Allohexaploid wheat (Triticum aestivum L.) is tolerant to halauxifen-methyl (HM) via rapid detoxification of the phytotoxic form of HM, halauxifen acid (HA), to non-phytotoxic metabolites. Previous research utilizing ‘Chinese Spring’ (CS) wheat, alien substitution (i.e., endogenous chromosome pair substituted with a homoeologous pair from diploid Sears’ goatgrass (Aegilops searsii M. Feldman & M. Kislev) (AS), or nullisomic-tetrasomic (NT) lines indicated plants lacking chromosome 5A are more sensitive than CS to HM. We hypothesized the increased HM sensitivity of these plants results from losing gene(s) on chromosome 5A associated with HA metabolism, which leads to a reduced HA detoxification rate relative to CS. To compare HA abundance among AS, CS, alien substitution, and NT lines during a time course, two excised-leaf studies using unlabeled HM and liquid chromatography–mass spectrometry analyses were performed. Aegilops searsii accumulated more HA than CS did, and each substitution line at 8, 12, and 24 h after treatment (HAT). Furthermore, only the wheat substitution line lacking chromosome 5A displayed greater abundance of HA relative to CS (2.4- to 3.8-fold, depending on the time point). In contrast, HA abundances in lines possessing chromosome 5A were not different than HA abundances in CS at all time points. When NT lines were compared with CS, the nullisomic 5D-tetrasomic 5A (N5D-T5A) line displayed similar HA abundance, whereas the nullisomic 5A-tetrasomic 5D (N5A-T5D) accumulated approximately 3-fold more HA at 12 and 24 HAT. These results biochemically support the hypothesis that genes encoding HA-detoxifying enzyme(s) are located on wheat chromosome 5A and corroborate findings from previous greenhouse phenotypic experiments. Future experimentation is needed to identify and characterize genes and enzymes on wheat chromosome 5A involved with HA detoxification, which may include cytochrome P450 monooxygenases, unknown oxidases, UDP-dependent glucosyltransferases, or, potentially, transcription factors that regulate expression of these genes associated with HA detoxification.

The South American tomato moth, Phthorimaea absoluta (Meyrick), is one of the key pests of tomato in India. Since its report in 2014, chemical control has been the main means of tackling this pest, both in the open field and protected cultivation. Despite regular insecticidal sprays, many outbreaks were reported from major tomato-growing regions of South India during 2019–2020. A study was conducted to investigate the effect of insecticide resistance on biology, biochemical enzymes, and gene expression in various P. absoluta field populations viz., Bangalore, Kolar, Madurai, Salem, and Anantapur to commonly used insecticides such as flubendiamide, cyantraniliprole, and indoxacarb. Increased levels of insecticide resistance ratios (RR) were recorded in P. absoluta populations of different locations. A significant increase in cytochrome P450 monooxygenase (CYP/MFO) and esterase levels was noticed in the resistant population compared to susceptible one. Through molecular studies, we identified four new CYP genes viz., CYP248f (flubendiamide), CYP272c, CYP724c (cyantraniliprole), and CYP648i (indoxacarb). The expression levels of these genes significantly increased as the folds of resistance increased from G1 to G20 (generation), indicating involvement of the identified genes in insecticide resistance development in P. absoluta. In addition, the resistant populations showed decreased fecundity, increased larval development period, and adult longevity, resulting in more crop damage. The information generated in the present study thus helps in understanding the development of insecticide resistance by P. absoluta and suggests the farmers and researchers to use insecticides wisely by adopting insecticide resistance management as a strategy under integrated pest management.

The basis for 2,4-D [(2,4-dichlorophenoxy)acetic acid] antagonism of diclofop-methyl {methyl 2-[4-(2,4-dichlorophenoxy) phenoxy] propanoate} toxicity to wild oat (Avena fatua L.) was investigated by studying changes in the metabolism of diclofop-methyl in vitro. An esterase from wild oat, which hydrolyzes diclofop-methyl to the acid diclofop, was extracted, partially purified, and the reaction characterized. The rate of hydrolysis of 14C-diclofop-methyl was 0.14 ηmoles/2 h at standard assay conditions of 0.25 mg lyophilized enzyme preparation (19.6% protein) in 0.1 ml phosphate buffer (0.1 M, pH 7.0), substrate 5 μM. The addition of 2,4-D to this reaction did not inhibit 14C-diclofop formation. Higher levels of 2,4-D stimulated enzymic hydrolysis. 14C-diclofop-methyl was rapidly metabolized to 14C-diclofop and polar 14C-conjugates when vacuum-infiltrated into wild oat leaf segments. The addition of 2,4-D caused small increases in the rates of both 14C-diclofop-methyl de-esterification and 14C-diclofop conjugation. It is concluded that 2,4-D does not inhibit the in vitro de-esterification of diclofop-methyl.

Esterase-based resistance in the peach-potato aphid, Myzus persicae (Sulzer), is generally due to one of two alternative amplified carboxylesterase genes, E4 or FE4 (fast E4). The E4 amplified form is distributed worldwide and it is correlated with a particular translocation between autosomes 1 and 3, whereas the FE4 form, which has hitherto not been found to be associated with chromosomal rearrangements, is typical of the Mediterranean regions. In this study, we present for the first time cytogenetic and molecular data on some M. persicae parthenogenetic lineages, which clearly show a chromosomal A1-3 translocation associated with esterase FE4 genes and unrelated to high levels of esterase-based resistance.

The effect of accelerated aging (AA) and seed priming on specific seed enzymes was studied in soybeans. Aging seeds for 48 and 96 h at 41°C and 100% humidity reduced germination from 92% to 68% and 0%, respectively. Malate dehydrogenase (MDH) glutamate dehydrogenase (GDH), and esterase were assayed in extracts from dry and imbibing seeds. Malate dehydrogenase activity was the least affected by AA treatments. Certain GDH isozyme activity was reduced by 17% after 48 h of AA and almost completely lost after 96 h. Imbibing for 24 h increased GDH activity in 48 h-accelerated-aged seeds but not the 96 h-aged seeds. Two esterase isozymes in dry seeds showed a 77% loss of activity after 48 h of AA. This treatment increased the total esterase activity, with the greatest activity observed after 96 h of AA. This was due to the appearance of a diffuse high molecular weight (HMW) region of activity observed on native polyacrylamide gels and specifically induced in the embryonic axes. Imbibing 48-h-accelerated-aged seeds for 48 h caused a reappearance of the two esterase enzymes lost during aging. The HMW esterase staining region also appeared after imbibing unaged seeds for 24 h. Seed priming, caused a response similar to imbibition for each enzyme except the HMW esterase activity. Priming 48-h-accelerated-aged seeds reduced the HMW esterase activity. This was not observed during imbibition of aged seeds and indicates the first identification of a specific enzymatic response unique to a seed priming treatment.

An enzyme found previously in extracts of adult Schistosoma mansoni worms, that hydrolysed the chromogenic substrate N-acetyl-DL-phenylalanine β-naphthyl-ester, has here been further investigated and characterized. Evidence that the molecule found in the parasite was antigenically and enzymatically homologous with a constituent of normal mouse plasma has been consolidated using a monospecific serum in immunoelectrophoresis and Western immunoblotting. The molecular size of the enzyme was found to be approximately 70 kDa and it was inhibited by a serine protease inhibitor, but not by inhibitors of other classes of protease. The enzymatic activity found in normal mouse serum was also found in normal rat serum, but not in sera from several other mammalian species.

The aim of this study was to develop and optimize silverleaf bioassay, esterase analysis and PCR-based techniques to distinguish quickly and reliably biotype B of the whitefly, Bemisia tabaci (Gennadius), from Indian indigenous biotypes. Zucchini and squash readily develop silverleaf symptoms upon feeding by the B biotype, but they are not readily available in Indian markets. A local pumpkin variety ‘Big’ was, therefore, used in silverleaf assay, which developed symptoms similar to those on zucchini and squash and can be used reliably to detect B biotype. Analysis of non-specific esterases of B and the indigenous biotypes indicated both quantitative and qualitative differences in esterase patterns. Two high molecular weight bands were unique to B biotype and they occurred in abundance. These esterases were used to develop quick and field-based novel detection methods for differentiating B from the indigenous biotypes. Development of these simple and cost-effective protocols has wider application as they can be potentially used to identify other agricultural pests. Mitochondrial cytochrome oxidase I gene sequences and randomly amplified polymorphic DNA (RAPD) polymorphisms, generated using the primer OpB11, were also found useful for detecting B. tabaci biotypes. A B biotype-specific RAPD band of 800 bp was sequenced, which was used to a develop sequence characterized amplified region (SCAR) marker. The SCAR marker involved the development of B biotype-specific primers that amplified 550 bp PCR products only from B biotype genomic DNA. Silverleaf assay, esterases, RAPDs or a SCAR marker were used in combination to analyse whitefly samples collected from selected locations in India, and it was found that any of these techniques can be used singly or in combination to detect B biotype reliably. The B biotype was found in southern parts of India but not in the north in 2004–06.

Two forms of Myzus persicae (Sulzer), F-15 and R306N′, resembling M. antirrhinii (Macchiati) in esterase profile have been found in Japan. To determine the genetic relationship of F-15 and R306N′ with M. antirrhinii, clonal lineages of F-15, R306N′ and those of 14 other forms of M. persicae collected in Japan were analysed for 17 polymorphic microsatellite DNA loci. Two microsatellite multi-locus (MS) genotypes of M. persicae and two MS genotypes, Ma3 and Ma4, of M. antirrhinii from Australia were used as references. The 16 Japanese forms were represented by different MS genotypes. F-15 and R306N′ had, respectively, 44.1% and 35.3% of their alleles at the 17 loci in common with Ma3 and Ma4, and some of the common alleles were peculiar in size. F-15 and R306N′ shared 22.6% and 29.6%, respectively, of their alleles with the other 16 MS genotypes of M. persicae. The corresponding values for Ma3 and Ma4 were both 27.4%. F-15 and R306N′ were grouped reliably with the two Australian MS genotypes of M. antirrhinii both by neighbour-joining method based on shared allele distance and by a Bayesian method with Markov Chain Monte Carlo algorithm. These results suggest that F-15 and R306N′ are genetically closely related to M. antirrhinii and isolated from the gene pool of M. persicae despite their ability to produce sexual morphs. It is therefore proposed that F-15 and R306N′ should be classified as M. antirrhinii.

Genetic diversity in four populations of the Eurasian badger Meles meles from Denmark was estimated by allozyme electrophoresis. Low genetic variability was found within populations as compared to the variability found in badger populations from other countries, whereas significant genetic differentiation was found between populations. The low genetic variability and the differentiation between populations is explained by the fragmentation of the Danish landscape, which reduces the effective population size of local populations and the gene flow between them. Low genetic variability found for Danish badgers also supports the hypothesis of a rapid recolonization following the last glaciation.

Bile-salt activated lipase (BAL) is a pancreatic enzyme that digests a variety of lipids in the small intestine. A distinct property of BAL is its dependency on bile salts in hydrolyzing substrates of long acyl chains or bulky alcoholic motifs. A crystal structure of the catalytic domain of human BAL (residues 1–538) with two surface mutations (N186D and A298D), which were introduced in attempting to facilitate crystallization, has been determined at 2.3 Å resolution. The crystal form belongs to space group P212121 with one monomer per asymmetric unit, and the protein shows an α/β hydrolase fold. In the absence of bound bile salt molecules, the protein possesses a preformed catalytic triad and a functional oxyanion hole. Several surface loops around the active site are mobile, including two loops potentially involved in substrate binding (residues 115–125 and 270–285).

Intensive investigations of genetic variation within and among populations of the whitefly, Bemisia tabaci, are under way in Israel and in Colombia. A polymorphic esterase (EST), which seems to be controlled by a Mendelian locus, has two alleles in sufficiently high frequencies to be useful as a genetic marker. The analysis of more than 10,000 individual adult whiteflies shows genetic differences among localities in the frequencies of the fast (F) and slow (S) alleles. These differences seem to be related to the intensity of insecticide use against the whitefly. The frequency of F is high in unsprayed areas and in control laboratory populations, but is close to zero in heavily-sprayed agricultural areas. The frequency of Fon cotton in agricultural areas is high in winter and spring, but decreases dramatically in the summer (coincidental with the onset of pest-control activities on cotton). A similar temporal pattern on untreated Lantana may be due to migration of adults from agricultural fields. There was no indication of differentiation of populations on different host plants. In Colombia, the difference in EST patterns in different States are very pronounced, suggestive of geographical races. Small samples from Florida, California and Kenya indicate that B. tabaci in different countries may be genetically distinct.