Growth Stage of Alopecurus Myosuroides Huds. Determines the Efficacy of Pinoxaden
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plants Communication Growth Stage of Alopecurus myosuroides Huds. Determines the Efficacy of Pinoxaden Ana Pintar 1,*, Zlatko Sveˇcnjak 2 , Valentina Šoštarˇci´c 1, Josip Laki´c 1, Klara Bari´c 1, Dragojka Brzoja 1 and Maja Š´cepanovi´c 1 1 Department of Weed Science, University of Zagreb Faculty of Agriculture, Svetošimunska Cesta 25, 10 000 Zagreb, Croatia; [email protected] (V.Š.); [email protected] (J.L.); [email protected] (K.B.); [email protected] (D.B.); [email protected] (M.Š.) 2 Department of Field Crops, Forages and Grasslands, University of Zagreb Faculty of Agriculture, Svetošimunska Cesta 25, 10 000 Zagreb, Croatia; [email protected] * Correspondence: [email protected]; Tel.: +385-1-2393-606 Abstract: Alopecurus myosuroides Huds. is an important pinoxaden-resistant grass weed in many countries of Europe. Recently, the low efficacy of pinoxaden was reported in winter cereals in Croatia, but a preliminary dose–response trial showed no herbicide resistance for the investigated weed pop- ulation. Therefore, a two-year experiment was conducted under greenhouse conditions to determine the efficacy of various pinoxaden doses (20, 40 and 80 g a.i. ha−1) on weed visual injuries and biomass reduction after herbicide application at different growth stages. As expected, the maximum weed biomass reduction (97.3%) was achieved by applying the highest dose (80 g a.i. ha−1) at the earliest −1 growth stage (ZCK 12–14). A pinoxaden dose of 20 g a.i. ha resulted in satisfactory weed biomass reduction (88.9%) only when applied at ZCK 12–14. The recommended dose (40 g a.i. ha−1) also Citation: Pintar, A.; Sveˇcnjak,Z.; provided sufficient weed control up to the growth stage ZCK 21–25. Slightly delayed (ZCK 31–32) Šoštarˇci´c,V.; Laki´c,J.; Bari´c,K.; application of the recommended dose brought about a low weed biomass reduction (60.1%). Double Brzoja, D.; Š´cepanovi´c,M. Growth than the recommended dose also failed to provide satisfactory weed control at the advanced weed Stage of Alopecurus myosuroides Huds. growth stages (ZCK 31–32 and ZCK 37–39). Thus, reported low efficacy of pinoxaden is most likely Determines the Efficacy of Pinoxaden. because of delayed herbicide application when A. myosuroides is overgrown. Plants 2021, 10, 732. https://doi.org/ 10.3390/plants10040732 Keywords: black–grass; growth stage; pinoxaden dose; delayed application; biomass reduction Academic Editor: Nicholas Emmanuel Korres 1. Introduction Received: 10 February 2021 Alopecurus myosuroides Huds. (black–grass) is a common annual grass weed in win- Accepted: 6 April 2021 Published: 9 April 2021 ter cereal crops in north-western Europe [1]. It is widespread in regions with climates influenced by the Atlantic [2], but it is also locally found in other north-western European Publisher’s Note: MDPI stays neutral countries on wet loam and clay soils [3]. Black-grass is a highly competitive species that with regard to jurisdictional claims in causes significant yield losses in winter cereals [4]. It has been found that the economic 2 published maps and institutional affil- threshold for A. myosuroides is 5.5 to 15 plants per m [5–7], and in most cases, more than 2 iations. 10 plants per m will cause significant yields losses [8]. Additionally, the high reproduction rate [9] and high seed vitality under the high-density conditions require efficient weed control methods [10]. The predicted percentage control of A. myosuroides required to main- tain a static population in the continuous winter cereals is >90% to achieve its long-term control [11]. The low base temperature (0–1 ◦C) for germination [1,12], combined with Copyright: © 2021 by the authors. a short primary dormancy, gives this weed species a broad emergence spectrum, which Licensee MDPI, Basel, Switzerland. This article is an open access article further increases its competitiveness [13]. As a weed species whose life cycle reveals a wide distributed under the terms and range of adaptation to conventional cropping systems [14], A. myosuroides peak emergence conditions of the Creative Commons occurs in early autumn, coinciding with the sowing time and early development stage Attribution (CC BY) license (https:// of winter cereals. Weed emergence is most commonly observed between October and creativecommons.org/licenses/by/ December (80% of the population), and usually hibernates in two developed leaves or in 4.0/). the tillering phase. Plants appearing in autumn are more advanced compared to those Plants 2021, 10, 732. https://doi.org/10.3390/plants10040732 https://www.mdpi.com/journal/plants Plants 2021, 10, 732 2 of 9 emerging in spring, mainly because A. myosuroides can undergo a vernalization period during the cold winter months. Vernalization allows plants to enter the reproductive stage earlier, compared to spring individuals that do not go through the vernalization period and, therefore, enter the reproductive stage later. In addition, spring individuals develop fewer shoots compared to individuals developed in autumn [1]. However, this annual weed may produce seeds in both winter and spring crops under a wide range of growing degree days [10]. Therefore, controlling A. myosuroides has always been a major challenge, even though several different herbicide groups are approved for use in winter cereals. Pinoxaden [15] (Axial® 50 EC, Syngenta Crop Protection AG, Basel, Switzerland) is a selective herbicide for the control of annual grass weeds in winter cereals [16]. It belongs to the phenylpyrazoline chemical class, and its mode of action is based on the inhibition of acetyl-CoA carboxylase (ACCase) [17]. The first symptoms of inhibition appear within the first week, such as initial chlorosis, followed by necrosis and death of rapidly growing meristematic tissue. A time period of two to three weeks is usually required for complete control of susceptible species [16]. The repeated use of ACCase inhibitors since the late 1980s has led to resistance issues in different crops. Resistance to pinoxaden has been identified in more than 15 countries, and the most prominent species is A. myosuroides, whose resistant biotypes have so far been identified in Germany, Italy, Poland, Spain and the United Kingdom [18]. In Croatia, Apera spica-venti is the most common, and A. myosuroides the second most common grass weed in winter cereals [19] as well as the most wide-spread on heavy and clayey soils [20] in the area around Slavonski Brod (45◦9014” N, 18◦1024” E). Cereal producers in this area regularly use herbicide pinoxaden to control A. myosuroides. The official recommendation in Croatia is that pinoxaden is to be applied between the 3-leaf (ZCK 13) and first node (ZCK 31) stage of the grass weeds with the dose of 40 g a.i. ha−1 [15]. However, in recent years, producers of winter cereals have complained about the poor or no efficacy of this herbicide, which was also discussed at the annual Croatian Crop Protection Seminar in 2020 [21]. There is no herbicide monitoring program in Croatia, as is the case in many European countries [18]. Consequently, we conducted a bioassay to evaluate the potential resistance of A. myosuroides to pinoxaden [22] and results showed that the investigated population was susceptible to pinoxaden (Figure S1). Therefore, a low efficacy of pinoxaden reported by Croatian farmers is to be attributed to other factors. The main objective of this research was to determine the efficacy of various pinoxaden doses on visual injuries and biomass reduction of A. myosuroides after herbicide application at different growth stages. 2. Results 2.1. Visual Injuries at 7, 14 and 21 Days after Treatment The symptoms of visual injuries were affected by the application doses of pinoxaden and the growth stage of A. myosuroides. The progression of injury symptoms was ob- served with increasing days after treatment (DAT) with pinoxaden for all growth stages of A. myosuroides. At 7 DAT, visual injuries were observed on plants treated at ZCK 12–14 and ZCK 21–25, with a maximum of 48.8% and 25.0%, respectively, on plants treated with the double dose (80 g a.i. ha−1) of pinoxaden (Figure1a). Sensitivity of A. myosuroides to pinox- aden was significantly reduced when plants were treated at ZCK 31–32 and ZCK 37–39; only about 15% of plants were injured when the double dose of pinoxaden was applied. Plants 2021, 10, x FOR PEER REVIEW 3 of 9 (ZCK 21–25) tillering (Figure 1b). Plants treated at ZCK 12–14 showed the highest sensi- tivity to all applied doses of pinoxaden; injury symptoms ranged from 62.5% after apply- ing pinoxaden at 20 g a.i. ha−1 to 77.5% following herbicide application of 80 g a.i. ha−1. Plants treated at ZCK 21–25 also showed injury symptoms, with a maximum of 62.5% after the application of double dose of pinoxaden. In contrast, only 42.5% and 35.0% of plants treated at ZCK 31–32 and ZCK 37–39, respectively, were visually injured after ap- plication of the double dose of pinoxaden. The highest visual injuries were determined at 21 DAT (Figure 1c) and were the most visible on plants treated at ZCK 12–14. At this earliest weed growth stage, plants were damaged 90.5, 96.0 and 97.3% after pinoxaden application at 20, 40 and 80 g a.i. ha−1, re- spectively. When pinoxaden was applied at ZCK 21–25, injury symptoms higher than 90% Plants 2021, 10, 732 were determined only when the recommended or double dose were applied. When plants 3 of 9 were treated at the more advanced growth stages (ZCK 31–32 and ZCK 37–39), visual injuries were less than 80% even when applying the double dose. 100 80 ZCK 12–14 60 ZCK 21–25 40 ZCK 31–32 20 ZCK 37–39 (%untreated of control) Visual injuries at 7 DAT DAT 7 at injuries Visual 0 20 40 80 Pinoxaden dose (g a.i.