Spectral Discrimination of Two Pigweeds from Cotton with Different Leaf Colors

To implement strategies to control Palmer amaranth (Amaranthus palmeri S. Wats.) and redroot pigweed (Amaranthus retroflexus L.) infestations in cotton (Gossypium hirsutum L.) production systems, managers need effective techniques to identify the weeds. Leaf light reflectance measurements have shown promise as a tool to distinguish crops from weeds. Studies have targeted plants with green leaves. This study focused on using leaf hyperspectral reflectance data to develop spectral profiles of Palmer amaranth, redroot pigweed, and cotton and to determine regions of the light spectrum most sensitive for pigweed and cotton discrimination. The study focused on cotton near-isogenic lines created to have bronze, green, or yellow colored leaves. Reflectance measurements within the 400 to 2500 nm spectral range were obtained from cotton and weed plants grown in a greenhouse in 2015 and 2016. Two scenarios were evaluated for the comparison: (1) Palmer amaranth versus cotton lines and (2) redroot pigweed versus cotton lines. Statistical significance (p ≤ 0.05) was determined with analysis of variance (ANOVA) and Dunnett’s test. Sensitivity measurements were tabulated to determine the optimal region of the light spectrum for weed and cotton line discrimination. Optimal bands for weed and cotton separation were 600 to 700 nm (both weeds versus cotton bronze and cotton yellow), 710 nm (Palmer amaranth versus cotton green), and 1460 nm (redroot pigweed versus cotton green). Spectral bands were identified for separating Palmer amaranth and redroot pigweed from cotton lines with bronze, green, and yellow leaves. Ground-based and airborne sensors can be tuned into the regions of spectrum identified, facilitating using remote sensing technology for Palmer amaranth and redroot pigweed identification in cotton production systems.

Interaction of the Bioherbicide <i>Myrothecium verrucaria</i>with Technical-Grade Glyphosate on Glyphosate-Susceptible and -Resistant Palmer Amaranth

Previously we found that a strain of Myrothecium verrucaria (MV) exhibited bioherbicidal activity against several important weeds, and that some commercial formulations of glyphosate applied with MV resulted in synergistic interactions that improved weed control efficacy. We also found that MV had bioherbicidal activity against glyphosate-resistant Palmer amaranth. We have also reported that some commercial formulations are inhibitory to MV. Our objectives were to test the effect of unformulated glyphosate (high purity, technical-grade glyphosate) alone and in combination with MV for bioherbicidal activity on glyphosate-susceptible and -resistant Palmer amaranth biotypes under greenhouse conditions and to examine technical-grade glyphosate on the growth of this bioherbicide. High purity glyphosate (without adjuvants/surfactants) was not toxic to MV growth and sporulation at concentrations up to 2.0 mM when grown on agar supplemented with the herbicide. Both biotypes were injured by MV and MV plus glyphosate treatments as early as 19 h after application (3 h after a dew period of 16 h). These injury effects increased and were more evident through the 6-day time course, when after 120 h the MV plus glyphosate treatment had killed all glyphosate-susceptible and -resistant plants. The interaction of glyphosate plus MV was synergistic toward the control of Palmer amaranth. Data strongly suggest that the active ingredient is responsible for the synergy previously found when this bioherbicide was combined with some commercial formulations of glyphosate. Results demonstrated that MV can control both glyphosate-resistant and -susceptible Palmer amaranth seedlings and act synergistically with high-purity glyphosate to provide improved weed control.

Bioassay and Characterization of Several Palmer Amaranth (<i>Amaranthus palmeri</i>) Biotypes with Varying Tolerances to Glyphosate

The wide distribution of Palmer amaranth (Amaranthus palmeri) in the southern US became a serious weed control problem prior to the extensive use of glyphosate-resistant crops. Currently glyphosate-resistant populations of Palmer amaranth occur in many areas of this geographic region creating an even more serious threat to crop production. Investigations were undertaken using four biotypes (one glyphosate-sensitive, one resistant from Georgia and two of unknown tolerance from Mississippi) of Palmer amaranth to assess bioassay techniques for the rapid detection and level of resistance in populations of this weed. These plants were characterized with respect to chlorophyll, betalain, and protein levels and immunological responses to an antibody of 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) the target site of glyphosate. Only slight differences were found in four biotypes grown under greenhouse conditions regarding extractable soluble protein and chlorophyll content, but one biotype was found to be devoid of the red pigment, betalain. Measurement of early growth (seedling shoot elongation) of seedlings was a useful detection tool to determine glyphosate resistance. A leaf disc bioassay (using visual ratings and/or chlorophyll analysis) and an assay for shikimate accumulation were effective methods for determining herbicide resistance levels. The two unknown biotypes were found to be resistant to this herbicide. Some differences were found in the protein profiles of the biotypes, and western blots demonstrated a weak labeling of antibody in the glyphosate-sensitive biotype, whereas strong labeling occurred in the resistant plants. This latter point supports research by others, that increased copy number of the EPSPS gene (and increased EPSPS protein levels) is the resistance mechanism in this species. Results indicate the utility of certain bioassays for the determination of resistance and provide useful comparative information on the levels of inherent constituents among closely related plants.

Using Vegetation Indices as Input into Random Forest for Soybean and Weed Classification

Weed management is a major component of a soybean (Glycine max L.) production system;thus, managers need tools to help them distinguish soybean from weeds. Vegetation indices derived from light reflectance properties of plants have shown promise as tools to enhance differences among plants. The objective of this study was to evaluate normalized difference vegetation indices derived from multispectral leaf reflectance data as input into random forest machine learner to differentiate soybean and three broad leaf weeds: Palmer amaranth (Amaranthus palmeri L.), redroot pigweed (A. retroflexus L.), and velvetleaf (Abutilon theophrasti Medik). Leaf reflectance measurements were acquired from plants grown in two separate greenhouse experiments conducted in 2014. Twelve normalized difference vegetation indices were derived from the reflectance measurements, including advanced, green, greenred, green-blue, and normalized difference vegetation indices, shortwave infrared water stress indices, normalized difference pigment and red edge indices, and structure insensitive pigment index. Using the twelve vegetation indices as input variables, the conditional inference version of random forest (cforest) readily distinguished soybean and velvetleaf from the two pigweeds (Palmer amaranth and redroot pigweed) and from each other with classification accuracies ranging from 93.3% to 100%. The greatest errors were observed between the two pigweed classes, with classification accuracies ranging from 70% to 93.3%. Results suggest combining them into one class to increase classification accuracy. Vegetation indices results were equivalent to or slightly better than results obtained with sixteen multispectral bands used as input data into cforest. This research further supports using vegetation indices and machine learning algorithms such as cforest as decision support tools for weed identification.

Effects of <i>Myrothecium verrucaria</i>on Two Glyphosate-Resistant <i>Amaranthus palmeri</i>Biotypes Differing in Betacyanin Content

Previously we found two biotypes of Amaranthus palmeri (Palmer amaranth) in a population of this economically important weed that were resistant to glyphosate but differed with respect to pigmentation. One biotype was typically red-pigmented (betacyanin) while the other was green, with no visual appearance of red hue on any plant part at any growth stage. We have also reported that a strain of Myrothecium verrucaria (MV) exhibited bioherbicidal activity against several important weeds including glyphosate-resistant Palmer amaranth. In greenhouse tests, MV was applied to these two biotypes (red and green) at two ages (3-week- and 6-week-old) and effects of this fungus monitored over a 5-day time course. Initial symptoms of MV (16 to 24 h after inoculation) were: epinastic curvature, wilting and development of lesions on leaves and stems. Generally, the younger plants tended to be more sensitive to MV than older plants. Bioherbicidal damage increased with time leading to necrosis and plant mortality and increasing disease progress. Severe loss of fresh weight occurred in both biotypes as compared to untreated plants. Results indicated that MV was effective on both biotypes, but effects on growth reduction and disease progression were more rapid and generally greater in the green biotype, suggesting that compounds responsible for red pigmentation may be more potent as defense against pathogen attack.

Genomic Stability of Palmer amaranth Plants Derived by Macro-Vegetative Propagation

qPCR (quantitative polymerase chain reaction) and random amplified polymorphic DNA (RAPD) were utilized to investigate genetic stability of Palmer amaranth cloned plants over 10 generations. DNA from original parent Palmer amaranth plants (grown from seeds) was re-analyzed using qPCR, and confidence levels for determining ΔΔCt (threshold crossing) values were established. ANOVA was used to determine variation (margin of error) of these ΔΔCt values. This margin of error was applied to qPCR analysis of DNA from eight individual parent plants and their descendants (10th generation) so that possible differences in EPSPS (5-enolpyruvylshikimate-3-phosphate synthase) gene copy number could be ascertained. This method (and the associated error) indicated a lack of agreement in ΔΔCt values of DNA from plants of these two generations. qPCR analysis showed that in five out of eight clones, EPSPS gene copy number varied more than the calculated error (P = 0.05). A second technique to monitor genetic stability, RAPD was used to determine possible changes in genomic DNA due to extended cloning of these regenerated plants. RAPD analysis showed that four out of the eight clones differed when the profiles of the two generations were compared. Results show that qPCR and RAPD analysis point to the fact that several Palmer amaranth clones experienced changes in genome structure over 10 generations. Although the glyphosate resistance trait was retained, results suggest that during cloning studies, the genetic stability of macro-vegetatively propagated lines should be monitored.

Have You Considered Eating Your Weeds?

In most parts of the developed world, Pigweed, Spider plant, Lambs amongst others are regarded as weeds. But in Africa and other developing countries, these plants form part of the daily diets of many rural households. The oldest inhabitants of South Africa have harvested leaves from wild plants to supplement the meat from hunted animals. Over 100 different species of plants are cooked as a potherb/relish with corn meal. These species include indigenous species as well as indigenized, mostly weedy, species. These species became part of the African culture and heritage and were collectively known as morogo or imifino. The popularity of specific species is a function of many factors, including availability, ease of preparation, taste, consistency and appearance. Some popular genera are Amaranthus, Cleome, Solanum and Corchorus. Micronutrient malnutrition is widespread in South Africa with vitamin A and iron as the major concern for micronutrient deficiency. Morogo can contribute to alleviating these micronutrient deficiencies. It was found that for the species tested, that morogo are low in energy and that leaves of nightshade, pigweed and spider flower provided more than 50% of the RDA for vitamin A.

Varying Tolerance to Glyphosate in a Population of Palmer Amaranth with Low EPSPS Gene Copy Number

A Palmer amaranth population (seeds collected in the year 2000;Washington Co., MS) suspected to be susceptible to glyphosate was examined as a population and as individual plants and found to exhibit varying tolerance or resistance to glyphosate. Whole plant spraying of glyphosate (0.84 kg·ha?1) to the population revealed that approximately 40% of this population were resistant to glyphosate and an LD50 of 0.75 kg·ha?1 was determined. Spray application of glyphosate indicated that some plants displayed varying degrees of resistance 14 days after treatment. Initial tests using leaf disc bioassays on 10 individual plants selected randomly from the population, allowed characterization of glyphosate resistance using both visual ratings of injury and quantitative measurement via chlorophyll content analysis. After initial bioassays and spray application, five plants with a range of tolerance to glyphosate were selected for cloning so that further studies could be accomplished on these individuals. Q-PCR analysis of these clones showed that resistance was not due to elevated EPSPS gene copy number. Shikimate levels were lower in the resistant and higher in the susceptible clones which correlated with varying degrees of resistance demonstrated in bioassays and spray application of glyphosate of these clones. Results demonstrate that individuals in a population can vary widely with respect to herbicide resistance and suggest that uptake, translocation, sequestration, metabolism or altered target site may contribute to the resistance in some individuals of this population.