The Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Liviidae), is considered the main vector of the bacteria associated with huanglongbing, a very serious disease that has threatened the world citrus industry. The absence of efficient control management protocols, including a lack of resistant cultivars, has led to the development of different approaches to study this pathosystem. The production of resistant genotypes relies on D. citri gene expression analyses by RT-qPCR to assess control of the vector population. High-quality, reliable RT-qPCR analyses depend upon proper reference gene selection and validation. However, adequate D. citri reference genes have not yet been identified. In the present study, we evaluated the genes EF 1-α, ACT, GAPDH, RPL7, RPL17, and TUB as candidate reference genes for this insect. Gene expression stability was evaluated using the mathematical algorithms deltaCt, NormFinder, BestKeeper, and geNorm, at five insect developmental stages, grown on two different plant hosts [Citrus sinensis (L.) Osbeck (Sapindales: Rutaceae) and Murraya paniculata (L.) Jack (Sapindales: Rutaceae)]. The final gene ranking was calculated using RefFinder software, and the V-ATPase-A gene was selected for validation. According to our results, two reference genes are recommended when different plant hosts and developmental stages are considered. Considering gene expression studies in D. citri grown on M. paniculata, regardless of the insect developmental stage, GAPDH and RPL7 have the best fit as reference genes in RT-qPCR analyses, whereas GAPDH and EF 1-α are recommended as reference genes in insect studies using C. sinensis.

Citrus species, including sweet oranges, grapefruits, pomelos, and lemons, are the most widely cultivated trees and consumed fruits worldwide. In citrus orchard management, the control of Huanglongbing (HLB) disease and its insect vector Diaphorina citri (Asian citrus psyllid, ACP) represents a major global challenge. Consumers have been increasingly pushing the citrus production chain toward a more sustainable system, including stringent measures to prevent the use of chemical pesticides. In recent years, biotechnological advances have offered safe and environmentally friendly alternatives for crop production. Technologies such as RNA interference (RNAi)-mediated gene silencing have emerged as innovative tools for agricultural pest management. Here, we provide an overview of RNAi as a promising approach for ACP control and discuss the associated challenges. Despite the availability of specific silencing sequences aimed at a target gene of the insect pest, the uptake of double-stranded RNA is limited in hemipteran insects. In this context, improved delivery methods, stability maintenance, and RNAi response are considered the factors contributing to the increased effectiveness of exogenous RNAi against hemipteran pests. These approaches can serve as potential tools for efficient ACP control.

The objective of this study was to investigate the effects of the application of indole-3-butyric acid (IBA) at concentrations of 0, 30, 60, and 90 mg·L ⁻¹ , for 24 hours, on rooting of softwood and semihardwood cuttings of tea ( Camellia sinensis var. sinensis ‘Yabukita’ and C. sinensis var. assamica ‘IAC-259’) collected in winter and summer. In the summer, IBA increased root percentage of softwood cuttings from ‘Yabukita’ compared with the control. However, the rooting of semihardwood cuttings was unaffected by this growth regulator. In winter, application of 90 mg·L ⁻¹ IBA increased the rooting regardless of the type of ‘Yabukita’ cuttings. In addition, in ‘IAC-259’, there was an increase in dry weight, number of roots (NOR), and rooting percentage of softwood cuttings collected in summer with application of 90 mg·L ⁻¹ IBA compared with control. In contrast, during the same period of the year, the semihardwood cuttings of ‘IAC-259’ were unaffected by the IBA. In winter, the percentage of cutting survival, rooting, the number, and length of roots were unaffected by IBA in ‘IAC-259’. Overall, we would recommend the use of exogenous IBA for rooting of cuttings collected in the summer or winter.

Fertilizer management is an essential step in the production process, as it allows the plant to use its productive capacity to the fullest extent possible. Researchers have tested maximum nutrient use with reduced losses to the environment aiming to increase productivity with fewer environmental impacts. This study compared the effects of controlled-release fertilizers (CRFs) with water-soluble fertilizer (WSF) and clear water (control) on the growth and nutrient uptake of croton ( Codiaeum variegatum L.) and nitrogen leaching. The experiment was conducted with three replications and six treatments: two rates (1.5 g and 3.0 g per liter of substrate) of two CRFs [Osmocote Plus (15% N, 3.93% P, and 9.96% K) and Basacote (15% N, 3.49% P, and 9.96% K)], WSF, and clean water as control. All CRFs were applied before planting and WSF was supplied as nutrient solution through automated moisture sensor activated irrigation system. Plant growth (number of leaves, leaf area, stem height, root volume, and shoot and root dry weights) and total nutrient contents in the leaf tissue were evaluated every 30 days. Electrical conductivity (EC), pH, nitrate, ammonium, and total nitrogen contents were measured in the leached solution. Indeed, results showed that CRFs at a low rate provided similar development and quality of croton plants compared with WSF. Plant growth indicators were similar until 90 days after transplanting (DAT). After that, at 150 DAT, the highest values to number of leaves and leaf area occurred with WSF and with the lowest CRF rate as compared with the other treatments and control. The highest root volume was found with the WSF, which resulted in larger roots compared with the other treatments. These results showed WSF can be replaced by CRFs at low rates on croton growth. Moreover, according to the visual scale, the best treatments were WSF and Basacote at the low rate, where plants were bright, with multicolored leaves with prominent orange shades. However, CRFs maintained pH and EC within the recommended range for the growth of croton and reduced the nitrogen leaching from the pots.

The objective of this work was to develop a method to create and validate CRISPR-Cas systems and different gRNAs in soybean (Glycine max) embryos. Two model genes were used for simple mutation with one gRNA or partial gene deletion with two guides. The gRNAs were inserted into the CRISPR transformation vectors by a type IIS restriction enzyme or by subcloning and inserting the promoter + gRNA2 in the final transformation vector using the classic restriction enzyme cloning method. The vectors were successfully constructed for one and two gRNAs. Agrobacterium-mediated transient transformation in soybean was carried out to test the quality of gRNAs and of the system itself (expression cassette). Simple mutation and gene deletion were detected in the embryos transformed after DNA enrichment by enzyme digestion followed by polymerase chain reaction and sequencing, which indicates that the CRISPR-Cas system and guides were working. This protocol can be used to accelerate CRISPR-based genome editing strategies for genetic transformation in soybean.