Jerry M. Green

Review of Glyphosate and ALS-Inhibiting Herbicide Crop Resistance and Resistant Weed Management

Weed management is a perennial challenge for growers, and continual innovation is essential to maintain the effectiveness of management technologies. The first generation of herbicide-resistant crops revolutionized weed control. However, weeds are adapting to crop systems that rely on a single mode of herbicide action. Crops with resistance to multiple modes of herbicide action could help maintain weed management. GAT/HRA is a new multiple herbicide–resistance technology for corn, soybean, and other crops. GAT/HRA combines metabolic glyphosate inactivation with an acetolactate synthase (ALS) enzyme that is insensitive to ALS-inhibiting herbicides. The mechanism to inactivate glyphosate is the glyphosate N-acetyltransferase enzyme, which transforms glyphosate into a nonphytotoxic metabolite. The gat gene is derived from a naturally occurring soil bacterium and optimized by repetitive gene shuffling and screening. The resistance mechanism to ALS-inhibiting herbicides is a double-mutant, highly resistant ALS (HRA) that is insensitive to all five classes of ALS herbicides. GAT/HRA crops will maintain natural tolerance to selective herbicides and thus provide more weed management options for growers to help deter weed spectrum shifts and delay the evolution of herbicide-resistant weeds. Nomenclature: Glyphosate; corn, Zea mays L; soybean, Glycine max (L.) Merr.

The benefits of herbicide‐resistant crops

Since 1996, genetically modified herbicide-resistant crops, primarily glyphosate-resistant soybean, corn, cotton and canola, have helped to revolutionize weed management and have become an important tool in crop production practices. Glyphosate-resistant crops have enabled the implementation of weed management practices that have improved yield and profitability while better protecting the environment. Growers have recognized their benefits and have made glyphosate-resistant crops the most rapidly adopted technology in the history of agriculture. Weed management systems with glyphosate-resistant crops have often relied on glyphosate alone, have been easy to use and have been effective, economical and more environmentally friendly than the systems they have replaced. Glyphosate has worked extremely well in controlling weeds in glyphosate-resistant crops for more than a decade, but some key weeds have evolved resistance, and using glyphosate alone has proved unsustainable. Now, growers need to renew their weed management practices and use glyphosate with other cultural, mechanical and herbicide options in integrated systems. New multiple-herbicide-resistant crops with resistance to glyphosate and other herbicides will expand the utility of existing herbicide technologies and will be an important component of future weed management systems that help to sustain the current benefits of high-efficiency and high-production agriculture.

Current state of herbicides in herbicide-resistant crops.

Current herbicide and herbicide trait practices are changing in response to the rapid spread of glyphosate-resistant weeds. Growers urgently needed glyphosate when glyphosate-resistant crops became available because weeds were becoming widely resistant to most commonly used selective herbicides, making weed management too complex and time consuming for large farm operations. Glyphosate made weed management easy and efficient by controlling all emerged weeds at a wide range of application timings. However, the intensive use of glyphosate over wide areas and concomitant decline in the use of other herbicides led eventually to the widespread evolution of weeds resistant to glyphosate. Today, weeds that are resistant to glyphosate and other herbicide types are threatening current crop production practices. Unfortunately, all commercial herbicide modes of action are over 20 years old and have resistant weed problems. The severity of the problem has prompted the renewal of efforts to discover new weed management technologies. One technology will be a new generation of crops with resistance to glyphosate, glufosinate and other existing herbicide modes of action. Other technologies will include new chemical, biological, cultural and mechanical methods for weed management. From the onset of commercialization, growers must now preserve the utility of new technologies by integrating their use with other weed management technologies in diverse and sustainable systems.

Identification of a Tall Waterhemp (Amaranthus tuberculatus) Biotype Resistant to HPPD-Inhibiting Herbicides, Atrazine, and Thifensulfuron in Iowa

Abstract Seeds of a putative 4-hydroxyphenylpyruvate dioxygenase (HPPD)-inhibiting herbicide–resistant tall waterhemp biotype from Henry County, IA, were collected from a seed corn field in fall 2009 after plants were not controlled following a POST application of mesotrione plus atrazine. The response of this biotype to various herbicide modes of action was evaluated in greenhouse and field tests. Under greenhouse conditions, the suspect biotype showed an eightfold decrease in sensitivity to mesotrione with a 50% control rate of 21 g ha−1 compared with 2.7 g ha−1 for the susceptible biotype. The biotype also had a 10-fold decrease in sensitivity to atrazine and a 28-fold decrease in sensitivity to thifensulfuron. Under field conditions, tall waterhemp was not controlled POST at the label rate of 1,100 g ha−1 of atrazine. Tall waterhemp control was less than 60% at the label rates of three commonly used POST HPPD-inhibiting herbicides in seed corn: 105 g ha−1 of mesotrione, 92 g ha−1 of tembotrione, or 18 g ha−1 of topramezone. Thus, this new tall waterhemp biotype is resistant to three herbicide modes of action: HPPD inhibitors, photosystem-II inhibitors, and acetolactate synthase (ALS) inhibitors. Nomenclature: Atrazine; mesotrione; tembotrione; thifensulfuron, topramezone, tall waterhemp, Amaranthus tuberculatus (Moq.) Sauer AMATU; corn, Zea mays L. ZEAMX

Enhancing the Biological Activity of Nicosulfuron with pH Adjusters

Adjuvants that increase the pH of the spray mixture and solubilize nicosulfuron can enhance biological activity under specific conditions. These conditions include high nicosulfuron rates, difficult-to-control weeds, low spray volumes, and initially acidic spray conditions. The most effective pH adjusters are tribasic potassium phosphate, sodium carbonate, and triethanolamine. In low spray volumes, these adjusters make the spray mixture alkaline and often enhance the activity of nicosulfuron on common cocklebur and large crabgrass. Alkaline conditions rapidly dissolve the sulfonylurea particles and enhance activity with crop oil concentrate, modified seed oil, and hydrophilic nonionic surfactants. pH adjusters did not enhance activity with lipophilic surfactants. Ammonium sulfate slightly increases the pH of spray mixtures and increases nicosulfuron activity depending on species, adjuvant type, and pH adjuster. These results generally support the concept that herbicide solubilization is necessary to maximize the foliar activity. Nomenclature: Nicosulfuron; common cocklebur, Xanthium strumarium L. #3 XANST; large crabgrass, Digitaria sanguinalis (L.) Scop. # DIGSA. Additional index words: Adjuvant, basic blend, buffer, crop oil concentrate, fertilizer, formulation, hydrophilic–lipophilic balance, methylated seed oil, modified seed oil, nonionic surfactant, potassium phosphate, soda ash, sulfonylurea, triethanolamine, water-dispersible granule. Abbreviations: AMS, ammonium sulfate; COC, crop oil concentrate; HLB, hydrophilic–lipophilic balance; MSO, modified seed oil; NIS, nonionic surfactant; NPE, nonylphenol ethoxylate; TEA, triethanolamine; WDG, water-dispersible granule.

Broad 4-Hydroxyphenylpyruvate Dioxygenase Inhibitor Herbicide Tolerance in Soybean with an Optimized Enzyme and Expression Cassette

A modified native promoter, dual protein localization, and an evolved desensitized maize protein variant enables field tolerance in soybean to multiple herbicides. With an optimized expression cassette consisting of the soybean (Glycine max) native promoter modified for enhanced expression driving a chimeric gene coding for the soybean native amino-terminal 86 amino acids fused to an insensitive shuffled variant of maize (Zea mays) 4-hydroxyphenylpyruvate dioxygenase (HPPD), we achieved field tolerance in transgenic soybean plants to the HPPD-inhibiting herbicides mesotrione, isoxaflutole, and tembotrione. Directed evolution of maize HPPD was accomplished by progressively incorporating amino acids from naturally occurring diversity and novel substitutions identified by saturation mutagenesis, combined at random through shuffling. Localization of heterologously expressed HPPD mimicked that of the native enzyme, which was shown to be dually targeted to chloroplasts and the cytosol. Analysis of the native soybean HPPD gene revealed two transcription start sites, leading to transcripts encoding two HPPD polypeptides. The N-terminal region of the longer encoded peptide directs proteins to the chloroplast, while the short form remains in the cytosol. In contrast, maize HPPD was found almost exclusively in chloroplasts. Evolved HPPD enzymes showed insensitivity to five inhibitor herbicides. In 2013 field trials, transgenic soybean events made with optimized promoter and HPPD variant expression cassettes were tested with three herbicides and showed tolerance to four times the labeled rates of mesotrione and isoxaflutole and two times the labeled rates of tembotrione.

Increasing and Decreasing pH to Enhance the Biological Activity of Nicosulfuron

Increasing the pH of the spray water to solubilize the weak acid herbicide nicosulfuron and then decreasing pH below its pKa so that it converts into a neutral form enhances biological activity under some conditions. The water-dispersible granule formulation of nicosulfuron starts as dispersed particles. Adding 1% wt/wt K3PO4 solubilizes nicosulfuron and increases its activity compared to its dispersion without base. The type of buffer and the surfactant HLB or hydrophilic lipophilic balance, a measure of the molecular balance of the hydrophilic and lipophilic groups, altered the activity of nicosulfuron. Adding 1% wt/wt K3PO4 increases the pH, and the optimum HLB ranged from 13 to 17 on large crabgrass. Adding 1% wt/wt H3PO4 reduces the pH and lowers the optimum HLB range from 10 to 14 on large crabgrass. Adding the acidic buffer converts the solubilized nicosulfuron into its neutral form and increases activity under some surfactant conditions. Thus, neutral nicosulfuron is more active with lipophilic surfactants, while ionic nicosulfuron is more active with hydrophilic surfactants. When tested on other species, low HLB surfactants are the most active at low pH. These results support the concept that the physicochemical properties of the herbicide, adjuvants, and weed species should be matched for optimum activity. Nomenclature: Common cocklebur, Xanthium strumarium L. #3 XANST; common lambsquarters, Chenopodium album L. # CHEAL; giant foxtail, Setaria faberi Herrm. # SETFA; large crabgrass, Digitaria sanguinalis (L.) Scop. # DIGSA; purple nutsedge, Cyperus rotundus L. # CYPRO; sicklepod, Senna obtusifolia (L.) Irwin and Barneby # CASOB; velvetleaf, Abutilon theophrasti Medicus # ABUTH. Additional index words: Acid, adjuvant, base, basic blend, buffer, formulation, hydrophilic–lipophilic balance, nicosulfuron, nonionic surfactant, phosphoric acid, potassium phosphate, sulfonylurea, water-dispersible granule, weak acid. Abbreviations: HLB, hydrophilic–lipophilic balance; NIS, nonionic surfactant; NPE, nonylphenol ethoxylate; WDG, water-dispersible granule.

Response of 98140 Corn With Gat4621 and hra Transgenes to Glyphosate and ALS-Inhibiting Herbicides

Abstract The transgenic corn line 98140 has a high level of resistance to glyphosate and all five chemical classes of herbicides that inhibit acetolactate synthase (ALS). The dual herbicide resistance is due to a molecular stack of two constitutively expressed genes: gat4621, which produces a glyphosate acetyltransferase that rapidly inactivates glyphosate, and hra, which produces a highly resistant ALS. On a rate basis, the positive 98140 isoline with a single copy of the gat4621 gene is over 1,000-fold more resistant to glyphosate than a negative isoline without the transgene. Similarly, the positive 98140 isoline with the hra gene is over 1,000-fold more resistant to ALS-inhibiting herbicides such as chlorimuron and sulfometuron at the whole-plant and enzyme level. The gat4621 and hra genes do not change the natural tolerance of corn to selective herbicides, so new corn hybrids based on 98140 will give growers more options to manage weeds and delay the evolution of herbicide-resistant weeds. Nomenclature: Chlorimuron, glyphosate, sulfometuron, corn, Zea mays L

The rise and future of glyphosate and glyphosate-resistant crops

Glyphosate and glyphosate-resistant crops had a revolutionary impact on weed management practices, but the epidemic of glyphosate-resistant (GR) weeds is rapidly decreasing the value of these technologies. In areas that fully adopted glyphosate and GR crops, GR weeds evolved and glyphosate and glyphosate traits now must be combined with other technologies. The chemical company solution is to combine glyphosate with other chemicals, and the seed company solution is to combine glyphosate resistance with other traits. Unfortunately, companies have not discovered a new commercial herbicide mode-of-action for over 30 years and have already developed or are developing traits for all existing herbicide types with high utility. Glyphosate mixtures and glyphosate trait combinations will be the mainstays of weed management for many growers, but are not going to be enough to keep up with the capacity of weeds to evolve resistance. Glufosinate, auxin, HPPD-inhibiting and other herbicide traits, even when combined with glyphosate resistance, are incremental and temporary solutions. Herbicide and seed businesses are not going to be able to support what critics call the chemical and transgenic treadmills for much longer. The long time without the discovery of a new herbicide mode-of-action and the epidemic of resistant weeds is forcing many growers to spend much more to manage weeds and creating a worst of times, best of times predicament for the crop protection and seed industry.

Adjuvants: Test Design, Interpretation, and Presentation of Results1

Abstract: Adjuvant research contributes much to the knowledge and practice of weed science though the scientific process of systematically asking precise questions and subsequently making distinctions among alternative explanations. The purpose of adjuvant experimentation is to answer these questions and the purpose of associated papers and presentations is to communicate the new information. These purposes are self-evident, but are difficult to perfect. Some factors are particularly difficult for adjuvant researchers and require that researchers plan thoroughly from the formulation of the experimental question to final presentation of results. Adjuvant research requires both chemical and biological expertise that is traditionally separated in most organizations. Scientists from other disciplines or weed scientists not primarily concerned with adjuvants often direct adjuvant studies. This paper discusses mistakes that are commonly made in test design, interpretation, and presentation and suggests guidelines to improve the quality of adjuvant research.