Unlocking the interaction of organophosphorus pesticide residues with ecosystem: Toxicity and bioremediation.

Organophosphorus adulteration in the environment creates terrestrial and aquatic pollution. It causes acute and subacute toxicity in plants, humans, insects, and animals. Due to the excessive use of organophosphorus pesticides, there is a need to develop environmentally friendly, economical, and bio-based strategies. The microbiomes, that exist in the soil, can reduce the devastating effects of organophosphates. The use of cell-free enzymes and yeast is also an advanced method for the degradation of organophosphates. Plant-friendly bacterial strains, that exist in the soil, can help to degrade these contaminants by oxidation-reduction reactions, enzymatic breakdown, and adsorption. The bacterial strains mostly from the genus Bacillus, Pseudomonas, Acinetobacter, Agrobacterium, and Rhizobium have the ability to hydrolyze the bonds of organophosphate compounds like profenofos, quinalphos, malathion, methyl-parathion, and chlorpyrifos. The native bacterial strains also promote the growth abilities of plants and help in detoxification of organophosphate residues. This bioremediation technique is easy to use, relatively cost-effective, very efficient, and ensures the safety of the environment. This review covers the literature gap by describing the major effects of organophosphates on the ecosystem and their bioremediation by using native bacterial strains.

IAA and ACC deaminase producing-bacteria isolated from the rhizosphere of pineapple plants grown under different abiotic and biotic stresses.

The roles of plant growth-promoting rhizobacteria in promoting plant growth and soil health, including alteration in plant metabolism and production of phytohormones such as indole-3-acetic acid (IAA) and the enzyme 1-aminocyclopropane-1-carboxylate (ACC) deaminase, are indisputable. This study aimed to isolate and characterize beneficial bacteria isolated from the rhizosphere of pineapple from distinct stress-inducing habitats, including water excess-, herbicide-over-treated-, and pathogen-infected areas at PT Great Giant Foods located in Lampung, Indonesia. The isolated bacteria were screened based on IAA production and ACC deaminase activities. Six selected isolates produced IAA with concentrations of up to 36.93 mgL-1. The highest value belongs to Bacillus sp. NCTB5I, followed by Brevundimonas sp. CHTB 2C (13.13 mgL-1) and Pseudomonas sp. CHTB 5B (6.65 mgL-1). All isolates were detected with ACC deaminase activities with Brevundimonas sp. CHTJ 5H consuming 88% of ACC over 24 h, the highest among all. Brevundimonas sp. CHTB 2C was detected with the highest ACC deaminase activity with the value of 13,370 nm α-ketobutyrate mg-1h-1. In another experiment, it was revealed that all selected isolates promote soybean growth. These bacteria are potential to be developed in the future as bioagents to promote plant growth, especially under stressful environmental conditions.

Co-inoculation of Mycorrhiza and methyl jasmonate regulates morpho-physiological and antioxidant responses of Crocus sativus (Saffron) under salinity stress conditions.

Salinity stress is the second most devastating abiotic factor limiting plant growth and yields. Climate changes have significantly increased salinity levels of soil. Besides improving the physiological responses under stress conditions, jasmonates modulate Mycorrhiza-Plant relationships. The present study aimed to evaluate the effects of methyl jasmonate (MeJ) and Funneliformis mosseae (Arbuscular mycorrhizal (AM) on morphology and improving antioxidant mechanisms in Crocus sativus L. under salinity stress. After inoculation with AM, pre-treated C. sativus corms with MeJ were grown under low, moderate, and severe salinity stress. Intense salinity levels damaged the corm, root, total leaf dry weight, and area. Salinities up to 50 mM increased Proline content and Polyphenol oxidase (PPO) activity, but MeJ increased this trend in proline. Generally, MeJ increased anthocyanins, total soluble sugars, and PPO. Total chlorophyll and superoxide dismutase (SOD) activity increased by salinity. The maximum catalase and SOD activities in + MeJ + AM were 50 and 125 mM, respectively, and the maximum total chlorophyll in -MeJ + AM treatment was 75 mM. Although 20 and 50 mM increased plant growth, using mycorrhiza and jasmonate enhanced this trend. Moreover, these treatments reduced the damage of 75 and 100 mM salinity stress. Using MeJ and AM can improve the growth of saffron under various ranges of salinity stress levels; however, in severe levels like 120 mM, this phytohormone and F. mosseae effects on saffron could be adverse.

Optimization and identification of siderophores produced by Pseudomonas monteilii strain MN759447 and its antagonism toward fungi associated with mortality in Dalbergia sissoo plantation forests.

Siderophore-positive bacteria present in the rhizosphere and in bulk soil assist plants by either inhibiting phytopathogen proliferation or increasing plant growth. The bacterial diversity of the Shisham forest ecosystem in the Tarai region of the Western Himalayas was studied and used for siderophore production, taking into account the large-scale dieback and wilt-induced mortality in Dalbergia sissoo (common name: shisham) plantation forests and the importance of soil microbes in tree health. In addition, Pseudomonas, Burkholderia, and Streptomyces were prominent siderophore-positive bacteria in Shisham forests. Pseudomonas species are known for their remarkable siderophore-producing ability. Bacterial siderophores inhibit pathogen growth by rapidly lowering the number of ferric ions in the rhizosphere. The Pseudomonas monteilii strain MN759447 was isolated from a D. sissoo plantation forest at the Agroforestry Research Centre, Pantnagar, Uttarakhand (28°58'N 79°25'E/28.97°N 79.41°E). It produces a significant number of siderophore units (80.36% in total). A two-stage optimization of growth factors was attempted in the strain MN759447 for better siderophore recovery. In the first-stage single-factor experiment, among the five variables studied, only pH, NH4NO3 concentration, and Fe concentration affected siderophore synthesis. In the second stage, an optimization of pH, NH4NO3 concentration, and Fe concentration for improved growth and enhanced siderophore production was carried out using a Box-Behnken design with response surface methodology. By using LC-MS, two derivatives of pseudomonine, salicylic acid, and kynurenic acid were detected as siderophores in the purified XAD-2 methanol extract of the P. monteilii strain MN759447. In addition to siderophore production, the P. monteilii strain MN759447 also exhibited a broad range of antagonistic activity against Aspergillus calidoustus (65%), Fusarium oxysporum (41.66%), Talaromyces pinophilus (65%), and Talaromyces verruculosus (65.1%) that are linked to sissoo mortality. To our knowledge, this is the first report on siderophore-producing bacteria isolated, identified, and characterized from the D. sissoo Roxb. forest habitat. This strain can also be developed as a commercial product.

Pseudomonas aeruginosa inhibits quorum-sensing mechanisms of soft rot pathogen Lelliottia amnigena RCE to regulate its virulence factors and biofilm formation.

The quorum-sensing (QS) cascade is responsible for the colonization and phenotypic behavior of the pathogenic organism and the regulation of diverse signal molecules. The disruption of the quorum-sensing system is an effective strategy to overcome the possibility of antibiotic resistance development in the pathogen. The quorum quenching does not kill the microbes. Instead, it hinders the expression of pathogenic traits. In the present experiment, Pseudomonas aeruginosa RKC1 was used to extract the metabolites responsible for quorum-sensing inhibition in soft rot pathogen Lelliottia amnigena RCE. During the initial screening, P. aeruginosa RKC1 was found to be most promising and inhibits violacein of Chromobacterium violaceum MTCC2656 pyocyanin, swarming-swimming motility of P. aeruginosa MTCC2297. The characterization of metabolites produced by the microbes which are responsible for quorum-sensing inhibition through GC-MS is very scarce in scientific literature. The ethyl acetate extract of P. aeruginosa RKC1 inhibits biofilm formation of L. amnigena RCE while inhibiting growth at higher concentrations. The GC-MS analysis suggested that Cyclic dipeptides (CDPs) such as Cyclo (L-prolyl-L-valine), Cyclo (Pro-Leu), and Cyclo(D-phenylalanyl-L-prolyl) were predominantly found in the ethyl acetate extract of the P. aeruginosa RKC1 (93.72%). This diketopiperazine (DKPs) exhibited quorum-sensing inhibition against the pathogen in liquid media during the active growth phase and regulated diverse metabolites of the pathogen. Moreover, the metabolites data from the clear zone around wells showed a higher concentration of DKSs (9.66%) compared to other metabolites. So far, very few reports indicate the role of DKPs or CDPs in inhibiting the quorum-sensing system in plant pathogenic bacteria. This is one such report that exploits metabolites of P. aeruginosa RKC1. The present investigation provided evidence to use quorum-sensing inhibitor metabolites, to suppress microbes' pathogenesis and thus develop an innovative strategy to overcome antibiotic resistance.

Environmental DNA Metabarcoding: A Novel Contrivance for Documenting Terrestrial Biodiversity.

The dearth of cardinal data on species presence, dispersion, abundance, and habitat prerequisites, besides the threats impeded by escalating human pressure has enormously affected biodiversity conservation. The innovative concept of eDNA, has been introduced as a way of overcoming many of the difficulties of rigorous conventional investigations, and is hence becoming a prominent and novel method for assessing biodiversity. Recently the demand for eDNA in ecology and conservation has expanded exceedingly, despite the lack of coordinated development in appreciation of its strengths and limitations. Therefore it is pertinent and indispensable to evaluate the extent and significance of eDNA-based investigations in terrestrial habitats and to classify and recognize the critical considerations that need to be accounted before using such an approach. Presented here is a brief review to summarize the prospects and constraints of utilizing eDNA in terrestrial ecosystems, which has not been explored and exploited in greater depth and detail in such ecosystems. Given these obstacles, we focused primarily on compiling the most current research findings from journals accessible in eDNA analysis that discuss terrestrial ecosystems (2012-2022). In the current evaluation, we also review advancements and limitations related to the eDNA technique.

Insight into Recent Progress and Perspectives in Improvement of Antioxidant Machinery upon PGPR Augmentation in Plants under Drought Stress: A Review.

Agriculture has a lot of responsibility as the rise in the world's population demands more food requirements. However, more than one type of biotic and abiotic stress continually impacts agricultural productivity. Drought stress is a major abiotic stress that significantly affects agricultural productivity every year as the plants undergo several morphological, biochemical, and physiological modifications, such as repressed root and shoot growth, reduced photosynthesis and transpiration rate, excessive production of reactive oxygen species (ROS), osmotic adjustments, and modified leaf senescence regulating and stress signaling pathways. Such modifications may permanently damage the plants; therefore, mitigation strategies must be developed. The use of drought resistant crop cultivars is more expensive and labor-intensive with few advantages. However, exploiting plant growth promoting rhizobacteria (PGPR) is a proven alternative with numerous direct and indirect advantages. The PGPR confers induced systemic tolerance (IST) mechanisms in plants in response to drought stress via multiple mechanisms, including the alteration of root architecture, maintenance of high relative water content, improvement of photosynthesis rate, production of phytohormones, exopolysaccharides, ACC deaminase, carotenoids and volatiles, induction of antioxidant defense system, and alteration in stress-responsive gene expression. The commercial application of PGPR as bioinoculants or biostimulants will remain contingent on more robust strain selection and performance under unfavorable environmental conditions. This review highlights the possible mechanisms of PGPR by activating the plant adaptive defense systems for enhancing drought tolerance and improving overall growth and yield.

Evaluation of Plant Growth-Promoting and Salinity Ameliorating Potential of Halophilic Bacteria Isolated From Saline Soil.

Among the biotic and abiotic stress affecting the physical, chemical, and biological properties of soil, salinity is a major threat that leads to the desertification of cultivable land throughout the world. The existence of diverse and versatile microbial populations inhabiting the nutrient-rich soil and varied soil conditions affects the soil dynamism. A normal soil constitutes 600 million bacteria belonging to about 20,000 species, which is reduced to 1 million with 5,000-8,000 species in stress conditions. Plant growth-promoting rhizobacteria (PGPR) are in symbiotic association with the plant system, which helps in combating the abiotic stress and increases the overall productivity and yield. These microorganisms are actively associated with varied cellular communication processes through quorum sensing and secondary metabolites such as the production of Indole-3-acetic acid (IAA), exopolysaccharide (EPS) siderophore, ammonia, ACC deaminase, and solubilization of phosphate. The present study focused on the isolation, identification, and characterization of the microorganisms isolated from the seacoast of Dandi, Navsari. Twelve isolates exhibited PGP traits at a high salt concentration of 15-20%. AD9 isolate identified as Bacillus halotolerans showed a higher ammonia production (88 ± 1.73 µg/mL) and phosphate solubilization (86 ± 3.06 µg/mL) at 15% salt concentration, while AD32* (Bacillus sp. clone ADCNO) gave 42.67 ±1.20 µg/mL IAA production at 20% salt concentration. AD2 (Streptomyces sp. clone ADCNB) and AD26 (Achromobacter sp. clone ADCNI) showed ACC deaminase activity of 0.61 ± 0.12 and 0.60 ± 0.04 nM α-ketobutyrate/mg protein/h, respectively. AD32 (Bacillus sp. clone ADCNL) gave a high siderophore activity of 65.40 ± 1.65%. These isolates produced salinity ameliorating traits, total antioxidant activities, and antioxidant enzymes viz. superoxide dismutase (SOD), Glutathione oxidase (GSH), and catalase (CAT). Inoculation of the multipotent isolate that produced PGP traits and salinity ameliorating metabolites promoted the plant growth and development in rice under salinity stress conditions. These results in 50% more root length, 25.00% more plant dry weight, and 41% more tillers compared to its control.

Plant-Derived Protectants in Combating Soil-Borne Fungal Infections in Tomato and Chilli.

Fungal infections transmitted through the soil continue to pose a threat to a variety of horticultural and agricultural products, including tomato and chilli. The indiscriminate use of synthetic pesticides has resulted in a slew of unintended consequences for the surrounding ecosystem. To achieve sustainable productivity, experts have turned their attention to natural alternatives. Due to their biodegradability, varied mode of action, and minimal toxicity to non-target organisms, plant-derived protectants (PDPs) are being hailed as a superior replacement for plant pesticides. This review outlines PDPs' critical functions (including formulations) in regulating soil-borne fungal diseases, keeping tomato and chilli pathogens in the spotlight. An in-depth examination of the impact of PDPs on pathogen activity will be a priority. Additionally, this review emphasises the advantages of the in silico approach over conventional approaches for screening plants' secondary metabolites with target-specific fungicidal activity. Despite the recent advances in our understanding of the fungicidal capabilities of various PDPs, it is taking much longer for that information to be applied to commercially available pesticides. The restrictions to solving this issue can be lifted by breakthroughs in formulation technology, governmental support, and a willingness to pursue green alternatives among farmers and industries.

Zinc nutrition and arbuscular mycorrhizal symbiosis effects on maize (Zea mays L.) growth and productivity.

Zinc (Zn) is an essential micronutrient required to enhance crop growth and yield. In the arid - semiarid region, Zn deficiency is expected due to alkaline calcareous soil. Contrarily, Zn toxicity is also becoming an environmental concern due to increasing anthropogenic activities (metal smelting, copper industry, etc.). Therefore, balanced Zn application is necessary to save resources and achieve optimum crop growth and yield. Most scientists suggest biological approaches to overcome the problem of Zn toxicity and deficiency. These biological approaches are mostly environment-friendly and cost-effective. In these biological approaches, the use of arbuscular mycorrhizae fungi (AMF) symbiosis is becoming popular. It can provide tolerance to the host plant against Zn-induced stress. Inoculation of AMF helps in balance uptake of Zn and enhances the growth and yield of crops. On the other hand, maize (Zea mays L.) is an important cereal crop due to its multifarious uses. As maize is an effective host for mycorrhizae symbiosis, that's why this review was written to elaborate on the beneficial role of arbuscular mycorrhizal fungi (AMF). The review aimed to glance at the recent advances in the use of AMF to enhance nutrient uptake, especially Zn. It was also aimed to discuss the mechanism of AMF to overcome the toxic effect of Zn. We have also discussed the detailed mechanism and physiological improvement in the maize plant. In conclusion, AMF can play an imperative role in improving maize growth, yield, and balance uptake of Zn by alleviating Zn stress and mitigating its toxicity.

Co-inoculation of rhizobacteria promotes growth, yield, and nutrient contents in soybean and improves soil enzymes and nutrients under drought conditions.

Drought stress is the major abiotic factor limiting crop production. Co-inoculating crops with nitrogen fixing bacteria and plant growth-promoting rhizobacteria (PGPR) improves plant growth and increases drought tolerance in arid or semiarid areas. Soybean is a major source of high-quality protein and oil for humans. It is susceptible to drought stress conditions. The co-inoculation of drought-stressed soybean with nodulating rhizobia and root-colonizing, PGPR improves the root and the shoot growth, formation of nodules, and nitrogen fixation capacity in soybean. The present study was aimed to observe if the co-inoculation of soybean (Glycine max L. (Merr.) nodulating with Bradyrhizobium japonicum USDA110 and PGPR Pseudomonas putida NUU8 can enhance drought tolerance, nodulation, plant growth, and nutrient uptake under drought conditions. The results of the study showed that co-inoculation with B. japonicum USDA110 and P. putida NUU8 gave more benefits in nodulation and growth of soybean compared to plants inoculated with B. japonicum USDA110 alone and uninoculated control. Under drought conditions, co-inoculation of B. japonicum USDA 110 and P. putida NUU8 significantly enhanced the root length by 56%, shoot length by 33%, root dry weight by 47%, shoot dry weight by 48%, and nodule number 17% compared to the control under drought-stressed. Co-inoculation with B. japonicum, USDA 110 and P. putida NUU8 significantly enhanced plant and soil nutrients and soil enzymes compared to control under normal and drought stress conditions. The synergistic use of B. japonicum USDA110 and P. putida NUU8 improves plant growth and nodulation of soybean under drought stress conditions. The results suggested that these strains could be used to formulate a consortium of biofertilizers for sustainable production of soybean under drought-stressed field conditions.

Optimizing nutrient use efficiency, productivity, energetics, and economics of red cabbage following mineral fertilization and biopriming with compatible rhizosphere microbes.

Conventional agricultural practices and rising energy crisis create a question about the sustainability of the present-day food production system. Nutrient exhaustive crops can have a severe impact on native soil fertility by causing nutrient mining. In this backdrop, we conducted a comprehensive assessment of bio-priming intervention in red cabbage production considering nutrient uptake, the annual change in soil fertility, nutrient use efficiency, energy budgeting, and economic benefits for its sustainable intensification, among resource-poor farmers of Middle Gangetic Plains. The compatible microbial agents used in the study include Trichoderma harzianum, Pseudomonas fluorescens, and Bacillus subtilis. Field assays (2016-2017 and 2017-2018) of the present study revealed supplementing 75% of recommended NPK fertilizer with dual inoculation of T. harzianum and P. fluorescens increased macronutrient uptake (N, P, and K), root length, heading percentage, head diameter, head weight, and the total weight of red cabbage along with a positive annual change in soil organic carbon. Maximum positive annual change in available N and available P was recorded under 75% RDF + P. fluorescens + B. subtilis and 75% RDF + T. harzianum + B. subtilis, respectively. Bio-primed plants were also higher in terms of growth and nutrient use efficiency (agronomic efficiency, physiological efficiency, apparent recovery efficiency, partial factor productivity). Energy output (26,370 and 26,630 MJ ha-1), energy balance (13,643 and 13,903 MJ ha-1), maximum gross return (US $ 16,030 and 13,877 ha-1), and net return (US $ 15,966 and 13,813 ha-1) were considerably higher in T. harzianum, and P. fluorescens treated plants. The results suggest the significance of the bio-priming approach under existing integrated nutrient management strategies and the role of dual inoculations in producing synergistic effects on plant growth and maintaining the soil, food, and energy nexus.

Pythium Damping-Off and Root Rot of Capsicum annuum L.: Impacts, Diagnosis, and Management.

Capsicum annuum L. is a significant horticulture crop known for its pungent varieties and used as a spice. The pungent character in the plant, known as capsaicinoid, has been discovered to have various health benefits. However, its production has been affected due to various exogenous stresses, including diseases caused by a soil-borne pathogen, Pythium spp. predominantly affecting the Capsicum plant in younger stages and causing damping-off, this pathogen can incite root rot in later plant growth stages. Due to the involvement of multiple Pythium spp. and their capability to disperse through various routes, their detection and diagnosis have become crucial. However, the quest for a point-of-care technology is still far from over. The use of an integrated approach with cultural and biological techniques for the management of Pythium spp. can be the best and most sustainable alternative to the traditionally used and hazardous chemical approach. The lack of race-specific resistance genes against Pythium spp. can be compensated with the candidate quantitative trait loci (QTL) genes in C. annuum L. This review will focus on the epidemiological factors playing a major role in disease spread, the currently available diagnostics in species identification, and the management strategies with a special emphasis on Pythium spp. causing damping-off and root rot in different cultivars of C. annuum L.

Multifarious Indigenous Diazotrophic Rhizobacteria of Rice (Oryza sativa L.) Rhizosphere and Their Effect on Plant Growth Promotion.

A diverse group of rhizobacteria persists in the rhizospheric soil, on the surface of roots, or in association with rice plants. These bacteria colonize plant root systems, enhance plant growth and crop yield. Indigenous rhizobacteria are known to promote soil health, grain production quality and serve as sustainable bioinoculant. The present study was aimed to isolate, identify and characterize indigenous plant growth promoting (PGP) diazotrophic bacteria associated with the rhizosphere of rice fields from different areas of Jammu and Kashmir, India. A total of 15 bacteria were isolated and evaluated for various PGP traits, antagonistic activity against phytopathogens, production of hydrolytic enzymes and biofilm formation under in-vitro conditions. The majority of the isolated bacteria were Gram-negative. Out of 15 bacterial isolates, nine isolates produced IAA (12.24 ± 2.86 to 250.3 ± 1.15 µg/ml), 6 isolates exhibited phosphate solubilization activity (36.69 ± 1.63 to 312.4 ± 1.15 µg/ml), 7 isolates exhibited rock phosphate solubilization while 5 isolates solubilized zinc (10-18 mm), 7 isolates showed siderophore production, 8 isolates exhibited HCN production, 6 isolates exhibited aminocyclopropane-1-carboxylate (ACC) deaminase activity, 13 isolates exhibited cellulase activity, nine isolates exhibited amylase and lipase activity and six isolates exhibited chitinase activity. In addition, 5 isolates showed amplification with the nifH gene and showed a significant amount of nitrogenase activity in a range of 0.127-4.39 µmol C2H4/mg protein/h. Five isolates viz., IHK-1, IHK-3, IHK-13, IHK-15 and IHK-25 exhibited most PGP attributes and successfully limited the mycelial growth of Rhizoctonia solani and Fusarium oxysporum in-vitro. All the five bacterial isolates were identified based on morphological, biochemical and 16S rDNA gene sequencing study, as Stenotrophomonas maltophilia, Enterobacter sp., Bacillus sp., Ochrobactrum haematophilum and Pseudomonas aeruginosa. Rice plants developed from seeds inoculated with these PGP strains individually had considerably higher germination percentage, seed vigor index and total dry biomass when compared to control. These findings strongly imply that the PGP diazotrophic bacteria identified in this work could be employed as plant growth stimulators in rice.

Sustainability appraisal of shifting cultivation in the Chittagong Hill Tracts of Bangladesh.

An integrated socioeconomic and erosion study on the sustainability of traditional shifting cultivation (Jhum) carried out in 1998 and 1999 in the Chittagong Hill Tracts (CHT) of Bangladesh showed the system to be nonsustainable under the current conditions with fallow periods of only 3-5 years and lack of land rights. An estimated input (mainly labor) of USD 380 ha(-1) yr(-1) results in only a total output of USD 360 ha(-1) yr(-1) and Jhum cultivated areas suffer severe loss of soil and valuable plant nutrients along with runoff during the rainy season. To compensate these losses by commercial fertilizers will cost nearly USD 2 million yr(-1) for CHT. To ensure long-term productivity of the soils, Jhum should therefore be adjusted to a tolerable level and farmers should be given rights and title to the land to motivate them to switch to improved, settled farming systems.

Molecular analysis and characterization of the Cochliobolus heterostrophus beta-tubulin gene and its possible role in conferring resistance to benomyl.

Cochliobolus heterostrophus Tub1 described here is the first beta-tubulin gene characterized from a naturally occurring benomyl-resistant ascomycete plant pathogen. The gene encodes a protein of 447 amino acids. The coding region of Tub1 is interrupted by three introns, of 116, 55, and 56 nt, situated after codons 4, 12, and 53, respectively. As a result of the preference for pyrimidines in the third position of the codons when a choice exists between purines and pyrimidines, codon usage in the Tub1 gene is biased. Tub1 shows high homology with beta-tubulin genes of other ascomycete species. However, Tub1 is exceptional in having Tyr(167), compared with Phe(167), possessed by beta-tubulin genes of other ascomycetes sequenced thus far. The Tyr(167) residue has been associated with benomyl resistance in other organisms. In contrast, all other benomyl-implicated residues of Tub1 correspond to sensitivity. Based on these results, we suggest that benomyl resistance in the fungus probably is attributed to Tyr(167).