Transformation of Rhodococcus Pigment Production Hydroxylase (PPH) gene into Camelina sativa: an alternative marker for the detection of transgenic plants / Transformação do gene Rhodococcus Pigment Production Hydroxylase (PPH) em Camelina sativa: um marcador alternativo para a detecção de plantas transgênicas

Production of transgenic plants with desired agronomic and horticultural traits has gained great importance to fulfill demands of the growing population. Genetic transformation is also a fundamental step to study basics of plant sciences. Different transformation protocols have been developed and used which are reliable and efficient. These protocols used antibiotic or herbicide resistance genes incorporated along with gene of interest to identify transformed plants from non-transformed ones. These marker genes may pose a threat to human and environment. Use of visual markers enables direct and easier observation of transformed plants with more precision. In current study a gene cassette with 'pigment production hydroxylase (PPH) gene under fiber specific promoter (GhSCFP) and downstream Nos-terminator was designed. After checking the structural and functional efficiency of codon optimized gene using bioinformatics tools, the cassette was sent for chemical synthesis from commercial source. The pigment gene cassette (PPH_CEMB), cloned in pCAMBIA-1301, was transformed into Agrobacterium through electroporation. Agrobacterium-mediated floral dip method was used to transform Camelina sativa inflorescence. After seed setting a total of 600 seed were observed for change in color and out of these, 19 seeds developed a reddish-brown coloration, while the remaining 581 seeds remained yellow. The transformation efficiency calculated on basis of color change was 1.0%. PCR analysis of leaves obtained after sowing reddish seeds confirmed the transformation of pigment production gene, while no PCR amplification was observed in leaves of plants from wild type seeds. From the results it is evident that Agrobacterium-mediated transformation of C. sativa inflorescence is very efficient and environment friendly technique not only for detection of transformed plants but also to study basic cellular processes.

A produção de plantas transgênicas com características agronômicas e hortícolas desejadas vem ganhando grande importância por atender às demandas da crescente população. A transformação genética também é um passo fundamental para estudar os fundamentos das ciências das plantas. Foram desenvolvidos e utilizados diferentes protocolos de transformação, ​​que são confiáveis ​​e eficientes. Esses protocolos usaram genes de resistência a antibióticos ou herbicidas incorporados ao gene de interesse para identificar plantas transformadas e não transformadas. Esses genes marcadores podem representar uma ameaça ao ser humano e ao meio ambiente. O uso de marcadores visuais permite a observação direta e fácil de plantas transformadas com mais precisão. No estudo atual, foi projetado um cassete de genes com o gene da hidroxilase de produção de pigmentos (PPH) sob promotor específico de fibra (GhSCFP) e terminador NOS a jusante. Após verificar a eficiência estrutural e funcional do gene otimizado por códons usando ferramentas de bioinformática, o cassete foi enviado para síntese química a partir de fonte comercial. O cassete do gene do pigmento (PPH_CEMB), clonado em pCAMBIA-1301, foi transformado em Agrobacterium por eletroporação. O método de imersão floral mediado por Agrobacterium foi usado para transformar a inflorescência de Camelina sativa. Após a formação de sementes, foi observado um total de 600 sementes com mudança de cor, das quais 19 desenvolveram uma coloração marrom-avermelhada, enquanto as 581 restantes permaneceram amarelas. A eficiência de transformação calculada com base na mudança de cor foi de 1%. A análise de PCR das folhas obtidas após a semeadura de sementes avermelhadas confirmou a transformação do gene produtor de pigmentos, enquanto não foi observada amplificação por PCR em folhas de plantas de sementes do tipo selvagem. A partir dos resultados, é evidente que a transformação da inflorescência de C. sativa mediada por Agrobacterium é uma técnica muito eficiente e favorável ao ambiente não só para a detecção de plantas transformadas, mas também para estudar processos celulares básicos.

Expression of Mn-sod, PAL1, aos1 and HPL genes in soybean plants overexpressing the NmDef02 defensin.

Plant defensins are a potential tool in crop improvement programs through biotechnology. Their antifungal action makes them attractive molecules for the production of transgenic plants. Information is currently lacking on what happens to the expression of defense genes in transgenic plants that overexpress a defensin. Here we show the relative expression of four defense-related genes: Mn-sod, PAL1, aos1 and HPL evaluated in two transgenic soybean events (Def1 and Def17) constitutively expressing the NmDef02 defensin gene from Nicotiana megalosiphon. The expression of these defense genes showed a differential profile in the transgenic events, with the increased expression of the aos1 gene and the repression of the Mn-sod gene in both events, when compared to the non-transgenic control. Furthermore, the expression of the PAL1 gene only increased in the Def17 event. The results indicate that although there were some changes in the expression of defense genes in transgenic plants overexpressing the defensin NmDef02; the morphoagronomic parameters evaluated were similar to the non-transgenic control. Understanding the molecular changes that occur in these transgenic plants could be of interest in the short, medium and long term.

The role of silver nanoparticles effects in the homeostasis of metals in soybean cultivation through qualitative and quantitative laser ablation bioimaging.

BACKGROUND: Nanoparticles (NPs) are currently found in the world in the form of natural colloids and volcanic ash, as well as in anthropogenic sources, such as nanofertilizers; however, in the literature, there is still a lack of toxicological evidence, risk assessment, and regulations about the use and environmental impact of NPs in the agroindustrial system. Therefore, the aim of this work was to evaluate alterations caused by the presence of AgNPs during the development of soybean plants. METHODS: The BRS232 non-transgenic (NT) soybean plant and 8473RR (TRR) and INTACTA RR2 PRO (TIntacta) transgenic soybean plants were irrigated for 18 days under controlled conditions with deionized water (control), AgNPs, and AgNO3. The isotopes 107Ag+, 55Mn+, 57Fe+, 63Cu+, and 64Zn+ were mapped in leaves, using 13C+ as an internal standard (IS), and carried out using a laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) technique with a Nd:YAG (213 nm) laser source in the imagagin mode using the LA-iMageS software and also Mathlab. RESULTS: Leaf images showed a low Ag translocation, indicated by the basal signal of this ion. Additionally, the presence of Ag in the ionic form and as NPs altered the homeostasis of 112Cd+, 64Zn+, 55Mn+, 63Cu+, and 57Fe+ in different ways. Quantitative image analysis was performed for Cu. CONCLUSION: The behavior of TRR and TIntacta plants was different in the presence of ionic silver or AgNPs, confirming that the metabolism of these two plants, despite both being transgenic, are different. Through the images, it was observed that the response of plants was different in the face of the same stress conditions during their development.

Residual Effects of Transgenic Cotton on the Intestinal Microbiota of Dysdercus concinnus.

The interaction among plants, insects, and microbes (PIM) is a determinant factor for the assembly and functioning of natural and anthropic ecosystems. In agroecosystems, the relationships among PIM are based on the interacting taxa, environmental conditions, and agricultural management, including genetically modified (GM) organisms. Although evidence for the unintended effects of GM plants on non-target insects is increasingly robust, our knowledge remains limited regarding their impact on gut microbes and their repercussions on the host's ecology, especially in the wild. In this study, we compared the gut microbial community of Dysdercus concinnus bugs collected on wild cotton (Gossypium hirsutum), with and without insecticidal transgenes (cry1ab/ac), in its center of origin and diversity. By sequencing the V4-V5 region of 16S rRNA, we show differences in the diversity, structure, and topology of D. concinnus gut microbial interactions between specimens foraging cotton plants with and without transgenes. Identifying unintended residual effects of genetic engineering in natural ecosystems will provide first-line knowledge for informed decision-making to manage genetic, ecological, and evolutionary resources. Thus, determining which organisms interact with GM plants and how is the first step toward conserving natural ecosystems with evidence of transgenic introgression.

The Ecotoxicology of Microbial Insecticides and Their Toxins in Genetically Modified Crops: An Overview.

The use of microbial insecticides and their toxins in biological control and transgenic plants has increased their presence in the environment. Although they are natural products, the main concerns are related to the potential impacts on the environment and human health. Several assays have been performed worldwide to investigate the toxicity or adverse effects of these microbial products or their individual toxins. This overview examines the published data concerning the knowledge obtained about the ecotoxicity and environmental risks of these natural pesticides. The data presented show that many results are difficult to compare due to the diversity of measurement units used in the different research data. Even so, the products and toxins tested present low toxicity and low risk when compared to the concentrations used for pesticide purposes. Complementary studies should be carried out to assess possible effects on human health.

Antisense Transcription in Plants: A Systematic Review and an Update on cis-NATs of Sugarcane.

Initially, natural antisense transcripts (NATs, natRNAs, or asRNAs) were considered repressors; however, their functions in gene regulation are diverse. Positive, negative, or neutral correlations to the cognate gene expression have been noted. Although the first studies were published about 50 years ago, there is still much to be investigated regarding antisense transcripts in plants. A systematic review of scientific publications available in the Web of Science databases was conducted to contextualize how the studying of antisense transcripts has been addressed. Studies were classified considering three categories: "Natural antisense" (208), artificial antisense used in "Genetic Engineering" (797), or "Natural antisense and Genetic Engineering"-related publications (96). A similar string was used for a systematic search in the NCBI Gene database. Of the 1132 antisense sequences found for plants, only 0.8% were cited in PubMed and had antisense information confirmed. This value was the lowest when compared to fungi (2.9%), bacteria (2.3%), and mice (54.1%). Finally, we present an update for the cis-NATs identified in Saccharum spp. Of the 1413 antisense transcripts found in different experiments, 25 showed concordant expressions, 22 were discordant, 1264 did not correlate with the cognate genes, and 102 presented variable results depending on the experiment.

Calcium-dependent protein kinase 2 plays a positive role in the salt stress response in potato.

KEY MESSAGE: StCDPK2 is an early player in the salt stress response in potato plants; its overexpression promoted ROS scavenging, chlorophyll stability, and the induction of stress-responsive genes conferring tolerance to salinity. The salinity of soils affects plant development and is responsible for great losses in crop yields. Calcium-dependent protein kinases (CDPKs) are sensor-transducers that decode Ca2+ signatures triggered by abiotic stimuli and translate them into physiological responses. Histochemical analyses of potato plants harboring StCDPK2 promoter fused to the reporter gene ß-glucuronidase (ProStCDPK2:GUS) revealed that GUS activity was high in the leaf blade and veins, it was restricted to root tips and lateral root primordia, and was observed upon stolon swelling. Comparison with ProStCDPK1:GUS and ProStCDPK3:GUS plants revealed their differential activities in the plant tissues. ProStCDPK2:GUS plants exposed to high salt presented enhanced GUS activity in roots which correlated with the numerous stress-responsive sites predicted in its promoter sequence. Moreover, StCDPK2 expression increased in in vitro potato plants after 2 h of high salt exposure and in greenhouse plants exposed to a dynamic stress condition. As inferred from biometric data and chlorophyll content, plants that overexpress StCDPK2 were more tolerant than wild-type plants when exposed to high salt. Overexpressing plants have a more efficient antioxidant system; they showed reduced accumulation of peroxide and higher catalase activity under salt conditions, and enhanced expression of WRKY6 and ERF5 transcription factors under control conditions. Our results indicate that StCDPK2 is an early player in the salt stress response and support a positive correlation between StCDPK2 overexpression and tolerance towards salt stress.

Methods for Delivery of dsRNAs for Agricultural Pest Control: The Case of Lepidopteran Pests.

RNA interference (RNAi) is a natural mechanism of gene regulation, highly conserved in eukaryotes. Since the elucidation of the gene silencing mechanism, RNAi became an important tool used in insect reverse genetics. The demonstration of effective target-gene silencing by ingestion of double-stranded RNA (dsRNA) produced by transgenic plants indicated the RNAi potential to be used in insect pest management, particularly in agriculture. However, the efficiency of gene silencing by RNAi in insects may vary according to the target taxa, and lepidopteran species have been shown to be quite recalcitrant to RNAi. Developing transgenic plants is a time-consuming and labor-intensive process, so alternative oral delivery systems are required to develop and optimize RNAi settings, such as selecting an efficient target gene, and dsRNA design, length, and stability, among other features. We have developed delivery systems to evaluate dsRNAs to silence genes from two important lepidopteran crop pests of tomato (Solanum lycopersicum) and sugarcane (Saccharum × officinarum): Tuta absoluta (Meyrick), the South American Tomato Pinworm, and Diatraea saccharalis (Fabricius), the Sugarcane Borer, respectively. The protocol described here can be used in similar species and includes (a) direct oral delivery by droplets containing dsRNA; (b) oral delivery by tomato leaflets that absorbed dsRNA solution; (c) delivery by Escherichia coli expressing dsRNA; and (d) delivery by transgenic plants expressing dsRNA.

Evaluation of the Chilli veinal mottle virus CP gene expressing transgenic Nicotiana benthamiana plants for disease resistance against the virus / Avaliação do gene CP do vírus Chilli veinal mottle que expressa plantas transgênicas Nicotiana benthamiana quanto à resistência a doenças contra o vírus

Vegetables are an important source of income and high-value crops for small farmers. Chilli (Capsicum spp.) is one of the most economically important vegetables of Pakistan and it is grown throughout the country. It is a rich source of nutrition especially vitamins A, B, C and E along with minerals as folic acid, manganese (Mn), potassium (K) and molybdenum (Mo). Chilli possesses seven times more amount of vitamin C than an orange. Vitamin A, C and betacarotenoids are strong antioxidants to scavenge the free radicals. Chilli production is restricted due to various biotic factors. Among these viruses, Chilli veinal mottle virus (ChiVMV) is one of the most destructive and menacing agents that inflicts heavy and colossal losses that accounted for 50% yield loss both in quality and quantity. Pathogen-Derived Resistance (PDR) approach is considered one of the effective approaches to manage plant viruses. In this study, ChiVMV was characterized on a molecular level, the coat protein (CP) gene of the virus was stably transformed into Nicotiana benthamiana plants using Agrobacterium tumefaciens. The transgenic plants were challenged with the virus to evaluate the level of resistance of plants against the virus. It was observed that the plants expressing CP gene have partial resistance against the virus in terms of symptoms' development and virus accumulation. Translation of this technique into elite chilli varieties will be resulted to mitigate the ChiVMV in the crop as well as an economic benefit to the farmers.

Vegetais são uma importante fonte de renda e culturas de alto valor para os pequenos agricultores. A pimenta-malagueta (Capsicum spp.) é uma das hortaliças mais importantes economicamente do Paquistão e é cultivada em todo o país. É uma rica fonte de nutrição, especialmente vitaminas A, B, C e E com minerais como ácido fólico, manganês (Mn), potássio (K) e molibdênio (Mo). O pimentão possui sete vezes mais vitamina C do que a laranja. Vitaminas A e C e betacarotenoides são antioxidantes fortes para eliminar os radicais livres. A produção de pimenta é restrita devido a vários fatores bióticos. Entre esses vírus, o ChiVMV é o agente mais destrutivo e ameaçador que inflige perdas pesadas e colossais que representam 50% da perda de rendimento, tanto em qualidade quanto em quantidade. A abordagem de resistência derivada de patógenos (PDR) é considerada uma das abordagens eficazes para gerenciar os vírus de plantas. Neste estudo, ChiVMV foi caracterizado em nível molecular e o gene CP do vírus foi transformado de forma estável em plantas Nicotiana benthamiana usando Agrobacterium tumefaciens. As plantas transgênicas foram desafiadas com o vírus para avaliar seu nível de resistência contra o vírus. Observou-se que as plantas que expressam o gene CP apresentam resistência parcial ao vírus em termos de desenvolvimento de sintomas e acúmulo de vírus. A tradução dessa técnica em variedades de pimenta de elite resultará na mitigação do ChiVMV na safra, bem como em benefícios econômicos para os agricultores em termos de melhor rendimento e baixo custo de produção.

Evaluation of the Chilli veinal mottle virus CP gene expressing transgenic Nicotiana benthamiana plants for disease resistance against the virus / Avaliação do gene CP do vírus Chilli veinal mottle que expressa plantas transgênicas Nicotiana benthamiana quanto à resistência a doenças contra o vírus

Vegetables are an important source of income and high-value crops for small farmers. Chilli (Capsicum spp.) is one of the most economically important vegetables of Pakistan and it is grown throughout the country. It is a rich source of nutrition especially vitamins A, B, C and E along with minerals as folic acid, manganese (Mn), potassium (K) and molybdenum (Mo). Chilli possesses seven times more amount of vitamin C than an orange. Vitamin A, C and beta carotenoids are strong antioxidants to scavenge the free radicals. Chilli production is restricted due to various biotic factors. Among these viruses, Chilli veinal mottle virus (ChiVMV) is one of the most destructive and menacing agents that inflicts heavy and colossal losses that accounted for 50% yield loss both in quality and quantity. Pathogen-Derived Resistance (PDR) approach is considered one of the effective approaches to manage plant viruses. In this study, ChiVMV was characterized on a molecular level, the coat protein (CP) gene of the virus was stably transformed into Nicotiana benthamiana plants using Agrobacterium tumefaciens. The transgenic plants were challenged with the virus to evaluate the level of resistance of plants against the virus. It was observed that the plants expressing CP gene have partial resistance against the virus in terms of symptoms' development and virus accumulation. Translation of this technique into elite chilli varieties will be resulted to mitigate the ChiVMV in the crop as well as an economic benefit to the farmers.(AU)

Vegetais são uma importante fonte de renda e culturas de alto valor para os pequenos agricultores. A pimenta-malagueta (Capsicum spp.) é uma das hortaliças mais importantes economicamente do Paquistão e é cultivada em todo o país. É uma rica fonte de nutrição, especialmente vitaminas A, B, C e E com minerais como ácido fólico, manganês (Mn), potássio (K) e molibdênio (Mo). O pimentão possui sete vezes mais vitamina C do que a laranja. Vitaminas A e C e betacarotenoides são antioxidantes fortes para eliminar os radicais livres. A produção de pimenta é restrita devido a vários fatores bióticos. Entre esses vírus, o ChiVMV é o agente mais destrutivo e ameaçador que inflige perdas pesadas e colossais que representam 50% da perda de rendimento, tanto em qualidade quanto em quantidade. A abordagem de resistência derivada de patógenos (PDR) é considerada uma das abordagens eficazes para gerenciar os vírus de plantas. Neste estudo, ChiVMV foi caracterizado em nível molecular e o gene CP do vírus foi transformado de forma estável em plantas Nicotiana benthamiana usando Agrobacterium tumefaciens. As plantas transgênicas foram desafiadas com o vírus para avaliar seu nível de resistência contra o vírus. Observou-se que as plantas que expressam o gene CP apresentam resistência parcial ao vírus em termos de desenvolvimento de sintomas e acúmulo de vírus. A tradução dessa técnica em variedades de pimenta de elite resultará na mitigação do ChiVMV na safra, bem como em benefícios econômicos para os agricultores em termos de melhor rendimento e baixo custo de produção.(AU)

Evaluation of the Chilli veinal mottle virus CP gene expressing transgenic Nicotiana benthamiana plants for disease resistance against the virus / Avaliação do gene CP do vírus Chilli veinal mottle que expressa plantas transgênicas Nicotiana benthamiana quanto à resistência a doenças contra o vírus

Vegetables are an important source of income and high-value crops for small farmers. Chilli (Capsicum spp.) is one of the most economically important vegetables of Pakistan and it is grown throughout the country. It is a rich source of nutrition especially vitamins A, B, C and E along with minerals as folic acid, manganese (Mn), potassium (K) and molybdenum (Mo). Chilli possesses seven times more amount of vitamin C than an orange. Vitamin A, C and beta carotenoids are strong antioxidants to scavenge the free radicals. Chilli production is restricted due to various biotic factors. Among these viruses, Chilli veinal mottle virus (ChiVMV) is one of the most destructive and menacing agents that inflicts heavy and colossal losses that accounted for 50% yield loss both in quality and quantity. Pathogen-Derived Resistance (PDR) approach is considered one of the effective approaches to manage plant viruses. In this study, ChiVMV was characterized on a molecular level, the coat protein (CP) gene of the virus was stably transformed into Nicotiana benthamiana plants using Agrobacterium tumefaciens. The transgenic plants were challenged with the virus to evaluate the level of resistance of plants against the virus. It was observed that the plants expressing CP gene have partial resistance against the virus in terms of symptoms' development and virus accumulation. Translation of this technique into elite chilli varieties will be resulted to mitigate the ChiVMV in the crop as well as an economic benefit to the farmers.

Vegetais são uma importante fonte de renda e culturas de alto valor para os pequenos agricultores. A pimenta-malagueta (Capsicum spp.) é uma das hortaliças mais importantes economicamente do Paquistão e é cultivada em todo o país. É uma rica fonte de nutrição, especialmente vitaminas A, B, C e E com minerais como ácido fólico, manganês (Mn), potássio (K) e molibdênio (Mo). O pimentão possui sete vezes mais vitamina C do que a laranja. Vitaminas A e C e betacarotenoides são antioxidantes fortes para eliminar os radicais livres. A produção de pimenta é restrita devido a vários fatores bióticos. Entre esses vírus, o ChiVMV é o agente mais destrutivo e ameaçador que inflige perdas pesadas e colossais que representam 50% da perda de rendimento, tanto em qualidade quanto em quantidade. A abordagem de resistência derivada de patógenos (PDR) é considerada uma das abordagens eficazes para gerenciar os vírus de plantas. Neste estudo, ChiVMV foi caracterizado em nível molecular e o gene CP do vírus foi transformado de forma estável em plantas Nicotiana benthamiana usando Agrobacterium tumefaciens. As plantas transgênicas foram desafiadas com o vírus para avaliar seu nível de resistência contra o vírus. Observou-se que as plantas que expressam o gene CP apresentam resistência parcial ao vírus em termos de desenvolvimento de sintomas e acúmulo de vírus. A tradução dessa técnica em variedades de pimenta de elite resultará na mitigação do ChiVMV na safra, bem como em benefícios econômicos para os agricultores em termos de melhor rendimento e baixo custo de produção.

Evaluation of the Chilli veinal mottle virus CP gene expressing transgenic Nicotiana benthamiana plants for disease resistance against the virus

Abstract Vegetables are an important source of income and high-value crops for small farmers. Chilli (Capsicum spp.) is one of the most economically important vegetables of Pakistan and it is grown throughout the country. It is a rich source of nutrition especially vitamins A, B, C and E along with minerals as folic acid, manganese (Mn), potassium (K) and molybdenum (Mo). Chilli possesses seven times more amount of vitamin C than an orange. Vitamin A, C and beta-carotenoids are strong antioxidants to scavenge the free radicals. Chilli production is restricted due to various biotic factors. Among these viruses, Chilli veinal mottle virus (ChiVMV) is one of the most destructive and menacing agents that inflicts heavy and colossal losses that accounted for 50% yield loss both in quality and quantity. Pathogen-Derived Resistance (PDR) approach is considered one of the effective approaches to manage plant viruses. In this study, ChiVMV was characterized on a molecular level, the coat protein (CP) gene of the virus was stably transformed into Nicotiana benthamiana plants using Agrobacterium tumefaciens. The transgenic plants were challenged with the virus to evaluate the level of resistance of plants against the virus. It was observed that the plants expressing CP gene have partial resistance against the virus in terms of symptoms development and virus accumulation. Translation of this technique into elite chilli varieties will be resulted to mitigate the ChiVMV in the crop as well as an economic benefit to the farmers.

Resumo Vegetais são uma importante fonte de renda e culturas de alto valor para os pequenos agricultores. A pimenta-malagueta (Capsicum spp.) é uma das hortaliças mais importantes economicamente do Paquistão e é cultivada em todo o país. É uma rica fonte de nutrição, especialmente vitaminas A, B, C e E com minerais como ácido fólico, manganês (Mn), potássio (K) e molibdênio (Mo). O pimentão possui sete vezes mais vitamina C do que a laranja. Vitaminas A e C e betacarotenoides são antioxidantes fortes para eliminar os radicais livres. A produção de pimenta é restrita devido a vários fatores bióticos. Entre esses vírus, o ChiVMV é o agente mais destrutivo e ameaçador que inflige perdas pesadas e colossais que representam 50% da perda de rendimento, tanto em qualidade quanto em quantidade. A abordagem de resistência derivada de patógenos (PDR) é considerada uma das abordagens eficazes para gerenciar os vírus de plantas. Neste estudo, ChiVMV foi caracterizado em nível molecular e o gene CP do vírus foi transformado de forma estável em plantas Nicotiana benthamiana usando Agrobacterium tumefaciens. As plantas transgênicas foram desafiadas com o vírus para avaliar seu nível de resistência contra o vírus. Observou-se que as plantas que expressam o gene CP apresentam resistência parcial ao vírus em termos de desenvolvimento de sintomas e acúmulo de vírus. A tradução dessa técnica em variedades de pimenta de elite resultará na mitigação do ChiVMV na safra, bem como em benefícios econômicos para os agricultores em termos de melhor rendimento e baixo custo de produção.

Antisense transcription in plants: a systematic review and an update on cis-NATs of sugarcane

Initially, natural antisense transcripts (NATs, natRNAs, or asRNAs) were considered repressors; however, their functions in gene regulation are diverse. Positive, negative, or neutral correlations to the cognate gene expression have been noted. Although the first studies were published about 50 years ago, there is still much to be investigated regarding antisense transcripts in plants. A systematic review of scientific publications available in the Web of Science databases was conducted to contextualize how the studying of antisense transcripts has been addressed. Studies were classified considering three categories: “Natural antisense” (208), artificial antisense used in “Genetic Engineering” (797), or “Natural antisense and Genetic Engineering”-related publications (96). A similar string was used for a systematic search in the NCBI Gene database. Of the 1132 antisense sequences found for plants, only 0.8% were cited in PubMed and had antisense information confirmed. This value was the lowest when compared to fungi (2.9%), bacteria (2.3%), and mice (54.1%). Finally, we present an update for the cis-NATs identified in Saccharum spp. Of the 1413 antisense transcripts found in different experiments, 25 showed concordant expressions, 22 were discordant, 1264 did not correlate with the cognate genes, and 102 presented variable results depending on the experiment.

Citrus Genetic Transformation: An Overview of the Current Strategies and Insights on the New Emerging Technologies.

Citrus are among the most prevailing fruit crops produced worldwide. The implementation of effective and reliable breeding programs is essential for coping with the increasing demands of satisfactory yield and quality of the fruit as well as to deal with the negative impact of fast-spreading diseases. Conventional methods are time-consuming and of difficult application because of inherent factors of citrus biology, such as their prolonged juvenile period and a complex reproductive stage, sometimes presenting infertility, self-incompatibility, parthenocarpy, or polyembryony. Moreover, certain desirable traits are absent from cultivated or wild citrus genotypes. All these features are challenging for the incorporation of the desirable traits. In this regard, genetic engineering technologies offer a series of alternative approaches that allow overcoming the difficulties of conventional breeding programs. This review gives a detailed overview of the currently used strategies for the development of genetically modified citrus. We describe different aspects regarding genotype varieties used, including elite cultivars or extensively used scions and rootstocks. Furthermore, we discuss technical aspects of citrus genetic transformation procedures via Agrobacterium, regular physical methods, and magnetofection. Finally, we describe the selection of explants considering young and mature tissues, protoplast isolation, etc. We also address current protocols and novel approaches for improving the in vitro regeneration process, which is an important bottleneck for citrus genetic transformation. This review also explores alternative emerging transformation strategies applied to citrus species such as transient and tissue localized transformation. New breeding technologies, including cisgenesis, intragenesis, and genome editing by clustered regularly interspaced short palindromic repeats (CRISPR), are also discussed. Other relevant aspects comprising new promoters and reporter genes, marker-free systems, and strategies for induction of early flowering, are also addressed. We provided a future perspective on the use of current and new technologies in citrus and its potential impact on regulatory processes.

Does refuge spillover affect arthropod food webs associated with Bt maize?

BACKGROUND: A high dose/refuge combination is the main tactic recommended for mitigating resistance selection of target herbivore species in crops expressing insecticidal proteins of the bacterium Bacillus thuringiensis (i.e. Bt proteins). The tactic consists of the simultaneous use of Bt crops expressing high levels of the Bt protein associated with neighboring areas of refuge of the same non-Bt crop species. Nonetheless, the approach faces controversy regarding its effectiveness and scale of adoption, at least in some regions. One concern focuses on its potential impact on the arthropod community, including its short-term and spatially dependent impact considering the likely biota spillover effect between Bt and non-Bt neighboring areas. Thus, the eventual spillover of Bt maize targeted and non-targeted arthropods was surveyed along transects extending from the refuge border to the center of the Bt maize area. RESULTS: Arthropods were collected throughout the maize vegetative and reproductive stages. A total of 85 arthropod species were collected, but their richness and abundance did not vary with distance from the refuge. By contrast, cultivation season played a significant role in distinguishing the arthropod communities. Refuge distance from the sampling point within Bt-fields did not significantly affect the food web metrics, unlike season, which affected the number of nodes integrating each food web. Winter maize cultivation exhibited higher arthropod diversity and combined values of species numeric abundance and biomass at each trophic level. CONCLUSIONS: No arthropod spillover was evident between the refuge edge and Bt maize, adding further controversy to the tactic currently subjected to lower usage in the region with a disputed cost-benefit relationship, because not even the target and its interdependent species were affected. © 2021 Society of Chemical Industry.

Citrus reticulata CrRAP2.2 Transcriptional Factor Shares Similar Functions to the Arabidopsis Homolog and Increases Resistance to Xylella fastidiosa.

Xylella fastidiosa is a worldwide multihost pathogen that causes diseases in different crops. It is considered a new global threat and substantial efforts have been made in order to identify sources of resistance. Indeed, many genes have been associated with resistance to X. fastidiosa, but without functional validation. Here, we describe a C. reticulata gene homologous to the transcriptional factor RAP2.2 from Arabidopsis thaliana that increases resistance to citrus variegated chlorosis (CVC). This gene was previously detected in C. reticulata challenged with X. fastidiosa. Bioinformatics analysis together with subcellular localization and auto-activation assays indicated that RAP2.2 from C. reticulata (CrRAP2.2) is a transcriptional factor orthologous to AtRAP2.2. Thus, we used A. thaliana as a model host to evaluate the functional role of CrRAP2.2 in X. fastidiosa resistance. The inoculation of X. fastidiosa in the A. thaliana rap2.2 mutant resulted in a larger bacterial population, which was complemented by CrRAP2.2. In addition, symptoms of anthocyanin accumulation were higher in the mutant, whose phenotype was restored by CrRAP2.2, indicating that they have conserved functions in plant defense response. We therefore transformed C. sinensis with CrRAP2.2 and verified a positive correlation between CVC resistance and gene expression in transgenic lines. This is the first study using A. thaliana as model host that characterizes the function of a gene related to X. fastidiosa defense response and its application in genetic engineering to obtain citrus resistance to CVC.

Biological parameters of Spodoptera cosmioides Walker (Lepidoptera: Noctuidae) in cotton varieties / Parâmetros biológicos de Spodoptera cosmioides Walker (Lepidoptera: Noctuidae) em variedades de algodoeiro

Spodoptera cosmioides Walker is a polyphagous insecticide-resistant species found in various regions of Brazil. Despite occurring at low densities, it is considered a potential pest of cotton, soybean, and bean crops in the Brazilian cerrado. This work investigated the comparative biology of S. cosmioides in the commercial transgenic cotton NuOPAL (Bollgard I, Evento 531) and the conventional isoline variety DeltaOPAL. The experiment was conducted under laboratory conditions (26 ± 1°C; relative humidity: 70 ± 10%; photoperiod: 12h), using newly-hatched and individualized larvae. We evaluated the duration and viability of immature stages, duration of the biological cycle, pupal weight, sex ratio, percentage of deformed adults and of adults trapped in the pupal case, adult longevity, fecundity, and egg viability. The biological parameters observed showed no significant differences between the two varieties studied, which enabled high consumption, high assimilation, high fecundity, and viability for S. cosmioides.(AU)

Spodoptera cosmioides Walker é uma espécie polífaga, resistente a inseticidas em várias regiões do país e, apesar de ocorrer em baixas densidades, é considerada uma praga potencial para as culturas de algodão, soja e feijão no cerrado. Neste trabalho, estudou-se a biologia comparada de S. cosmioides sobre o cultivar transgênico comercial de algodoeiro NuOPAL (Bollgard I, Evento 531) e sobre o cultivar isolinha convencional DeltaOPAL. O experimento foi desenvolvido em condições de laboratório (26 ± 1°C; UR: 70 ± 10%; fotofase: 12h) a partir de lagartas recém-eclodidas e individualizadas. Foram avaliados a duração e a viabilidade das fases imaturas, a duração do ciclo biológico, o peso de pupas, a razão sexual, a porcentagem de deformação de adultos e de adultos não liberados dos invólucros pupais, a longevidade de adultos, a fecundidade e viabilidade de ovos. Os parâmetros biológicos observados não diferiram significativamente em relação às duas cultivares, que proporcionaram alto consumo, alta assimilação, alta fecundidade e viabilidade à S. cosmioides.(AU)

Biological parameters of Spodoptera cosmioides Walker (Lepidoptera: Noctuidae) in cotton varieties / Parâmetros biológicos de Spodoptera cosmioides Walker (Lepidoptera: Noctuidae) em variedades de algodoeiro

Spodoptera cosmioides Walker is a polyphagous insecticide-resistant species found in various regions of Brazil. Despite occurring at low densities, it is considered a potential pest of cotton, soybean, and bean crops in the Brazilian cerrado. This work investigated the comparative biology of S. cosmioides in the commercial transgenic cotton NuOPAL (Bollgard I, Evento 531) and the conventional isoline variety DeltaOPAL. The experiment was conducted under laboratory conditions (26 ± 1°C; relative humidity: 70 ± 10%; photoperiod: 12h), using newly-hatched and individualized larvae. We evaluated the duration and viability of immature stages, duration of the biological cycle, pupal weight, sex ratio, percentage of deformed adults and of adults trapped in the pupal case, adult longevity, fecundity, and egg viability. The biological parameters observed showed no significant differences between the two varieties studied, which enabled high consumption, high assimilation, high fecundity, and viability for S. cosmioides.(AU)

Spodoptera cosmioides Walker é uma espécie polífaga, resistente a inseticidas em várias regiões do país e, apesar de ocorrer em baixas densidades, é considerada uma praga potencial para as culturas de algodão, soja e feijão no cerrado. Neste trabalho, estudou-se a biologia comparada de S. cosmioides sobre o cultivar transgênico comercial de algodoeiro NuOPAL (Bollgard I, Evento 531) e sobre o cultivar isolinha convencional DeltaOPAL. O experimento foi desenvolvido em condições de laboratório (26 ± 1°C; UR: 70 ± 10%; fotofase: 12h) a partir de lagartas recém-eclodidas e individualizadas. Foram avaliados a duração e a viabilidade das fases imaturas, a duração do ciclo biológico, o peso de pupas, a razão sexual, a porcentagem de deformação de adultos e de adultos não liberados dos invólucros pupais, a longevidade de adultos, a fecundidade e viabilidade de ovos. Os parâmetros biológicos observados não diferiram significativamente em relação às duas cultivares, que proporcionaram alto consumo, alta assimilação, alta fecundidade e viabilidade à S. cosmioides.(AU)

Expression of a Chloroplast-Targeted Cyanobacterial Flavodoxin in Tomato Plants Increases Harvest Index by Altering Plant Size and Productivity.

Tomato is the most important horticultural crop worldwide. Domestication has led to the selection of highly fruited genotypes, and the harvest index (HI), defined as the ratio of fruit yield over total plant biomass, is usually employed as a biomarker of agronomic value. Improvement of HI might then result from increased fruit production and/or lower vegetative growth. Reduction in vegetative biomass has been accomplished in various plant species by expression of flavodoxin, an electron shuttle flavoprotein that interacts with redox-based pathways of chloroplasts including photosynthesis. However, the effect of this genetic intervention on the development of reproductive organs has not been investigated. We show herein that expression of a plastid-targeted cyanobacterial flavodoxin in tomato resulted in significant reduction of plant size affecting stems, leaves, and fruit. Decreased size correlated with smaller cells and was accompanied by higher pigment contents and photosynthetic activities per leaf cross-section. Flavodoxin accumulated in green fruit but declined with ripening. Significant increases in HI were observed in flavodoxin-expressing lines due to the production of higher fruit number per plant in smaller plants. Therefore, overall yields can be enhanced by increasing plant density in the field. Metabolic profiling of ripe red fruit showed that levels of sugars, organic acids, and amino acids were similar or higher in transgenic plants, indicating that there was no trade-off between increased HI and fruit metabolite contents in flavodoxin-expressing plants. Taken together, our results show that flavodoxin has the potential to improve major agronomic traits when introduced in tomato.