized AR accessions showed higher winter biomass production. Detected variation in the quantitative assessment of populations could be useful for selection in breeding for traits that convey favorable functions within specific contexts.

Phenotypic selection under two contrasting environments in wild sunflower and its crop-wild hybrid.

Hybridization is a common phenomenon in plants and can lead to the introgression of alleles from one population into another, generate new hybrid lineages, or cause species extinction. The environmental conditions and the genetic background of the participating populations may influence these outcomes since they can affect the fitness of hybrids, thereby increasing or decreasing the chances of introgression. Thus, it is important to understand the context-dependent prospects for introgression of alleles into diverse populations and under multiple ecological environments. Crop-wild hybridization presents an opportunity to explore these dynamics in agroecosystems. To this end, we used diverse wild and hybrid sunflowers from across the northern United States as a basis for evaluating variation in morphological traits and assessing context-dependent selection. These crop-wild hybrids and their wild counterparts were grown under agricultural conditions in the field with and without wheat competition. Interactions between origin and cross type affected expression of early functional traits, while interactions between competition and cross type acted on reproductive traits. A smattering of early and reproductive traits was affected by interactions between cross type and competition that varied by origin (i.e., 3-way interactions). Seven functional traits, especially number of branches and tertiary head diameter, underwent net and direct directional selection, while six out of these seven traits appear to also be experiencing nonlinear selection dynamics. In general, wild-like traits were favored under both sets of conditions, while, under wheat competition, some crop-like traits related to fast growth and primary head diameter became important. These data reaffirm the hypothesis that stressful conditions establish a scenario more suitable for crop introgression and clarify that nonlinear selection dynamics may play a role in this process.

Genetic Diversity and Population Structure of Wild Sunflower (Helianthus annuus L.) in Argentina: Reconstructing Its Invasion History.

Studying the levels and patterns of genetic diversity of invasive populations is important to understand the evolutionary and ecological factors promoting invasions and for better designing preventive and control strategies. Wild sunflower (Helianthus annuus L.) is native to North America and was introduced, and has become invasive, in several countries, including Argentina (ARG). Here, using classical population genetic analyses and approximate Bayesian computation (ABC) modeling, we studied the invasion history of wild sunflower in ARG. We analyzed 115 individuals belonging to 15 populations from ARG (invasive range) and United States (US, native range) at 14 nuclear and 3 chloroplast simple sequence repeat markers along with 23 phenotypic variables. Populations from ARG showed similar levels of nuclear genetic diversity to US populations and higher genetic diversity in the chloroplast genome, indicating no severe genetic bottlenecks during the invasion process. Bayesian clustering analysis, based on nuclear markers, suggests the presence of 3 genetic clusters, all present in both US and ARG. Discriminant analysis of principal components (DAPC) detected an overall low population structure between central US and ARG populations but separated 2 invasive populations from the rest. ABC modeling supports multiple introductions but also a southward dispersal within ARG. Genetic and phenotypic data support the central US as a source of introduction while the source of secondary introductions could not be resolved. Finally, using genetic markers from the chloroplast genome, we found lower population structure in ARG when compared with US populations, suggesting a role for seed-mediated gene flow in Argentina.

AHAS Trp574Leu substitution in Raphanus sativus L.: screening, enzyme activity and fitness cost.

BACKGROUND: Feral radish (Raphanus sativus L.) is a problematic weed that has become resistant to acetohydroxyacid synthase (AHAS) inhibitor herbicides due to the Trp574Leu mutation. An AHAS gene mutation that causes herbicide resistance may have negative pleiotropic effects on plant fitness. This study reports the effects of the Trp574Leu mutation on AHAS activity and reproductive traits of R. sativus. RESULTS: Eight of 17 feral radish accessions presented individuals resistant to metsulfuron-methyl at 0.5% to >90.0% and all the resistant individuals analyzed showed the Trp574Leu mutation. Without herbicide selection, the AHAS activity was 3.2-fold higher in the susceptible accession than in the resistant one. The resistant accession was >9000-fold more resistant to metsulfuron-methyl and imazethapyr than the susceptible accession. Under low intraspecific competition during two growing seasons, AHAS-resistant feral radish accessions showed 22-38% and 21-47% lower seed numbers and yield per plant than the susceptible accession. CONCLUSION: This is the first report of fitness cost associated with the AHAS Trp574Leu mutation in R. sativus populations. This fitness cost could reduce frequency of the resistant allele without herbicide selection. © 2018 Society of Chemical Industry.

Transgene escape and persistence in an agroecosystem: the case of glyphosate-resistant Brassica rapa L. in central Argentina.

Brassica rapa L. is an annual Brassicaceae species cultivated for oil and food production, whose wild form is a weed of crops worldwide. In temperate regions of South America and especially in the Argentine Pampas region, this species is widely distributed. During 2014, wild B. rapa populations that escaped control with glyphosate applications by farmers were found in this area. These plants were characterized by morphology and seed acidic profile, and all the characters agreed with B. rapa description. The dose-response assays showed that the biotypes were highly resistant to glyphosate. It was also shown that they had multiple resistance to AHAS-inhibiting herbicides. The transgenic origin of the glyphosate resistance in B. rapa biotypes was verified by an immunological test which confirmed the presence of the CP4 EPSPS protein and by an event-specific GT73 molecular marker. The persistence of the transgene in nature was confirmed for at least 4 years, in ruderal and agrestal habitats. This finding suggests that glyphosate resistance might come from GM oilseed rape crops illegally cultivated in the country or as a seed contaminant, and it implies gene flow and introgression between feral populations of GM B. napus and wild B. rapa. The persistence and spread of the resistance in agricultural environments was promoted by the high selection pressure imposed by intensive herbicide usage in the prevalent no-till farming systems.

Transgenic glyphosate-resistant oilseed rape (Brassica napus) as an invasive weed in Argentina: detection, characterization, and control alternatives.

The presence of glyphosate-resistant oilseed rape populations in Argentina was detected and characterized. The resistant plants were found as weeds in RR soybeans and other fields. The immunological and molecular analysis showed that the accessions presented the GT73 transgenic event. The origin of this event was uncertain, as the cultivation of transgenic oilseed rape cultivars is prohibited in Argentina. This finding might suggest that glyphosate resistance could come from unauthorized transgenic oilseed rape crops cultivated in the country or as seed contaminants in imported oilseed rape cultivars or other seed imports. Experimentation showed that there are alternative herbicides for controlling resistant Brassica napus populations in various situations and crops. AHAS-inhibiting herbicides (imazethapyr, chlorimuron and diclosulam), glufosinate, 2,4-D, fluroxypyr and saflufenacil proved to be very effective in controlling these plants. Herbicides evaluated in this research were employed by farmers in one of the fields invaded with this biotype and monitoring of this field showed no evidence of its presence in the following years.

Broad resistance to acetohydroxyacid-synthase-inhibiting herbicides in feral radish (Raphanus sativus L.) populations from Argentina.

BACKGROUND: Soon after the commercial release of sunflower cultivars resistant to imidazolinone herbicides, several uncontrolled feral radish (Raphanus sativus L.) populations were found in south-eastern Buenos Aires, Argentina. These populations were studied in field, glasshouse and laboratory experiments aiming to characterise their resistance profile and to develop management tools. RESULTS: Three feral radish accessions were highly resistant to ten active ingredients of five families of acetohydroxyacid synthase (AHAS)-inhibiting herbicides. Sequence analysis of the AHAS gene detected a Trp574Leu mutation in all resistant accessions. One accession with an intermediate level of resistance was heterozygous for this mutation, probably owing to gene exchange with a susceptible subpopulation located in the field margin. Herbicide-resistant and herbicide-susceptible radish could be controlled in sunflower by alternative herbicides. CONCLUSION: This is the first report of feral radish with resistance to herbicides belonging to all the AHAS-inhibiting herbicide families, conferred by Trp574Leu mutation in the AHAS gene. An appropriate herbicide rotation with alternative herbicides such as fluorochloridone or aclonifen and an increase in the diversity of cropping systems are important for minimising the prevalence of these biotypes.