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Relationships between the development of adventitious roots and the biosynthesis of anthocyanins in first internodes of sorghum.

The initiation and subsequent growth of adventitious roots in excised first internodes of Sorghum vulgare var. Wheatland milo were studied to determine the effect of these processes on anthocyanin biosyntheses. Segmentation of the internodes inhibited both adventitious root growth and accumulation of cyanidin equally in all segments; these results can be interpreted as a common requirement for bidirectional longitudinal transport. The presence of the coleoptile, especially in the absence of the base of the internode, inhibited the growth of the roots, but increased the number of root initials. High intensities of white and blue light which induced cyanidin synthesis slightly decreased adventitious root growth. Anaerobic conditions produced by solution infiltration strongly inhibited the growth of adventitious roots and greatly increased the accumulation of apigeninidin and luteolinidin. Addition of indoleacetic acid, kinetin and cofactors such as pyridoxine produced effects on the initiation and subsequent growth of these roots similar to those effects reported in the literature. But unlike root formation in hypocotyls, the initiation of adventitious roots in Sorghum internodes was not always directly correlated with the accumulation of anthocyanins, and the subsequent growth of these roots was frequently inversely correlated with some of the anthocyanin biosyntheses. The possible nature of these correlations is discussed. Comparisons are made with related Sorghum lines and mutants.

Journal Article↗

Cytokinin receptors are required for normal development of auxin-transporting vascular tissues in the hypocotyl but not in adventitious roots.

Plants alter the architecture of their root systems to adapt to the environment by modulating post-embryonic (lateral and adventitious) root formation and growth. To understand better the genetic basis of this regulation, we screened ethylmethane sulfonate-mutagenized lines of Arabidopsis thaliana for adventitious rooting mutants. One mutant showed retardation of the primary root growth, no production of lateral roots and enhanced formation of adventitious roots. Mapping and genetic complementation revealed that this mutant named wooden leg-3 (wol-3) was an allele of ARABIDOPSIS HISTIDINE KINASE 4 (AHK4), a locus known to encode a cytokinin receptor. Although the vascular system of the primary root and hypocotyl in the wol-3 mutant was aborted, that of the adventitious roots was normally developed. In the hypocotyl of the wol-3 mutant, auxin signals accumulated around the aborted vascular system. The application of auxin to primary roots induced lateral root formation in the wol-3 mutant. Transport of radiolabeled auxin from the top of the hypocotyl to the primary root was inhibited in wol-3. Although only a single amino acid alteration had occurred in AHK4, the root morphology in the wol-3 mutant was quite similar to that in the ahk2 ahk3 ahk4 triple mutant, which is a loss-of-function mutant of the three cytokinin receptors. This implies that the functional disturbance of AHK4 affects the function of the other receptors. Our results suggest that cytokinin receptors are necessary for the formation of auxin-transporting vascular tissues in the hypocotyl, but not in adventitious roots.

Alleles↗

Dynamics of Aerenchyma distribution in the cortex of sulfate-deprived adventitious roots of maize.

BACKGROUND AND AIMS: Aerenchyma formation in maize adventitious roots is induced in nutrient solution by the deprivation of sulfate (S) under well-oxygenated conditions. The aim of this research was to examine the extent of aerenchyma formation in the cortex of sulfate-deprived adventitious roots along the root axis, in correlation with the presence of reactive oxygen species (ROS), calcium levels and pH of cortex cells and root lignification. METHODS: The morphometry of the second whorl of adventitious (W2) roots, subject to S-deprivation conditions throughout development, was recorded in terms of root length and lateral root length and distribution. W2 roots divided into sectors according to the mean length of lateral roots, and cross-sections of each were examined for aerenchyma. In-situ detection of alterations in ROS presence, calcium levels and pH were performed by means of fluorescence microscopy using H(2)DCF-DA, fluo-3AM and BCECF, respectively. Lignification was detected using the Wiesner test. KEY RESULTS: S-deprivation reduced shoot growth and enhanced root proliferation. Aerenchyma was found in the cortex of 77 % of the root length, particularly in the region of emerging or developing lateral roots. The basal and apical sectors had no aerenchyma and no aerenchyma connection was found with the shoot. S-deprivation resulted in alterations of ROS, calcium levels and pH in aerenchymatous sectors compared with the basal non-aerenchymatous region. Lignified epidermal layers were located at the basal and the proximal sectors. S-deprivation resulted in shorter lateral roots in the upper sectors and in a limited extension of the lignified layers towards the next lateral root carrying sector. CONCLUSIONS: Lateral root proliferation is accompanied by spatially localized induced cell death in the cortex of developing young maize adventitious roots during S-deprivation.

Calcium↗

Ethylene induces epidermal cell death at the site of adventitious root emergence in rice.

In deepwater rice (Oryza sativa), adventitious root primordia initiate at the nodes as part of normal development. Emergence of the roots is dependent on flooding of the plant and is mediated by ethylene action. Root growth was preceded by the induced death of epidermal cells of the node external to the tip of the root primordium. Cell death proceeded until the epidermis split open. Through this crack the root eventually emerged. Induced death was confined to nodal epidermal cells covering the tip of the primordia. Our results suggest that this process facilitates adventitious root emergence and prevents injury to the growing root. Cell death was inducible not only by submergence but also by application of 1-aminocyclopropane-1-carboxylic acid, the natural precursor of ethylene and it was suppressed in the presence of 2,5-norbornadiene (bicyclo[2.2.1]hepta-2,5-diene), an inhibitor of ethylene action. Adventitious root growth and epidermal cell death are therefore linked to the ethylene signaling pathway, which is activated in response to low oxygen stress.

Amino Acids, Cyclic↗

Partial flooding enhances aeration in adventitious roots of black willow (Salix nigra) cuttings.

Black willow (Salix nigra) cuttings are used for streambank stabilization where they are subjected to a range of soil moisture conditions including flooding. Flooding has been shown to adversely impact cutting performance, and improved understanding of natural adaptations to flooding might suggest handling and planting techniques to enhance success. However, data assessing the root aeration in adventitious roots that are developed on cuttings of woody species are scant. In addition, it appears that no data are available regarding aeration of the root system under partially flooded conditions. This experiment was designed to examine the effects of continuous flooding (CF) and partial flooding (PF) on aerenchyma formation and radial oxygen loss (ROL) in black willow cuttings. Photosynthetic and growth responses to these conditions were also investigated. Under laboratory condition, replicated potted cuttings were subjected to three treatments: no flooding (control, C), CF, and PF. Water was maintained above the soil surface in CF and at 10 cm depth in PF. Results indicated that after the 28-d treatments, root porosity ranged between 28.6% and 33.0% for the CF and C plants but was greater for the PF plants (39.2% for the drained and 37.2% for the flooded portions). A similar response pattern was found for ROL. In addition, CF treatment led to decreases in final root biomass and root/shoot ratio. Neither CF nor PF had any detectable adverse effects on plant gas exchange or photosystem II functioning. Our results indicated that S. nigra cuttings exhibited avoidance mechanisms in response to flooding, especially the partially flooded condition which is the most common occurrence in riparian systems.

Oxygen↗

Activity and isoforms of peroxidases, lignin and anatomy, during adventitious rooting in cuttings of Ebenus cretica L.

Adventitious rooting of Ebenus cretica cuttings was studied in order to examine a) the rooting ability of different genotypes in relation to electrophoretic patterns of peroxidases. b) the activity and electrophoretic patterns of soluble and wall ionically bound peroxidases, the lignin content and anatomical changes in the control and IBA treated cuttings of and genotypes in the course of adventitious root formation. In addition, a fraction of soluble cationic peroxidases was separated by gel filtration chromatography from the total soluble peroxidases of a genotype. No rooting occurred in cuttings without IBA-treatment. In both genotypes, electrophoretic patterns of soluble anionic peroxidases revealed two common peroxidase isoforms, while a fast-migrating anionic peroxidase isoform (A3) appeared only in genotypes. Both genotypes showed similar patterns of soluble, as well as wall ionically bound cationic peroxidase isoforms. The number of isoforms was unchanged during the rooting process (induction, initiation and expression phase) but an increase in peroxidase activity (initiation phase) followed by decrease has been found in IBA-treated cuttings. During initiation phase the lignin content was almost similar to that on day 0 in genotype while it was reduced at by about 50% in genotype at the respective time. Microscopic observations revealed anatomical differences between genotypes. According to this study, the and genotypes display differences in anatomy, lignin content, activity of soluble peroxidases and the electrophoretic patterns of soluble anionic peroxidase isoforms. The A3-anionic peroxidase isoform could be used as biochemical marker to distinguish and genotypes of E. cretica and seems to be correlated to lignin synthesis in rooting process.

Ebenaceae↗

[Ethylene-induced activation of xylanase in adventitious roots of maize as a response to the stress effect of root submersion].

Submersion of roots of ten-day-old maize (Zea mays L.) seedlings was accompanied by a decrease in pO2 and an increase in pCO2 of the medium adjacent to roots. These changes stimulated ethylene evolution in intact plants. Enhanced biosynthesis of ethylene was accompanied by xylanase activation in adventitious roots. As a result, an enhanced formation of aerenchyma was observed in the cortex of adventitious roots. Therefore, these processes resulted in the development of a ventilation system by which O2 can reach the root system exposed to hypoxia. The volume of aerenchyma was assessed by the volume of gas cavities (porosity). In contrast to the main root, the growth of adventitious roots was not inhibited under these conditions. Enlargement of the stem base and increase in the number of aerenchymatous adventitious roots facilitated the oxygen supply to submerged organs of plants.

Adaptation, Physiological↗

[Effects of mineral cations on the accumulation of tanshinone II A and protocatechuic aldehyde in the adventitious root culture of Salvia niltiorrhiza].

OBJECTIVE: To study the effects of mineral cations on the growth of Salvia niltiorrhiza adventitious roots and the accumulation of tanshinone II A and protocatechuic aldehyde. METHOD: The adventitious roots were cultured under different concentrations of metal cations and the contents of tanshinone II A and protocatechuic aldehyde were determined by HPLC. RESULT: Fe2 + and Mn2+ in MS medium are suitable for adventitious root growth, but Cu2+ and Mg2+ in high concentration could improve root proliferation, and Zn2+ has no obvious effect on root growth. As for tanshinone II A and protocatechuic aldehyde biosynthesis, Cu2+ and Zn2+ would inhibit the biosynthesis of protocatechuic aldehyde, furthermore, proper Fe2+ and Mg2+ concentration could advance the biosynthesis of protocatechuic aldehyde, whereas, Mn2+ in high concentration could accelerate the biosynthesis of protocatechuic aldehyde. Cu2+, Zn2+, Mg2+ in low concentration and Fe2+ and Mn2+ in high concentration would all advance the biosynthesis of tanshinone II A. CONCLUSION: The mineral cations have obvious effects on the secondary metabolites biosynthesis in adventitious root culture of S. niltiorrhiza.

Abietanes↗

Adventitious roots, leaf abscission and nutrient status of flooded Gmelina and Tectona seedlings.

When flooded, seedlings of Gmelina arborea Roxb. produced more adventitious roots, had lower foliar Mn concentrations and lost fewer leaves than seedlings of Tectona grandis L.f. Severing the adventitious roots produced by flooded Gmelina seedlings increased leaf Mn concentration and leaf abscission and reduced whole-plant dry matter production. Flooded Gmelina cuttings, which do not produce adventitious roots, abscised few leaves until foliar concentrations of Mn and Fe had risen substantially above those of unflooded cuttings, at which time most leaves were shed. The results indicate that the development of adventitious roots in flooded seedlings of Gmelina suppressed uptake of Mn thereby minimizing leaf abscission.

Journal Article↗

Nitric oxide mediates the indole acetic acid induction activation of a mitogen-activated protein kinase cascade involved in adventitious root development.

Recently, it was demonstrated that nitric oxide (NO) and cGMP are involved in the auxin response during the adventitious rooting process in cucumber (Cucumis sativus; Pagnussat et al., 2002, 2003). However, not much is known about the complex molecular network operating during the cell proliferation and morphogenesis triggered by auxins and NO in that process. Anatomical studies showed that formation of adventitious root primordia was clearly detected in indole acetic acid (IAA)- and NO-treated cucumber explants, while neither cell proliferation nor differentiation into root primordia could be observed in control explants 3 d after primary root was removed. In order to go further with signal transduction mechanisms that operate during IAA- and NO-induced adventitious root formation, experiments were designed to test the involvement of a mitogen-activated protein kinase (MAPK) cascade in that process. Cucumber explants were treated with the NO-donor sodium nitroprusside (SNP) or with SNP plus the specific NO-scavenger cPTIO. Protein extracts from those explants were assayed for protein kinase (PK) activity by using myelin basic protein (MBP) as substrate in both in vitro and in-gel assays. The activation of a PK of approximately 48 kD could be detected 1 d after NO treatment with a maximal activation after 3 d of treatment. In control explants, a PK activity was detected only after 4 d of treatment. The MBP-kinase activity was also detected in extracts from IAA-treated explants, while no signal was observed in IAA + cPTIO treatments. The PK activity could be inhibited by the cell-permeable MAPK kinase inhibitor PD098059, suggesting that the NO-dependent MBP-kinase activity is a MAPK. Furthermore, when PD098059 was administered to explants treated with SNP or IAA, it produced a delay in root emergence and a dose-dependent reduction in root number. Altogether, our results suggest that a MAPK signaling cascade is activated during the adventitious rooting process induced by IAA in a NO-mediated but cGMP-independent pathway. The activation of MAPKs is discussed in relation to the cell responses modulating mitotic process.

Cucumis sativus↗

[Effects of IAA and stimulated microgravity on formation of adventitious roots of Chinese cabbage].

Under the induction of indole-3-acetic acid (IAA), adventitious roots were differentiated on hypocotyl segments derived from seedlings of Chinese cabbage (Brassica campestris spp. pekinensis). IAA at concentration of 0.4-1.0 mg/L in solid MS medium incited many adventitious roots on hypocotyl segments. The earliest anatomic changes were observed on cut surface of hypocotyl segments under optical microscope 24 hours after IAA treatment: cytoplasmic and nuclear density became higher in a few of parenchytmatous cells adjacent to phloem in tissue of pericycle, followed by cell divisions. Lately, the dividing cells expanded and developed into root primordium from which root cap was differentiated. After five days, most roots protruded through hypocotyl cortex and appeared just below the cut surface. The rooting capacity of the segments derived from three regions of each hypocotyl was different. High level of IAA modified the polarity of root formation on segment inserted upside down and sucrose increased the function of IAA. Additionally, microgravity did not significantly change the rooting polarity under the condition of stimulated microgravity, but it increased the competence of explants to IAA treatment. The results presented here provided an experimental system for further investigation of molecular events associated with adventitious root initiation.

Brassica↗

Tropane alkaloids in adventitious root cultures of Physochlaina physaloides.

Adventitious root cultures of Physochlaina physaloides were established and their growth and tropane alkaloid production were investigated. Although the roots cultured in Murashige-skoog liquid medium containing 1.0-3.0 mg/l IBA grew well, the addition of lower concentrations of auxins was more suitable for alkaloid production. The best alkaloid yield was obtained with 0.01 mg/l NAA in 1/2 MS liquid medium. In addition, the alkaloid differences between the cultured roots and the regenerated plant cultivated in the soil was studied. Cultured roots in this condition showed higher level of alkaloids when compared to leaf, stem and roots from the regenerated plant (3 months in field conditions).

Culture Media↗

Two S-adenosylmethionine synthetase-encoding genes differentially expressed during adventitious root development in Pinus contorta.

Two S-adenosylmethionine synthetase (SAMS) cDNAs, PcSAMS1 and PcSAMS2, have been identified in Pinus contorta. We found that the two genes are differentially expressed during root development. Thus, PcSAMS1 is preferentially expressed in roots and exhibits a specific expression pattern in the meristem at the onset of adventitious root development, whereas PcSAMS2 is expressed in roots as well as in shoots and is down-regulated during adventitious root formation. The expression of the two SAMS genes is different from the SAMS activity levels during adventitious root formation. We conclude that other SAMS genes that remain to be characterized may contribute to the observed SAMS activity, or that the activities of PcSAMS1 and PcSAMS2 are affected by post-transcriptional regulation. The deduced amino acid sequences of PcSAMS1 and PcSAMS2 are highly divergent, suggesting different functional roles. However, both carry the two perfectly conserved motifs that are common to all plant SAMS. At the protein level, PcSAMS2 shares about 90% identity to other isolated eukaryotic SAMS, while PcSAMS1 shares less than 50% identity with other plant SAMS. In a phylogenetic comparison, PcSAMS1 seems to have diverged significantly from all other SAMS genes. Nevertheless, PcSAMS1 was able to complement a Saccharomyces cerevisiae sam1 sam2 double mutant, indicating that it encodes a functional SAMS enzyme.

Amino Acid Sequence↗

IAA synergism and vitamin B1 antagonism with calcium for induction and growth of adventitious roots in branch cuttings of Dalbergia sissoo Roxb.

IAA induced adventitious roots in branch cuttings of D. sissoo, whereas vit. B1 promoted growth of these roots. A strong IAA synergism and vit. B1 antagonism with calcium was observed in induction and growth of adventitious roots. Treatments with IAA (571 microM) and calcium (134 microM) proved to be an ideal combination, exhibiting 87% rooting and 0.87 R/S ratio in branch cuttings, coupled with an increase of root length, root number, root fresh weight and root dry weight as compared to control.

Calcium↗

Nuclear localization and interaction of RolB with plant 14-3-3 proteins correlates with induction of adventitious roots by the oncogene rolB.

The rooting-locus gene B (rolB) on the T-DNA of the root-inducing (Ri) plasmid in Agrobacterium rhizogenes is responsible for the induction of transformed adventitious roots, although the root induction mechanism is unknown. We report here that the RolB protein of pRi1724 (1724RolB) is associated with Nicotianatabacum14-3-3-like protein omegaII (Nt14-3-3 omegaII) in tobacco bright yellow (BY)-2 cells. Nt14-3-3 omegaII directly interacts with 1724RolB protein. Green fluorescent protein (GFP)-fused 1724RolB is localized to the nucleus. GFP-fused mutant 1724RolB proteins having a deletion or amino acid substitution are unable to interact with Nt14-3-3 omegaII and also show impaired nuclear localization. Moreover, these 1724RolB mutants show decreased capacity for adventitious root induction. These results suggest that adventitious root induction by 1724RolB protein correlates with its interaction with Nt14-3-3 omegaII and the nuclear localization of 1724RolB protein.

14-3-3 Proteins↗

A PIN1 family gene, OsPIN1, involved in auxin-dependent adventitious root emergence and tillering in rice.

Auxin transport affects a variety of important growth and developmental processes in plants, including the regulation of shoot and root branching. The asymmetrical localization of auxin influx and efflux carriers within the plasma membrane establishes the auxin gradient and facilitates its transport. REH1, a rice EIR1 (Arabidopsis ethylene insensitive root 1)-like gene, is a putative auxin efflux carrier. Phylogenetic analysis of 32 members of the PIN family, taken from across different species, showed that in terms of evolutionary relationship, OsPIN1 is closer to the PIN1 family than to the PIN2 family. It is, therefore, renamed as OsPIN1 in this study. OsPIN1 was expressed in the vascular tissues and root primordial in a manner similar to AtPIN1. Adventitious root emergence and development were significantly inhibited in the OsPIN1 RNA interference (RNAi) transgenic plants, which was similar to the phenotype of NPA (N-1-naphthylphalamic acid, an auxin-transport inhibitor)-treated wild-type plants. alpha-naphthylacetic acid (alpha-NAA) treatment was able to rescue the mutated phenotypes occurring in the RNAi plants. Overexpression or suppression of the OsPIN1 expression through a transgenic approach resulted in changes of tiller numbers and shoot/root ratio. Taken together, these data suggest that OsPIN1 plays an important role in auxin-dependent adventitious root emergence and tillering.

Base Sequence↗

The CsTBH-CsROP2 Module Regulates Waterlogging Tolerance via Auxin-Mediated Adventitious Root Formation in Cucumber.

Cucumber (Cucumis sativus L.) requires frequent irrigation due to its shallow root system and high transpiration rate of the aboveground parts. However, it is also prone to waterlogging damage. Therefore, understanding its response to waterlogging is crucial for breeding waterlogging-tolerant varieties. Although Rho of Plants GTPases play well-established roles in regulating development and stress signalling, their functions in plant adaptation to waterlogging stress has yet to be fully elucidated. Here, we identified nine CsROP genes in the cucumber genome, which exhibit evolutionary diversification but retain conserved functional domains. Functional analysis revealed that CsROP2 acts as a negative regulator of adventitious root formation. It modulates auxin accumulation in hypocotyl vascular bundles, thereby suppressing adventitious root development and enhancing waterlogging sensitivity. The HD-Zip I transcription factor CsTBH directly binds the CsROP2 promoter and activates its expression. Our study uncovers a CsTBH-CsROP2 module that governs adventitious rooting and waterlogging tolerance by modulating auxin homeostasis. These findings provide new insights into the crosstalk between developmental programmes and stress signalling pathways and offer potential genetic targets for improving stress resilience in cucumber and other crops.

CsROP2↗

Comparative study of cellulases associated with adventitious root initiation, apical buds, and leaf, flower, and pod abscission zones in soybean.

Cellulase activity was measured in soybean (Glycine max) leaf abscission zones, flower abscission zones, pod abscission zones, apical buds, and adventitious rooting hypocotyls. Immunoprecipitation data showed that a cellulase immunologically similar to the bean abscission cellulase (isoelectric point 9.5) is present in soybean leaf, flower, and pod abscission zones, but is not present in soybean apical buds or rooting hypocotyls. cDNA and genomic clones for two different soybean genes were identified and show sequence similarity with the bean abscission cellulase clone pBAC10. The cDNA clone pSAC1, isolated from a soybean abscission cDNA library, hybridized to transcripts in soybean leaf, flower, and pod abscission zones. Although ethylene has been shown to play a role in the increase in cellulase activity associated with both abscission and adventitious root initiation, no signal was seen for hybridization of the soybean abscission cellulase clone, pSAC1, to RNA from soybean adventitious rooting hypocotyls. In addition, no soybean abscission cellulase transcripts were detected in apical buds. Transcripts for a second soybean cellulase gene (SC2) were not detected in any of the tissues surveyed.

Amino Acid Sequence↗