Hugh J. Beckie

Hybridization between transgenic Brassica napus L. and its wild relatives: Brassica rapa L., Raphanus raphanistrum L., Sinapis arvensis L., and Erucastrum gallicum (Willd.) O.E. Schulz

Abstract. The frequency of gene flow from Brassica napus L. (canola) to four wild relatives, Brassica rapa L., Raphanus raphanistrum L., Sinapis arvensis L. and Erucastrum gallicum (Willd.) O.E. Schulz, was assessed in greenhouse and/or field experiments, and actual rates measured in commercial fields in Canada. Various marker systems were used to detect hybrid individuals: herbicide resistance traits (HR), green fluorescent protein marker (GFP), species-specific amplified fragment length polymorphisms (AFLPs) and ploidy level. Hybridization between B. rapa and B. napus occurred in two field experiments (frequency approximately 7%) and in wild populations in commercial fields (approximately 13.6%). The higher frequency in commercial fields was most likely due to greater distance between B. rapa plants. All F1 hybrids were morphologically similar to B. rapa, had B. napus- and B. rapa-specific AFLP markers and were triploid (AAC, 2n = 29 chromosomes). They had reduced pollen viability (about 55%) and segregated for both self-incompatible and self-compatible individuals (the latter being a B. napus trait). In contrast, gene flow between R. raphanistrum and B. napus was very rare. A single R. raphanistrum × B. napus F1 hybrid was detected in 32,821 seedlings from the HR B. napus field experiment. The hybrid was morphologically similar to R. raphanistrum except for the presence of valves, a B. napus trait, in the distorted seed pods. It had a genomic structure consistent with the fusion of an unreduced gamete of R. raphanistrum and a reduced gamete of B. napus (RrRrAC, 2n = 37), both B. napus- and R. raphanistrum-specific AFLP markers, and had <1% pollen viability. No hybrids were detected in the greenhouse experiments (1,534 seedlings), the GFP field experiment (4,059 seedlings) or in commercial fields in Québec and Alberta (22,114 seedlings). No S. arvensis or E. gallicum × B. napus hybrids were detected (42,828 and 21,841 seedlings, respectively) from commercial fields in Saskatchewan. These findings suggest that the probability of gene flow from transgenic B. napus to R. raphanistrum, S. arvensis or E. gallicum is very low (<2–5 × 10–5). However, transgenes can disperse in the environment via wild B. rapa in eastern Canada and possibly via commercial B. rapa volunteers in western Canada.

Herbicide-Resistant Weeds: Management Tactics and Practices1

In input-intensive cropping systems around the world, farmers rarely proactively manage weeds to prevent or delay the selection for herbicide resistance. Farmers usually increase the adoption of integrated weed management practices only after herbicide resistance has evolved, although herbicides continue to be the dominant method of weed control. Intergroup herbicide resistance in various weed species has been the main impetus for changes in management practices and adoption of cropping systems that reduce selection for resistance. The effectiveness and adoption of herbicide and nonherbicide tactics and practices for the proactive and reactive management of herbicide-resistant (HR) weeds are reviewed. Herbicide tactics include sequences and rotations, mixtures, application rates, site-specific application, and use of HR crops. Nonherbicide weed-management practices or nonselective herbicides applied preplant or in crop, integrated with less-frequent selective herbicide use in diversified cropping systems, have mitigated the evolution, spread, and economic impact of HR weeds. Additional index words: Herbicide resistance, integrated weed management. Abbreviations: ACCase, acetyl-CoA carboxylase; ALS, acetolactate synthase; APP, aryloxyphenoxypropionate; CHD, cyclohexanedione; DSS, decision-support system; EPSPS, enolpyruvylshikimate-3-phosphate synthase; HR, herbicide resistant; HS, herbicide susceptible; IWM, integrated weed management.

GENE FLOW IN COMMERCIAL FIELDS OF HERBICIDE‐RESISTANT CANOLA (BRASSICA NAPUS)

Multiple herbicide resistance to glyphosate, glufosinate, bromoxynil, or imidazolinone in volunteer plants of canola (Brassica napus) has been attributed to pollen flow among cultivars with different resistance traits. A study was conducted in Saskatchewan, Canada, in 1999 and 2000 to assess gene flow in space and time in adjacent commercial fields of glyphosate- and glufosinate-resistant canola, including (1) estimation of gene flow with distance; (2) frequency and distribution of volunteers, and effect on gene flow; (3) effect of adventitious double herbicide-resistant seed presence in seedlots planted; and (4) a comparison of various marker systems to track gene flow events. At 11 sites in 1999, gene flow was determined by sampling seeds from plants located at 0, 50, 100, 200, 400, 600, or 800 m along a transect perpendicular to the common border in the paired fields, spraying seedlings with glyphosate and glufosinate, and confirming the presence of the transgenes using commercial test strips and PCR analysis. In the spring of 2000, putative double herbicide-resistant volunteers that survived sequential herbicide applications were mapped at three of the sites using GPS and resistance in sampled plants was characterized. In 1999, gene flow between the paired fields was detected to a maximum distance of 400 m. Values ranged from 1.4% outcrossing at the border common to the paired fields to 0.04% at 400 m. In 2000, gene flow as a result of pollen flow in 1999 was detected to the limits of the study areas (800 m). Large variation in gene flow levels and patterns among the three sites was evident. Adventitious presence of double herbicide-resistant seed in glyphosate-resistant seedlots planted at two of the sites in 1999 contributed to the occurrence of double herbicide-resistant volunteers in 2000. The results of this study suggest that gene stacking in B. napus canola volunteers in western Canada may be common, and reflects pollen flow between different herbicide-resistant canola, presence of double herbicide-resistant off-types in seedlots, and/or agronomic practices typically employed by Canadian growers.

Screening for Herbicide Resistance in Weeds1

Abstract: Diagnosing herbicide-resistant weeds as a first step in resistance management and monitoring their nature, distribution, and abundance demands efficient and effective screening tests. This review summarizes and recommends appropriate seed sampling techniques, protocols for screening weeds for resistance to herbicides of different sites of action, interpretation of results, and information given to the grower. Elements common to all screening procedures are reviewed. Choosing appropriate discriminating doses to distinguish between resistant and susceptible weed biotypes is the most important factor in achieving accurate and consistent results. Interpretation of results is also critical because resistant weeds may comprise a small portion of the population in suspected accessions or biotypes. Additional index words: Bioassay, discriminating dose, seed sampling, site of action, surveys. Abbreviations: ACCase, acetyl-CoA carboxylase (EC 6.4.1.2); ALS, acetolactate synthase (EC 4.1.3.18); AOPP, aryloxyphenoxy propionate; CHD, cyclohexanedione; DAT, days after treatment; EPSP synthase, 5-enolpyruvylshikimate-3-phosphate synthase (EC 2.5.1.19); KARI, ketol-acid reductoisomerase (EC 1.1.1.86); POST, postemergence; PRE, preemergence; R, resistant; S, susceptible.

A decade of herbicide-resistant crops in Canada

This review examines some agronomic, economic, and environmental impacts of herbicide-resistant (HR) canola, soybean, corn, and wheat in Canada after 10 yr of growing HR cultivars. The rapid adoption of HR canola and soybean suggests a net economic benefit to farmers. HR crops often have improved weed management, greater yields or economic returns, and similar or reduced environmental impact compared with their non-HR crop counterparts. There are no marked changes in volunteer weed problems associated with these crops, except in zero-tillage systems when glyphosate is used alone to control canola volunteers. Although gene flow from glyphosate-HR canola to wild populations of bird’s rape (Brassica rapa L.) in eastern Canada has been measured, enrichment of hybrid plants in such populations should only occur when and where herbicide selection pressure is applied. Weed shifts as a consequence of HR canola have been documented, but a reduction in weed species diversity has not been demonstrated. However, reli...

Gene Flow, Invasiveness, and Ecological Impact of Genetically Modified Crops

The main environmental concerns about genetically modified (GM) crops are the potential weediness or invasiveness in the crop itself or in its wild or weedy relatives as a result of transgene movement. Here we briefly review evidence for pollen‐ and seed‐mediated gene flow from GM crops to non‐GM or other GM crops and to wild relatives. The report focuses on the effect of abiotic and biotic stress‐tolerance traits on plant fitness and their potential to increase weedy or invasive tendencies. An evaluation of weediness and invasive traits that contribute to the success of agricultural weeds and invasive plants was of limited value in predicting the effect of biotic and abiotic stress‐tolerance GM traits, suggesting context‐specific evaluation rather than generalizations. Fitness data on herbicide, insect, and disease resistance, as well as cold‐, drought‐, and salinity‐tolerance traits, are reviewed. We describe useful ecological models predicting the effects of gene flow and altered fitness in GM crops and wild/weedy relatives, as well as suitable mitigation measures. A better understanding of factors controlling population size, dynamics, and range limits in weedy volunteer GM crop and related host or target weed populations is necessary before the effect of biotic and abiotic stress‐tolerance GM traits can be fully assessed.

Selecting for weed resistance: herbicide rotation and mixture.

Abstract Herbicide rotations and mixtures are widely recommended to manage herbicide resistance. However, little research has quantified how these practices actually affect the selection of herbicide resistance in weeds. A 4-yr experiment was conducted in western Canada from 2004 to 2007 to examine the impact of herbicide rotation and mixture in selecting for acetolactate synthase (ALS) inhibitor resistance in the annual broadleaf weed, field pennycress, co-occurring in wheat. Treatments consisted of the ALS-inhibitor herbicide, ethametsulfuron, applied in a mixture with bromoxynil/MCPA formulated herbicide (photosystem-II inhibitor/synthetic auxin), or in rotation with the non-ALS inhibitor at an ALS-inhibitor application frequency of 0, 25, 50, 75, and 100% (i.e., zero to four applications, respectively) over the 4-yr period. The field pennycress seed bank at the start of the experiment contained 5% ethametsulfuron-resistant seed. Although weed control was only marginally reduced, resistance frequency of progeny of survivors increased markedly after one ALS-inhibitor application. At the end of the experiment, the level of resistance in the seed bank was buffered by susceptible seed, increasing from 29% of recruited seedlings after one application to 85% after four applications of the ALS inhibitor. The level of resistance in the seed bank for the mixture treatment after 4 yr remained similar to that of the nontreated (weedy) control or 0% ALS-inhibitor rotation frequency treatment. The results of this study demonstrate how rapidly ALS-inhibitor resistance can evolve as a consequence of repeated application of herbicides with this site of action, and supports epidemiological information from farmer questionnaire surveys and modeling simulations that mixtures are more effective than rotations in mitigating resistance evolution through herbicide selection. Nomenclature: Bromoxynil; ethametsulfuron; MCPA; field pennycress, Thlaspi arvense L. THLAR; wheat, Triticum aestivum L. ‘AC Barrie’.

Herbicide‐resistant weed management: focus on glyphosate

This review focuses on proactive and reactive management of glyphosate-resistant (GR) weeds. Glyphosate resistance in weeds has evolved under recurrent glyphosate usage, with little or no diversity in weed management practices. The main herbicide strategy for proactively or reactively managing GR weeds is to supplement glyphosate with herbicides of alternative modes of action and with soil-residual activity. These herbicides can be applied in sequences or mixtures. Proactive or reactive GR weed management can be aided by crop cultivars with alternative single or stacked herbicide-resistance traits, which will become increasingly available to growers in the future. Many growers with GR weeds continue to use glyphosate because of its economical broad-spectrum weed control. Government farm policies, pesticide regulatory policies and industry actions should encourage growers to adopt a more proactive approach to GR weed management by providing the best information and training on management practices, information on the benefits of proactive management and voluntary incentives, as appropriate. Results from recent surveys in the United States indicate that such a change in grower attitudes may be occurring because of enhanced awareness of the benefits of proactive management and the relative cost of the reactive management of GR weeds.

The biology of Canadian weeds. 138. Kochia scoparia (L.) Schrad.

Kochia [Kochia scoparia (L.) Schrad.] is an annual broadleaf weed species native to Eurasia and introduced as an ornamental to the Americas by immigrants in the mid- to late 1800s. Although sometimes categorized in the genus Bassia, there is no compelling reason for this classification. This naturalized species is a common and economically important weed in crop production systems and ruderal areas in semiarid to arid regions of North America, and has expanded northward in the Canadian Prairies during the past 30 yr. Although primarily self-pollinated, substantial pollen-mediated gene flow and efficient seed dispersal aids both short- and long-distance spread. The weed is morphologically highly variable, and its growth and development are markedly affected by environmental conditions. Kochia, a C4 species, is highly competitive in cropping systems because of its ability to germinate at low soil temperatures and emerge early, grow rapidly, tolerate heat, drought and salinity, and exert allelopathic effects...

Basis for Herbicide Resistance in Canadian Populations of Wild Oat (Avena fatua)

Abstract Wild oat is the second-most abundant, but most economically important, weed across the Canadian Prairies of western Canada. Despite the serious economic effects of resistance to acetyl-CoA carboxylase (ACC) or acetolactate synthase (ALS) inhibitors or both in this weed throughout the Northern Great Plains of North America, little research has examined the basis for herbicide resistance. We investigated target-site and nontarget-site mechanisms conferring ACC- and ALS-inhibitor resistance in 16 wild oat populations from across western Canada (four ACC-inhibitor resistant, four ALS-inhibitor resistant, and eight ACC- and ALS-inhibitor resistant). The ACC1 mutations were found in 8 of the 12 ACC inhibitor-resistant populations. The Ile1781Leu mutation was detected in three populations, the Trp2027Cys and Asp2078Gly mutations were in two populations each, and the Trp1999Cys, Ile2041Asn, Cys2088Arg, and Gly2096Ser substitutions were in one population each. Three populations had two ACC1 mutations. Only 2 of the 12 ALS inhibitor-resistant populations had an ALS target-site mutation—Ser653Thr and Ser653Asn substitutions. This is the first global report of ALS target-site mutations in Avena spp. and four previously undocumented ACC1 mutations in wild oat. Based on these molecular analyses, seedlings of five ACC + ALS inhibitor-resistant populations (one with an ACC1 mutation; four with no ACC or ALS mutations) were treated with malathion, a known cytochrome P450 monooxygenase inhibitor, followed by application of one of four ACC- or ALS-inhibiting herbicides. Malathion treatment often resulted in control or suppression of these populations, suggesting involvement of this enzyme system in contributing to resistance to both ACC and ALS inhibitors. Nomenclature: Wild oat, Avena fatua L. AVEFA.