Lateral Root Length Research Articles

A root-localized gene in normal apples is ectopically expressed in aerial parts of columnar apples

The columnar apple ‘McIntosh Wijcik’, which is a mutation of ‘McIntosh’ shoots, has short internodes, thick stems, upright growth, poor lateral branches, and increased spur density. These columnar traits are controlled by a single dominant gene known as Co. We previously identified a putative dioxygenase gene (designated as 91071) as a promising Co candidate (expressed in the shoot apices of ‘McIntosh Wijcik’). However, tissue expression and function of the 91071 gene in noncolumnar apples is still not clear. In this study, we used reverse transcription polymerase chain reaction to demonstrate that the 91071 gene is mainly expressed in the roots of noncolumnar apples, whereas it is also expressed in shoot apices and leaves of columnar apples. In situ hybridization revealed that the 91071 gene is expressed at the primordium of lateral roots and root tips of both noncolumnar and columnar apple trees and in the shoot meristem and leaf primordium in the columnar apple ‘McIntosh Wijcik’. Grafting experiments of noncolumnar scion onto columnar rootstocks revealed that the columnar growth phenotype is not transmissible from rootstock to scion. These results indicated that ectopic expression of the 91071 gene in aerial parts causes columnar growth, whereas the expression of the 91071 gene in roots does not produce columnar growth. Furthermore, transgenic apples overexpressing the 91071 gene showed larger median adventitious root length and higher median number of lateral roots than control apples. Our result suggests that the 91071 gene may be related to root development.

Real-time detection of the nanoparticle induced phytotoxicity in rice root tip through the visible red emissions of Eu3+ ions.

Phytotoxicity is one of the most important factors involved in the reduction of crop production. With the introduction of NaBiF4 nanoparticles, the effect of the particle size (>50 nm) on rice development was systematically studied. Through the exogenous treatment of multiple concentrations of nanoparticles, the primary root length, lateral root number, and lateral root length were significantly inhibited under higher content of nanoparticles, but more crown root formation was induced, which might be due to phytotoxicity. With the help of the red emission of the Eu3+-activated NaBiF4 nanoparticles, we could infer that the nanoparticles were accumulated in the root tip cells in the division and elongation zone but not in the mature region. Additionally, the investigation on the influence of the studied nanoparticles on the gene level and the expression of phytotoxicity related genes was performed to further identify the effect of the nanoparticles on the rice root development. These results potentially explain the effect of larger nanoparticles on phytotoxicity in the plant roots.

Responses of grapevine rootstocks to drought through altered root system architecture and root transcriptomic regulations

Roots are the major interface between the plant and various stress factors in the soil environment. Alteration of root system architecture (RSA) (root length, spread, number and length of lateral roots) in response to environmental changes is known to be an important strategy for plant adaptation and productivity. In light of ongoing climate changes and global warming predictions, the breeding of drought-tolerant grapevine cultivars is becoming a crucial factor for developing a sustainable viticulture. Root-trait modeling of grapevine rootstock for drought stress scenarios, together with high-throughput phenotyping and genotyping techniques, may provide a valuable background for breeding studies in viticulture. Here, tree grafted grapevine rootstocks (110R, 5BB and 41B) having differential RSA regulations and drought tolerance were investigated to define their drought dependent root characteristics. Root area, root length, ramification and number of root tips reduced less in 110R grafted grapevines compared to 5BB and 41B grafted ones during drought treatment. Root relative water content as well as total carbohydrate and nitrogen content were found to be much higher in the roots of 110R than it was in the roots of other rootstocks under drought. Microarray-based root transcriptome profiling was also conducted on the roots of these rootstocks to identify their gene regulation network behind drought-dependent RSA alterations. Transcriptome analysis revealed totally 2795, 1196 and 1612 differentially expressed transcripts at the severe drought for the roots of 110R, 5BB and 41B, respectively. According to this transcriptomic data, effective root elongation and enlargement performance of 110R were suggested to depend on three transcriptomic regulations. First one is the drought-dependent induction in sugar and protein transporters genes (SWEET and NRT1/PTR) in the roots of 110R to facilitate carbohydrate and nitrogen accumulation. In the roots of the same rootstock, expression increase in osmolyte producer genes revealed another transcriptomic regulation enabling effective root osmotic adjustment under drought stress. The third mechanism was linked to root suberization with upregulation of transcripts functional in wax producing enzymes (Caffeic acid 3-O-methyltransferase, Eceriferum3, 3-ketoacyl-CoAsynthase). These three transcriptomic regulations were suggested to provide essential energy and water preservation to the roots of 110R for its effective RSA regulation under drought. This phenotypic and genotypic knowledge could be used to develop root-dependent drought tolerant grapevines in breeding programs and could facilitate elucidation of genetic regulations behind RSA alteration in other plants.

Root plasticity under fluctuating soil moisture stress exhibited by backcross inbred line of a rice variety, Nipponbare carrying introgressed segments from KDML105 and detection of the associated QTLs

In rainfed lowland rice ecosystem, rice plants are often exposed to alternating recurrences of waterlogging and drought due to erratic rainfall. Such soil moisture fluctuation (SMF) which is completely different from simple or progressive drought could be stressful for plant growth, thereby causing reduction in yield. Root plasticity is one of the key traits that play important roles for plant adaptation under such conditions. This study aimed to evaluate root plasticity expression and its functional roles in dry matter production and yield under SMF using Nipponbare, KDML 105 and three backcross inbred lines (BILs) and to identify QTL(s) associated with root traits in response to SMF at two growth stages using Nipponbare/KDML105 F2 plants. A BIL, G3-3 showed higher shoot dry matter production and yield than Nipponbare due to its greater ability to maintain stomatal conductance concomitant with greater root system development caused by promoted production of nodal and lateral roots under SMF. QTLs were identified for total nodal root length, total lateral root length, total root length, number of nodal roots, and branching index under SMF at vegetative and reproductive stages. The QTLs detected at vegetative and reproductive stages were different. We discuss here that relationship between root system of G3-3 and the detected QTLs. Therefore, G3-3 and the identified QTLs could be useful genetic materials in breeding program for improving the adaptation of rice plants in target rainfed lowland areas.

Foxtail Millet [Setaria italica (L.) Beauv.] Grown under Low Nitrogen Shows a Smaller Root System, Enhanced Biomass Accumulation, and Nitrate Transporter Expression.

Foxtail millet (FM) [Setaria italica (L.) Beauv.] is a grain and forage crop well adapted to nutrient-poor soils. To date little is known how FM adapts to low nitrogen (LN) at the morphological, physiological, and molecular levels. Using the FM variety Yugu1, we found that LN led to lower chlorophyll contents and N concentrations, and higher root/shoot and C/N ratios and N utilization efficiencies under hydroponic culture. Importantly, enhanced biomass accumulation in the root under LN was in contrast to a smaller root system, as indicated by significant decreases in total root length; crown root number and length; and lateral root number, length, and density. Enhanced carbon allocation toward the root was rather for significant increases in average diameter of the LN root, potentially favorable for wider xylem vessels or other anatomical alterations facilitating nutrient transport. Lower levels of IAA and CKs were consistent with a smaller root system and higher levels of GA may promote root thickening under LN. Further, up-regulation of SiNRT1.1, SiNRT2.1, and SiNAR2.1 expression and nitrate influx in the root and that of SiNRT1.11 and SiNRT1.12 expression in the shoot probably favored nitrate uptake and remobilization as a whole. Lastly, more soluble proteins accumulated in the N-deficient root likely as a result of increases of N utilization efficiencies. Such “excessive” protein-N was possibly available for shoot delivery. Thus, FM may preferentially transport carbon toward the root facilitating root thickening/nutrient transport and allocate N toward the shoot maximizing photosynthesis/carbon fixation as a primary adaptive strategy to N limitation.

Diversity in root growth responses to moisture deficit in young faba bean (Vicia faba L.) plants.

BackgroundSoil moisture deficiency causes yield reduction and instability in faba bean (Vicia faba L.) production. The extent of sensitivity to drought stress varies across accessions originating from diverse moisture regimes of the world. Hence, we conducted successive greenhouse experiments in pots and rhizotrons to explore diversity in root responses to soil water deficit.MethodsA set of 89 accessions from wet and dry growing regions of the world was defined according to the Focused Identification of Germplasm Strategy and screened in a perlite-sand medium under well watered conditions in a greenhouse experiment. Stomatal conductance, canopy temperature, chlorophyll concentration, and root and shoot dry weights were recorded during the fifth week of growth. Eight accessions representing the range of responses were selected for further investigation. Starting five days after germination, they were subjected to a root phenotyping experiment using the automated phenotyping platform GROWSCREEN-Rhizo. The rhizotrons were filled with peat-soil under well watered and water limited conditions. Root architectural traits were recorded five, 12, and 19 days after the treatment (DAT) began.ResultsIn the germplasm survey, accessions from dry regions showed significantly higher values of chlorophyll concentration, shoot and root dry weights than those from wet regions. Root and shoot dry weight as well as seed weight, and chlorophyll concentration were positively correlated with each other. Accession DS70622 combined higher values of root and shoot dry weight than the rest. The experiment in GROWSCREEN-Rhizo showed large differences in root response to water deficit. The accession by treatment interactions in taproot and second order lateral root lengths were significant at 12 and 19 DAT, and the taproot length was reduced up to 57% by drought. The longest and deepest root systems under both treatment conditions were recorded by DS70622 and DS11320, and total root length of DS70622 was three times longer than that of WS99501, the shortest rooted accession. The maximum horizontal distribution of a root system and root surface coverage were positively correlated with taproot and total root lengths and root system depth. DS70622 and WS99501 combined maximum and minimum values of these traits, respectively. Thus, roots of DS70622 and DS11320, from dry regions, showed drought-avoidance characteristics whereas those of WS99501 and Mèlodie/2, from wet regions, showed the opposite.DiscussionThe combination of the germplasm survey and use of GROWSCREEN-Rhizo allowed exploring of adaptive traits and detection of root phenotypic markers for potential drought avoidance. The greater root system depth and root surface coverage, exemplified by DS70622 and DS11320, can now be tested as new sources of drought tolerance.

Natural variation of hormone levels in Arabidopsis roots and correlations with complex root architecture.

Studies on natural variation are an important tool to unravel the genetic basis of quantitative traits in plants. Despite the significant roles of phytohormones in plant development, including root architecture, hardly any studies have been done to investigate natural variation in endogenous hormone levels in plants. Therefore, in the present study a range of hormones were quantified in root extracts of thirteen Arabidopsis thaliana accessions using a ultra performance liquid chromatography triple quadrupole mass spectrometer. Root system architecture of the set of accessions was quantified, using a new parameter (mature root unit) for complex root systems, and correlated with the phytohormone data. Significant variations in phytohormone levels among the accessions were detected, but were remarkably small, namely less than three‐fold difference between extremes. For cytokinins, relatively larger variations were found for ribosides and glucosides, as compared to the free bases. For root phenotyping, length‐related traits—lateral root length and total root length—showed larger variations than lateral root number‐related ones. For root architecture, antagonistic interactions between hormones, for example, indole‐3‐acetic acid to trans‐zeatin were detected in correlation analysis. These findings provide conclusive evidence for the presence of natural variation in phytohormone levels in Arabidopsis roots, suggesting that quantitative genetic analyses are feasible.

Root hair abundance impacts cadmium accumulation in Arabidopsis thaliana shoots.

Root hairs increase the contact area of roots with soil and thereby enhance the capacity for solute uptake. The strict hair/non-hair pattern of Arabidopsis thaliana can change with nutrient deficiency or exposure to toxic elements, which modify root hair density. The effects of root hair density on cadmium (Cd) accumulation in shoots of arabidopsis genotypes with altered root hair development and patterning were studied. Arabidopsis mutants that are unable to develop root hairs (rhd6-1 and cpc/try) or produce hairy roots (wer/myb23) were compared with the ecotype Columbia (Col-0). Plants were cultivated on nutrient agar for 2 weeks with or without Cd. Cadmium was applied as Cd(NO3)2 at two concentrations, 10 and 100 µm. Shoot biomass, root characteristics (primary root length, lateral root number, lateral root length and root hair density) and Cd concentrations in shoots were assessed. Anatomical features (suberization of the endodermis and development of the xylem) that might influence Cd uptake and translocation were also examined. Cadmium inhibited plant growth and reduced root length and the number of lateral roots and root hairs per plant. Suberin lamellae in the root endodermis and xylem differentiation developed closer to the root apex in plants exposed to 100 µm Cd. The latter effect was genotype dependent. Shoot Cd accumulation was correlated with root hair abundance when plants were grown in the presence of 10 µm Cd, but not when grown in the presence of 100 µm Cd, in which treatment the development of suberin lamellae closer to the root tip appeared to restrict Cd accumulation in shoots. Root hair density can have a large effect on Cd accumulation in shoots, suggesting that the symplasmic pathway might play a significant role in the uptake and accumulation of this toxic element.

OsNRT2.4 encodes a dual-affinity nitrate transporter and functions in nitrate-regulated root growth and nitrate distribution in rice

Plant NRT2 nitrate transporters commonly require a partner protein, NAR2, for transporting nitrate at low concentrations, but their role in plants is not well understood. In this study, we characterized the gene for one of these transporters in the rice genome, OsNRT2.4, in terms of its activity and roles in rice grown in environments with different N supply. In Xenopus oocytes, OsNRT2.4 alone without OsNAR2 co-expression facilitated nitrate uptake showing biphasic kinetics at a wide concentration range, with high- and low-affinity KM values of 0.15 and 4 mM, respectively. OsNRT2.4 did not have nitrate efflux or IAA influx activity. In rice roots, OsNRT2.4 was expressed mainly in the base of lateral root primordia. Knockout of OsNRT2.4 decreased lateral root number and length, and the total N uptake per plant at both 0.25 and 2.5 mM NO3- levels. In the shoots, OsNRT2.4 was expressed mainly in vascular tissues, and its knockout decreased the growth and NO3--N distribution. Knockout of OsNRT2.4, however, did not affect rice growth and N uptake under conditions without N or with only NH4+ supply. We conclude that OsNRT2.4 functions as a dual-affinity nitrate transporter and is required for nitrate-regulated root and shoot growth of rice.

Does a Specific Location of Composted Poultry Litter in Soil Influence Nutrient Use Efficiency and Vegetable Production? A Mesocosm Experiment

Animal wastes may be promoted as an alternative to mineral fertilizers that remain unaffordable to the overwhelming part of smallholder farmers in Sub-saharan Africa. However for an efficient use, mechanisms that underly their impact on crops should be well understood. This study was conducted in mesocosm to evaluate impacts of two ways of composted poultry litter (CPL) addition on growth and nutrient use efficiency by cucumber. It included three treatments with five-bucket replicates each: Control, CPL applied on soil surface (CS) or buried to 10 cm-depth (CB). Dry CPL was added at the rate of 0.5 kg bucket-1. At harvest, root distribution was examined in the 0-5, 5-10 and 10-20 cm depths. Dry biomasses of roots, shoot and fruits were also determined and allowed for calculation of diverse indexes of biomass allocation (root:shoot ratio, root weight ratio, stem weight ratio, leaf weight ratio) and nutrient use efficiency (factor productivity of the compost, partial factor productivity of nutrients, agronomic efficiency of compost, and apparent agronomic efficiency of nutrients). The results showed that application of CPL led to a significant improvement of all considered parameters except for the leaf weight ratio which was higher in the control (44.1±3.3) than in CS (28.1±1.9) and CB 31.2±3.5). Total lateral root length was significantly higher in CS than in CB (113.5±10.7 cm vs. 75.5±9.0 cm). The number of lateral roots per plant in the 0-5 cm soil layer was higher in CS than in CB (5.4 vs. 1 root plant-1); the reverse was observed in 5-10 cm (1.2 vs. 4.4 root plant-1). Both fresh fruit yield and total dry mass were positively correlated to root attributes. These were themselves negatively impacted by soil acidity. All nutrient use efficiency indexes were higher in CS than CB. The CPL improved the agronomic performance of cucumber particularly when applied at soil surface.

Quantitative trait locus analysis of adventitious and lateral root morphology of barley grown at low and high P.

Barley (Hordeum vulgare L) may alter its root morphology to improve P acquisition efficiency under low-P (LP) stress. This research studied the variations in adventitious and lateral root morphological traits of barley and mapped their quantitative trait loci (QTLs) under LP and high P (HP). The recombinant inbred lines were derived from the F1 population of a cross between CN4027 and Baudin. Two experiments aimed to identify QTLs related to adventitious and lateral root morphological traits under LP and HP. The length, surface area and volume of adventitious and lateral roots were measured. Under HP, Baudin had larger root morphology, especially lateral root morphology, than CN4027. LP stress induced lateral root growth but inhibited adventitious root growth. Nineteen QTLs for root morphological traits were detected. These QTLs clustered within four regions (Cl-2H, Cl-3H, Cl-4H and Cl-7H) on chromosomes 2H, 3H, 4H and 7H, with corresponding contributions of 12.0-42.9%. Some QTLs are linked with the QTLs for P efficiency detected previously, demonstrating the role of root morphological traits in P efficiency. The Cl-2H region was identified in the interval bPb3927665-bPb3265744 on chromosome 2H and had major effects on lateral root growth, especially under LP. Lateral root length and surface area increased when alleles from Baudin were present at the QTLs in Cl-2H. This study demonstrated the patterns of growth among root types and the role of lateral roots in barley's adaption to LP stress. The QTL clusters, especially Cl-2H, may offer clues for fine mapping and map-based cloning.

Phenotyping Root System Architecture of Cotton (Gossypium barbadense L.) Grown Under Salinity

Abstract Soil salinity causes an annual deep negative impact to the global agricultural economy. In this study, the effects of salinity on early seedling physiology of two Egyptian cotton (Gossypium barbadense L.) cultivars differing in their salinity tolerance were examined. Also the potential use of a low cost mini-rhizotron system to measure variation in root system architecture (RSA) traits existing in both cultivars was assessed. Salt tolerant cotton cultivar ‘Giza 90’ produced significantly higher root and shoot biomass, accumulated lower Na+/K+ ratio through a higher Na+ exclusion from both roots and leaves as well as synthesized higher proline contents compared to salt sensitive ‘Giza 45’ cultivar. Measuring RSA in mini-rhizotrons containing solid MS nutrient medium as substrate proved to be more precise and efficient than peat moss/sand mixture. We report superior values of main root growth rate, total root system size, main root length, higher number of lateral roots and average lateral root length in ‘Giza 90’ under salinity. Higher lateral root density and length together with higher root tissue tolerance of Na+ ions in ‘Giza 90’ give it an advantage to be used as donor genotype for desirable root traits to other elite cultivars.

PRODUCTIVITY AND QUALITY OF AQUAPONICALLY GROWN TOMATO (Solanum lycopersicum L.) SUPPLEMENTED WITH DIFFERENT NUTRIENT SOLUTIONS

Tomato (Solanum lycopersicum L.) is an important vegetable in the country which is consumed both raw and as processed product. This study aimed to evaluate the growth, yield, postharvest qualities and profitability of aquaponically grown tomato supplemented with different nutrient solutions. The study was laid out in Randomized Complete Block Design with 8 treatments replicated 3 times. Tomato fruits were first harvested 60 days after transplanting with 100 percent survival rate. Early flowering of tomato plant was observed when applied with VSU liquid nutrient formulation (VSU-LNF) with node number ranging from 13.33-14.00 bearing the first flower. Growth parameters such as plant height, number of lateral shoot, root length and fruit size are stimulated by the application of VSU liquid nutrient formulation and in combination with ferments. This also resulted to high number and weight of marketable fruit in tomato plant. Carotenoid and chlorophyll contents of the fruits were higher in aquaponically grown tomato supplemented with fermented kuhol (Pomacea canaliculata L.). Free radical scavenging activity was found highest in the fruit of tomato grown in aquaponic system supplemented with combined VSU liquid nutrient formulation with fermented malunggay (Moringa olifera). However, vitamin Cand sugar content of tomato fruits were found highest when supplemented with combine VSU liquid nutrient formulation and fermented kuhol. No significant differences were obtained on firmness, percent weight loss, respiration rate at 2 and 4 weeks after storage, titratable acidity, fruit nitrogen, pH of fruit, oxidation reduction potential, electrical conductivity and total dissolved solids. On the other hand, higher moisture content of the tomato fruits were observed with the application of VSU liquid nutrient formulation and in combination with ferments. Therefore, high yield was produced with the application of VSU liquid nutrient formulation which resulted to higher net return. Aquaponically grown tomato supplemented with VSU-LNF produced lesser yield but with considerable net return.

Hydrogen Sulphide Improves Iron Homeostasis in Wheat Under Iron-Deficiency

Hydrogen sulphide (H2S) is emerging as an important gaseous molecule involved in various plant developmental processes and plant stress responses. In this study, exogenous H2S donor (sodium hydrosulfide, NaHS) treated wheat plants were used to investigate the role of H2S in response to iron-deficiency. The results showed that H2S significantly alleviated leaf chlorosis under iron-deficient conditions, and thus improved photosynthesis. Moreover, H2S increased the lateral root (LR) number, density and length of wheat seedlings grown in iron-sufficient and deficient culture solution, and promoted phytosiderophores (PSs) secretion from roots simultaneously, which eventually led to an increase in iron uptake. Taken together, these results indicate that H2S improved iron uptake by regulating root development and PSs secretion, and consequently increased chlorophyll biosynthesis and photosynthesis in plants under iron-deficiency.

Nitric Oxide Accelerates Mycorrhizal Effects on Plant Growth and Root Development of Trifoliate Orange

Arbuscular mycorrhizal fungi (AMF) actively colonize plant roots and thus enhance plant growth through different mechanisms. In the present study, trifoliate orange (Poncirus trifoliata) seedlings inoculated with Diversispora versiformis were subjected to 0 and 0.2 mmol/L sodium nitroprusside (SNP, a nitric oxide donor) treatments. After eight weeks, exogenous SNP considerably increased root mycorrhizal colonization by 25%, showing a positive stimulating effect of NO on mycorrhizal formation. Mycorrhizal inoculation significantly increased plant growth performance (height, stem diameter, leaf number and shoot and root dry weight) and root traits (length, projected area, surface area, volume and number of 2nd and 3rd order lateral roots) than non-mycorrhizal treatment and NO (exogenous SNP treatment) heavily strengthened the mycorrhizal effects. Moreover, NO and mycorrhization induced more fine root (0-0.5 cm) formation. There was an opposite changed trend in root sucrose and leaf and root glucose contents by SNP in AMF versus non-AMF seedlings. All these results implied that NO plays important roles in mycorrhizal formation and development and also accelerates mycorrhizal effects on plant growth and root development of trifoliate orange.

Chloroplast protein PLGG1 is involved in abscisic acid-regulated lateral root development and stomatal movement in Arabidopsis

The plant hormone abscisic acid (ABA) plays a crucial role in root architecture; however, the molecular mechanism of ABA-regulated lateral root (LR) growth is not well known. We screened an Arabidopsis thaliana mutant with LR growth that was sensitive to ABA from a T-DNA insertion mutant library, which was an allelic mutant of plgg1-1, termed plgg1-2. PLGG1 encodes a chloroplast protein that transports plastidic glycolate and glycerate. The length and number of LRs at the root-hypocotyl junction of plgg1-1 and plgg1-2 were significantly impaired under exogenous ABA treatment, and the transgenic plant complementary lines of plgg1-2 restored LR growth in response to ABA. In addition, we found that PLGG1 is involved in other major ABA responses, including ABA-inhibited seed germination, ABA-mediated stomatal movement, and drought tolerance. These findings open new perspectives on elucidating the mechanism of ABA response, and provide clues for analysing the functions of chloroplast proteins in regulating root growth.

Effects of soil compaction on growth variables in Cappadocian maple (Acer cappadocicum) seedlings

This study investigates the effects of increasing soil penetration resistance (SPR) on seedling morphology and seedling architecture. When seedlings of deciduous Cappadocian maple (Acer cappadocicum Gled.) were grown in a greenhouse in a loamy soil under a wide range of soil compactions, all morphological variables studied changed significantly with increasing SPR. The relationships between increasing SPR and all morphological responses except lateral root length followed a negative quadratic curve. All biomass variables except lateral root biomass showed a bell-shaped response with respect to SPR, with a maximum biomass variable between 0.6 and 1.2 MPa, decreasing at higher soil compaction values. All allocation ratios were significantly affected by soil penetration resistance. Biomass allocation to roots was also affected by soil compaction. There was not a significant relationship between the specific stem length and increasing soil penetration resistance. The specific root length showed two trends to increasing SPR; it first decreased in response to the moderate compaction treatment (up to about 1.2 MPa), then increased significantly. We concluded that increasing soil compaction caused morphological changes to root and shoot sections of A. cappadocicum seedlings.

Hydrogen Gas Is Involved in Auxin-Induced Lateral Root Formation by Modulating Nitric Oxide Synthesis.

Metabolism of molecular hydrogen (H2) in bacteria and algae has been widely studied, and it has attracted increasing attention in the context of animals and plants. However, the role of endogenous H2 in lateral root (LR) formation is still unclear. Here, our results showed that H2-induced lateral root formation is a universal event. Naphthalene-1-acetic acid (NAA; the auxin analog) was able to trigger endogenous H2 production in tomato seedlings, and a contrasting response was observed in the presence of N-1-naphthyphthalamic acid (NPA), an auxin transport inhibitor. NPA-triggered the inhibition of H2 production and thereafter lateral root development was rescued by exogenously applied H2. Detection of endogenous nitric oxide (NO) by the specific probe 4-amino-5-methylamino-2′,7′-difluorofluorescein diacetate (DAF-FM DA) and electron paramagnetic resonance (EPR) analyses revealed that the NO level was increased in both NAA- and H2-treated tomato seedlings. Furthermore, NO production and thereafter LR formation induced by auxin and H2 were prevented by 2-4-carboxyphenyl-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO; a specific scavenger of NO) and the inhibitor of nitrate reductase (NR; an important NO synthetic enzyme). Molecular evidence confirmed that some representative NO-targeted cell cycle regulatory genes were also induced by H2, but was impaired by the removal of endogenous NO. Genetic evidence suggested that in the presence of H2, Arabidopsis mutants nia2 (in particular) and nia1 (two nitrate reductases (NR)-defective mutants) exhibited defects in lateral root length. Together, these results demonstrated that auxin-induced H2 production was associated with lateral root formation, at least partially via a NR-dependent NO synthesis.

Involvement of jasmonates in responses of sunflower (Helianthus annuus) seedlings to moderate water stress

Jasmonates (JAs), a type of phytohormone, are involved in sensing and signaling of several environmental stresses (biotic and abiotic). Jasmonic acid (JA) has been suggested to function in plant responses to drought, because this type of stress induces expression of several genes that also respond to JA. We investigated the involvement of JA and its precursor (12-oxo-phytodienoic acid; OPDA) on seedling morphological and physiological characteristics of two sunflower (Helianthus annuus) inbred lines with contrasting responses (sensitive vs. tolerant) to water stress. Our experimental treatments were based on moderate water stress (simulated by application of mannitol 400 mM) and on blocking of JA biosynthesis (by the chemical inhibitor salicylhydroxamic acid; SHAM). Water stress resulted in reduction of primary root (PR) growth and lateral root (LR) growth, but in increased LR number. SHAM treatment increased PR length, LR number, and LR length, thus strongly affecting root architecture. Water stress had differential effects on various physiological parameters, including relative water content (RWC), stomatal conductance, and content of photosynthetic pigments (chlorophylls, carotenoids). OPDA and JA accumulation in aerial part and roots induced by water stress was reversed by combined water stress plus SHAM treatment at day 14. Our findings suggest that SHAM effectively inhibits de novo JA biosynthesis induced by water stress, and that JAs play a protective role in responses of sunflower seedlings to this stress. JAs, particularly OPDA, are highly effective signaling molecules in mediation of sunflower seedling responses to water stress.

Is there genetic variation in mycorrhization of Medicago truncatula?

Differences in the plant’s response among ecotypes or accessions are often used to identify molecular markers for the respective process. In order to analyze genetic diversity of Medicago truncatula in respect to interaction with the arbuscular mycorrhizal (AM) fungus Rhizophagus irregularis, mycorrhizal colonization was evaluated in 32 lines of the nested core collection representing the genetic diversity of the SARDI collection. All studied lines and the reference line Jemalong A17 were inoculated with R. irregularis and the mycorrhization rate was determined at three time points after inoculation. There were, however, no reliable and consistent differences in mycorrhization rates among all lines. To circumvent possible overlay of potential differences by use of the highly effective inoculum, native sandy soil was used in an independent experiment. Here, significant differences in mycorrhization rates among few of the lines were detectable, but the overall high variability in the mycorrhization rate hindered clear conclusions. To narrow down the number of lines to be tested in more detail, root system architecture (RSA) of in vitro-grown seedlings of all lines under two different phosphate (Pi) supply condition was determined in terms of primary root length and number of lateral roots. Under high Pi supply (100 µM), only minor differences were observed, whereas in response to Pi-limitation (3 µM) several lines exhibited a drastically changed number of lateral roots. Five lines showing the highest alterations or deviations in RSA were selected and inoculated with R. irregularis using two different Pi-fertilization regimes with either 13 mM or 3 mM Pi. Mycorrhization rate of these lines was checked in detail by molecular markers, such as transcript levels of RiTubulin and MtPT4. Under high phosphate supply, the ecotypes L000368 and L000555 exhibited slightly increased fungal colonization and more functional arbuscules, respectively. To address the question, whether capability for mycorrhizal colonization might be correlated to general invasion by microorganisms, selected lines were checked for infection by the root rot causing pathogen, Aphanoymces euteiches. The mycorrhizal colonization phenotype, however, did not correlate with the resistance phenotype upon infection with two strains of A. euteiches as L000368 showed partial resistance and L000555 exhibited high susceptibility as determined by quantification of A. euteiches rRNA within infected roots. Although there is genetic diversity in respect to pathogen infection, genetic diversity in mycorrhizal colonization of M. truncatula is rather low and it will be rather difficult to use it as a trait to access genetic markers.