Effect of temperature on the biological parameters of pink bollworm, Pectinophora gossypiella Saunders (Lepidoptera: Gelechiidae).

Pink bollworm (PBW) Pectinophora gossypiella is an important pest cotton worldwide. There are multiple factors which determines the occurrence and distribution of P. gossypiella across different cotton growing regions of the world, and one such key factor is 'temperature'. The aim was to analyze the life history traits of PBW across varying temperature conditions. We systematically explored the biological and demographic parameters of P. gossypiella at five distinct temperatures; 20, 25, 30, 35 and 40 ± 1 °C maintaining a photoperiod of LD 16:8 h. The results revealed that the total developmental period of PBW shortens with rising temperatures, and the highest larval survival rates were observed between 30 °C and 35 °C, reaching 86.66% and 80.67%, respectively. Moreover, significant impacts were observed as the pupal weight, percent mating success, and fecundity exhibited higher values at 30 °C and 35 °C. Conversely, percent egg hatching, larval survival, and adult emergence were notably lower at 20 °C and 40 °C, respectively. Adult longevity decreased with rising temperatures, with females outliving males across all treatments. Notably, thermal stress had a persistent effect on the F1 generation, significantly affecting immature stages (egg and larvae), while its impact on reproductive potential was minimal. These findings offer valuable insights for predicting the population dynamics of P. gossypiella at the field level and developing climate-resilient management strategies in cotton.

Investigating the second whitefly population outbreak within a decade in the cotton growing zone of North India.

The whitefly, Bemisia tabaci (Gennadius), is a polyphagous and major pest of cotton worldwide. Both adults and nymphs of B. tabaci affect the crop by causing direct and indirect damage. A severe whitefly outbreak was experienced during 2015 on cotton in North India and this was followed by a profound infestation during 2022. The present research rigorously examined whether the proliferation in the whitefly population was an outbreak or the result of a multi factor resurgence. During 2015, whitefly counts remained above the economic threshold level (ETL) between 28th and 35th Standard Meteorological Week (SMW). However, during 2022 above ETL population was observed in 27th SMW and it persisted until 36th SMW. The peak incidence of the whitefly was noticed during 31st and 29th SMW in 2015 and 2022, respectively. The early pest build up in 2022 and longer persistence (≥10 weeks) over the cotton season resulted in more damage to cotton crop. Additionally, pest survillence across the zone on the farmers' fields during 2022 revealed 44.4 per cent spots (585 out of 1,317 locations) above ETL while the corresponding locations in 2015 was 57% (620 out of 1,089). Thus, in 2022 infestation was not uniform in the entire zone wherein only few blocks of Punjab, Haryana and Rajasthan states of India experienced severe infestations of the whitefly. This study reports the complex of factors including weather, delayed sowing, use of tank mixtures/ subleathal doses of insecticides, pest resurgence etc. that might have possibly contributed to these upsurges in whitefly on cotton in north India.

Effectiveness and residual activity of four common insecticides used in the Mississippi Delta to control tarnished plant bugs in cotton.

The tarnished plant bug, (TPB) Lygus lineolaris Palisot de Beauvois (Hemiptera: Miridae) is a key pest of cotton in the midsouth region and some areas of the eastern United States. Its control methods have been solely based on chemical insecticides which has contributed to insecticidal resistance and shortened residual periods for control of this insect pest. This study was conducted over a two-year period and examined the efficacy and residual effect of four commercial insecticides including lambda-cyhalothrin (pyrethroid), acephate (organophosphate), imidacloprid (neonicotinoid), and sulfoxaflor (sulfoxamine). The effectiveness and residual effects of these insecticides were determined by application on cotton field plots on four different dates during each season using three different concentrations (high: highest labeled commercial dose (CD), medium: 1/10 of the CD, low: 1/100 of the CD) on field cotton plots. Four groups of cotton leaves were randomly pulled from each treated plot and control 0-, 2-, 4-, 7-, and 9-days post treatment (DPT) and exposed to a lab colony of TPB adults. One extra leaf sample/ plot/ spray /DPT interval (0-2-4-7-9-11) during 2016 was randomly collected from the high concentration plots and sent to Mississippi State Chemical Laboratory for residual analysis. Mortality of TPB adults was greatest for those placed on leaves sprayed with the organophosphate insecticide with mortalities (%) of 81.7±23.4 and 63.3±28.8 (SE) 1-day after exposure (DAE) on leaves 0-DPT with the high concentration for 2016 and 2017, respectively, reaching 94.5±9.5 and 95.4±7.6 6-DAE each year. Mortality to all insecticides continued until 9 and 4-DPT for high and medium concentrations, respectively. However, organophosphate (39.4±28.6) and pyrethroid (24.4±9.9) exhibited higher mortality than sulfoxamine (10.6±6.6) and the neonicotinoid (4.0±1.5) 7-DAE on 9-DPT leaves with the high concentration. Based on our results using the current assay procedure, TPB adults were significantly more susceptible to contact than systemic insecticides and due to its residual effect, organophosphate could kill over 80% of the TPB population 7-DPT.

Ecotoxicity of Cadmium along the Soil-Cotton Plant-Cotton Bollworm System: Biotransfer, Trophic Accumulation, Plant Growth, Induction of Insect Detoxification Enzymes, and Immunocompetence.

Cadmium (Cd) is a hazardous element that may jeopardize environmental safety and human health through biotransfer and trophic accumulation. Here, we tested Cd toxicity on cotton plants, cotton bollworms, and their responses. Results demonstrated that Cd accumulated in plant roots, aerial parts, insect larvae, pupae, and frass in a dose-dependent pattern. The ∼9.35 mg kg-1 of Cd in plant aerial parts, ∼3.68 in larvae, ∼6.43 in pupae, and high transfer coefficient (∼5.59) indicate significant mobility. The ∼19.61 mg kg-1 of Cd in larvae frass suggests an effective detoxification strategy, while BAFcotton (∼1.14) and BAFworm (∼0.54) indicated low bioaccumulation. Cadmium exposure resulted in compromised plant growth and yield as well as alterations in photosynthetic pigment contents, antioxidant enzyme activities, and certain life history traits of cotton bollworms. Furthermore, carboxylesterase activity and encapsulation rates of insect larvae decreased with increasing Cd concentrations, whereas acetylcholinesterase, phenol oxidase, glutathione S-transferase, and multifunctional oxidase exhibited hormesis responses.

GhHAM regulates GoPGF-dependent gland development and contributes to broad-spectrum pest resistance in cotton.

Cotton is a globally cultivated crop, producing 87% of the natural fiber used in the global textile industry. The pigment glands, unique to cotton and its relatives, serve as a defense structure against pests and pathogens. However, the molecular mechanism underlying gland formation and the specific role of pigment glands in cotton's pest defense are still not well understood. In this study, we cloned a gland-related transcription factor GhHAM and generated the GhHAM knockout mutant using CRISPR/Cas9. Phenotypic observations, transcriptome analysis, and promoter-binding experiments revealed that GhHAM binds to the promoter of GoPGF, regulating pigment gland formation in cotton's multiple organs via the GoPGF-GhJUB1 module. The knockout of GhHAM significantly reduced gossypol production and increased cotton's susceptibility to pests in the field. Feeding assays demonstrated that more than 80% of the cotton bollworm larvae preferred ghham over the wild type. Furthermore, the ghham mutants displayed shorter cell length and decreased gibberellins (GA) production in the stem. Exogenous application of GA3 restored stem cell elongation but not gland formation, thereby indicating that GhHAM controls gland morphogenesis independently of GA. Our study sheds light on the functional differentiation of HAM proteins among plant species, highlights the significant role of pigment glands in influencing pest feeding preference, and provides a theoretical basis for breeding pest-resistant cotton varieties to address the challenges posed by frequent outbreaks of pests.

Sublethal effects of a commercial Bt product and Bt cotton flowers on the bollworm (Helicoverpa zea) with impacts to predation from a lady beetle (Hippodamia convergens).

Insecticidal Bacillus thuringiensis Berliner (Bt) toxins produced by transgenic cotton (Gossypium hirsutum L.) plants have become an essential component of cotton pest management. Bt toxins are the primary management tool in transgenic cotton for lepidopteran pests, the most important of which is the bollworm (Helicoverpa zea Boddie) (Lepidoptera: Noctuidae) in the United States (U.S.). However, bollworm larvae that survive after consuming Bt toxins may experience sublethal effects, which could alter interactions with other organisms, such as natural enemies. Experiments were conducted to evaluate how sublethal effects of a commercial Bt product (Dipel) incorporated into artificial diet and from Bt cotton flowers impact predation from the convergent lady beetle (Hippodamia convergens Guérin-Méneville) (Coleoptera: Coccinellidae), common in cotton fields of the mid-southern U.S. Sublethal effects were detected through reduced weight and slower development in bollworm larvae which fed on Dipel incorporated into artificial diet, Bollgard II, and Bollgard 3 cotton flowers. Sublethal effects from proteins incorporated into artificial diet were found to significantly alter predation from third instar lady beetle larvae. Predation of bollworm larvae also increased significantly after feeding for three days on a diet incorporated with Bt proteins. These results suggest that the changes in larval weight and development induced by Bt can be used to help predict consumption of bollworm larvae by the convergent lady beetle. These findings are essential to understanding the potential level of biological control in Bt cotton where lepidopteran larvae experience sublethal effects.

Monitoring cotton squares with opened and/or yellowed bracts to determine the need and efficency of chemical control against the cotton boll weevil.

The technique of terrestrial sampling of boll weevil (BW) populations is expensive and inefficient over large areas, but may be cheaper and more efficient without involving the manipulation of cotton squares. The aim of this study was to develop a technique to sampling cotton squares based on the observation of opened and/or yellowing bracts to determine the need and efficacy of chemical control of BW in cotton crops. The first experiment aimed to estimate the ratio between the number of cotton squares with opened and/or yellowed bracts and that of squares with BW oviposition punctures. The second experiment, aimed to determine the efficacy of chemical control for BW by sampling cotton squares with opened and/or yellowed bracts. The ratio between the number of opened and/or yellowed bracts and the number of cotton squares with oviposition punctures was 2:1. The level and efficiency of chemical control of BW, based on the percentage and sampling of cotton plants with opened and/or yellowed bracts, was 5% and did not differ from the one based on the observation of cotton plants with 10% cotton squares with oviposition punctures by BW females. The control level based on sampling cotton plants with open and/or yellowing bracts was 5%. The efficiency of chemical insecticides using this economic threshold against the BW did not differ from that based on sampling cotton plants with 10% of cotton squares with oviposition punctures by BW females. This indicates that the chemical control of cotton boll weevil can be carried out based on cotton squares with open and/or yellowed bracts.

Cotton plants overexpressing the Bacillus thuringiensis Cry23Aa and Cry37Aa binary-like toxins exhibit high resistance to the cotton boll weevil (Anthonomus grandis).

The cotton boll weevil (CBW, Anthonomus grandis) stands as one of the most significant threats to cotton crops (Gossypium hirsutum). Despite substantial efforts, the development of a commercially viable transgenic cotton event for effective open-field control of CBW has remained elusive. This study describes a detailed characterization of the insecticidal toxins Cry23Aa and Cry37Aa against CBW. Our findings reveal that CBW larvae fed on artificial diets supplemented exclusively with Cry23Aa decreased larval survival by roughly by 69%, while supplementation with Cry37Aa alone displayed no statistical difference compared to the control. However, the combined provision of both toxins in the artificial diet led to mortality rates approaching 100% among CBW larvae (LC50 equal to 0.26 PPM). Additionally, we engineered transgenic cotton plants by introducing cry23Aa and cry37Aa genes under control of the flower bud-specific pGhFS4 and pGhFS1 promoters, respectively. Seven transgenic cotton events expressing high levels of Cry23Aa and Cry37Aa toxins in flower buds were selected for greenhouse bioassays, and the mortality rate of CBW larvae feeding on their T0 and T1 generations ranged from 75% to 100%. Our in silico analyses unveiled that Cry23Aa displays all the hallmark characteristics of ß-pore-forming toxins (ß-PFTs) that bind to sugar moieties in glycoproteins. Intriguingly, we also discovered a distinctive zinc-binding site within Cry23Aa, which appears to be involved in protein-protein interactions. Finally, we discuss the major structural features of Cry23Aa that likely play a role in the toxin's mechanism of action. In view of the low LC50 for CBW larvae and the significant accumulation of these toxins in the flower buds of both T0 and T1 plants, we anticipate that through successive generations of these transgenic lines, cotton plants engineered to overexpress cry23Aa and cry37Aa hold promise for effectively managing CBW infestations in cotton crops.

iJAZ-based approach to engineer lepidopteran pest resistance in multiple crop species.

The fall armyworm (FAW) poses a significant threat to global crop production. Here we showed that overexpression of jasmonate ZIM-domain (JAZ) protein GhJAZ24 confers resistance to cotton bollworm and FAW, while also causing sterility in transgenic cotton by recruiting TOPLESS and histone deacetylase 6. We identified the NGR motif of GhJAZ24 that recognizes and binds the aminopeptidase N receptor, enabling GhJAZ24 to enter cells and disrupt histone deacetylase 3, leading to cell death. To overcome plant sterility associated with GhJAZ24 overexpression, we developed iJAZ (i, induced), an approach involving damage-induced expression and a switch from intracellular to extracellular localization of GhJAZ24. iJAZ transgenic cotton maintained fertility and showed insecticidal activity against cotton bollworm and FAW. In addition, iJAZ transgenic rice, maize and tobacco plants showed insecticidal activity against their lepidopteran pests, resulting in an iJAZ-based approach for generating alternative insecticidal proteins with distinctive mechanisms of action, thus holding immense potential for future crop engineering.

Identification of Crucial Modules and Genes Associated with Bt Gene Expression in Cotton.

The expression of Bacillus thuringiensis (Bt) toxins in transgenic cotton confers resistance to insect pests. However, it has been demonstrated that its effectiveness varies among cotton cultivars and different tissues. In this study, we evaluated the expression of Bt protein in 28 cotton cultivars and selected 7 cultivars that differed in Bt protein expression for transcriptome analysis. Based on their Bt protein expression levels, the selected cultivars were categorized into three groups: H (high Bt protein expression), M (moderate expression), and L (low expression). In total, 342, 318, and 965 differentially expressed genes were detected in the H vs. L, M vs. L, and H vs. M comparison groups, respectively. And three modules significantly associated with Bt protein expression were identified by weighted gene co-expression network analysis. Three hub genes were selected to verify their relationships with Bt protein expression using virus-induced gene silencing (VIGS). Silencing GhM_D11G1176, encoding an MYC transcription factor, was confirmed to significantly decrease the expression of Bt protein. The present findings contribute to an improved understanding of the mechanisms that influence Bt protein expression in transgenic cotton.

Prey-mediated effects of Mpp51Aa2-producing cotton on longevity and reproduction of Orius majusculus.

Genetically engineered (GE) cotton event MON 88702, producing Mpp51Aa2 (previously mCry51Aa2) from Bacillus thuringiensis (Bt), controls sucking pests, such as Lygus spp. (Hemiptera: Miridae) and thrips (Thysanoptera). Ingesting high doses of the insecticidal protein resulted in adverse effects on life table parameters of beneficial, predatory Orius spp. (Hemiptera: Anthocoridae). This triggered laboratory studies with more realistic food treatments, including different combinations of prey types with and without Bt protein to further characterize risks to this important group of non-target organisms. In this work, exclusive feeding of frozen spider mites (Tetranychus urticae, Acari: Tetranychidae) from Bt cotton confirmed adverse effects on longevity and fecundity of O. majusculus adults. Alternate feeding of Bt protein-containing spider mites and Bt-free Ephestia kuehniella (Lepidoptera: Pyralidae) eggs mitigated effects on longevity, but not on fecundity. When living larvae of Spodoptera littoralis (Lepidoptera: Noctuidae) from Bt cotton were fed to the predators, however, no effects on longevity and reproduction of female O. majusculus were observed, despite the fact that Bt protein concentrations in larvae were almost as high as concentrations in spider mites. When a diverse mix of prey species with various Bt protein concentrations is consumed in the field, it is unlikely that exposure of Orius spp. to Mpp51Aa2 is high enough to exert adverse effects on predator populations. MON 88702 cotton may thus be a valuable tool for integrated management of sucking pests.

Rapid detection of multi-scale cotton pests based on lightweight GBW-YOLOv5 model.

BACKGROUND: Pest infestation is one of the primary causes of decreased cotton yield and quality. Rapid and accurate identification of cotton pest categories is essential for producers to implement effective and expeditious control measures. Existing multi-scale cotton pest detection technology still suffers from poor accuracy and rapidity of detection. This study proposed the pruned GBW-YOLOv5 (Ghost-BiFPN-WIoU You Only Look Once version 5), a novel model for the rapid detection of cotton pests. RESULTS: The detection performance of the pruned GBW-YOLOv5 model for cotton pests was evaluated based on the self-built cotton pest dataset. In comparison with the original YOLOv5 model, the pruned GBW-YOLOv5 model demonstrated significant reductions in complexity, size, and parameters by 68.4%, 66.7%, and 68.2%, respectively. Remarkably, the mean average precision (mAP) decreased by a mere 3.8%. The pruned GBW-YOLOv5 model outperformed other classic object detection models, achieving an outstanding detection speed of 114.9 FPS. CONCLUSION: The methodology proposed by our research enabled rapid and accurate identification of cotton pests, laying a solid foundation for the implementation of precise pest control measures. The pruned GBW-YOLOv5 model provided theoretical research and technical support for detecting cotton pests under field conditions. © 2024 Society of Chemical Industry.

Interaction of Fusarium Wilt Race 4 with Root-Knot Nematode Increases Disease Severity in Cotton.

Fusarium wilt, caused by Fusarium oxysporum f. sp. vasinfectum, is a severe disease of cotton (Gossypium spp.). Strains of the wilt pathogen in the United States, such as race 1, require the presence of nematodes such as southern root-knot nematode (Meloidogyne incognita) to cause appreciable disease. The exception is the race 4 strain of the wilt pathogen, which can attack cotton without concomitant infection by plant-parasitic nematodes and was first identified in California in 2001 and in Texas and New Mexico since 2017. The effects of the interaction between M. incognita and race 1 or race 4 on wilt severity and nematode reproduction on two Gossypium hirsutum cultivars, Acala 44 and FM 966, and a G. barbadense cultivar, Pima S-4, were directly compared in growth chamber assays. All three cultivars were susceptible to M. incognita. Suppression of nematode reproduction by the wilt pathogen was detected only for race 4 on all three cultivars on a per plant basis but not on a per gram root tissue basis. The control, M. incognita alone, and race 1 alone treatments caused no symptoms. Inoculation with race 1 and M. incognita caused moderate wilt symptoms in 'Acala 44' and 'FM 966' and mild symptoms in 'Pima S-4'. However, race 4 treatment caused severe wilt in 'Pima S-4' and moderate wilt severity in 'Acala 44' and 'FM 966'. The symptom severity of 'Acala 44' and 'FM 966' further increased in the presence of M. incognita. Thus, race 4 is not only capable of causing wilt in the absence of M. incognita but can also interact with the nematode to further increase disease severity. Though control of wilt caused by race 1 can be achieved mainly through breeding for nematode resistance, it will be imperative to incorporate both southern root-knot nematode and race 4 resistance to effectively control the disease should race 4 expand into southern root-knot nematode-infested fields.

Mutation in the Cadherin Gene Is a Key Factor for Pink Bollworm Resistance to Bt Cotton in China.

Transgenic crops producing Bacillus thuringiensis (Bt) toxins are widely planted for insect control, but their efficacy may decrease as insects evolve resistance. Understanding the genetic basis of insect resistance is essential for developing an integrated strategy of resistance management. To understand the genetic basis of resistance in pink bollworm (Pectinophora gossypiella) to Bt cotton in the Yangtze River Valley of China, we conducted an F2 screening for alleles associated with resistance to the Bt (Cry1Ac) protein for the first time. A total of 145 valid single-paired lines were screened, among which seven lines were found to carry resistance alleles. All field parents in those seven lines carried recessive resistance alleles at the cadherin locus, including three known alleles, r1, r13 and r15, and two novel alleles, r19 and r20. The overall frequency of resistance alleles in 145 lines was 0.0241 (95% CI: 0.0106-0.0512). These results demonstrated that resistance was rare and that recessive mutation in the cadherin gene was the primary mechanism of pink bollworm resistance to Bt cotton in the Yangtze River Valley of China, which will provide a scientific basis for implementing targeted resistance management statics of pink bollworm in this region.

Plant age, crop stage and surrounding habitats: their impact on sucking pests and predators complex in cotton (Gossypium hirsutum L) field plots in arid climate at district Layyah, Punjab, Pakistan / Idade da planta, estágio da cultura e habitats circundantes: seu impacto no complexo de pragas sugadoras e predadores em parcelas de algodão (Gossypium hirsutum L) em clima árido no distrito de Layyah, Punjab, Paquistão

Sucking pests are major threat to cotton field crop which cause unbearable losses to the crop yield. Aim of the current study was to record seasonal dynamics of major sucking insect pests including whitefly, jassid, thrips and their natural arthropod predators i.e. green lacewings and spiders in cotton field plots. The effects of surrounding field crops on pests' density and predatory efficiency of predators were also recorded. For sampling and survey of insects, the visual counting was found to be the most efficient method for recording the abundance of insects, trailed by net sweeping and tapping. Whitefly was the most dominant sucking pest found on the vegetative stage of cotton, followed by jassid and thrips. Fluctuated populations of predatory arthropods, spiders and green lacewings were also recorded during whole cropping season however, the densities of pests and predators varied with crop phenology. Spiders' population was encouraging at both vegetative and flowering stage and also the same trend of jassid and whitefly were observed at both stages of the crop. Surrounding habitats showed non-significant effect on population densities of insect pests and predators. For abiotic factors, the spiders showed strong positive correlation with humidity and temperature. However, green lacewing was only positively correlated with humidity. On the other hand, the populations of whitefly, jassid and thrips showed non-significant correlation with both temperature and humidity. Overall densities of sucking insect pests were found above economic threshold level. The plant age, crop stage and surrounding habitats effect on the population fluctuation of pests as well as the predators' abundance. The future studies are also warranted to investigate the altered habitats and multiple trap cropping to find out their impact on unattended insect predators and parasitoids in cotton crop.

As pragas sugadoras são uma grande ameaça para a cultura do algodão, causando perdas insuportáveis no rendimento da cultura. O objetivo do estudo atual foi registrar a dinâmica sazonal das principais pragas de insetos sugadores, incluindo mosca-branca, jassid, tripes e seus artrópodes predadores naturais, ou seja, crisopídeos e aranhas verdes em parcelas de algodão. Os efeitos das plantações circundantes na densidade de pragas e na eficiência predatória de predadores também foram registrados. Para amostragem e pesquisa de insetos, a contagem visual foi considerada o método mais eficiente para registrar a abundância de insetos, seguido por varredura e batida de rede. A mosca-branca foi a praga sugadora mais dominante encontrada na fase vegetativa do algodoeiro, seguida pelo jassid e tripes. Populações flutuantes de artrópodes predadores, aranhas e crisálidas também foram registradas durante toda a safra, no entanto as densidades de pragas e predadores variaram com a fenologia da cultura. A população de aranhas foi encorajadora tanto na fase vegetativa como na floração e também a mesma tendência de jassid e mosca-branca foi observada em ambas as fases da cultura. Os habitats circundantes mostraram efeito não significativo nas densidades populacionais de insetos-praga e predadores. Para os fatores abióticos, as aranhas apresentaram forte correlação positiva com umidade e temperatura. No entanto, lacewing verde foi apenas positivamente correlacionado com a umidade. Por outro lado, as populações de mosca-branca, jassid e tripes apresentaram correlação não significativa com temperatura e umidade. As densidades gerais de pragas sugadoras de insetos foram encontradas acima do nível do limiar econômico. A idade da planta, o estágio da cultura e os habitats circundantes afetam a flutuação populacional de pragas, bem como a abundância de predadores. Os estudos futuros também são necessários para investigar os habitats alterados e cultivo com armadilhas múltiplas [...].

Path analysis based on genetic association of yield components and insects pest in upland cotton varieties.

Gossypium hirsutum L. is also called upland cotton or Mexican cotton. It is the most widely cultivated species of cotton in the whole world. Globally, about 90% of all cotton production comes from the cultivars derived from this species. Some genetic parameters like monopodial branches per plant, sympodial branches per plant, sympodial branch length, bolls per plant, boll weight, sympo-boll distance, Ginning Out Turn%, staple length (rg = 0.9199**), and fiber strength along with seed cotton yield were evaluated for their potential utilization via selection in seed cotton yield improvement. Significant positive genetic correlations were estimated for monopodial branches per plant (rg = 0.9722**), sympodial branches per plant (rg = 0.7098**), sympodial branch length (rg = 0.617**), bolls per plant (rg = 0.8271**), boll weight (rg = 0.8065**), sympo-boll distance (rg = 0.6507**), Ginning Out Turn (GOT)% (rg = 0.7541**), staple length (rg = 0.9199**), and fiber strength (rg = 0.7534**) with seed cotton yield. A path analysis of all the yield traits under study revealed strong positive direct effects of monopodial branch length (1.1556), sympo-boll distance (0.8173) and staple length (0.7633), while plant height exerted a highly strong direct negative effect (-1.2096) on yield. It is concluded that a direct selection based on monopodial branch length and sympo-boll distance, and staple length is effective, whereas, monopodial branch length, and sympodial branch length are good selection indicators via bolls per plant for yield improvement in cotton.

Foliar herbivory increases sucrose concentration in bracteal extrafloral nectar of cotton.

Cultivated cotton, such as Gossypium hirsutum L., produces extrafloral (EF) nectar on leaves (foliar) and reproductive structures (bracteal) as an indirect anti-herbivore defense. In exchange for this carbohydrate-rich substance, predatory insects such as ants protect the plant against herbivorous insects. Some EF nectar-bearing plants respond to herbivory by increasing EF nectar production. For instance, herbivore-free G. hirsutum produces more bracteal than foliar EF nectar, but increases its foliar EF nectar production in response to herbivory. This study is the first to test for systemically induced changes to the carbohydrate composition of bracteal EF nectar in response to foliar herbivory on G. hirsutum. We found that foliar herbivory significantly increased the sucrose content of bracteal EF nectar while glucose and fructose remained unchanged. Sucrose content is known to influence ant foraging behavior and previous studies of an herbivore-induced increase to EF nectar caloric content found that it led to increased ant activity on the plant. As a follow-up to our finding, ant recruitment to mock EF nectar solutions that varied in sucrose content was tested in the field. The ants did not exhibit any preference for either solution, potentially because sucrose is a minor carbohydrate component in G. hirsutum EF nectar: total sugar content was not significantly affected by the increase in sucrose. Nonetheless, our findings raise new questions about cotton's inducible EF nectar responses to herbivory. Further research is needed to determine whether an herbivore-induced increase in sucrose content is typical of Gossypium spp., and whether it constitutes a corollary of systemic sucrose induction, or a potentially adaptive mechanism which enhances ant attraction to the plant.

Engineering tolerance to CLCuD in transgenic Gossypium hirsutum cv. HS6 expressing Cotton leaf curl Multan virus-C4 intron hairpin.

Cotton leaf curl disease (CLCuD), caused by begomoviruses in combination with betasatellite molecule, has adversely affected cotton industry of Indian subcontinent. To devise a CLCuD-control strategy, RNAi-mediated approach was followed in this study. Gossypium hirsutum cv. HS6 plants were transformed with intron-hairpin RNAi (ihpRNAi-C4) construct carrying silencing suppressor C4 gene of Cotton leaf curl Multan virus (CLCuMuV). Efficacy of the construct in imparting CLCuD resistance was evaluated in transgenic (T0, T1) cotton lines. Accumulation of CLCuMuV/betasatellite and attenuation of CLCuD symptoms in the transgenic lines were monitored at different times interval after virus inoculation. Northern hybridization revealed the expression of C4-gene derived siRNA. Expression of the ihpRNAi transcript was recorded higher in transgenic lines expressing siRNA which supposedly targeted the C4 gene. A significant delay in detection of virus as well as betasatellite was observed in the transgenic lines. At 30 days post inoculation (dpi), none of the lines tested positive. At 45 dpi, however, it could be detected in few lines having much lower titre as compared to non-transformed control plants. Notably, till 60 dpi, no significant progression of the virus/betasatellite DNA was observed and the plants did not exhibit any characteristic CLCuD symptoms. A tolerance phenomenon leading to escape of CLCuD symptoms in the transformed cotton was described.

CRISPR-mediated mutations in the ABC transporter gene ABCA2 confer pink bollworm resistance to Bt toxin Cry2Ab.

Crops genetically engineered to produce insecticidal proteins from Bacillus thuringiensis (Bt) have many benefits and are important globally for managing insect pests. However, the evolution of pest resistance to Bt crops reduces their benefits. Understanding the genetic basis of such resistance is needed to better monitor, manage, and counter pest resistance to Bt crops. Previous work shows that resistance to Bt toxin Cry2Ab is associated with mutations in the gene encoding the ATP-binding cassette protein ABCA2 in lab- and field-selected populations of the pink bollworm (Pectinophora gossypiella), one of the world's most destructive pests of cotton. Here we used CRISPR/Cas9 gene editing to test the hypothesis that mutations in the pink bollworm gene encoding ABCA2 (PgABCA2) can cause resistance to Cry2Ab. Consistent with this hypothesis, introduction of disruptive mutations in PgABCA2 in a susceptible strain of pink bollworm increased the frequency of resistance to Cry2Ab and facilitated creation of a Cry2Ab-resistant strain. All Cry2Ab-resistant individuals tested in this study had disruptive mutations in PgABCA2. Overall, we found 17 different disruptive mutations in PgABCA2 gDNA and 26 in PgABCA2 cDNA, including novel mutations corresponding precisely to single-guide (sgRNA) sites used for CRISPR/Cas9. Together with previous results, these findings provide the first case of practical resistance to Cry2Ab where evidence identifies a specific gene in which disruptive mutations can cause resistance and are associated with resistance in field-selected populations.

Field efficacy of Bt cotton containing events DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 against lepidopteran pests and impact on the non-target arthropod community in Brazil.

The efficacy and non-target arthropod effects of transgenic DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 Bt cotton, expressing proteins Cry1Ac, Cry1F and Vip3Aa19, was examined through field trials in Brazil. Fifteen field efficacy experiments were conducted from 2014 through the 2020 growing season across six different states in Brazil to evaluate performance against key lepidopteran pests through artificial infestations of Chrysodeixis includens (Walker), Spodoptera frugiperda (J.E. Smith,1797), Spodoptera cosmioides (Walker, 1858) and Chloridea virescens (F., 1781), and natural infestations of Alabama argillacea (Hübner) and S. frugiperda. The impact of this Bt cotton technology on the non-target arthropod community in Brazilian cotton production systems was also assessed in a multi-site experiment. DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 cotton significantly reduced the feeding damage caused by S. frugiperda, S. cosmioides, C. includens, C. virescens and A. argillacea, causing high levels of mortality (greater than 99%) to all target lepidopteran pests evaluated during vegetative and/or reproductive stages of crop development. Non-target arthropod community-level analyses confirmed no unintended effects on the arthropod groups monitored. These results demonstrate the value of transgenic Bt cotton containing event DAS-21023-5 × DAS-24236-5 × SYN-IR102-7 for consideration as part of an integrated approach for managing key lepidopteran pests in Brazilian cotton production systems.