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All components of the rhizosphere can be engineered to promote plant health and growth, two features that strongly depend upon the interactions of living organisms with their environment. This review describes the progress in plant and microbial molecular genetics and ecology that has led to a wealth of potential applications. Recent efforts especially deal with the plant defense machinery that is instrumental in engineering plant resistance to biotic stresses. Another approach involves microbial population engineering rather than single strain engineering. More generally, the plants (and the associated microbes) are no longer seen as 'individual' but rather as a holobiont, in other words a unit of selection in evolution, a concept that holds great promise for future plant breeding programs., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Abstract Single-cell transcriptome sequencing (scRNA-seq) is a powerful tool for describing the transcriptome dynamics of plant development but has not yet been utilized to analyze the tissue ontology of sweetpotato. This study established a stable method for isolating single protoplast cells for scRNA-seq to reveal the cell heterogeneity of sweetpotato root tip meristems at the single-cell level. The study analyzed 12,172 single cells and 27,355 genes in the root tips of the sweetpotato variety Guangshu 87, which were distributed into 15 cell clusters. Pseudo-time analysis showed that there were transitional cells in the apical development trajectory of mature cell types from stem cell niches. Furthermore, we identified novel development regulators of sweetpotato tubers via trajectory analysis. The transcription factor IbGATA4 was highly expressed in the adventitious roots during the development of sweetpotato root tips, where it may regulate the development of sweetpotato root tips. In addition, significant differences were observed in the transcriptional profiles of cell types between sweetpotato, Arabidopsis thaliana, and maize. This study mapped the single-cell transcriptome of sweetpotato root tips, laying a foundation for studying the types, functions, differentiation, and development of sweetpotato root tip cells.

Abstract Background Soybean establishes a mutualistic interaction with nitrogen-fixing rhizobacteria, acquiring most of its nitrogen requirements through symbiotic nitrogen fixation. This crop is susceptible to water deficit; evidence suggests that its nodulation status—whether it is nodulated or not—can influence how it responds to water deficit. The translational control step of gene expression has proven relevant in plants subjected to water deficit. Results Here, we analyzed soybean roots’ differential responses to water deficit at transcriptional, translational, and mixed (transcriptional + translational) levels. Thus, the transcriptome and translatome of four combined-treated soybean roots were analyzed. We found hormone metabolism-related genes among the differentially expressed genes (DEGs) at the translatome level in nodulated and water-restricted plants. Also, weighted gene co-expression network analysis followed by differential expression analysis identified gene modules associated with nodulation and water deficit conditions. Protein-protein interaction network analysis was performed for subsets of mixed DEGs of the modules associated with the plant responses to nodulation, water deficit, or their combination. Conclusions Our research reveals that the stand-out processes and pathways in the before-mentioned plant responses partially differ; terms related to glutathione metabolism and hormone signal transduction (2 C protein phosphatases) were associated with the response to water deficit, terms related to transmembrane transport, response to abscisic acid, pigment metabolic process were associated with the response to nodulation plus water deficit. Still, two processes were common: galactose metabolism and branched-chain amino acid catabolism. A comprehensive analysis of these processes could lead to identifying new sources of tolerance to drought in soybean.

Abstract Peanut (Arachis hypogaea L.) is an important oilseed crop worldwide. However, soil salinization becomes one of the main limiting factors of peanut production. Therefore, developing salt-tolerant varieties and understanding the molecular mechanisms of salt tolerance is important to protect peanut yield in saline areas. In this study, we selected four peanut varieties with contrasting response to salt challenges with T1 and T2 being tolerance and S1 and S2 being susceptible. High-throughput RNA sequencing resulted in more than 314.63 Gb of clean data from 48 samples. We identified 12,057 new genes, 7,971of which have functional annotations. KEGG pathway enrichment analysis of uniquely expressed genes in salt-tolerant peanut revealed that upregulated genes in the root are involved in the MAPK signaling pathway, fatty acid degradation, glycolysis/gluconeogenesis, and upregulated genes in the shoot were involved in plant hormone signal transduction and the MAPK signaling pathway. Na+ content, K+ content, K+/ Na+, and dry mass were measured in root and shoot tissues, and two gene co-expression networks were constructed based on weighted gene co-expression network analysis (WGCNA) in root and shoot. In this study, four key modules that are highly related to peanut salt tolerance in root and shoot were identified, plant hormone signal transduction, phenylpropanoid biosynthesis, starch and sucrose metabolism, flavonoid biosynthesis, carbon metabolism were identified as the key biological processes and metabolic pathways for improving peanut salt tolerance. The hub genes include genes encoding ion transport (such as HAK8, CNGCs, NHX, NCL1) protein, aquaporin protein, CIPK11 (CBL-interacting serine/threonine-protein kinase 11), LEA5 (late embryogenesis abundant protein), POD3 (peroxidase 3), transcription factor, and MAPKKK3. There were some new salt-tolerant genes identified in peanut, including cytochrome P450, vinorine synthase, sugar transport protein 13, NPF 4.5, IAA14, zinc finger CCCH domain-containing protein 62, beta-amylase, fatty acyl-CoA reductase 3, MLO-like protein 6, G-type lectin S-receptor-like serine/threonine-protein kinase, and kinesin-like protein KIN-7B. The identification of key modules, biological pathways, and hub genes in this study enhances our understanding of the molecular mechanisms underlying salt tolerance in peanuts. This knowledge lays a theoretical foundation for improving and innovating salt-tolerant peanut germplasm.

Abstract Understanding the molecular processes and hormonal signals that govern root growth is of paramount importance for effective forest management. While Arabidopsis studies have shed light on the role of the primary root in root system development, the structure of root systems in trees is considerably more intricate, posing challenges to comprehend taproot growth in acorn-sown and nursery-cultivated seedlings. In this study, we investigated Quercus robur seedlings using rhizotrons, containers, and transplanted containers to rhizotrons, aiming to unravel the impact of forest nursery practices on processes governing taproot growth and root system development. Root samples were subjected to RNA-seq analysis to identify gene expression patterns and perform differential gene expression and phytohormone analysis. Among studied cultivation systems, differentially expressed genes (DEGs) exhibited significant diversity, where the number of co-occurring DEGs among cultivation systems was significantly smaller than the number of unique DEGs in different cultivation systems. Moreover, the results imply that container cultivation triggers the activation of several genes associated with linolenic acid and peptide synthesis in root growth. Upon transplantation from containers to rhizotrons, rapid enhancement in gene expression occurs, followed by gradual reduction as root growth progresses, ultimately reaching a similar expression pattern as observed in the taproot of rhizotron-cultivated seedlings. Phytohormone analysis revealed that taproot growth patterns under different cultivation systems are regulated by the interplay between auxin and cytokinin concentrations. Moreover, the diversification of hormone levels within the root zone and cultivation systems allows for taproot growth inhibition and prompt recovery in transplanted seedlings. Our study highlights the crucial role of hormone interactions during the early stages of taproot elongation, influencing root system formation across.

Abstract Background Phosphorus (P) and salt stress are common abiotic stressors that limit crop growth and development, but the response mechanism of soybean to low phosphorus (LP) and salt (S) combined stress remains unclear. Results In this study, two soybean germplasms with similar salt tolerance but contrasting P-efficiency, A74 (salt-tolerant and P-efficient) and A6 (salt-tolerant and P-inefficient), were selected as materials. By combining physiochemical and transcriptional analysis, we aimed to elucidate the mechanism by which soybean maintains high P-efficiency under salt stress. In total, 14,075 differentially expressed genes were identified through pairwise comparison. PageMan analysis subsequently revealed several significantly enriched categories in the LP vs. control (CK) or low phosphorus + salt (LPS) vs. S comparative combination when compared to A6, in the case of A74. These categories included genes involved in mitochondrial electron transport, secondary metabolism, stress, misc, transcription factors and transport. Additionally, weighted correlation network analysis identified two modules that were highly correlated with acid phosphatase and antioxidant enzyme activity. Citrate synthase gene (CS), acyl-coenzyme A oxidase4 gene (ACX), cytokinin dehydrogenase 7 gene (CKXs), and two-component response regulator ARR2 gene (ARR2) were identified as the most central hub genes in these two modules. Conclusion In summary, we have pinpointed the gene categories responsible for the LP response variations between the two salt-tolerant germplasms, which are mainly related to antioxidant, and P uptake process. Further, the discovery of the hub genes layed the foundation for further exploration of the molecular mechanism of salt-tolerant and P-efficient in soybean.

Heavy metals are widely available in the environment due to the natural processes, industrial, anthropogenic activities and ultimately in the results contaminate the immediate environment. The presence of the heavy metals such as Pb, Cd, Cr, Ni, Fe and Cu in environment disturb the quality of ecosystems, soils, water, air and vegetation. The chromium metal at higher level in ecosystem is an alarming signal for both developed and developing countries. Pennisetum glaucum is an annual grass which is widely cultivated in drought, rain fed and high temperature areas. The limited amount of literature available on the impact of chromium stress on P. glaucum. This study was conducted to investigate the beneficial or harmful effect of chromium stress on seed germination and seedling growth performances of P. glaucum in in vitro conditions. The different concentration of chromium 0, 25, 50, 75 and 100 ppm was applied. In present study, the overall results suggests the variable response on the rate of seed germination percentage and plant growth of P. glaucum to chromium stress was recorded. Results showed that the chromium treatment at 25 ppm significantly (p

The wet (rain, snow, fog, sleet, dew) and dry (transport of aerosol, particles and gases) deposition of acidic substance in environment results due to human, automobile, fossil fuel burning and industrial activities. Acid deposition is worldwide environmental degradation problems and in recent years these acidic materials are increasing at alarming scale in the environment both in developed and developing countries, including Pakistan. Some scientific literature survey reports suggest that plant growth and agriculture yield decrease due to consequence of acid rain. In addition, acid rain is found responsible for producing toxic effects on the morphological parameters of agricultural crop. The evidence collected from last more than fifty years showed the common significant effects of acid rain on seed germination percentage, seedling height, root hair and structure, alteration in leaf anatomy, size and area, stomatal structure, size, pollen germination, photosynthetic pigments and physiological changes in herbs, shrubs and trees. Still, little is known on the impact of acid rain on plant growth. This study was aimed to review the effects of acid mist on growth performances of some selected plant species. This review is contributed with the help of literature survey, research work published on the impact of acid rain on the plant growth.

Several studies have been devoted to seeking some beneficial plant-related microorganisms for a long time, and on this basis, it has been found that arbuscular mycorrhizal fungi (AMF) have a considerable positive impact on plant health as a biological fungal agent. In this study, we focused on the effects of different AMF on the growth dynamics and root configuration of licorice under saline and alkali conditions. The metabolites of licorice under different AMF were assessed using liquid chromatography–tandem mass spectrometry (LC-MS/MS). Funneliformis mosseae (Fm) and Rhizophagus intraradices (Ri) were added as different AMF treatments, while the sterilized saline–alkali soil was treated as a control. Samples were taken in the R1 period (15 d after AMF treatment) and the R2 period (45 d after AMF treatment). The results showed that the application of AMF significantly increased the root growth of licorice and significantly increased the biomass of both shoot and root. A total of 978 metabolites were detected and divided into 12 groups including lipids, which accounted for 15.44%; organic acids and their derivatives, at 5.83%; benzene compounds and organic heterocyclic compounds, at 5.42%; organic oxides, at 3.78%; and ketones, accounting for 3.17%. Compared with the control, there were significant changes in the differential metabolites with treatment inoculated with AMF; the metabolic pathways and biosynthesis of secondary metabolites were the main differential metabolite enrichment pathways in the R1 period, and those in the R2 period were microbial metabolism in diverse environments and the degradation of aromatic compounds. In conclusion, the use of AMF as biofertilizer can effectively improve the growth of licorice, especially in terms of the root development and metabolites, in saline–alkali soil conditions.

Abstract Background Leymus chinensis (L. chinensis) is a perennial native forage grass widely distributed in the steppe of Inner Mongolia as the dominant species. Calcium (Ca) is an essential mineral element important for plant adaptation to the growth environment. Ca limitation was previously shown to strongly inhibit Arabidopsis (Arabidopsis thaliana) seedling growth and disrupt plasma membrane stability and selectivity, increasing fluid-phase-based endocytosis and contents of all major membrane lipids. Results In this study, we investigated the significance of Ca for L. chinensis growth and membrane stability relative to Arabidopsis. Our results showed that Ca limitation did not affect L. chinensis seedling growth and endocytosis in roots. Moreover, the plasma membrane maintained high selectivity. The lipid phosphatidylcholine (PC): phosphatidylethanolamine (PE) ratio, an indicator of the membrane stability, was five times higher in L. chinensis than in Arabidopsis. Furthermore, in L. chinensis, Ca limitation did not affect the content of any major lipid types, decreased malondialdehyde (MDA) content, and increased superoxide dismutase (SOD) activity, showing an opposite pattern to that in Arabidopsis. L. chinensis roots accumulated higher contents of PC, phosphatidylinositol (PI), monogalactosyldiacylglycerol (MGDG), phosphatidylglycerol (PG), cardiolipin (CL), digalactosyldiacylglycerol (DGDG), and lysophosphatidylcholine (LPC) but less phosphatidylethanolamine (PE), diacylglycerol (DAG), triacylglycerolv (TAG), phosphatidylserine (PS), lysobisphosphatidic acids (LPAs), lysophosphatidylethanolamine (LPE), and lysophosphatidylserine (LPS) than Arabidopsis roots. Moreover, we detected 31 and 66 unique lipids in L. chinensis and Arabidopsis, respectively. Conclusions This study revealed that L. chinensis roots have unique membrane lipid composition that was not sensitive to Ca limitation, which might contribute to the wider natural distribution of this species.

Litter and root decomposition is an important source of soil organic matter and nutrients. To ascertain the contribution of litter and root to natural grassland nutrients in rocky desertification areas, from March 2017 to January 2018, the continuous soil column method, collector method, and litter decomposition method were used to study the soil nutrients, litter and root biomass, decomposition, and nutrient release of potential, moderate, and severe rocky desertification grasslands, as well as their responses to rocky desertification. The results showed that the litter and root decomposition rate showed a trend of being first fast and then slow, and the decomposition rate of litter and root was greater than 50% after 300 days. The annual litter decomposition rates of potential, moderate, and severe rocky desertification grasslands were 69.98%, 62.14%, and 49.79%, respectively, and the annual decomposition rates of root were 73.64%, 67.61%, and 64.09%, respectively. With a deepening degree of rocky desertification, the litter and root decomposition rate decreased. The decomposition coefficients, k, of litter in potential, moderate, and severe rocky desertification grasslands were 1.128, 0.896, and 0.668, respectively, and the decomposition coefficients, k, of root were 1.152, 1.018, and 0.987, respectively. The nutrient release processes of litter and root were different, and the release mode ultimately manifests as “release”. In rocky desertification grasslands, the organic carbon (OC), total nitrogen (TN), total phosphorus (TP), and total potassium (TK) released by litter and root decomposition were 18.93–263.03 g·m−2·yr−1, 1.79–5.59 g·m−2·yr−1, 0.18–0.47 g·m−2·yr−1, and 0.66–3.70 g·m−2·yr−1, respectively. The contribution of root to soil nutrients was greater than that of litter. The degree of rocky desertification was negatively correlated with the biomass, decomposition rate, and nutrient return amount of litter and root. The results of this study provide direct field evidence and illustrate the contribution of litter and root decomposition in rocky desertification grasslands to soil nutrients.

Proso millet (Panicum miliaceum L.) is resilient to abiotic stress, especially to drought. However, the mechanisms by which its roots adapt and tolerate salt stress are obscure. In this study, to clarify the molecular mechanism of proso millet in response to drought stress, the physiological indexes and transcriptome in the root of seedlings of the proso millet cultivar ‘Yumi 2’ were analyzed at 0, 0.5, 1.0, 1.5, and 3.0 h of stimulated drought stress by using 20% PEG-6000 and after 24 h of rehydration. The results showed that the SOD activity, POD activity, soluble protein content, MDA, and O2−· content of ‘Yumi 2’ increased with the time of drought stress, but rapidly decreased after rehydration. Here, 130.46 Gb of clean data from 18 samples were obtained, and the Q30 value of each sample exceeded 92%. Compared with 0 h, the number of differentially expressed genes (DEGs) reached the maximum of 16,105 after 3 h of drought, including 9153 upregulated DEGs and 6952 downregulated DEGs. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analyses revealed that upregulated DEGs were mainly involved in ATP binding, nucleus, protein serine/threonine phosphatase activity, MAPK signaling pathway–plant, plant–pathogen interactions, and plant hormone signal transduction under drought stress, while downregulated DEGs were mainly involved in metal ion binding, transmembrane transporter activity, and phenylpropanoid biosynthesis. Additionally, 1441 TFs screened from DEGs were clustered into 64 TF families, such as AP2/ERF-ERF, bHLH, WRKY, NAC, MYB, and bZIP TF families. Genes related to physiological traits were closely related to starch and sucrose metabolism, phenylpropanoid biosynthesis, glutathione metabolism, and plant hormone signal transduction. In conclusion, the active oxygen metabolism system and the soluble protein of proso millet root could be regulated by the activity of protein serine/threonine phosphatase. AP2/ERF-ERF, bHLH, WRKY, NAC, MYB, and bZIP TF families were found to be closely associated with drought tolerance in proso millet root. This study will provide data to support a subsequent study on the function of the drought tolerance gene in proso millet.

Soil salinization poses a significant threat to agricultural productivity, necessitating innovative agronomic strategies to mitigate its impact. This study focuses on improving salt stress resistance in tomato plants through the application of silicon (Si) in roots. A greenhouse experiment was carried out under normal conditions (control, and 1 and 4 mM Si) and under salinity stress (salt control, and 1 and 4 mM Si). Various parameters were analyzed in leaves and roots. Under normal conditions, tomato plants grown in non-saline conditions exhibited some toxicity when exposed to Na2SiO3. As for the experiments under salt stress conditions, Si mitigated oxidative damage, preserving root cell membrane integrity. The concentration of malondialdehyde was reduced by 69.5%, that of proline was reduced by 56.4% and there was a 57.6% decrease in catalase activity for tomato plants treated with 1 mM Si under salt stress. Furthermore, Fe uptake and distribution, under salt conditions, increased from 91 to 123 mg kg−1, the same concentration as that obtained for the normal control. In all cases, the lower dose produced better results under normal conditions than the 4 mM dose. In summary, this research provides a potential application of Si in non-fertigated crop systems through a radicular pathway.

Abstract Exogenous GAs have an indeterminate effect on root development. Our current study used female papaya to reveal how the roots and rhizosphere respond to the exogenous application of GA3 by investigating the transcriptome profile in roots, metabolic profile and microbial community in both roots and rhizosphere of GA3-treated and control female papaya. The results demonstrated that exogenous GA3 treatment enhanced female papaya lateral root development, which gave plants physical advantages of water and nutrient uptake. In addition, it was likely that GA3 spraying in papaya shoot apices increased the level of auxin, which was transported to roots by CpPIN1, where auxin upregulated CpLBD16 and repressed CpBP to promote the lateral root initiation and development. In papaya roots, corresponding transporters (CpTMT3, CpNRT1:2, CpPHT1;4, CpINT2, CpCOPT2, CpABCB11, CpNIP4;1) were upregulated and excretion transporters were downregulated such as CpNAXT1 for water and nutrients uptake with exogenous GA3 application. Moreover, in GA3-treated papaya roots, CpALS3 and CpMYB62 were downregulated, indicating a stronger abiotic resistance to aluminum toxic and phosphate starvation. On the other hand, BRs and JAs, which involve in defense responses, were enriched in the roots and rhizosphere of GA3-treated papayas. The upregulation of the two hormones might result in the reduction of pathogens in roots and rhizosphere such as Colletotrichum and Verticillium. GA3-treated female papaya increased the abundance of beneficial bacteria species including Mycobacterium, Mitsuaria, and Actinophytocola, but decreased that of the genera Candidatus and Bryobacter for that it required less nitrate. Overall, the roots and rhizosphere of female papaya positively respond to exogenous application of GA3 to promote development and stress tolerance. Treatment of female papaya with GA3 might result in the promotion of lateral root formation and development by upregulating CpLBD16 and downregulating CpBP. GA3-treated papaya roots exhibited feedback control of brassinolide and jasmonate signaling in root development and defense. These findings revealed complex response to a growth hormone treatment in papaya roots and rhizosphere and will lead to investigations on the impact of other plant hormones on belowground development in papaya.

Soil salinization is the main abiotic stressor faced by crops. An improved understanding of the transcriptional response to salt stress in roots, the organ directly exposed to a high salinity environment, can inform breeding strategies to enhance tolerance and increase crop yield. Here, RNA-sequencing was performed on the roots of salt-tolerant wheat breeding line CH7034 at 0, 1, 6, 24, and 48 h after NaCl treatment. Based on transcriptome data, a weighted gene co-expression network analysis (WGCNA) was constructed, and five gene co-expression modules were obtained, of which the blue module was correlated with the time course of salt stress at 1 and 48 h. Two GO terms containing 249 differentially expressed genes (DEGs) related to osmotic stress response and salt-stress response were enriched in the blue module. These DEGs were subsequently used for association analysis with a set of wheat germplasm resources, and the results showed that four genes, namely a Walls Are Thin 1-related gene (TaWAT), an aquaporin gene (TaAQP), a glutathione S-transfer gene (TaGST), and a zinc finger gene (TaZFP), were associated with the root salt-tolerance phenotype. Using the four candidate genes as hub genes, a co-expression network was constructed with another 20 DEGs with edge weights greater than 0.6. The network showed that TaWAT and TaAQP were mainly co-expressed with fifteen interacting DEGs 1 h after salt treatment, while TaGST and TaZFP were mainly co-expressed with five interacting DEGs 48 h after salt treatment. This study provides key modules and candidate genes for understanding the salt-stress response mechanism in wheat roots.

Timothy (Phleum pratense) is a cool-season perennial forage grass widely grown for silage and hay production in northern regions. Climate change scenarios predict an increase in extreme weather events with fluctuating periods of high rainfall, requiring new varieties adapted to waterlogging (WL). Wild accessions could serve as germplasm for breeding, and we evaluated the responses of 11 wild and 8 domesticated accessions of timothy, P. nodosum and P. alpinum from different locations in northern Europe. Young plants at tillering stage were exposed to WL for 21 days in a greenhouse, and responses in growth allocation and root anatomy were studied. All accessions produced adventitious roots and changed allocation of growth between shoot and root as a response to WL, but the magnitude of these responses varied among species and among accessions. P. pratense responded less in these traits in response to WL than the other two species. The ability to form aerenchyma in the root cortex in response to WL was found for all species and also varied among species and among accessions, with the highest induction in P. pratense. Interestingly, some accessions were able to maintain and even increase root growth, producing more leaves and tillers, while others showed a reduction in the root system. Shoot dry weight (SDW) was not significantly affected by WL, but some accessions showed different and significant responses in the rate of production of leaves and tillers. Overall correlations between SDW and aerenchyma and between SDW and adventitious root formation were found. This study identified two wild timothy accessions and one wild P. nodosum accession based on shoot and root system growth, aerenchyma formation and having a root anatomy considered to be favorable for WL tolerance. These accessions are interesting genetic resources and candidates for development of climate-resilient timothy varieties.

Cichorium intybus is a valuable crop due to its high nutritional and pharmaceutical value. In this work, the study of the effect of harvesting time on the biometric and biochemical properties of Cichorium intybus L. var. foliosum (chicory salad witloof) was carried out on five varieties. The period of vegetation affects rosette diameter, number of leaves and root weight. A strong correlation between the weight of roots before laying for forcing and the weight of forcing heads (r = 0.79) was revealed. The roots of variety Conus, managed to accumulate a sufficient amount of nutrients for the formation of heads in a 98 days. The accumulation of sugars in forcing heads depended on their initial content in roots with a 75% reliability (r = 0.75). The results showed that in the northern latitudes the forcing can be carried out in winter in any room without light at a temperature of 10 to 17°C. In addition, subsurface heating of the substrate or maintaining water in the containers with roots provided a larger yield of heads obtained in a shorter time.

Abstract Premise Plant disease severity assessments are used to quantify plant–pathogen interactions and identify disease‐resistant lines. One common method for disease assessment involves scoring tissue manually using a semi‐quantitative scale. Automating assessments would provide fast, unbiased, and quantitative measurements of root disease severity, allowing for improved consistency within and across large data sets. However, using traditional Root System Markup Language (RSML) software in the study of root responses to pathogens presents additional challenges; these include the removal of necrotic tissue during the thresholding process, which results in inaccurate image analysis. Methods Using PlantCV, we developed a Python‐based pipeline, herein called RootDS, with two main objectives: (1) improving disease severity phenotyping and (2) generating binary images as inputs for RSML software. We tested the pipeline in common bean inoculated with Fusarium root rot. Results Quantitative disease scores and root area generated by this pipeline had a strong correlation with manually curated values (R2 = 0.92 and 0.90, respectively) and provided a broader capture of variation than manual disease scores. Compared to traditional manual thresholding, images generated using our pipeline did not affect RSML output. Discussion Overall, the RootDS pipeline provides greater functionality in disease score data sets and provides an alternative method for generating image sets for use in available RSML software.

Arbuscular mycorrhizal fungi (AMF) have the function of promoting water absorption for the host plant, whereas the role of easily extractable glomalin-related soil protein (GRSP), an N-linked glycoprotein secreted by AMF hyphae and spores, is unexplored for citrus plants. In this study, the effects on plant growth performance, root system characteristics, and leaf water status, along with the changes of mineral element content and relative expressions of tonoplast intrinsic protein (TIP) genes in lemon (Citrus limon L.) seedlings, were investigated under varying strengths of exogenous EE-GRSP application under potted conditions. The results showed that 1/2, 3/4, and full-strength exogenous EE-GRSP significantly promoted plant growth performance, as well as increased the biomass and root system architecture traits including root surface area, volume, taproot length, and lateral root numbers of lemon seedlings. The four different strengths of exogenous GRSP displayed differential effects on mineral element content: notably increased the content of phosphorus (P) and iron (Fe) in both leaves and roots, as well as magnesium (Mg) and zinc (Zn) content in the roots, but dramatically decreased the content of calcium (Ca) and manganese (Mn) in the roots, as well as Zn and Mn in the leaves. Exogenous EE-GRSP improved leaf water status, manifesting as decreases in leaf water potential, which was associated with the upregulated expressions of tonoplast intrinsic proteins (TIPs), including ClTIP1;1, ClTIP1;2, ClTIP1;3, ClTIP2;1, ClTIP2;2, ClTIP4;1, and ClTIP5;1 both in leaves and roots, and TIPs expressions exhibited diverse responses to EE-GRSP application. It was concluded that exogenous EE-GRSP exhibited differential responses on plant growth performance, which was related to its strength, and the effects were associated with nutrient concentration and root morphology, especially in the improvement in water status related to TIPs expressions. Therefore, EE-GRSP can be used as a biological promoter in plant cultivation, especially in citrus.

CRISPR/Cas9 (hereafter Cas9)-mediated gene knockout is one of the most important tools for studying gene function. However, many genes in plants play distinct roles in different cell types. Engineering the currently used Cas9 system to achieve cell-type-specific knockout of functional genes is useful for addressing the cell-specific functions of genes. Here we employed the cell-specific promoters of the WUSCHEL RELATED HOMEOBOX 5 (WOX5), CYCLIND6;1 (CYCD6;1), and ENDODERMIS7 (EN7) genes to drive the Cas9 element, allowing tissue-specific targeting of the genes of interest. We designed the reporters to verify the tissue-specific gene knockout in vivo. Our observation of the developmental phenotypes provides strong evidence for the involvement of SCARECROW (SCR) and GIBBERELLIC ACID INSENSITIVE (GAI) in the development of quiescent center (QC) and endodermal cells. This system overcomes the limitations of traditional plant mutagenesis techniques, which often result in embryonic lethality or pleiotropic phenotypes. By allowing cell-type-specific manipulation, this system has great potential to help us better understand the spatiotemporal functions of genes during plant development.

Cannabis sativa L. is typically propagated through micropropagation or vegetative cuttings, but the use of root-inducing hormones, such as indole-3-butyric acid (IBA), is not allowed for growing medicinal cannabis in Denmark. This study examined alternative rooting treatments, including Rhizobium rhizogenes inoculation, water-only as well as IBA treatments, in eight cannabis cultivars. PCR on root tissue suggested that 19% of R. rhizogenes-inoculated cuttings were transformed. These were derived from “Herijuana”, “Wild Thailand”, “Motherlode Kush”, and “Bruce Banner”, indicating a variation in cultivar susceptibility toward R. rhizogenes. A 100% rooting success was achieved regardless of cultivar and treatment, suggesting that alternative rooting agents are not required for efficient vegetative propagation. However, rooted cuttings differed in shoot morphology with improved shoot growth in cuttings treated with R. rhizogenes (195 ± 7 mm) or water (185 ± 7 mm) while inhibited shoot growth under IBA treatment (123 ± 6 mm). This could have advantageous economic implications should cuttings not treated with hormone reach maturity faster than those exposed to the hormone, thereby contributing to completing a full growing cycle more effectively. IBA exposure increased root length, root dry weight, and root/shoot dry weight ratio compared to cuttings treated with R. rhizogenes or water but simultaneously inhibited shoot growth compared to these.

Abstract Background Phosphorus (P) is an essential macronutrient for plant growth and development. Upon P shortage, plant responds with massive reprogramming of transcription, the Phosphate Starvation Response (PSR). In parallel, the production of strigolactones (SLs)—a class of plant hormones that regulates plant development and rhizosphere signaling molecules—increases. It is unclear, however, what the functional link is between these two processes. In this study, using tomato as a model, RNAseq was used to evaluate the time-resolved changes in gene expression in the roots upon P starvation and, using a tomato CAROTENOID CLEAVAGE DIOXYGENASES 8 (CCD8) RNAi line, what the role of SLs is in this. Results Gene ontology (GO)-term enrichment and KEGG analysis of the genes regulated by P starvation and P replenishment revealed that metabolism is an important component of the P starvation response that is aimed at P homeostasis, with large changes occurring in glyco-and galactolipid and carbohydrate metabolism, biosynthesis of secondary metabolites, including terpenoids and polyketides, glycan biosynthesis and metabolism, and amino acid metabolism. In the CCD8 RNAi line about 96% of the PSR genes was less affected than in wild-type (WT) tomato. For example, phospholipid biosynthesis was suppressed by P starvation, while the degradation of phospholipids and biosynthesis of substitute lipids such as sulfolipids and galactolipids were induced by P starvation. Around two thirds of the corresponding transcriptional changes depend on the presence of SLs. Other biosynthesis pathways are also reprogrammed under P starvation, such as phenylpropanoid and carotenoid biosynthesis, pantothenate and CoA, lysine and alkaloids, and this also partially depends on SLs. Additionally, some plant hormone biosynthetic pathways were affected by P starvation and also here, SLs are required for many of the changes (more than two thirds for Gibberellins and around one third for Abscisic acid) in the gene expression. Conclusions Our analysis shows that SLs are not just the end product of the PSR in plants (the signals secreted by plants into the rhizosphere), but also play a major role in the regulation of the PSR (as plant hormone).

Abstract Background Despite its good salt-tolerance level, key genes and pathways involved with temporal salt response of common bermudagrass (Cynodon dactylon (L.) Pers.) have not been explored. Therefore, in this study, to understand the underlying regulatory mechanism following the different period of salt exposure, a comprehensive transcriptome analysis of the bermudagrass roots was conducted. Results The transcripts regulated after 1 h, 6 h, or 24 h of hydroponic exposure to 200 mM NaCl in the roots of bermudagrass were investigated. Dataset series analysis revealed 16 distinct temporal salt-responsive expression profiles. Enrichment analysis identified potentially important salt responsive genes belonging to specific categories, such as hormonal metabolism, secondary metabolism, misc., cell wall, transcription factors and genes encoded a series of transporters. Weighted gene co-expression network analysis (WGCNA) revealed that lavenderblush2 and brown4 modules were significantly positively correlated with the proline content and peroxidase activity and hub genes within these two modules were further determined. Besides, after 1 h of salt treatment, genes belonging to categories such as signalling receptor kinase, transcription factors, tetrapyrrole synthesis and lipid metabolism were immediately and exclusively up-enriched compared to the subsequent time points, which indicated fast-acting and immediate physiological responses. Genes involved in secondary metabolite biosynthesis such as simple phenols, glucosinolates, isoflavones and tocopherol biosynthesis were exclusively up-regulated after 24 h of salt treatment, suggesting a slightly slower reaction of metabolic adjustment. Conclusion Here, we revealed salt-responsive genes belonging to categories that were commonly or differentially expressed in short-term salt stress, suggesting possible adaptive salt response mechanisms in roots. Also, the distinctive salt-response pathways and potential salt-tolerant hub genes investigated can provide useful future references to explore the molecular mechanisms of bermudagrass.

Inappropriate spatial distribution of canopy and roots limits further improvements to the grain yield of maize with increased planting density. We explored an integrated management practice called strip deep rotary with staggered planting (SRS) which includes comprehensive technology for both canopy layers and topsoil. Here, field experiments were conducted under two maize cropping systems (spring maize and summer maize) to evaluate the effect of SRS on the spatial distribution of the canopy and roots for maize under high planting density (90,000 plants ha−1) and to determine the physiological factors involved in yield formation. Compared with conventional management practices (no-tillage with single planting, NTS), SRS decreased the LAI of the middle to top layers while improving the light distribution of the middle and lower layers by 72.99% and 84.78%, respectively. Meanwhile, SRS increased the root dry weight density and root sap bleeding by 51.26% and 21.77%, respectively, due to the reduction in soil bulk density by an average of 5.08% in the 0–40 cm soil layer. SRS improved the SPAD in the ear and lower leaves and maximized the LAD, which was conducive to dry matter accumulation (DMA), increasing it by 14.02–24.16% compared to that of NTS. As a result, SRS increased maize grain yield by 6.71–25.44%. These results suggest that strip deep rotary combined with staggered planting noticeably optimized the distribution of light in the canopy and reduced the soil bulk density to promote root vitality and growth, to maintain canopy longevity, and to promote the accumulation of dry matter, which eventually increased the grain yield of the maize under high planting density conditions. Therefore, SRS can be considered a better choice for the sustainable high yield of maize under high-density planting conditions in the NCP and similar areas throughout the world.

Cabbage grown in contaminated soils can accumulate high levels of arsenic (As) in the edible parts, posing serious health risks. The efficiency of As uptake varies drastically among cabbage cultivars, but the underlying mechanisms are not clear. We screened out low (HY, Hangyun 49) and high As accumulating cultivars (GD, Guangdongyizhihua) to comparatively study whether the As accumulation is associated with variations in root physiological properties. Root biomass and length, reactive oxygen species (ROS), protein content, root activity, and ultrastructure of root cells of cabbage under different levels of As stress (0 (control), 1, 5, or 15 mg L−1) were measured As results, at low concentration (1 mg L−1), compared to GD, HY reduced As uptake and ROS content, and increased shoot biomass. At a high concentration (15 mg L−1), the thickened root cell wall and higher protein content in HY reduced arsenic damage to root cell structure and increased shoot biomass compared to GD. In conclusion, our results highlight that higher protein content, higher root activity, and thickened root cell walls result in lower As accumulation properties of HY compared to GD.

High voltage electrical discharge (HVED) is an eco-friendly low-cost method based on the creation of plasma-activated water (PAW) through the release of electrical discharge in water which results in the formation of reactive particles. Recent studies have reported that such novel plasma technologies promote germination and growth but their hormonal and metabolic background is still not known. In the present work, the HVED-induced hormonal and metabolic changes were studied during the germination of wheat seedlings. Hormonal changes including abscisic acid (ABA), gibberellic acids (GAs), indol acetic acid (IAA) and jasmonic acid (JA) and the polyphenol responses were detected in the early (2nd day) and late (5th day) germination phases of wheat as well as their redistribution in shoot and root. HVED treatment significantly stimulated germination and growth both in the shoot and root. The root early response to HVED involved the upregulation of ABA and increased phaseic and ferulic acid content, while the active form of gibberellic acid (GA1) was downregulated. In the later phase (5th day of germination), HVED had a stimulatory effect on the production of benzoic and salicylic acid. The shoot showed a different response: HVED induced the synthesis of JA_Le_Ile, an active form of JA, and provoked the biosynthesis of cinnamic, p-coumaric and caffeic acid in both phases of germination. Surprisingly, in 2-day-old shoots, HVED decreased the GA20 levels, being intermediate in the synthesis of bioactive gibberellins. These HVED-provoked metabolic changes indicated a stress-related response that could contribute to germination in wheat.

Abstract Background Potassium (K+) is an essential ion for most plants, as it is involved in the regulation of growth and development. K+ homeostasis in plant cells has evolved to facilitate plant adaptation to K+-deficiency stress. Argonaute1 (AGO1) is regulated by miR168 to modulate the small RNA regulatory pathway by RNA silencing complex (RISC) in tomatoes. However, the role of miR168-mediated regulation of AGO1 in the context of K+ deficiency stress in tomatoes has not been elucidated yet. Results SlmiR168 and its target gene SlAGO1A were differentially expressed among low-K+-tolerant JZ34 and low-K+-sensitive JZ18 tomato plants. Transgenic tomato plants constitutively expressing pri-SlmiR168a showed stronger root system growth, better leaves development, and higher K+ contents in roots under K+-deficiency stress than those of the transgenic tomato lines expressing rSlAGO1A (SlmiR168-resistant) and the wild type (WT). Deep sequencing analysis showed that 62 known microRNAs (miRNAs) were up-regulated in 35S:rSlAGO1 compared with WT tomatoes. The same miRNAs were down-regulated in 35S:SlmiR168a compared with WT plants. The integrated analysis found 12 miRNA/mRNA pairs from the 62 miRNAs, including the root growth and cytokinin (CTK)/abscisic acid (ABA) pathways. Conclusions The regulation mediated by SlmiR168 of SlAGO1A contributes to the plant development under low-K+ stress. Moreover, this regulation mechanism may influence downstream miRNA pathways in response to low-K+ stress through the CTK/ABA and root growth modulation pathways.

Abstract Background Non-photosynthetic plastids of plants are known to be involved in a range of metabolic and biosynthetic reactions, even if they have been difficult to study due to their small size and lack of color. The morphology of root plastids is heterogeneous and also the plastid size, density and subcellular distribution varies depending on the cell type and developmental stage, and therefore the functional features have remained obscure. Although the root plastid proteome is likely to reveal specific functional features, Arabidopsis thaliana root plastid proteome has not been studied to date. Results In the present study, we separated Arabidopsis root protein fraction enriched with plastids and mitochondria by 2D-PAGE and identified 84 plastid-targeted and 77 mitochondrion-targeted proteins using LC-MS/MS. The most prevalent root plastid protein categories represented amino acid biosynthesis, carbohydrate metabolism and lipid biosynthesis pathways, while the enzymes involved in starch and sucrose metabolism were not detected. Mitochondrion-targeted proteins were classified mainly into the energetics category. Conclusions This is the first study presenting gel-based map of Arabidopsis thaliana root plastid and mitochondrial proteome. Our findings suggest that Arabidopsis root plastids have broad biosynthetic capacity, and that they do not play a major role in a long-term storage of carbohydrates. The proteomic map provides a tool for further studies to compare changes in the proteome, e.g. in response to environmental cues, and emphasizes the role of root plastids in nitrogen and sulfur metabolism as well as in amino acid and fatty acid biosynthesis. The results enable taking a first step towards an integrated view of root plastid/mitochondrial proteome and metabolic functions in Arabidopsis thaliana roots.

Abstract Background Nanoparticle causes soil pollution, which affected plant development and then resulted in biomass decreased, especially in crops. However, little is known how sodium nanoparticles affect wheat root development at plant physiological level. Results We used NaBiF4 (size of 50–100 nm) to analyze the effect in wheat development at plant physiological level. Under exogenous application of 50 μM NaBiF4 for treatment, wheat root elongation was inhibited, but fresh weight and dry weight were increased. We also found that NaBiF4 induced that the plant had lower content of sodium than negative control. Used no-sodium nanoparticle of BiF3 for another negative control, it was also supported that NaBiF4 entered into cell to replace of sodium and exported sodium out of plant. These results implied NaBiF4 might induce sodium export to maintain the balance between sodium and potassium elements. Additionally, metabolism analysis demonstrated that SOD activity was increased, but CAT and POD activity reduced under exogenous treatment of NaBiF4 nanoparticles. Conclusions Sodium nanoparticles (NaBiF4) inhibited plant development by nanoparticle accumulation and sodium homeostasis broken, and then involved reactive oxygen species (ROS) signaling system response. These results provided more sights of sodium nanoparticle effect in plant development.

Abstract Background The basic helix-loop-helix (bHLH) gene family is one of the largest transcription factor families in plants and is functionally characterized in diverse species. However, less is known about its functions in the economically important allopolyploid oil crop, Brassica napus. Results We identified 602 potential bHLHs in the B. napus genome (BnabHLHs) and categorized them into 35 subfamilies, including seven newly separated subfamilies, based on phylogeny, protein structure, and exon-intron organization analysis. The intron insertion patterns of this gene family were analyzed and a total of eight types were identified in the bHLH regions of BnabHLHs. Chromosome distribution and synteny analyses revealed that hybridization between Brassica rapa and Brassica oleracea was the main expansion mechanism for BnabHLHs. Expression analyses showed that BnabHLHs were widely in different plant tissues and formed seven main patterns, suggesting they may participate in various aspects of B. napus development. Furthermore, when roots were treated with five different hormones (IAA, auxin; GA3, gibberellin; 6-BA, cytokinin; ABA, abscisic acid and ACC, ethylene), the expression profiles of BnabHLHs changed significantly, with many showing increased expression. The induction of five candidate BnabHLHs was confirmed following the five hormone treatments via qRT-PCR. Up to 246 BnabHLHs from nine subfamilies were predicted to have potential roles relating to root development through the joint analysis of their expression profiles and homolog function. Conclusion The 602 BnabHLHs identified from B. napus were classified into 35 subfamilies, and those members from the same subfamily generally had similar sequence motifs. Overall, we found that BnabHLHs may be widely involved in root development in B. napus. Moreover, this study provides important insights into the potential functions of the BnabHLHs super gene family and thus will be useful in future gene function research.

The bidirectional fluxes of cadmium and calcium across the plasma membrane were assessed and compared in subapical maize root segments. This homogeneous material provides a simplified system for investigating ion fluxes in whole organs. The kinetic profile of cadmium influx was characterized by a combination of a saturable rectangular hyperbola (Km = 30.15) and a straight line (k = 0.0013 L h−1 g−1 fresh weight), indicating the presence of multiple transport systems. In contrast, the influx of calcium was described by a simple Michaelis–Menten function (Km = 26.57 µM). The addition of calcium to the medium reduced cadmium influx into the root segments, suggesting a competition between the two ions for the same transport system(s). The efflux of calcium from the root segments was found to be significantly higher than that of cadmium, which was extremely low under the experimental conditions used. This was further confirmed by comparing cadmium and calcium fluxes across the plasma membrane of inside-out vesicles purified from maize root cortical cells. The inability of the root cortical cells to extrude cadmium may have driven the evolution of metal chelators for detoxifying intracellular cadmium ions.

Polygonatum odoratum (Mill.) Druce is an essential Chinese herb, but continuous cropping (CC) often results in a serious root rot disease, reducing the yield and quality. Phenolic acids, released through plant root exudation, are typical autotoxic substances that easily cause root rot in CC. To better understand the phenolic acid biosynthesis of P. odoratum roots in response to CC, this study performed a combined microRNA (miRNA)-seq and RNA-seq analysis. The phenolic acid contents of the first cropping (FC) soil and CC soil were determined by HPLC analysis. The results showed that CC soils contained significantly higher levels of p-coumaric acid, phenylacetate, and caffeic acid than FC soil, except for cinnamic acid and sinapic acid. Transcriptome identification and miRNA sequencing revealed 15,788 differentially expressed genes (DEGs) and 142 differentially expressed miRNAs (DEMs) in roots from FC and CC plants. Among them, 28 DEGs and eight DEMs were involved in phenolic acid biosynthesis. Meanwhile, comparative transcriptome and microRNA-seq analysis demonstrated that eight miRNAs corresponding to five target DEGs related to phenolic acid synthesis were screened. Among them, ath-miR172a, ath-miR172c, novel_130, sbi-miR172f, and tcc-miR172d contributed to phenylalanine synthesis. Osa-miR528-5p and mtr-miR2673a were key miRNAs that regulate syringyl lignin biosynthesis. Nta-miR156f was closely related to the shikimate pathway. These results indicated that the key DEGs and DEMs involved in phenolic acid anabolism might play vital roles in phenolic acid secretion from roots of P. odoratum under the CC system. As a result of the study, we may have a better understanding of phenolic acid biosynthesis during CC of roots of P. odoratum.

Reactive oxygen species (ROS), a type of oxygen monoelectronic reduction product, play integral roles in root growth and development. The epigenetic mechanism plays a critical role in gene transcription and expression; however, its regulation of ROS metabolism in root development is still limited. We found that the chromatin remodeling complex SWR1 regulates root length and lateral root formation in Arabidopsis. Our transcriptome results and gene ontology (GO) enrichment analysis showed that the oxidoreductase activity-related genes significantly changed in mutants for the Arabidopsis SWR1 complex components, such as arp6 and pie1, and histone variant H2A.Z triple mutant hta8 hta9 hta11. The three encoding genes in Arabidopsis are the three H2A.Z variants hta8, hta9, and hta11. Histochemical assays revealed that the SWR1 complex affects ROS accumulation in roots. Furthermore, chromatin immunoprecipitation quantitative real-time PCR (ChIP-qPCR) analysis showed that the reduced H2A.Z deposition in oxidoreductase activity-related genes caused ROS to accumulate in arp6, pie1, and hta8 hta9 hta11. H2A.Z deposition-deficient mutants decreased after the trimethylation of lysine 4 on histone H3 (H3K4me3) modifications and RNA polymerase II (Pol II) enrichment, and increased after the trimethylation of lysine 27 on histone H3 (H3K27me3) modifications, which may account for the expression change in oxidoreductase activity-related genes. In summary, our results revealed that the chromatin complex SWR1 regulates ROS accumulation in root development, highlighting the critical role of epigenetic mechanisms.

trans-Cinnamic acid is a phenolic compound widely studied in plant metabolism due to its importance in regulating different plant processes. Previous studies on maize plants showed that this compound could affect plant growth and causes metabolic changes in the leaves when applied. However, its effects on root metabolism are not well known. This study analyses the short-term effect of trans-cinnamic acid on the morphology of vascular bundle elements and metabolism in maize roots. At short times (between 6 and 12 h), there is a reduction in the content of many amino acids which may be associated with the altered nitrogen uptake observed in earlier work. In addition, the compound caused an alteration of the vascular bundles at 48 h and seemed to have changed the metabolism in roots to favor lignin and galactose synthesis. The results obtained complement those previously carried out on maize plants, demonstrating that in the short term trans-cinnamic acid can trigger stress-coping processes in the treated plants.

We conducted a genome-wide transcriptomic analysis of three drought tolerant and sensitive genotypes of common bean to examine their transcriptional responses to terminal drought stress. We then conducted pairwise comparisons between the root and leaf transcriptomes from the resulting tissue based on combined transcriptomic data from the tolerant and sensitive genotypes. Our transcriptomic data revealed that 491 (6.4%) DEGs (differentially expressed genes) were upregulated in tolerant genotypes, whereas they were downregulated in sensitive genotypes; likewise, 396 (5.1%) DEGs upregulated in sensitive genotypes were downregulated in tolerant genotypes. Several transcription factors, heat shock proteins, and chaperones were identified in the study. Several DEGs in drought DB (data Base) overlapped between genotypes. The GO (gene ontology) terms for biological processes showed upregulation of DEGs in tolerant genotypes for sulfate and drug transmembrane transport when compared to sensitive genotypes. A GO term for cellular components enriched with upregulated DEGs for the apoplast in tolerant genotypes. These results substantiated the temporal pattern of root growth (elongation and initiation of root growth), and ABA-mediated drought response in tolerant genotypes. KEGG (kyoto encyclopedia of genes and genomes) analysis revealed an upregulation of MAPK (mitogen activated protein kinase) signaling pathways and plant hormone signaling pathways in tolerant genotypes. As a result of this study, it will be possible to uncover the molecular mechanisms of drought tolerance in response to terminal drought stress in the field. Further, genome-wide transcriptomic analysis of both tolerant and sensitive genotypes will assist us in identifying potential genes that may contribute to improving drought tolerance in the common bean.

Abstract Background Modification of root architecture and improvement of root resistance to stresses can increase crop productivity. Functional analyses of root-specific genes are necessary for root system improvement, and root-specific promoters enable research into the regulation of root development and genetic manipulation of root traits. Maize is an important crop species; however, little systematic mining of root-specific genes and promoters has been performed to date. Results Genomic-scale mining based on microarray data sets followed by transcript detection resulted in the identification of 222 root-specific genes. Gene Ontology enrichment analyses revealed that these 222 root-specific genes were mainly involved in responses to chemical, biotic, and abiotic stresses. Of the 222 genes, 33 were verified by quantitative reverse transcription polymerase chain reaction, and 31 showed root-preferential activity. About 2 kb upstream 5 of the 31 identified root-preferential genes were cloned from the maize genome as putative promoters and named p8463, p5023, p1534, p8531 and p6629. GUS staining of transgenic maize-derived promoter-GUS constructs revealed that the five promoters drove GUS expression in a root-preferential manner. Conclusions We mined root-preferential genes and their promoters in maize and verified p8463, p5023, p1534, p8531 and p6629 as root-preferential promoters. Our research enables the identification of other tissue-specific genes and promoters in maize and other species. In addition, the five promoters may enable enhancement of target gene(s) of maize in a root-preferential manner to generate novel maize cultivars with resistance to water, fertilizer constraints, or biotic stresses.

Soil salinity negatively affects the growth, development and yield of plants. Acidification of the cytosol in cells of glycophytes was reported under salinity, while various types of plant cells can have a specific reaction under the same conditions. Transgenic Arabidopsis plants expressing the pH sensor Pt-GFP in the cytosol were used in this work for determination of morphometric changes and cytosolic pH changes in the superficial cells of Arabidopsis roots under chronic salinity in vitro. We did not find changes in the length of the root cap cells, while there was a decrease in the length of the differentiation zone under 50, 75 mM NaCl and the size of the epidermal cells of the differentiation zone under 75 mM NaCl. The most significant changes of cytosolic pH to chronic salinity was noted in columella (decrease by 1 pH unit at 75 mM NaCl) and epidermal cells of the differentiation zone (decrease by 0.6 and 0.4 pH units at 50 and 75 mM NaCl, respectively). In developed lateral root cap cells, acidification of cytosol by 0.4 units occurred only under 75 mM NaCl in the medium. In poorly differentiated lateral cells of the root cap, there were no changes in pH under chronic salinity.

During the growing season, potato plants are often exposed to soil drought, frequently accompanied by heat stress, which results in crop losses. In our experiment, the impact of these stresses, both separately and simultaneously, on the above-ground, on the root, and on the tuber mass was assessed. Four potato cultivars were tested. In vitro plants were planted in plastic tubes. Four treatments were used: control–optimal irrigation and temperature (22/18 °C), drought stress, high temperature stress (38/25 °C), and drought and high temperature stresses combined. The stresses were applied for two weeks during the tuberization phase. Both stresses caused changes in plant morphology. Drought stress had a greater impact on these changes than high temperatures. The biggest changes, however, took place when both stresses were applied simultaneously. Under all stresses, a decrease in tuber yield was found. The largest decrease was recorded in the case of applying both stresses simultaneously, while the smallest one was in the case of high temperature stress in relation to a control condition. Among the morphological parameters studied, the mass of the root system and its share in the entire biomass of the plant had the greatest impact on the decrease in yield. This mainly concerned drought stress.

Drought stress is one of the most devastating abiotic factors limiting plant growth and development. Devising an efficient and rapid screening method at the seedling stage is vital in identifying genotypes best suited under drought conditions. An experiment was conducted to assess 74 rice genotypes for drought tolerance using specially designed mini-hoop structures. Two treatments were imposed on rice seedlings, including 100% moisture and a 50% moisture regime. Several shoot morpho-physiological traits and root traits were measured and analyzed. The genotypes exhibited a wide range of variability for the measured traits, with the leaf area showing the most significant variation, followed by plant height, tiller number, and shoot dry weight. In contrast, the drought did not significantly affect most root traits. The germplasm was classified into different categories using cumulative drought stress response indices (CDSRI); 19 genotypes (26%) were identified as drought sensitive, and 33 (45%), 15 (20%), and 7 (9%) were determined as low, moderately, and highly drought-tolerant, respectively. Genotypes IR86638 and IR49830 were the most and least drought-tolerant, respectively. Overall, a poor correlation was observed between CDSRI, total shoot traits (R2 = 0.36), and physiological parameters (R2 = 0.10). A strong linear correlation was found between CDSRI and root traits (R2 = 0.81), suggesting that root traits are more crucial and better descriptors in screening for drought tolerance. This study can help rice breeders and scientists to accelerate breeding by adopting a mini-hoop rapid screening method. The tolerant genotypes could serve as appropriate donor parents, progenies, and potential genotypes for developing drought-tolerant commercial cultivars.

Abstract Background Barley is a low phosphorus (P) demand cereal crop. Tibetan wild barley, as a progenitor of cultivated barley, has revealed outstanding ability of tolerance to low-P stress. However, the underlying mechanisms of low-P adaption and the relevant genetic controlling are still unclear. Results We identified low-P tolerant barley lines in a doubled-haploid (DH) population derived from an elite Tibetan wild barley accession and a high-yield cultivar. The tolerant lines revealed greater root plasticity in the terms of lateral root length, compared to low-P sensitive lines, in response to low-P stress. By integrating the QTLs associated with root length and root transcriptomic profiling, candidate genes encoding isoflavone reductase, nitrate reductase, nitrate transporter and transcriptional factor MYB were identified. The differentially expressed genes (DEGs) involved the growth of lateral root, Pi transport within cells as well as from roots to shoots contributed to the differences between low-P tolerant line L138 and low-P sensitive lines L73 in their ability of P acquisition and utilization. Conclusions The plasticity of root system is an important trait for barley to tolerate low-P stress. The low-P tolerance in the elite DH line derived from a cross of Tibetan wild barley and cultivated barley is characterized by enhanced growth of lateral root and Pi recycling within plants under low-P stress.

Abstract Background Cold stress is the main factor that reduces rice yield in subtropical areas, especially at the seedling stage. Most of the current studies on cold stress focus the responses of rice shoots to cold stress. Limited studies are available on that of rice roots to cold stress. This study aimed to illustrate the biochemical responses of rice root under cold treatment, and subject to the establishment of cold stress-related biochemical traits for rice breeding or cropping-adjustment. Results Our results showed that the growth of rice seedling diminished under cold stress with difference extents among eight rice cultivars of most productive in Taiwan. Under cold treatments, the tested cultivars with higher growth rate had a higher level of hydrogen peroxide (H2O2) in the shoots but had a lower level in the roots. In contrast, the tested cultivates with low growth rate had higher levels of H2O2 in the roots but a lower level in the shoots. Meanwhile, higher MDA contents and higher cell-damage related electrolyte leakage were also found in the roots not in the shoots, suggesting that cold stress might induce oxidative stress in the roots, not in the shoots. Furthermore, the activity analysis of four antioxidant enzymes, namely superoxide dismutase (SOD), catalase (CAT), ascorbic peroxidase (APX), and glutathione reductase (GR), revealed that cold stress could increase SOD and CAT activities in the roots. Conclusions In summary, low H2O2 and low MDA contents along with lower SOD and CAT activities in rice root could be the biochemical traits of cold responses in rice seedlings. The results are hoping to have a contribution to the rice breeding or cropping-adjustment on cold tolerance.

Abstract Background Potassium is a nutrient element necessary for tobacco growth. Tobacco leaves with high potassium content are elastic and tough, rich in oil. And the same time, potassium can also improve the scent and aromatic value of flue-cured tobacco by regulating the synthesis of aromatic hydrocarbons in leaves.. It is an important quality indicator for flue-cured tobacco. However, the potassium concentration in tobacco leaves in most areas of China is generally lower than the global standard for high quality tobacco. Two tobacco genotypes were grafted to each other under different potassium levels to test whether potassium content and plant growth can be improved by grafting in tobacco. Results The growth of tobacco in all treatments was inhibited under potassium starvation, and grafting significantly alleviated this potassium stress in ‘Yunyan 87’. The trends in whole plant K+ uptake and K+ transfer efficiency to the leaves corresponded to the growth results of the different grafts. The nutrient depletion test results showed that the roots of ‘Wufeng No.2’ had higher K+ absorption potential, K+ affinity, and K+ inward flow rate. K+ enrichment circles appeared at the endoderm of the root section in the energy dispersive X-ray figure, indicating that the formation of Casparian strips may be partly responsible for the lower rate of lateral movement of K+ in the roots of ‘Yunyan 87’. Gene expression analysis suggested that energy redistribution at the whole plant level might constitute one strategy for coping with potassium starvation. The feedback regulation effects between scion ‘Wufeng No.2’ and rootstock ‘Yunyan 87’ indicated that the transmission of certain signaling substances had occurred during grafting. Conclusions ‘Wufeng No.2’ tobacco rootstock grafting can increase the K+ uptake and transport efficiency of ‘Yunyan 87’ and enhance plant growth under potassium stress. The physiological mechanism of the improved performance of grafted tobacco is related to higher K+ uptake and utilization ability, improved xylem K+ loading capacity, and up-regulated expression of genes related to energy supply systems.

Abstract Background MicroRNA (miRNA) are key players in regulating expression of target genes at post-transcriptional level. A number of miRNAs are implicated in modulating tolerance to various abiotic stresses. Waterlogging is an abiotic stress that deters plant growth and productivity by hypoxia. Dozens of reports mention about the miRNAs expressed in response to waterlogging and hypoxia. Despite the fact that tomato is a model vegetable but waterlogging sensitive crop, the role of miRNAs in hypoxia tolerance is poorly understood in tomato. Results In this study, we investigated the differentially expressed miRNAs between hypoxia-treated and untreated wild tomato root by using high-throughput sequencing technology. A total of 33 known miRNAs were lowly expressed, whereas only 3 miRNAs showed higher expression in hypoxia-treated wild tomato root compared with untreated wild tomato root. Then two conserved and lowly expressed miRNAs, miR171 and miR390, were deactivated by Short Tandem Target Mimic (STTM) technology in Arabidopsis. As the results, the number and length of lateral roots were more in STTM171 and STTM390 transgenic lines compared with that of wild type plant, which partly phenocopy the increase root number and shortening the root length in hypoxia-treated wild tomato root. Conclusions The differentially expressed miRNAs between hypoxia-treated wild tomato and control root, which contribute to the auxin homeostasis, morphologic change, and stress response, might result in reduction in the biomass and length of the root in hypoxiated conditions.

Soybean (Glycine max L.) is a crop native to Northeast Asia, including China, Korea, and Japan, but currently cultivated all over the world. The National Agrobiodiversity Center in Korea at the Rural Development Administration (RDA) conserves approximately 26,000 accessions and conducts characterizations of its accessions, to accumulate new information. Roots are essential organs of a plant, providing mechanical support, as well as aiding water and nutrient acquisition. Currently, not much information is available in international gene banks regarding root characterization. We studied the root phenotype of 374 soybean accessions, using a high-throughput method. Eight root morphological traits (RMT) were studied and we observed that the surface area (SA), number of forks (NF), and number of tips (NT) had a positive correlation with total length (LENGTH), and that link average length (LAL) and other traits all had a negative correlation. Additionally, the correlation between seed traits (height, width, and 100-seed weight) and root traits was confirmed for the first time in this experiment. The germplasms were divided into three clusters by k-means clustering, and orthogonal projections to latent structures discriminant analysis (OPLS-DA) was used to compare clusters. The most distinctive characteristics between clusters were total lateral average length (LAD) and total lateral average length (DIAM). Cluster 3 had the highest LENGTH, SA, NF, and NF, whereas cluster 1 had the smallest LENGTH, SA, and NF. We selected the top 10 accessions for each RMT, and IT208321, IT216313, and IT216137 were nominated as the best germplasms. These accessions can be recommended to breeders as materials for breeding programs. This is a preliminary report on the characterization of the root phenotype at an international gene bank and will open up the possibility of improving the available information on accessions in gene banks worldwide.

With global climate change, waterlogging stress is becoming more frequent. Waterlogging stress inhibits root growth and physiological metabolism, which ultimately leads to yield loss in wheat. Waterlogging priming has been proven to effectively enhance waterlogging tolerance in wheat. However, it is not known whether waterlogging priming can improve the offspring’s waterlogging resistance. Here, wheat seeds that applied waterlogging priming for one generation, two generations and three generations are separately used to test the hypoxia stress tolerance in wheat, and the physiological mechanisms are evaluated. Results found that progeny of primed plants showed higher plant biomass by enhancing the net photosynthetic rate and antioxidant enzyme activity. Consequently, more sugars are transported to roots, providing a metabolic substrate for anaerobic respiration and producing more ATP to maintain the root growth in the progeny of primed plants compared with non-primed plants. Furthermore, primed plants’ offspring promote ethylene biosynthesis and further induce the formation of a higher rate of aerenchyma in roots. This study provides a theoretical basis for improving the waterlogging tolerance of wheat.

The phytohormone salicylic acid (SA) can influence the polyamine metabolism in plants. Additionally, polyamines (PAs) can regulate the synthesis of SA, providing an exciting interplay between them not only in plant growth and development but also in biotic or abiotic stress conditions. The effect of SA on polyamine metabolism of leaves is well-studied but the root responses are rarely investigated. In this study, tomato roots were used to investigate the effect of short-term exposition of SA in two different concentrations, a sublethal 0.1 mM and a lethal 1 mM. To explore the involvement of SA in regulating PAs in roots, the degradation of PAs was also determined. As both SA and PAs can induce reactive oxygen species (ROS) and nitric oxide (NO) production, the balance of ROS and NO was analyzed in root tips. The results showed that 0.1 mM SA induced the production of higher PAs, spermidine (Spd), and spermine (Spm), while 1 mM SA decreased the PA contents by activating degrading enzymes. Studying the ROS and NO levels in root tips, the ROS production was induced earlier than NO, consistent with all the investigated zones of roots. This study provides evidence for concentration-dependent rapid effects of SA treatments on polyamine metabolism causing an imbalance of ROS–NO in root tips.

Abstract Background There are two plant phosphorus (P)-uptake pathways, namely the direct P uptake by roots and the indirect P uptake through arbuscular mycorrhizal fungi (AMF). Maximizing the efficiency of root and AMF processes associated with P acquisition by adjusting soil conditions is important for simultaneously ensuring high yields and the efficient use of available P. Results A root box experiment was conducted in 2015 and 2016. The aim was to investigate the effects of different P and soil water conditions on root/mycorrhizal growth and P uptake by cotton plants. Hyphal growth was induced in well-watered soil, but decreased with increasing P concentrations. Additionally, P fertilizers regulated root length only under well-watered conditions, with the longest roots observed in response to 0.2 g P2O5 kg− 1. In contrast, root elongation was essentially unaffected by P fertilizers under drought conditions. And soil water in general had more significant effects on root and hyphal growth than phosphorus levels. In well-watered soil, the application of P significantly increased the cotton plant P uptake, but there were no differences between the effects of 0.2 and 1 g P2O5 kg− 1. So optimizing phosphorus inputs and soil water can increase cotton growth and phosphorus uptake by maximizing the efficiency of phosphorus acquisition by roots/mycorrhizae. Conclusions Soil water and P contents of 19–24% and 20–25 mg kg− 1, respectively, simultaneously maximized root/mycorrhizal growth and P uptake by cotton plants.

Abstract Besides their ecological importance, epiphytic species of Orchidaceae play economic and social roles through their commercialization and some are at great risk of extinction. The objectives of this study were to characterize the leaf and root anatomy of fourteen epiphytic Orchidaceae species, which occur in indigenous territory in the Parque Estadual da Serra do Tabuleiro (P.E.S.T.), Santa Catarina, Brazil and to identify adaptive anatomical characteristics related to the epiphytic habit. The species are commercialized by the Guarani and were collected during interviews and guided tours with Guarani in the indigenous territory. The results reveal the species have many morphoanatomical structures that are useful during water shortages resulting from the epiphytic habit. Notable characteristics are related to reserving water (i.e., pseudobulbs and a hypodermis with water-storage cells) and resistance to desiccation in the leaf (i.e., conspicuous cuticle, suprastomatic chamber and extraxylary and pericyclic fibers) and root (i.e., tilosomes and/or exodermal thickening and cortex cells with phi thickenings or sclereids). Descriptions and the identification of adaptive characteristics of epiphytic plant species are useful for conservation and cultivation studies, especially for plants commercially used by the Guarani Indians.

The overall survival of a plant depends on the development, growth, and functioning of the roots. Root development and growth are not only genetically programmed but are constantly influenced by environmental factors, with the roots adapting to such changes. The peptide AEDL (alanine–glutamine acid–asparagine acid–leucine) at a concentration of 10−7 M had an elongating effect on the root cells of Nicotiana tabacum seedlings. The action of this peptide at such a low concentration is similar to that of peptide phytohormones. In the presence of 150 mM NaCl, a strong distortion in the development and architecture of the tobacco roots was observed. However, the combined presence of AEDL and NaCl resulted in normal root development. In the presence of AEDL, reactive oxygen species (ROS) were detected in the elongation and root hair zones of the roots. The ROS marker fluorescence intensity in plant cells grown with AEDL was much lower than that of plant cells grown without the peptide. Thus, AEDL protected the root tissue from damage by oxidative stress caused by the toxic effects of NaCl. Localization and accumulation of AEDL at the root were tissue-specific. Fluorescence microscopy showed that FITC-AEDL predominantly localized in the zones of elongation and root hairs, with insignificant localization in the meristem zone. AEDL induced a change in the structural organization of chromatin. Structural changes in chromatin caused significant changes in the expression of numerous genes associated with the development and differentiation of the root system. In the roots of tobacco seedlings grown in the presence of AEDL, the expression of WOX family genes decreased, and differentiation of stem cells increased, which led to root elongation. However, in the presence of NaCl, elongation of the tobacco root occurred via a different mechanism involving genes of the expansin family that weaken the cell wall in the elongation zone. Root elongation of plants is of fundamental importance in biology and is especially relevant to crop production as it can affect crop yields.

Nitrogen (N) is one of the important factors affecting maize root morphological construction and growth development. An association panel of 124 maize inbred lines was evaluated for root and shoot growth at seedling stage under normal N (CK) and low N (LN) treatments, using the paper culture method. Twenty traits were measured, including three shoot traits and seventeen root traits, a genome-wide association study (GWAS) was performed using the Bayesian-information and Linkage-disequilibrium Iteratively Nested Keyway (BLINK) methods. The results showed that LN condition promoted the growth of the maize roots, and normal N promoted the growth of the shoots. A total of 185 significant SNPs were identified, including 27 SNPs for shoot traits and 158 SNPs for root traits. Four important candidate genes were identified. Under LN conditions, the candidate gene Zm00001d004123 was significantly correlated with the number of crown roots, Zm00001d025554 was correlated with plant height. Under CK conditions, the candidate gene Zm00001d051083 was correlated with the length and area of seminal roots, Zm00001d050798 was correlated with the total root length. The four candidate genes all responded to the LN treatment. The research results provide genetic resources for the genetic improvement of maize root traits.