Stabilized Double-Stranded RNA Strategy Improves Cotton Resistance to CBW (Anthonomus grandis).

Cotton is the most important crop for fiber production worldwide. However, the cotton boll weevil (CBW) is an insect pest that causes significant economic losses in infested areas. Current control methods are costly, inefficient, and environmentally hazardous. Herein, we generated transgenic cotton lines expressing double-stranded RNA (dsRNA) molecules to trigger RNA interference-mediated gene silencing in CBW. Thus, we targeted three essential genes coding for chitin synthase 2, vitellogenin, and ecdysis-triggering hormone receptor. The stability of expressed dsRNAs was improved by designing a structured RNA based on a viroid genome architecture. We transformed cotton embryos by inserting a promoter-driven expression cassette that overexpressed the dsRNA into flower buds. The transgenic cotton plants were characterized, and positive PCR transformed events were detected with an average heritability of 80%. Expression of dsRNAs was confirmed in floral buds by RT-qPCR, and the T1 cotton plant generation was challenged with fertilized CBW females. After 30 days, data showed high mortality (around 70%) in oviposited yolks. In adult insects fed on transgenic lines, chitin synthase II and vitellogenin showed reduced expression in larvae and adults, respectively. Developmental delays and abnormalities were also observed in these individuals. Our data remark on the potential of transgenic cotton based on a viroid-structured dsRNA to control CBW.

Cotton fiber quality affected by water availability and silicon application / Qualidade da fibra de algodão afetada pela disponibilidade de água e aplicação de silício

The quality of cotton fiber is a fundamental criterion for determining the commercial value of the product, being influenced by the conditions of cultivation. The study aimed to evaluate irrigated and non-irrigated cultivation systems and the management of silicon fertilization on cotton fiber characteristics. The experiment was carried out in the field in Selvíria-MS, in the 2017/2018 agricultural crop. It was adopted a randomized block design, in a 2x6 factorial scheme, with two cultivationconditions (irrigated and rainfed) and six doses of Si (0, 50, 100, 150, 200 and 400 g ha-1), with four replications. Seeds of the genotype TMG 11 WS were used. Irrigation was carried out with sprinklers spaced at 6x6 m. The application of Si via leaf was carried out at 50 and 70 days after emergence. From thecotton fiber, the characteristics of length, micronaire, strength, uniformity, elongation, reflectance, yellowing, degree of leaves, impurity and impurity particles were analyzed. The data submitted to analysis of variance by the F test, the means compared for the cultivation condition, and regression analysis for the amount of Si. The cotton fiber in irrigated cultivation showed improvement in the characteristics resistance, reflectance and yellowing.The application of Si via leaf reduced the leaf grade in the cotton fibers.

A qualidade da fibra doalgodão é critério fundamental para determinação do valor comercial do produto, sendo influenciado pelas condições de cultivo. O estudo teve como objetivo avaliarsistemas de cultivo irrigado e não irrigado e o manejo da adubação de silício sobre características da fibra do algodão. O experimento foi desenvolvido a campo no município de Selvíria-MS, na safra 2017/2018. Foi adotado delineamento em blocos ao acaso, em esquema fatorial 2x6, sendo duas condições de cultivo (irrigado e sequeiro) e seis doses de Si (0, 50, 100, 150, 200 e 400 g ha-1), com quatro repetições. Foi utilizado sementes do genótipo TMG 11 WS. A irrigação foi realizada com aspersores espaçados em 6x6 m. A aplicação de Si via foliar foi realizado aos 50 e 70 dias após a emergência. A partir da fibra do algodão, foram analisadas as características de comprimento, micronaire, resistência, uniformidade, alongamento, reflectância, amarelecimento, graus de folhas, impureza e partículas de impureza.Os dados submetidos a análise de variância pelo teste F, as médias comparadas para a condição de cultivo, e análise de regressão para quantidade de Si. A fibra de algodão em cultivo irrigado apresentou melhoria nas características de resistência, reflectância e amarelamento. A aplicação de Si via foliar reduziu o grau de folhas nas fibras de algodão.

Cotton fiber quality affected by water availability and silicon application / Qualidade da fibra de algodão afetada pela disponibilidade de água e aplicação de silício

The quality of cotton fiber is a fundamental criterion for determining the commercial value of the product, being influenced by the conditions of cultivation. The study aimed to evaluate irrigated and non-irrigated cultivation systems and the management of silicon fertilization on cotton fiber characteristics. The experiment was carried out in the field in Selvíria-MS, in the 2017/2018 agricultural crop. It was adopted a randomized block design, in a 2x6 factorial scheme, with two cultivationconditions (irrigated and rainfed) and six doses of Si (0, 50, 100, 150, 200 and 400 g ha-1), with four replications. Seeds of the genotype TMG 11 WS were used. Irrigation was carried out with sprinklers spaced at 6x6 m. The application of Si via leaf was carried out at 50 and 70 days after emergence. From thecotton fiber, the characteristics of length, micronaire, strength, uniformity, elongation, reflectance, yellowing, degree of leaves, impurity and impurity particles were analyzed. The data submitted to analysis of variance by the F test, the means compared for the cultivation condition, and regression analysis for the amount of Si. The cotton fiber in irrigated cultivation showed improvement in the characteristics resistance, reflectance and yellowing.The application of Si via leaf reduced the leaf grade in the cotton fibers.(AU)

A qualidade da fibra doalgodão é critério fundamental para determinação do valor comercial do produto, sendo influenciado pelas condições de cultivo. O estudo teve como objetivo avaliarsistemas de cultivo irrigado e não irrigado e o manejo da adubação de silício sobre características da fibra do algodão. O experimento foi desenvolvido a campo no município de Selvíria-MS, na safra 2017/2018. Foi adotado delineamento em blocos ao acaso, em esquema fatorial 2x6, sendo duas condições de cultivo (irrigado e sequeiro) e seis doses de Si (0, 50, 100, 150, 200 e 400 g ha-1), com quatro repetições. Foi utilizado sementes do genótipo TMG 11 WS. A irrigação foi realizada com aspersores espaçados em 6x6 m. A aplicação de Si via foliar foi realizado aos 50 e 70 dias após a emergência. A partir da fibra do algodão, foram analisadas as características de comprimento, micronaire, resistência, uniformidade, alongamento, reflectância, amarelecimento, graus de folhas, impureza e partículas de impureza.Os dados submetidos a análise de variância pelo teste F, as médias comparadas para a condição de cultivo, e análise de regressão para quantidade de Si. A fibra de algodão em cultivo irrigado apresentou melhoria nas características de resistência, reflectância e amarelamento. A aplicação de Si via foliar reduziu o grau de folhas nas fibras de algodão.(AU)

Overexpression of the CaHB12 transcription factor in cotton (Gossypium hirsutum) improves drought tolerance.

The Coffea arabica HB12 gene (CaHB12), which encodes a transcription factor belonging to the HD-Zip I subfamily, is upregulated under drought, and its constitutive overexpression (35S:CaHB12OX) improves the Arabidopsis thaliana tolerance to drought and salinity stresses. Herein, we generated transgenic cotton events constitutively overexpressing the CaHB12 gene, characterized these events based on their increased tolerance to water deficit, and exploited the gene expression level from the CaHB12 network. The segregating events Ev8.29.1, Ev8.90.1, and Ev23.36.1 showed higher photosynthetic yield and higher water use efficiency under severe water deficit and permanent wilting point conditions compared to wild-type plants. Under well-irrigated conditions, these three promising transformed events showed an equivalent level of Abscisic acid (ABA) and decreased Indole-3-acetic acid (IAA) accumulation, and a higher putrescine/(spermidine + spermine) ratio in leaf tissues was found in the progenies of at least two transgenic cotton events compared to non-transgenic plants. In addition, genes that are considered as modulated in the A. thaliana 35S:CaHB12OX line were also shown to be modulated in several transgenic cotton events maintained under field capacity conditions. The upregulation of GhPP2C and GhSnRK2 in transgenic cotton events maintained under permanent wilting point conditions suggested that CaHB12 might act enhancing the ABA-dependent pathway. All these data confirmed that CaHB12 overexpression improved the tolerance to water deficit, and the transcriptional modulation of genes related to the ABA signaling pathway or downstream genes might enhance the defense responses to drought. The observed decrease in IAA levels indicates that CaHB12 overexpression can prevent leaf abscission in plants under or after stress. Thus, our findings provide new insights on CaHB12 gene and identify several promising cotton events for conducting field trials on water deficit tolerance and agronomic performance.

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.

Demographic Performance of Helicoverpa zea Populations on Dual and Triple-Gene Bt Cotton.

Insecticidal toxins from Bacillus thuringiensis (Bt) are valuable tools for pest management worldwide, contributing to the management of human disease insect vectors and phytophagous insect pests of agriculture and forestry. Here, we report the effects of dual and triple Bt toxins expressed in transgenic cotton cultivars on the fitness and demographic performance of Helicoverpa zea (Boddie)-a noctuid pest, known as cotton bollworm and corn earworm. Life-history traits were determined for individuals of three field populations from a region where H. zea overwintering is likely. Triple-gene Bt cotton cultivars that express Cry and Vip3Aa toxins killed 100% of the larvae in all populations tested. In contrast, dual-gene Bt cotton that express Cry1Ac+Cry1F and Cry1Ac+Cry2Ab allowed population growth with the intrinsic rate of population growth (rm) 38% lower than on non-Bt cotton. The insects feeding on Bt cotton plants that express Cry1Ac+Cry2Ab, Cry1Ac+Cry1F, or Cry1Ab+Cry2Ae exhibited reduced larval weight, survival rate, and increased development time. Additionally, fitness parameters varied significantly among the insect populations, even on non-Bt cotton plants, likely because of their different genetic background and/or previous Bt toxin exposure. This is the first report of the comparative fitness of H. zea field populations on dual-gene Bt cotton after the recent reports of field resistance to certain Bt toxins. These results document the population growth rates of H. zea from an agricultural landscape with 100% Bt cotton cultivars. Our results will contribute to the development and validation of resistance management recommendations.

Nutrient support via fertigation with domestic effluent and growth of cotton / Aporte de nutrientes via fertirrigação com efluente doméstico e crescimento do algodoeiro

Because of climatic problems in the Brazilian semi-arid region, it is of fundamental importance to seek a culture that considers water scarcity and available natural resources. Thus, a randomized block design, with subdivided plots and six replications, was carried out in the experimental area of the treatment and wastewater reuse station of the Milagres Settlement, Apodi, Brazil, with the objective of evaluating cultivation of herbaceous cotton (Gossipyum hirsutum) fertigated with domestic sewage treated in a semi-arid environment. Two cultivation cycles were carried out, and the cotton plants were fed with three different solutions containing treated domestic sewage (TDE) plus supply water (PSW), to supply the crop evapotranspiration (ETc) (100% PSW ­ control, 50% PSW + 50% TDE and 100% TDE), with and without phosphate fertilization in pre-planting. The contribution of nutrients to the soil and plant growth were analyzed. The results indicated that the wastewater presents potential fertilizer, reaching to the soil amounts of some nutrients exceeding the cotton's needs, when 100% of the ETc was supplied via fertigation with domestic effluent, resulting in an increase in the growth rates of the plants. There was no interaction between the percentages of wastewater and phosphate fertilization in pre-planting. Phosphorus alone promoted significant variation only in the first cycle, indicating that there is a residual fertigation effect with domestic effluent on the nutrient supply.(AU)

Devido aos problemas climáticos no semiárido brasileiro é de fundamental importância que se busque uma cultura que considere a escassez hídrica e os recursos naturais disponíveis. Deste modo, um experimento em delineamento de blocos ao acaso, com parcelas subdivididas e seis repetições foi conduzido na área experimental da estação de tratamento e reuso de águas residuais do Assentamento Milagres, Apodi - RN, com o objetivo de avaliar o cultivo de algodoeiro herbáceo (Gossipyum hirsutum) fertirrigado com efluente de esgoto doméstico tratado em ambiente semiárido. Foram realizados dois ciclos de cultivo e as plantas de algodão foram nutridas com três diferentes soluções contendo efluente de esgoto doméstico tratado (PSW) + água de abastecimento (TDE) para suprir a evapotranspiração da cultura (ETc) (100% PSW ­ control, 50% PSW + 50% TDE and 100% TDE) com e sem adubação fosfatada em pré-plantio. Analisou-se o aporte de nutrientes ao solo e o crescimento das plantas. Os resultados indicaram que o efluente doméstico tratado apresenta potencial fertilizante, chegando a aportar ao solo quantidades de alguns nutrientes superiores a necessidade do algodoeiro, quando 100% da ETc foi suprida via fertirrigação com efluente doméstico, resultando em incremento das taxas de crescimento das plantas. Não se constatou interação entre as porcentagens de efluente doméstico tratado e a adubação fosfatada em pré-plantio. O fósforo isoladamente promoveu variação significativa apenas no primeiro ciclo, indicando que há um efeito residual da fertirrigação com efluente doméstico no aporte do nutriente.(AU)

Metabolizable energy of rice bran, cottonseed meal and wheat bran for slow-growing broilers at two ages / Energia metabolizável dos farelos de arroz, algodão e trigo para frangos de crescimento lento em duas idades

The aim of this study was to evaluate bromatological composition and metabolizable energy values of cottonseed meal, rice meal, and wheat bran when provided to slow growth rate broilers at different ages. Energy values of 15- to 25- and 35- to 45-day-old broilers were determined by the total feces collection method using two digestibility bioassays. The birds were distributed in a completely randomized experimental design to evaluate the age effect, with six replicates of 10 chickens each, for each ingredient. Values of apparent metabolizable energy adjusted by nitrogen balance (kcal/kg), for the first and second ages were: 1,255 and 1,564 for cottonseed meal, 3,124 and 3,301 for rice meal, and, 2,211 and 2,383 for wheat bran, respectively. The ingredients showed some variations regarding chemical composition and energy values, which were higher (P < 0.05) in older broiler chickens.(AU)

O objetivo nesta pesquisa foi determinar a composição bromatológica e os valores de energia metabolizável para frangos de corte de crescimento lento em diferentes idades dos ingredientes: farelo de algodão, farelo de arroz e farelo de trigo. Para determinação dos valores energéticos foi utilizado o método de coleta total de excretas em dois ensaios de digestibilidade de 15 a 25 e de 35 a 45 dias de idade. As aves foram distribuídas em delineamento inteiramente casualizado, avaliando o efeito da idade, com seis repetições constituídas de 10 aves, para cada ingrediente. Os valores de energia metabolizável aparente corrigida pelo balanço de nitrogênio (kcal/kg), para primeira e segunda idade respectivamente, foram: farelo de algodão, 1.255 e 1.564; farelo de arroz, 3.124 e 3.301; farelo de trigo, 2.211 e 2.383. Os alimentos apresentaram variação quanto a composição química, bem como o valor energético foi superior (P<0,05) para os frangos de maior idade.(AU)

Effects of amount and recurrence of leaf herbivory on the induction of direct and indirect defences in wild cotton.

The induction of defences in response to herbivory is a key mechanism of plant resistance. While a number of studies have investigated the time course and magnitude of plant induction in response to a single event of herbivory, few have looked at the effects of recurrent herbivory. Furthermore, studies measuring the effects of the total amount and recurrence of herbivory on both direct and indirect plant defences are lacking. To address this gap, here we asked whether insect leaf herbivory induced changes in the amount and concentration of extrafloral nectar (an indirect defence) and concentration of leaf phenolic compounds (a direct defence) in wild cotton (Gossypium hirsutum). We conducted a greenhouse experiment where we tested single event or recurrent herbivory effects on defence induction by applying mechanical leaf damage and caterpillar (Spodoptera frugiperda) regurgitant. Single events of 25% and 50% leaf damage did not significantly influence extrafloral nectar production or concentration. Extrafloral nectar traits did, however, increase significantly relative to controls when plants were exposed to recurrent herbivory (two episodes of 25% damage). In contrast, phenolic compounds increased significantly in response to single events of  leaf damage but not to recurrent damage. In addition, we found. that local induction of extrafloral nectar production was stronger than systemic induction, whereas the reverse pattern was observed for phenolics. Together, these results reveal seemingly inverse patterns of induction of direct and indirect defences in response to herbivory in wild cotton.

Compensation to simulated insect leaf herbivory in wild cotton (Gossypium hirsutum): responses to multiple levels of damage and associated traits.

Identifying the mechanisms of compensation to insect herbivory remains a major challenge in plant biology and evolutionary ecology. Most previous studies have addressed plant compensatory responses to one or two levels of insect herbivory, and the underlying traits mediating such responses remain elusive in many cases. We evaluated responses associated with compensation to multiple intensities of leaf damage (0% control, 10%, 25%, 50%, 75% of leaf area removed) by means of mechanical removal of foliar tissue and application of a caterpillar (Spodoptera exigua) oral secretions in 3-month-old wild cotton plants (Gossypium hirsutum). Four weeks post-treatment, we measured plant growth and multiple traits associated with compensation, namely: changes in above- and belowground, biomass and the concentration of nutrients (nitrogen and phosphorus) and non-structural carbon reserves (starch and soluble sugars) in roots, stems and leaves. We found that wild cotton fully compensated in terms of growth and biomass allocation when leaf damage was low (10%), whereas moderate (25%) to high leaf damage in some cases led to under-compensation. Nonetheless, high levels of leaf removal (50% and 75%) in most cases did not cause further reductions in height and allocation to leaf and stem biomass relative to low and moderate damage. There were significant positive effects of leaf damage on P concentration in leaves and stems, but not roots, as well as a negative effect on soluble sugars in roots. These results indicate that wild cotton fully compensated for a low level of leaf damage but under-compensated under moderate to high leaf damage, but can nonetheless sustain growth despite increasing losses to herbivory. Such responses were possibly mediated by a re-allocation of carbohydrate reserves from roots to shoots.

Transgene-product expression levels in genetically engineered breeding stacks are equivalent to those of the single events.

Transgene product expression levels are measured in genetically engineered (GE) crops containing single transformation events and the measured expression levels are then utilized in food, feed, and environmental safety assessments as part of the requirements for de-regulation of the event. Many countries also require measurement of expression levels and safety assessments for GE breeding stacks, even though the breeding stacks are composed of single events that have been previously assessed. Transgene product expression levels were measured in tissues of maize, soybean, and cotton breeding stacks and each of their component single events. Expression levels in the breeding stacks were plotted against expression levels in the single events to quantify the ability of the single events to predict transgene product expression levels in the breeding stacks. These results indicate that transgene product expression levels in single events are a reliable indicator of expression levels in breeding stacks. Based on these results it is concluded that safety assessments for breeding stacks can be conducted using transgene product expression levels from single events.

Phosphorus-zinc interactions in cotton: consequences for biomass production and nutrient-use efficiency in photosynthesis.

The fragmentary information on phosphorus (P) × zinc (Zn) interactions in plants warrants further study, particularly in plants known for their high P and Zn requirements, such as cotton (Gossypium hirsutum L.). The objective of this study was to investigate the effect of P × Zn interactions in a modern cultivar of cotton grown hydroponically. Biomass, mineral nutrition and photosynthetic parameters were monitored in plants receiving contrasting combinations of P and Zn supply. Root biomass, length and surface area were similar in plants with low P and/or low Zn supply to those in plants grown with high P and high Zn supply, reflecting an increased root/shoot biomass quotient when plants lack sufficient P or Zn for growth. Increasing P supply and reducing Zn supply increased shoot P concentrations, whilst shoot Zn concentrations were influenced largely by Zn supply. A balanced P × Zn supply (4 mM P × 4 µM Zn) enabled greatest biomass accumulation, while an imbalanced supply of these nutrients led to Zn deficiency, P toxicity or Zn toxicity. Net photosynthetic rate, stomatal conductance, transpiration rate and instantaneous carboxylation efficiency increased as P or Zn supply increased. Although increasing P supply reduced the P-use efficiency in photosynthesis (PUEP) and increasing Zn supply reduced the Zn-use efficiency in photosynthesis (ZnUEP), increasing Zn supply at a given P supply increased PUEP and increasing P supply at a given Zn supply increased ZnUEP. These results suggest that agricultural management strategies should seek for balanced mineral nutrition to optimize yields and resource-use efficiencies.

Metabolizable energy of rice bran, cottonseed meal and wheat bran for slow-growing broilers at two ages / Energia metabolizável dos farelos de arroz, algodão e trigo para frangos de crescimento lento em duas idades

The aim of this study was to evaluate bromatological composition and metabolizable energy values of cottonseed meal, rice meal, and wheat bran when provided to slow growth rate broilers at different ages. Energy values of 15- to 25- and 35- to 45-day-old broilers were determined by the total feces collection method using two digestibility bioassays. The birds were distributed in a completely randomized experimental design to evaluate the age effect, with six replicates of 10 chickens each, for each ingredient. Values of apparent metabolizable energy adjusted by nitrogen balance (kcal/kg), for the first and second ages were: 1,255 and 1,564 for cottonseed meal, 3,124 and 3,301 for rice meal, and, 2,211 and 2,383 for wheat bran, respectively. The ingredients showed some variations regarding chemical composition and energy values, which were higher (P < 0.05) in older broiler chickens.

O objetivo nesta pesquisa foi determinar a composição bromatológica e os valores de energia metabolizável para frangos de corte de crescimento lento em diferentes idades dos ingredientes: farelo de algodão, farelo de arroz e farelo de trigo. Para determinação dos valores energéticos foi utilizado o método de coleta total de excretas em dois ensaios de digestibilidade de 15 a 25 e de 35 a 45 dias de idade. As aves foram distribuídas em delineamento inteiramente casualizado, avaliando o efeito da idade, com seis repetições constituídas de 10 aves, para cada ingrediente. Os valores de energia metabolizável aparente corrigida pelo balanço de nitrogênio (kcal/kg), para primeira e segunda idade respectivamente, foram: farelo de algodão, 1.255 e 1.564; farelo de arroz, 3.124 e 3.301; farelo de trigo, 2.211 e 2.383. Os alimentos apresentaram variação quanto a composição química, bem como o valor energético foi superior (P<0,05) para os frangos de maior idade.

Plant odor and sex pheromone are integral elements of specific mate recognition in an insect herbivore.

Specific mate recognition relies on the chemical senses in most animals, and especially in nocturnal insects. Two signal types mediate premating olfactory communication in terrestrial habitats: sex pheromones, which blend into an atmosphere of plant odorants. We show that host plant volatiles affect the perception of sex pheromone in males of the African cotton leafworm Spodoptera littoralis and that pheromone and plant volatiles are not perceived as independent messages. In clean air, S. littoralis males are attracted to single synthetic pheromone components or even the pheromone of a sibling species, oriental cotton leafworm S. litura. Presence of host plant volatiles, however, reduces the male response to deficient or heterospecific pheromone signals. That plant cues enhance discrimination of sex pheromone quality confirms the idea that specific mate recognition in noctuid moths has evolved in concert with adaptation to host plants. Shifts in either female host preference or sex pheromone biosynthesis give rise to new communication channels that have the potential to initiate or contribute to reproductive isolation.

Evaluation of sample extracting methods of FCSM by Lactobacillus acidophilus based on a UPLC-Q-TOF-MS global metabolomics analysis

Abstract The study of metabolomics requires extracting as many metabolites as possible from a biological sample. This study aimed to determine the optimal method for the extraction of metabolites from solid-state fermented cottonseed meal (FCSM). The UPLC-Q-TOF-MS global metabolomics technology was used to detect the metabolites in FCSM, and the extraction quantity and extraction efficiency of seven different extraction methods, specifically the WA, 50MeOH, 50MeOHB, 50MeCNB, 80MeOHB, 80MeOH and AMF methods were evaluated. The results showed that the number of VIP metabolites extracted by AMF method are 196 and 184 in ESI+ and ESI- mode respectively, it is the largest number of all exacted methods; and the AMF methods also provided a higher extraction efficiency compared with the other methods, especially in indoleacrylic acid, dl-tryptophan and epicatechin (p < 0.01). As a result, AMF/-4 °C method was identified as the best method for the extraction of metabolites from FCSM by Lactobacillus acidophilus. Our study establishes a technical basis for future metabolomics research of fermented feed.

Validating a probe from GhSERK1 gene for selection of cotton genotypes with somatic embryogenic capacity.

Substantial progress is being reported in the techniques for plant transformation, but successful regeneration of some genotypes remains a challenging step in the attempts to transform some recalcitrant species. GhSERK1 gene is involved on embryo formation, and its overexpression enhances the embryogenic competence. In this study we validate a short GhSERK1 probe in order to identify embryogenic cotton genotypes using RT-qPCR and blotting assays. Cotton genotypes with contrasting somatic embryogenic capacity were tested using in vitro procedures. High expression of transcripts was found in embryogenic genotypes, and the results were confirmed by the RT-PCR-blotting using a non-radioactive probe. The regeneration ability was confirmed in embryogenic genotypes. We confirmed that GhSERK1 can be used as marker for estimating the somatic embryogenesis ability of cotton plants.

Evaluation of sample extracting methods of FCSM by Lactobacillus acidophilus based on a UPLC-Q-TOF-MS global metabolomics analysis.

The study of metabolomics requires extracting as many metabolites as possible from a biological sample. This study aimed to determine the optimal method for the extraction of metabolites from solid-state fermented cottonseed meal (FCSM). The UPLC-Q-TOF-MS global metabolomics technology was used to detect the metabolites in FCSM, and the extraction quantity and extraction efficiency of seven different extraction methods, specifically the WA, 50MeOH, 50MeOHB, 50MeCNB, 80MeOHB, 80MeOH and AMF methods were evaluated. The results showed that the number of VIP metabolites extracted by AMF method are 196 and 184 in ESI+ and ESI- mode respectively, it is the largest number of all exacted methods; and the AMF methods also provided a higher extraction efficiency compared with the other methods, especially in indoleacrylic acid, dl-tryptophan and epicatechin (p<0.01). As a result, AMF/-4°C method was identified as the best method for the extraction of metabolites from FCSM by Lactobacillus acidophilus. Our study establishes a technical basis for future metabolomics research of fermented feed.

Larval Dispersal of Spodoptera frugiperda Strains on Bt Cotton: A Model for Understanding Resistance Evolution and Consequences for its Management.

High dispersal of Lepidoptera larvae between non-Bt and Bt cotton plants can favour the evolution of insect resistance; however, information on host acceptance of neonates in tropical transgenic crops is scarce. Therefore, the purposes of this study were as follows: (i) to investigate the feeding behaviour of susceptible and Cry1F-resistant strains of Spodoptera frugiperda (J.E. Smith) on Bt and non-Bt cotton (Gossypium hirsutum L.) varieties and (ii) to understand the possible effects of cotton field contamination on the dispersal and infestation capacity of S. frugiperda larvae by using an individual-based model. The main results of this paper are as follows: (1) the highest post-feeding larval dispersal of the Cry1F-resistant strain occurred at an exposure time of 18-24 h; (2) via video tracking assays, we found that the least distance moved was by larvae resistant to Cry1F on non-Bt cotton; and (3) the model indicated differences in mobility capacity between Bt and non-Bt cotton. We conclude that resistant neonates exhibit sedentary behaviour. Our report represents the first findings concerning the fitness cost of larval behaviour traits of S. frugiperda associated with Cry1F resistance in Brazilian populations.

Transgenic cotton expressing Cry10Aa toxin confers high resistance to the cotton boll weevil.

Genetically modified (GM) cotton plants that effectively control cotton boll weevil (CBW), which is the most destructive cotton insect pest in South America, are reported here for the first time. This work presents the successful development of a new GM cotton with high resistance to CBW conferred by Cry10Aa toxin, a protein encoded by entomopathogenic Bacillus thuringiensis (Bt) gene. The plant transformation vector harbouring cry10Aa gene driven by the cotton ubiquitination-related promoter uceA1.7 was introduced into a Brazilian cotton cultivar by biolistic transformation. Quantitative PCR (qPCR) assays revealed high transcription levels of cry10Aa in both T0 GM cotton leaf and flower bud tissues. Southern blot and qPCR-based 2-ΔΔCt analyses revealed that T0 GM plants had either one or two transgene copies. Quantitative and qualitative analyses of Cry10Aa protein expression showed variable protein expression levels in both flower buds and leaves tissues of T0 GM cotton plants, ranging from approximately 3.0 to 14.0 µg g-1 fresh tissue. CBW susceptibility bioassays, performed by feeding adults and larvae with T0 GM cotton leaves and flower buds, respectively, demonstrated a significant entomotoxic effect and a high level of CBW mortality (up to 100%). Molecular analysis revealed that transgene stability and entomotoxic effect to CBW were maintained in T1 generation as the Cry10Aa toxin expression levels remained high in both tissues, ranging from 4.05 to 19.57 µg g-1 fresh tissue, and the CBW mortality rate remained around 100%. In conclusion, these Cry10Aa GM cotton plants represent a great advance in the control of the devastating CBW insect pest and can substantially impact cotton agribusiness.

Influence of Two Acyclic Homoterpenes (Tetranorterpenes) on the Foraging Behavior of Anthonomus grandis Boh.

Previous studies have shown that the boll weevil, Anthonomus grandis, is attracted to constitutive and conspecific herbivore-induced cotton volatiles, preferring the blend emitted by cotton at the reproductive over the vegetative stage. Moreover, this preference was paralleled by the release of the acyclic homoterpenes (tetranorterpenes) (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT) and (E,E)-4,8,12-trimethyltrideca-1,3,7,11-tetraene (TMTT) in Delta Opal cotton being higher at the vegetative than at the reproductive stage. Here, we evaluated whether this difference in release of acyclic homoterpenes also occurred in other cotton varieties, and if boll weevils could recognize these compounds as indicators of a specific cotton phenological stage. Results showed that cotton genotypes CNPA TB-90, BRS-293 and Delta Opal all produced higher levels of DMNT and TMTT at the vegetative stage than at the reproductive stage and that these homoterpenes allowed for principal component analysis separation of volatiles produced by the two phenological stages. Electroantennograms confirmed boll weevil antennal responses to DMNT and TMTT. Behavioral assays, using Y-tube olfactometers, showed that adding synthetic homoterpenes to reproductive cotton volatiles (mimicking cotton at the vegetative stage in terms of homoterpene levels) resulted in reduced attraction to boll weevils compared to that to unmodified reproductive cotton. Weevils showed no preference when given a choice between plants at the vegetative stage and the vegetative stage-mimicked plant. Altogether, the results show that DMNT and TMTT are used by boll weevils to distinguish between cotton phenological stages.