Citrus leprosis (CL) is a re-emergent viral disease affecting citrus crops in the Americas, and citrus leprosis virus C (CiLV-C), belonging to the genus Cilevirus, is the main pathogen responsible for the disease. Despite the economic importance of CL to the citrus industry, very little is known about the performance of viral proteins. Here, we present a robust in vivo study around functionality of p29, p15, p61, MP, and p24 CiLV-C proteins in the host cells. The intracellular sub-localization of all those viral proteins in plant cells are shown, and their co-localization with the endoplasmic reticulum (ER), Golgi complex (GC) (p15, MP, p61 and p24), actin filaments (p29, p15 and p24), nucleus (p15), and plasmodesmata (MP) are described. Several features are disclosed, including i) ER remodeling and redistribution of GC apparatus, ii) trafficking of the p29 and MP along the ER network system, iii) self-interaction of the p29, p15, and p24 and hetero-association between p29-p15, p29-MP, p29-p24, and p15-MP proteins in vivo. We also showed that all proteins are associated with biological membranes; whilst p15 is peripherally associated, p29, p24, and MP are integrally bound to cell membranes. Furthermore, while p24 exposes an N-cytoplasm-C-lumen topology, p29, and p15 are oriented toward the cytoplasmic face of the biological membrane. Based on our findings, we discuss the possible performance of each protein in the context of infection and a hypothetical model encompassing the virus spread and sites for replication and particle assembly is suggested.

This review emphasizes the biotechnological potential of molecules implicated in the different layers of plant immunity, including, pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI), effector-triggered susceptibility (ETS), and effector-triggered immunity (ETI) that can be applied in the development of disease-resistant genetically modified (GM) plants. These biomolecules are produced by pathogens (viruses, bacteria, fungi, oomycetes) or plants during their mutual interactions. Biomolecules involved in the first layers of plant immunity, PTI and ETS, include inhibitors of pathogen cell-wall-degrading enzymes (CWDEs), plant pattern recognition receptors (PRRs) and susceptibility (S) proteins, while the ETI-related biomolecules include plant resistance (R) proteins. The biomolecules involved in plant defense PTI/ETI responses described herein also include antimicrobial peptides (AMPs), pathogenesis-related (PR) proteins and ribosome-inhibiting proteins (RIPs), as well as enzymes involved in plant defensive secondary metabolite biosynthesis (phytoanticipins and phytoalexins). Moreover, the regulation of immunity by RNA interference (RNAi) in GM disease-resistant plants is also considered. Therefore, the present review does not cover all the classes of biomolecules involved in plant innate immunity that may be applied in the development of disease-resistant GM crops but instead highlights the most common strategies in the literature, as well as their advantages and disadvantages.

Resumo -O objetivo deste trabalho foi estimar a divergência genética entre acessos de mandioca açucarados e não açucarados, por meio de marcadores moleculares, caracteres quantitativos e qualitativos, bem como determinar a correlação entre essas estimativas. Foram utilizados quatro acessos de mandioca açucarados e quatro não açucarados, com duas variedades locais e duas comerciais. Os acessos foram avaliados em campo, em laboratório, com marcadores RAPD, quanto a 12 caracteres quantitativos e 33 morfológicos. Foram estimadas as matrizes de dissimilaridade/distância genética entre os acessos, por meio dos caracteres qualitativos e quantitativos e da significância da correlação entre as matrizes. A divergência genética entre os acessos foi elevada e os acessos açucarados foram diferenciados das variedades não açucaradas locais e comerciais. As distâncias estimadas, por meio de marcadores moleculares e caracteres qualitativos, evidenciaram elevada associação entre si e associação moderada com a estimada por meio de caracteres quantitativos.Termos para indexação: Manihot esculenta, melhoramento genético, recursos genéticos, variabilidade genética. Genetic divergence among sugary and nonsugary cassava accessionsAbstract -The objective of this work was to estimate the genetic divergence among sugary and nonsugary cassava accessions, through molecular markers, and quantitative and qualitative characters, as well as to determine the correlation among these estimates. Four sugary cassava accessions and four nonsugary ones were used, including two landraces and two nonsugary improved varieties. The accessions were evaluated in field conditions and in laboratories, with RAPD markers, in 12 quantitative and 33 qualitative characters. Matrixes of genetic dissimilarity/distance among accessions were estimated, based on qualitative and quantitative characters, molecular markers, and on the correlation significance among matrixes. High genetic divergence among the evaluated accessions was observed, the sugary accessions were differentiated from the landraces and improved nonsugary varieties. The distances estimated through molecular markers and qualitative characters showed the high association among each other and moderate association with the distance estimated through quantitative characters.

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SummaryGenetically 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.

ResumoO objetivo do trabalho foi caracterizar e estimar a variabilidade genética entre acessos de mandioca de indústria, com potencial de adaptação às condições do Cerrado do Brasil Central, por meio de caracteres quantitativos, qualitativos e marcadores moleculares de forma isolada e conjunta, bem como estimar a correlação entre as estimativas obtidas com base em cada grupo de caracteres. Dezesseis acessos de mandioca de indústria com potencial de adaptação às condições do Cerrado do Brasil Central foram avaliados a campo quanto a 11 caracteres quantitativos e 33 caracteres qualitativos, em experimento conduzido na Embrapa Cerrados. Os acessos foram também avaliados quanto a marcadores RAPD em laboratório. Posteriormente, foram estimadas as matrizes de dissimilaridade/distância genética entre os acessos por meio dos caracteres qualitativos, quantitativos e marcadores moleculares e pela análise conjunta dos dados. Além disso, foi estimada a correlação entre as matrizes. No grupo de acessos de indústria avaliados existe ampla variabilidade quanto aos caracteres quantitativos, qualitativos e moleculares aferidos. As únicas correlações significativas encontradas foram entre (i) a matriz de dissimilaridade estimada por meio da análise conjunta e a matriz de dissimilaridade estimada por meio de dados qualitativos (r = 0,52) e; (ii) a matriz de dissimilaridade estimada por meio da análise conjunta e a matriz de dissimilaridade estimada por meio de marcadores moleculares (r= 0,75). A fraca associação entre essas medidas indicou que a melhor estratégia para orientar ações de conservação e uso de germoplasma de mandioca de indústria é por meio de estudos de divergência genética com o emprego de marcadores moleculares, caracteres qualitativos e caracteres quantitativos de forma conjunta e complementar.

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