Multiple resistant Brassica rapa (=B. campestris) from Argentina

INTERNATIONAL HERBICIDE-RESISTANT WEED DATABASE

MULTIPLE RESISTANT BIRDSRAPE MUSTARD
(Brassica rapa (=B. campestris))

Multiple Resistance: 2 Sites of Action
Inhibition of Acetolactate Synthase HRAC Group 2 (Legacy B)
Inhibition of Enolpyruvyl Shikimate Phosphate Synthase HRAC Group 9 (Legacy G)

Argentina

INTRODUCTION		BIRDSRAPE MUSTARD
Birdsrape Mustard (Brassica rapa (=B. campestris)) is a dicot weed in the Brassicaceae family. In Argentina this weed first evolved multiple resistance (to 2 herbicide sites of action) in 2012 and infests Soybean, and Wheat. Multiple resistance has evolved to herbicides in the Groups 2 (Legacy B), and Inhibition of Enolpyruvyl Shikimate Phosphate Synthase HRAC Group 9 (Legacy G). These particular biotypes are known to have resistance to diclosulam, glyphosate, imazapyr, and metsulfuron-methyl and they may be cross-resistant to other herbicides in the Groups 2 (Legacy B), and Inhibition of Enolpyruvyl Shikimate Phosphate Synthase HRAC Group 9 (Legacy G). The 'Group' letters/numbers that you see throughout this web site refer to the classification of herbicides by their site of action. To see a full list of herbicides and HRAC herbicide classifications click here.

QUIK STATS (last updated Jan 27, 2018 )
Common Name	Birdsrape Mustard
Species	Brassica rapa (=B. campestris)
Group	Inhibition of Acetolactate Synthase HRAC Group 2 (Legacy B) Inhibition of Enolpyruvyl Shikimate Phosphate Synthase HRAC Group 9 (Legacy G)
Herbicides	diclosulam, glyphosate, imazapyr, and metsulfuron-methyl
Location	Argentina
Year	2012
Situation(s)	Soybean, and Wheat
Contributors - (Alphabetically)	Miguel Cantamutto, Claudio Pandolfo, and Alejandro Presotto
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NOTES ABOUT THIS BIOTYPE

GENERAL

Claudio Pandolfo

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

Claudio E. Pandolfo& Alejandro Presotto & Francisco Torres Carbonell& Soledad Ureta& Mónica Poverene& Miguel Cantamutto

Environmental Science and Pollution Research, Published online 14th December 2017

https://doi.org/10.1007/s11356-017-0726-3

Abstract 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.

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MECHANISM

Claudio Pandolfo

This biotype has multiple herbicide resistance, to glyphosate and AHAS-inhibiting herbicides. However, we have probed that the glyphosate resistance was transgenic. We believe that the presence of the transgene in wild populations of B. rapa involved gene flow from transgenic oilseed rape cultivars or from feral populations having that event or through the introduction of these B. rapa biotypes as a seed contaminant from other countries (because transgenic oilseed rape is forbidden in Argentina). The multiple resistance to AHAS-inhibiting herbicides might been also due to gene flow with IMI-resistant (Clearfield) oilseed rape cultivars, but we couldn’t probed this hypothesis yet. We have probed that this Brassica rapa biotype has the Trp-574-Leu mutation in AHAS enzyme gene, as in other resistant weed cases (like Raphanus sativus from Argentina).

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ACADEMIC ASPECTS

Confirmation Tests

Greenhouse, and Laboratory trials comparing a known susceptible Birdsrape Mustard biotype with this Birdsrape Mustard biotype have been used to confirm resistance. For further information on the tests conducted please contact the local weed scientists that provided this information.

Genetics

Genetic studies on HRAC Group 2, 9 resistant Birdsrape Mustard have not been reported to the site. There may be a note below or an article discussing the genetics of this biotype in the Fact Sheets and Other Literature

Mechanism of Resistance

Studies on the mechanism of resistance of multiple resistant Birdsrape Mustard from Argentina indicate that resistance is due to an altered target site. There may be a note below or an article discussing the mechanism of resistance in the Fact Sheets and Other Literature

Relative Fitness

There is no record of differences in fitness or competitiveness of these resistant biotypes when compared to that of normal susceptible biotypes. If you have any information pertaining to the fitness of multiple resistant Birdsrape Mustard from Argentina please update the database.

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CONTRIBUTING WEED SCIENTISTS

MIGUEL CANTAMUTTO

Full Profesor Universidad Nacional Del Sur Departamento De Agronomía San Andrés 800 Bahía Blanca, 8000, Buenos Aires Argentina Email Miguel Cantamutto

CLAUDIO PANDOLFO

Dr. - Postdoc - T.A. Universidad Nacional Del Sur, CERZOS-CONICET Departamento De Agronomía San Andrés 800 Bahía Blanca, 8000, Buenos Aires Argentina Email Claudio Pandolfo Web : Web Site Link

ALEJANDRO PRESOTTO

Adjunct Researcher Universidad Nacional del Sur; CERZOS-CONICET Departamento de Agronomía San Andrés 800 Bahía Blanca, 8000, Buenos Aires Argentina Email Alejandro Presotto Web : Web Site Link

ACKNOWLEDGEMENTS
The Herbicide Resistance Action Committee, The Weed Science Society of America, and weed scientists in Argentina have been instrumental in providing you this information. Particular thanks is given to Miguel Cantamutto, Claudio Pandolfo, and Alejandro Presotto for providing detailed information.

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Herbicide Resistant Birdsrape Mustard Globally
(Brassica rapa (=B. campestris))

Country

FirstYear

Situation

Active Ingredients

Site of Action

Argentina

2012

Soybean, and Wheat

diclosulam, glyphosate, imazapyr, and metsulfuron-methyl

Multiple Resistance: 2 Sites of Action
Inhibition of Acetolactate Synthase ( HRAC Group 2 (Legacy B)
Inhibition of Enolpyruvyl Shikimate Phosphate Synthase ( HRAC Group 9 (Legacy G)

Argentina

2015

Soybean, Wheat, and Winter barley

2,4-D

Auxin Mimics ( HRAC Group 4 (Legacy O)

Canada (Quebec)

1977

Corn (maize)

atrazine

PSII inhibitors - Serine 264 Binders ( HRAC Group 5 (Legacy C1 C2)

Canada (Quebec)

2017

Corn (maize), and Soybean

glyphosate

Inhibition of Enolpyruvyl Shikimate Phosphate Synthase ( HRAC Group 9 (Legacy G)

Literature about Similar Cases

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Mithila, J. ; Hall, J. C.. 2013. Transfer of auxinic herbicide resistance from Brassica kaber to Brassica juncea and Brassica rapa through embryo rescue. In Vitro Cellular & Developmental Biology - Plant 49 : 461 - 467.

Auxinic herbicides are widely used in agriculture to selectively control broadleaf weeds. Prolonged use of auxinic herbicides has resulted in the evolution of resistance to these herbicides in some biotypes of Brassica kaber (wild mustard), a common weed in agricultural crops. In this study, auxinic herbicide resistance from B. kaber was transferred to Brassica juncea and Brassica rapa, two commercially important Brassica crops, by traditional breeding coupled with in vitro embryo rescue. A high frequency of embryo regeneration and hybrid plant establishment was achieved. Transfer of auxinic herbicide resistance from B. kaber to the hybrids was assessed by whole-plant screening of hybrids with dicamba, a widely used auxinic herbicide. Furthermore, the hybrids were tested for fertility (both pollen and pistil) and their ability to produce backcross progeny. The auxinic herbicide-resistant trait was introgressed into B. juncea by backcross breeding. DNA ploidy of the hybrids as well as of the backcross progeny was estimated by flow cytometry. Creation of auxinic herbicide-resistant Brassica crops by non-transgenic approaches should facilitate effective weed control, encourage less tillage, provide herbicide rotation options, minimize occurrence of herbicide resistance, and increase acceptance of these crops..

Konagaya, K. ; Tsuda, M. ; Okuzaki, A. ; Ando, S. ; Tabei, Y.. 2013. Application of the acetolactate synthase gene as a cisgenic selectable marker for Agrobacterium-mediated transformation in Chinese cabbage (Brassica rapa ssp. pekinensis). Plant Biotechnology 30 : 125 - 133.

Heading Chinese cabbage (Brassica rapa ssp. pekinensis) has been recalcitrant to regeneration and transformation. In particular, there are few reports concerning Japanese cultivars. We evaluated the factors that inhibit Agrobacterium-mediated transformation in heading Chinese cabbage. Then, we investigated the effects of selectable markers using heading Chinese cabbage cv. Chihiri 70 and compared the effects with those in broccoli cv. Ryokurei (B. oleracea var. italica) as a control for an easily transformable Brassica species. To utilize a selectable marker derived from a host plant cisgene, we cloned a genomic DNA fragment containing regulatory and coding sequences of the acetolactate synthase (ALS) gene from Chinese cabbage and mutagenized it to a herbicide resistant form. After transformation of Chinese cabbage and broccoli with this construct, transgenic plants were efficiently selected with the herbicide bispyribac sodium salt and screened by DNA gel blot analysis. The average transformation frequency of Chinese cabbage was 1.2±0.2%, which was similar to those in reports using antibiotic selectable markers and was lower than for broccoli (13.9±2.0%). Furthermore, the escape rate was restricted at a low level (about 35-50% lower than hygromycin selection), which is an advantage on practical transformation. We confirmed transgene inheritance and herbicide resistance of potted plants in the T₁ generation. This report is the first to describe a selection system for the transformation of a Brassica crop that uses a herbicide-tolerant selectable marker derived from a cisgene..

Collier, M. J. ; Mullins, E.. 2012. Assessing the impact of pollen-mediated gene flow from GM herbicide tolerant Brassica napus into common wild relatives in Ireland. Biology and Environment: Proceedings of the Royal Irish Academy, Section B 112B : 257 - 266.

Although now we have had many years of research completed on assessing the potential environmental impact of GM crops, concern remains over their potential impact on biodiversity in the rural landscape. In particular, issues have arisen in regards to the modification of crops with traits that could introgress into sexually compatible wild relatives. In contrast to wheat, barley, potato and maize, Brassica napus (oilseed rape) is the only commercial crop grown in Ireland at present with the potential to successfully transfer its DNA, via pollen-mediated gene flow, into inter-related weed species. This review details the species in question and by examining the relevant literature that relates to Irish agronomic conditions, demonstrates that gene flow is likely to occur, especially to an earlier used cultivar, Brassica rapa. However, the critical factor remains not that GM traits will flow from the commercial source but what might the consequences of said gene flow events be. This review indicates that the conferred trait in question (in this case, herbicide tolerance) can only impact on weed diversity in the presence of selecting herbicide action. In the absence of the herbicide, the GM traits will be lost from the wild species over time and will not confer any selective advantage that could facilitate population growth..

Aono, M. ; Wakiyama, S. ; Nagatsu, M. ; Kaneko, Y. ; Nishizawa, T. ; Nakajima, N. ; Tamaoki, M. ; Kubo, A. ; Saji, H.. 2011. Seeds of a possible natural hybrid between herbicide-resistant Brassica napus and Brassica rapa detected on a riverbank in Japan. GM Crops 2 : 201 - 210.

Transgenic herbicide-resistant varieties of Brassica napus, or oilseed rape, from which canola oil is obtained, are imported into Japan, where this plant is not commercially cultivated to a large extent. This study aimed to examine the distribution of herbicide-resistant B. napus and transgene flow to escaped populations of its closely related species, B. rapa and B. juncea. Samples were collected from 12 areas near major ports through which oilseed rape imports into Japan passed - Kashima, Chiba, Yokohama, Shimizu, Nagoya, Yokkaichi, Sakai-Senboku, Kobe, Uno, Mizushima, Kita-Kyushu, and Hakata - and the presence of glyphosate- and/or glufosinate-resistant B. napus was confirmed in all areas except Yokohama, Sakai-Senboku, Uno, and Kita-Kyushu. The Yokkaichi area was the focus because several herbicide-resistant B. napus plants were detected not only on the roadside where oilseed rape spilled during transportation but also on the riverbanks, where escaped populations of B. rapa and B. juncea grew. Samples of B. napus that were tolerant to both herbicides were detected in four continuous years (2005-2008) in this area, suggesting the possibility of intraspecific transgene flow within the escaped B. napus populations. Moreover, in 2008, seeds of a possible natural hybrid between herbicide-tolerant B. napus (2n=38) and B. rapa (2n=20) were detected; some seedlings derived from the seeds collected at a Yokkaichi site showed glyphosate resistance and had 2n=29 chromosomes. This observation strongly suggests the occurrence of hybridization between herbicide-resistant B. napus and escaped B. rapa and the probability of introgression of a herbicide-resistance gene into related escaped species..

Zhang JunJie ; Liu Fan ; Yao Lei ; Luo Chen ; Zhao Qing ; Huang YuBi. 2011. Vacuum infiltration transformation of non-heading Chinese cabbage (Brassica rapa L. ssp. chinensis) with the pinII gene and bioassay for diamondback moth resistance. Plant Biotechnology Reports 5 : 217 - 224.

Non-heading Chinese cabbage (Brassica rapa L. ssp. chinensis) is a popular vegetable in Asian countries. The diamondback moth (DBM), Plutella xylostella (L.), an insect with worldwide distribution, is a main pest of Brassicaceae crops and causes enormous crop losses. Transfer of the anti-insect gene into the plant genome by transgenic technology and subsequent breeding of insect-resistant varieties will be an effective approach to reducing the damage caused by this pest. We have produced transgenic non-heading Chinese cabbage plants expressing the potato proteinase inhibitor II gene (pinII) and tested the pest resistance of these transgenic plants. Non-heading Chinese cabbages grown for 45 days on which buds had formed were used as experimental materials for Agrobacterium-mediated vacuum infiltration transformation. Forty-one resistant plants were selected from 1166 g of seed harvested from the infiltrated plants based on the resistance of the young seedlings to the herbicide Basta. The transgenic traits were further confirmed by the Chlorophenol red test, PCR, and genomic Southern blotting. The results showed that the bar and pinII genes were co-integrated into the resistant plant genome. A bioassay of insect resistance in the second generation of individual lines of the transgenic plants showed that DBM larvae fed on transgenic leaves were severely stunted and had a higher mortality than those fed on the wild-type leaves..

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