Bioactivity of hydroalcoholic extracts from tropaeolum majus L. (tropaeolaceae) on the germination, initial plant development and cell cycle of Lactuca sativa L.

Natural products are usually considered harmless; however, these substances need to be consumed with caution. Biological assays with plant models are a suitable alternative for prospective studies to assess natural product-initiated toxicity. The aim of this study was to examine the toxic potential of leaf and flower extracts derived from Tropaeolum majus L. a widely used plant in traditional medicine. Seeds of Lactuca sativa L. were exposed to T. majus extracts and based upon the seedling growth curve values, the 50% Inhibition Concentration (IC50) was calculated and applied for cell cycle analysis exposure. Both extracts contained organic acids, proteins, amino acids, and terpene steroids. Sesquiterpene lactones and depside were detected in leaf extracts. The higher concentration tested exhibited a marked phytotoxic effect. The extracts induced clastogenic, aneugenic cytotoxic, and potential mutagenic effects. The possible relationships between the classes of compounds found in the extracts and effects on cells and DNA were determined.

Integrated multi-omic approach reveals the effect of a Graminaceae-derived biostimulant and its lighter fraction on salt-stressed lettuce plants.

Plant biostimulants are widely applied in agriculture for their ability to improve plant fitness. In the present work, the impact of Graminaceae-derived protein hydrolysate (P) and its lighter molecular fraction F3 (< 1 kDa) on lettuce plants, subjected to either no salt or high salt conditions, was investigated through the combination of metabolomics and transcriptomics. The results showed that both treatments significantly modulated the transcriptome and metabolome of plants under salinity stress, highlighting an induction of the hormonal response. Nevertheless, P and F3 also displayed several peculiarities. F3 specifically modulated the response to ethylene and MAPK signaling pathway, whereas P treatment induced a down-accumulation of secondary metabolites, albeit genes controlling the biosynthesis of osmoprotectants and antioxidants were up-regulated. Moreover, according with the auxin response modulation, P promoted cell wall biogenesis and plasticity in salt-stressed plants. Notably, our data also outlined an epigenetic control of gene expression induced by P treatment. Contrarily, experimental data are just partially in agreement when not stressed plants, treated with P or F3, were considered. Indeed, the reduced accumulation of secondary metabolites and the analyses of hormone pathways modulation would suggest a preferential allocation of resources towards growth, that is not coherent with the down-regulation of the photosynthetic machinery, the CO2 assimilation rate and leaves biomass. In conclusion, our data demonstrate that, although they might activate different mechanisms, both the P and F3 can result in similar benefits, as far as the accumulation of protective osmolytes and the enhanced tolerance to oxidative stress are concerned. Notably, the F3 fraction exhibits slightly greater growth promotion effects under high salt conditions. Most importantly, this research further corroborates that biostimulants' mode of action is dependent on plants' physiological status and their composition, underscoring the importance of investigating the bioactivity of the different molecular components to design tailored applications for the agricultural practice.

Heavy metals and potential health risk assessment of Lactuca sativa and Daucus carrota from soil treated with organic manures and chemical fertilizer.

The large-scale production of food crops with heavy application of chemical fertilizers in the effort to meet the astronomical increase in food demands may be counterproductive to the goal of food security. This study investigated the effect of different soil treatments on the levels of heavy metals (Cr, Cu, Fe, Ni, Pb, and Zn) in two types of vegetables Lactuca sativa (lettuce) and Daucus carrota (carrot). The potential carcinogenic and non-carcinogenic health risks from their consumption were also evaluated. Planting experiment was set up in a randomized block design, with different soil treatments of soil + cow dung (CD), soil + sewage sludge (SS), soil + chemical fertilizer (nitrogen-phosphorus-potassium (NPK)), and untreated soil (UNTRD). The vegetables were harvested at maturity, washed with distilled water, and subjected to an acid digestion process before the levels of heavy metals were measured by inductively coupled plasma spectrometry (ICP-MS). The mean concentrations of the metals in the vegetables across all treatments were below the maximum permissible limits. The pattern of heavy metal accumulation by the vegetables suggested that the lettuce from SS treatment accumulated higher concentrations of heavy metals like Cr (0.20 mg/kg), Cu (3.91 mg/kg), Ni (0.33 mg/kg), and Zn (20.44 mg/kg) than carrot, with highest concentrations of Fe (90.89 mg/kg) and Pb (0.16 mg/kg) recorded in lettuce from NPK treatment. The bioaccumulation factor (BAF) showed that lettuce, a leafy vegetable, has bioaccumulated more heavy metals than carrot, a root vegetable. The BAF was generally below the threshold value of 1 in both vegetables, except in lettuce from NPK and CD treatments and carrot from NPK treatments, with BAF values of 1.6, 1.69, and 1.39, respectively. The cancer risk assessment factors were well below the unacceptable maximum range of 10-4 suggesting that consuming these vegetables might not expose an individual to potential risk of cancer development. The hazard quotient estimations were below the threshold values of 1 for all heavy metals; however, the hazard index (HI) values of 1.27 and 1.58 for lettuce from NPK and SS treatments indicate a potential non-carcinogenic health risk to consumers from intake of all the heavy metals.

Uptake and Biotransformation of Guvermectin in Three Crops after In Vivo and In Vitro Exposure.

Guvermectin, as a novel nucleoside-like biopesticide, could increase the rice yield excellently, but the potential environmental behaviors remain unclear, which pose potential health risks. Therefore, the uptake and biotransformation of guvermectin in three types of crops (rice, lettuce, and carrot) were first evaluated with a hydroponic system. Guvermectin could be rapidly absorbed and reached equilibrium in roots (12-36 h) and shoots (24-60 h) in three plants, and guvermectin was also vulnerable to dissipation in roots (t1/2 1.02-3.65 h) and shoots (t1/2 9.30-17.91 h). In addition, 8 phase I and 2 phase II metabolites, transformed from guvermectin degradation in vivo and in vitro exposure, were identified, and one was confirmed as psicofuranine, which had antibacterial and antitumor properties; other metabolites were nucleoside-like chemicals. Molecular simulation and quantitative polymerase chain reaction further demonstrated that guvermectin was metabolized by the catabolism pathway of an endogenous nucleotide. Guvermectin had similar metabolites in three plants, but the biotransformation ability had a strong species dependence. In addition, all the metabolites exhibit neglectable toxicities (bioconcentration factor <2000 L/kg b.w., LC50,rat > 5000 mg/kg b.w.) by prediction. The study provided valuable evidence for the application of guvermectin and a better understanding of the biological behavior of nucleoside-like pesticides.

A low-cost spectroscopic nutrient management system for Microscale Smart Hydroponic system.

Hydroponics offers a promising approach to help alleviate pressure on food security for urban residents. It requires minimal space and uses less resources, but management can be complex. Microscale Smart Hydroponics (MSH) systems leverage IoT systems to simplify hydroponics management for home users. Previous work in nutrient management has produced systems that use expensive sensing methods or utilized lower cost methods at the expense of accuracy. This study presents a novel inexpensive nutrient management system for MSH applications that utilises a novel waterproofed, IoT spectroscopy sensor (AS7265x) in a transflective application. The sensor is submerged in a hydroponic solution to monitor the nutrients and MSH system predicts the of nutrients in the hydroponic solution and recommends an adjustment quantity in mL. A three-phase model building process was carried out resulting in significant MLR models for predicting the mL, with an R2 of 0.997. An experiment evaluated the system's performance using the trained models with a 30-day grow of lettuce in a real-world setting, comparing the results of the management system to a control group. The sensor system successfully adjusted and maintained nutrient levels, resulting in plant growth that outperformed the control group. The results of the models in actual deployment showed a strong, significant correlation of 0.77 with the traditional method of measuring the electrical conductivity of nutrients. This novel nutrient management system has the potential to transform the way nutrients are monitored in hydroponics. By simplifying nutrient management, this system can encourage the adoption of hydroponics, contributing to food security and environmental sustainability.

Using multi-omics to explore the effect of Bacillus velezensis SAAS-63 on resisting nutrient stress in lettuce.

To avoid the unreasonable use of chemical fertilizer, an environmentally friendly means of improving soil fertility is required. This study explored the role of the plant growth-promoting rhizosphere bacteria (PGPR) strain Bacillus velezensis SAAS-63 in improving nutrient stress in lettuce. Compared with no inoculation, B. velezensis SAAS-63 inoculants exhibited significantly increased fresh weight, root length, and shoot height under nutrient deficiency, as well as improved antioxidant activities and proline contents. The exogenous addition of B. velezensis SAAS-63 also significantly increased the accumulation of macroelements and micronutrients in lettuce. To elucidate the resistance mechanisms induced by B. velezensis SAAS-63 under nutrient stress, high-throughput sequencing and multi-omics analysis were performed. Inoculation with B. velezensis SAAS-63 altered the microbial community of the rhizosphere and increased the relative abundances of Streptomyces, Actinoallomurus, Verrucomicrobia, and Chloroflexi. It is worth noting that the inoculant SAAS-63 can affect plant rhizosphere metabolism. The inoculant changed the metabolic flow of phenylpropanoid metabolic pathway under nutrient deficiency and promoted phenylalanine to participate more in the synthesis of lignin precursors and coumarin substances by inhibiting the synthesis of flavone and isoflavone, thus improving plant resistance. This study showed that the addition of inoculant SAAS-63 could help plants recruit microorganisms to decompose and utilize trehalose and re-established the carbon metabolism of the plant rhizosphere. Additionally, microbes were found to be closely related to the accumulation of metabolites based on correlation analysis. The results indicated that the addition of PGPRs has an important role in regulating soil rhizosphere microbes and metabolism, providing valuable information for understanding how PGPRs affect complex biological processes and enhance plant adaptation to nutrient deficiency. KEY POINTS: • Inoculation with SAAS-63 significantly promoted plant growth under nutrient-deficient conditions • Inoculation with SAAS-63 affected rhizosphere microbial diversity and community structure • Inoculation with SAAS-63 affected plant rhizosphere metabolism and induced plants to synthesize substances that resist stress.

Iceberg lettuce cultivated in different systems of planting and sources of fertilizer / Alface americana cultivada em diferentes sistemas de plantio e fontes de fertilizantes

Organic fertilization is a cheaper and highly effective option for profitability and consequent improvement of the soil's physical, chemical, and biological structure. Thus, the objective of this work was to evaluate different types of fertilization: organic (poultry shed litter), mineral, and leaf path on yield parameters of lettuce grown in various types of planting. The treatments consisted of using two planting systems (P1 - Line and P2 - quincunxes) and mineral and organic fertilizers (A1 - mineral fertilization; A2 - mineral fertilization + leaf fertilization; A3 - organic fertilization with poultry shed litter and A4 - fertilization organic + mineral). The experimental units consisted of 36 and 52 plants, respectively, for treatments P1 and P2, and all central plants of the experimental unit were evaluated. Heart height, fresh mass, and leaf number were observed. The mineral and mineral + leaf treatments did not differentiate, either in line or in quincunxes. The treatment that stood out about the analyzed variables was the organic fertilization and quincunxes planting system, reflecting a more significant number of lettuce plants and better use of the area.

A adubação orgânica é uma opção mais barata e de grande eficácia em relação à rentabilidade e consequente melhoria da estrutura física, química e biológica do solo. Desta forma, o objetivo deste trabalho foi avaliar diferentes tipos de adubação: orgânica (cama de frango), mineral e via foliar sobre parâmetros de produtividade de alface cultivada em diferentes tipos de plantio. Os tratamentos consistiram na utilização de dois sistemas de plantio (P1 - Linha e P2 - Quincôncio) e adubações minerais e orgânicas (A1 - adubação mineral; A2 - adubação mineral + adubação foliar; A3 - adubação orgânica com cama de aviário e A4 - adubação orgânica + mineral). As unidades experimentais foram compostas por 36 e 52 plantas, respectivamente, para os tratamentos em linha e em quincôncio, sendo avaliadas todas as plantas centrais da unidade experimental. Foram observados a altura do coração, massa fresca e número de folhas. Os tratamentos mineral e mineral + foliar não diferenciaram entre si, tanto em linha quanto em quincôncio. O tratamento que se destacou em relação às variáveis analisadas foi aquele baseado na adubação orgânica e sistema de plantio em quincôncio, refletindo em maior número de pés de alface e melhor aproveitamento da área.

A Novel Method for Quantifying Plant Morphological Characteristics Using Normal Vectors and Local Curvature Data via 3D Modelling-A Case Study in Leaf Lettuce.

Three-dimensional measurement is a high-throughput method that can record a large amount of information. Three-dimensional modelling of plants has the possibility to not only automate dimensional measurement, but to also enable visual assessment to be quantified, eliminating ambiguity in human judgment. In this study, we have developed new methods that could be used for the morphological analysis of plants from the information contained in 3D data. Specifically, we investigated characteristics that can be measured by scale (dimension) and/or visual assessment by humans. The latter is particularly novel in this paper. The characteristics that can be measured on a scale-related dimension were tested based on the bounding box, convex hull, column solid, and voxel. Furthermore, for characteristics that can be evaluated by visual assessment, we propose a new method using normal vectors and local curvature (LC) data. For these examinations, we used our highly accurate all-around 3D plant modelling system. The coefficient of determination between manual measurements and the scale-related methods were all above 0.9. Furthermore, the differences in LC calculated from the normal vector data allowed us to visualise and quantify the concavity and convexity of leaves. This technique revealed that there were differences in the time point at which leaf blistering began to develop among the varieties. The precise 3D model made it possible to perform quantitative measurements of lettuce size and morphological characteristics. In addition, the newly proposed LC-based analysis method made it possible to quantify the characteristics that rely on visual assessment. This research paper was able to demonstrate the following possibilities as outcomes: (1) the automation of conventional manual measurements, and (2) the elimination of variability caused by human subjectivity, thereby rendering evaluations by skilled experts unnecessary.

Estimation of Greenhouse Lettuce Growth Indices Based on a Two-Stage CNN Using RGB-D Images.

Growth indices can quantify crop productivity and establish optimal environmental, nutritional, and irrigation control strategies. A convolutional neural network (CNN)-based model is presented for estimating various growth indices (i.e., fresh weight, dry weight, height, leaf area, and diameter) of four varieties of greenhouse lettuce using red, green, blue, and depth (RGB-D) data obtained using a stereo camera. Data from an online autonomous greenhouse challenge (Wageningen University, June 2021) were employed in this study. The data were collected using an Intel RealSense D415 camera. The developed model has a two-stage CNN architecture based on ResNet50V2 layers. The developed model provided coefficients of determination from 0.88 to 0.95, with normalized root mean square errors of 6.09%, 6.30%, 7.65%, 7.92%, and 5.62% for fresh weight, dry weight, height, diameter, and leaf area, respectively, on unknown lettuce images. Using red, green, blue (RGB) and depth data employed in the CNN improved the determination accuracy for all five lettuce growth indices due to the ability of the stereo camera to extract height information on lettuce. The average time for processing each lettuce image using the developed CNN model run on a Jetson SUB mini-PC with a Jetson Xavier NX was 0.83 s, indicating the potential for the model in fast real-time sensing of lettuce growth indices.

A PARTHENOGENESIS allele from apomictic dandelion can induce egg cell division without fertilization in lettuce.

Apomixis, the clonal formation of seeds, is a rare yet widely distributed trait in flowering plants. We have isolated the PARTHENOGENESIS (PAR) gene from apomictic dandelion that triggers embryo development in unfertilized egg cells. PAR encodes a K2-2 zinc finger, EAR-domain protein. Unlike the recessive sexual alleles, the dominant PAR allele is expressed in egg cells and has a miniature inverted-repeat transposable element (MITE) transposon insertion in the promoter. The MITE-containing promoter can invoke a homologous gene from sexual lettuce to complement dandelion LOSS OF PARTHENOGENESIS mutants. A similar MITE is also present in the promoter of the PAR gene in apomictic forms of hawkweed, suggesting a case of parallel evolution. Heterologous expression of dandelion PAR in lettuce egg cells induced haploid embryo-like structures in the absence of fertilization. Sexual PAR alleles are expressed in pollen, suggesting that the gene product releases a block on embryogenesis after fertilization in sexual species while in apomictic species PAR expression triggers embryogenesis in the absence of fertilization.

Mycetoindole, an N-acyl dehydrotryptophan with plant growth inhibitory activity from an actinomycete of the genus Actinomycetospora.

A rare actinomycetal strain of the genus Actinomycetospora was found to produce a new tryptophan derivative, designated mycetoindole (1). The structure of 1 was determined to be N-3-methylcrotonoyl (Z)-dehydrotryptophan by NMR and MS analytical methods. Compound 1 reduced the root growth of lettuce Lactuca sativa seedlings at concentrations above 0.1 µM and almost completely inhibited seed germination at 10 µM.

Alternative splicing triggered by the insertion of a CACTA transposon attenuates LsGLK and leads to the development of pale-green leaves in lettuce.

Lettuce (Lactuca sativa) is one of the most important vegetable crops in the world. As a leafy vegetable, the polymorphism of lettuce leaves from dark to pale green is an important trait. However, the genetic and molecular mechanisms underlying such variations remain poorly understood. In this study, one major locus controlling the polymorphism of dark- and pale-green leaves in lettuce was identified using genome-wide association studies (GWAS). This locus was then fine mapped to an interval of 5375 bp on chromosome 4 using a segregating population containing 2480 progeny. Only one gene, homologous to the GLK genes in Arabidopsis and other plants, is present in the candidate region. A complementation test confirmed that the candidate gene, LsGLK, contributes to the variation of dark- and pale-green leaves. Sequence analysis showed that a CACTA transposon of 7434 bp was inserted 10 bp downstream of the stop codon of LsGLK, followed by a duplication of a 1826-bp fragment covering exons 3-6 of the LsGLK gene. The transposon insertion did not change the expression level of the LsGLK gene. However, because of alternative splicing, only 6% of the transcripts produced from the transposon insertion were wild-type transcripts, which led to the production of pale-green leaves. An evolutionary analysis revealed that the insertion of the CACTA transposon occurred in cultivated lettuce and might have been selected in particular cultivars to satisfy the diverse demands of consumers. In this study, we demonstrated that a transposon insertion near a gene may affect its splicing and consequently generate phenotypic variations.

A Physiological behavior and tolerance of Lactuca sativa to lead nitrate and silver nitrate heavy metals / Comportamento fisiológico e tolerância de Lactuca sativa aos metais pesados nitrato de chumbo e nitrato de prata

This study goal to evaluate the effects of different concentrations of lead (Pb) and silver (Ag) on germination, initial growth and anatomical alterations of Lactuca sativa L. Plants use various mechanisms to reduce the impacts caused by anthropic action, such as xenobiotic elements of soils and water contaminated by heavy metals. These metals were supplied as lead nitrate and silver nitrate and the following treatments were established: control for both metals, maximum dose of heavy metals, for arable soils, allowed by the National Council of the Environment (Ag = 25 mg. Kg-¹, Pb = 180 mg. Kg-¹), double (Ag = 50 mg. Kg-¹, Pb = 360 mg. Kg-¹) and triple (Ag = 75 mg. Kg-¹, Pb = 540 mg. Kg-¹) of this dosage. Vigor and germination tests of the seeds and possible anatomical changes in the leaves and roots of lettuce plants were performed. The species showed a high capacity to germinate under Pb and Ag stress, and the germination was never completely inhibited; however, the germination decreased with increasing Pb concentrations, but not under Ag stress. The use of increasing doses of metals reduced seed vigor and increased chlorophyll content. An increase in biomass was also observed in plants from treatments submitted to Pb. The phytotoxic effects of metals were more pronounced at 15 days after sowing. Anatomically, L. sativa was influenced by metal concentrations, and had a reduction of up to 79.9% in root epidermis thickness at the highest Pb concentration, although some structures did not suffer significant changes. The results suggest that L. sativa presents tolerance to high concentrations of heavy metals, showing possible mechanisms to overcome the stress caused by these metals. In this research lettuce possibly used the mechanism of exclusion of metals retaining Pb and Ag in the roots preserving the photosynthetic apparatus in the aerial part of the plants. In general, the chemical element Pb was more toxic than Ag, in these experimental conditions.

Este estudo teve como objetivo avaliar os efeitos de diferentes concentrações de chumbo (Pb) e prata (Ag) na germinação, crescimento inicial e alterações anatômicas de Lactuca sativa L. As plantas utilizam vários mecanismos para reduzir os impactos causados pela ação antrópica, como elementos xenobióticos de solos e água contaminada por metais pesados. Esses metais foram fornecidos como nitrato de chumbo e nitrato de prata e foram estabelecidos os seguintes tratamentos:controle para ambos os metais, dose máxima de metais pesados, para solos cultiváveis, permitida pelo Conselho Nacional do Meio Ambiente (Ag = 25mg.Kg-¹, Pb = 180mg.Kg-¹), dobro (Ag = 50mg.Kg-¹, Pb = 360mg.Kg-¹) e triplo (Ag = 75mg.Kg-¹, Pb = 540 mg.Kg-¹) desta dosagem. Foram realizados testes de vigor e germinação das sementes e possíveis alterações anatômicas nas folhas e raízes das plantas de alface. A espécie apresentou alta capacidade de germinar sob estresse de Ag e Pb, e a germinação nunca foi completamente inibida; entretanto, a germinação diminuiu com o aumento das concentrações de Pb, mas não sob estresse de Ag. O uso de doses crescentes dos metais, reduziu o vigor das sementes e aumentou o teor de clorofila. Também foi observado aumento da biomassa nas plantas a partir dos tratamentos submetidos ao Pb. Os efeitos fitotóxicos dos metais foram mais acentuados aos 15 dias após a semeadura. Anatomicamente, L. sativa foi influenciada pelas concentrações de metais, e teve uma redução de até 79,9% na espessura da epiderme radicular na maior concentração de Pb, embora algumas estruturas não tenham sofrido alterações significativas. Os resultados sugerem que L. sativa apresenta tolerância a altas concentrações de metais pesados, mostrando possíveis mecanismos para superar o estresse causado por esses metais. Nesta pesquisa a alface possivelmente utilizou o mecanismo de exclusão de metais retendo Pb e Ag nas raízes preservando o aparato [...].

Short-term changes related to autotetraploidy in essential oil composition of Eucalyptus benthamii Maiden & Cambage and its applications in different bioassays.

Some forest trees have been polyploidized to improve their traits and to supply new germplasms for breeding programs. As trees have a long juvenile stage, the early characterization of the chromosome set doubling effects is crucial for previous selection. Thus, we aimed to characterize the chemical variability of essential oils from diploid and autotetraploid germplasms (autotetraploid A and B) of Eucalyptus benthamii, as well as to evaluate their larvicidal and allelopathic effects. Autotetraploid A showed a higher essential oil yield than diploid and autotetraploid B, which did not differ quantitatively. Aromadendrene, viridiflorol and α-pinene were the major compounds in the diploid essential oil. In contrast, compounds were present in autotetraploids, such as 1,8-cineole, limonene, α-terpineol, and α-terpinyl-acetate. Essential oils from the diploid at 50-200 ppm were twice as larvicidal than those from autotetraploids against Aedes aegypti larvae. Considering the phytotoxicity bioassays using Lactuca sativa, essential oils from both ploidy levels affected root growth. Moreover, the essential oils inhibited shoot growth at all concentrations tested (187.5; 375; 750; 1500; and 3000 ppm). Autotetraploid A and B had the same effect on shoot growth as glyphosate. The essential oils had no cytogenotoxic effect on root meristematic cells of L. sativa, whereas phytotoxic potential was identified mainly in shoot growth. This work demonstrated a dramatic change in secondary metabolism (terpene composition) related to an increase in the ploidy level in Eucalyptus germplasms. In addition, we report the novelty of the chemical composition of essential oils among germplasms and their potential use as larvicidal and post-emergence weed control agents.

Comparative Analysis of Major Flavonoids among Parts of Lactuca indica during Different Growth Periods.

L. indica L. cv. Mengzao, a medicinal plant of the Ixeris genus, is rich in flavonoids. In order to thoroughly analyze the the distribution and dynamic change of major flavonoids in its various parts from different growth periods, the flavonoids extracted from L. indica L. cv. Mengzao were identified and quantitatively analyzed by ultra-high-performance liquid chromatography mass spectrometer (LC-MS/MS). Results indicated that 15 flavonoids were identified from L. indica L. cv. Mengzao, and rutin, luteolin, luteolin-7-O-glucoside, kaempferol, quercetin, and apigenin are the major flavonoids in L. indica L. cv. Mengzao. In general, the total flavonoids' content in different parts of L. indica L. cv. Mengzao followed the order flowers > leaves > stems > roots. Flowers and leaves are the main harvesting parts of L. indica L. cv. Mengzao, and the flowering period is the most suitable harvesting period. This study provides valuable information for the development and utilization of L. indica L. cv. Mengzao and determined the best part to harvest and the optimal time for harvesting.

Leaf nodule endosymbiotic Burkholderia confer targeted allelopathy to their Psychotria hosts.

After a century of investigations, the function of the obligate betaproteobacterial endosymbionts accommodated in leaf nodules of tropical Rubiaceae remained enigmatic. We report that the α-D-glucose analogue (+)-streptol, systemically supplied by mature Ca. Burkholderia kirkii nodules to their Psychotria hosts, exhibits potent and selective root growth inhibiting activity. We provide compelling evidence that (+)-streptol specifically affects meristematic root cells transitioning to anisotropic elongation by disrupting cell wall organization in a mechanism of action that is distinct from canonical cellulose biosynthesis inhibitors. We observed no inhibitory or cytotoxic effects on organisms other than seed plants, further suggesting (+)-streptol as a bona fide allelochemical. We propose that the suppression of growth of plant competitors is a major driver of the formation and maintenance of the Psychotria-Burkholderia association. In addition to potential agricultural applications as a herbicidal agent, (+)-streptol might also prove useful to dissect plant cell and organ growth processes.

Dereplication of Bioactive Spirostane Saponins from Agave macroacantha.

A dereplication strategy using UPLC-QTOF/MSE, the HMAI method, and NMR spectroscopy led to the identification of five main steroidal saponins (1-5), including three previously unknown compounds named macroacanthosides A-C (3-5), in a bioactive fraction of Agave macroacantha. The major saponins were isolated, and some of them together with the saponin-rich fraction were then evaluated for phytotoxicity on a standard target species, Lactuca sativa. The inhibition values exhibited by the pure compounds were confirmed to be in agreement with the phytotoxicity of the saponin-rich fraction, which suggests that the saponin fraction could be applied successfully as an agrochemical without undergoing any further costly and/or time-consuming purification processes. The NMR data of the pure compounds as well as of those corresponding to the same compounds in the fraction were comparable, which indicated that the main saponins could be identified by means of this replication workflow and that no standards are required.

Identification of GROWTH-REGULATING FACTOR transcription factors in lettuce (Lactuca sativa) genome and functional analysis of LsaGRF5 in leaf size regulation.

BACKGROUND: GROWTH-REGULATING FACTORs (GRFs), a type of plant-specific transcription factors, play important roles in regulating plant growth and development. Although GRF gene family has been identified in various plant species, a genome-wide analysis of this family in lettuce (Lactuca sativa L.) has not been reported yet. RESULTS: Here we identified 15 GRF genes in lettuce and performed comprehensive analysis of them, including chromosomal locations, gene structures, and conserved motifs. Through phylogenic analysis, we divided LsaGRFs into six groups. Transactivation assays and subcellular localization of LsaGRF5 showed that this protein is likely to act as a transcriptional factor in the cell nucleus. Furthermore, transgenic lettuce lines overexpressing LsaGRF5 exhibited larger leaves, while smaller leaves were observed in LsaMIR396a overexpression lines, in which LsaGRF5 was down-regulated. CONCLUSIONS: These results in lettuce provide insight into the molecular mechanism of GRF gene family in regulating leaf growth and development and foundational information for genetic improvement of the lettuce variations specialized in leaf character.

Native Plant Growth-Promoting Rhizobacteria for Growth Promotion of Lettuce from Qassim, Saudi Arabia.

<b>Background and Objective:</b> Plant Growth-Promoting Rhizobacteria (PGPR) are a group of bacteria that colonize plant roots and enhance the growth and productivity of plants. However, only those PGPR that is acclimatized to the local soil conditions performs well. The present study aims to pick up effective PGPR isolates from local soil and utilize them as potential bio-inoculants to enhance lettuce plant growth. <b>Materials and Methods:</b> Rhizospheric soil samples were obtained from each of six desert plant species in the Qassim region and 45 bacterial isolates were obtained. Four of them were identified and tested for growth-promoting activities by application to the soil in which lettuce was grown under greenhouse conditions. <b>Results:</b> The selected bacterial isolates were identified as <i>Bacillus cereus</i> BW-201B, <i>Pseudomonas aeruginosa</i> AMU1, <i>Pseudomonas putida</i> CNE30 and <i>Enterobacter</i> sp. CZGRY7. Application of these four isolates to the soil in which lettuce was grown under greenhouse conditions resulted in significant increases in shoot height, shoot weight, chlorophyll levels and the percentages of N, P and K compared with those of control treatment. <b>Conclusion:</b> These findings suggest that local soil bacterial strains represent excellent bioinoculants for growth and yield increase in lettuce under local agro-climatic conditions in Saudi Arabia. Our approach might offer a good alternative for the chemical-free farming of lettuce.

TSA Promotes CRISPR/Cas9 Editing Efficiency and Expression of Cell Division-Related Genes from Plant Protoplasts.

Trichostatin A (TSA) is a representative histone deacetylase (HDAC) inhibitor that modulates epigenetic gene expression by regulation of chromatin remodeling in cells. To investigate whether the regulation of chromatin de-condensation by TSA can affect the increase in the efficiency of Cas9 protein-gRNA ribonucleoprotein (RNP) indel formation from plant cells, genome editing efficiency using lettuce and tobacco protoplasts was examined after several concentrations of TSA treatments (0, 0.1, 1 and 10 µM). RNP delivery from protoplasts was conducted by conventional polyethylene glycol (PEG) transfection protocols. Interestingly, the indel frequency of the SOC1 gene from TSA treatments was about 3.3 to 3.8 times higher than DMSO treatment in lettuce protoplasts. The TSA-mediated increase of indel frequency of the SOC1 gene in lettuce protoplasts occurred in a concentration-dependent manner, although there was not much difference. Similar to lettuce, TSA also increased the indel frequency by 1.5 to 1.8 times in a concentration-dependent manner during PDS genome editing using tobacco protoplasts. The MNase test clearly showed that chromatin accessibility with TSA treatments was higher than that of DMSO treatment. Additionally, TSA treatment significantly increased the level of histone H3 and H4 acetylation from lettuce protoplasts. The qRT-PCR analysis showed that expression of cell division-related genes (LsCYCD1-1, LsCYCD3-2, LsCYCD6-1, and LsCYCU4-1) was increased by TSA treatment. These findings could contribute to increasing the efficiency of CRISPR/Cas9-mediated genome editing. Furthermore, this could be applied for the development of useful genome-edited crops using the CRISPR/Cas9 system with plant protoplasts.