Different Ways of Formation of Root System in Plants,Maize and Flax Seedlings Taken as Examples

Comparison of the appearance and development of lateral roots in the flax and maize seedlings has shown the way of root branching in the flax, as distinct from that in most plants. Some primordia in the flax main root did not develop immediately into lateral roots, but remained quiescent, which determines different reactions of the maize and flax root systems to experimental influences. Decapitation of the main root in the maize did not leads to a significant increase in the number of lateral roots, while in the flax, their number noticeably increased due to the development of previously quiescent primordia into lateral roots. The treatment with synthetic auxin did not induce the formation of additional primordia and lateral roots in the maize roots. In the flax, the number of primordia increased significantly and that of lateral roots increased to a somewhat lesser extent. Apparently, the development of a primordium into a lateral root proceeds in two stages and they have different regulation.     

Different methods of forming root systems in plants using maize and flax seedlings as examples

Comparison of the appearance and development of lateral roots in the flax and maize seedlings has shown the way of root branching in the flax, as distinct from that in most plants. Some primordia in the flax main root did not develop immediately into lateral roots, but remained quiescent, which determines different reactions of the maize and flax root systems to experimental influences. Decapitation of the main root in the maize did not leads to a significant increase in the number of lateral roots, while in the flax, their number noticeably increased due to the development of previously quiescent primordia into lateral roots. The treatment with synthetic auxin did not induce the formation of additional primordia and lateral roots in the maize roots. In the flax, the number of primordia increased significantly and that of lateral roots increased to a somewhat lesser extent. Apparently, the development of a primordium into a lateral root proceeds in two stages and they have different regulation.     

Analysis of possible mechanisms of regulation of root branching

The formation of lateral roots under the influence of growth inhibitors and phytohormons were studied with germs of corn and flax. All corn primordiums develop into the lateral roots without dormancy period, while some flax primordiums are delayed in development. Removal of root apical meristem (decapitation) does not result in the increase in number of lateral roots of corn germs. Decapitation of flax germs induces the development of some dormant primordiums, that leads to the considerable increase in number of lateral roots. Formation of additional primordium under decapitation does not take place. Auxin stimulates the formation of additional primordiums in flax, but does not effect the number of mature lateral roots. In corn germs synthetic auxin stimulates the development of additional primordiums in root zone, where primordiums are forming after the disturbance. It is shown that the time of primordium development does not change under inhibitor influence, if they do not selectively effect mitoses. It seems that auxins regulate initiation of pericycle division and formation of primordiums of lateral roots on the stage of development of meristematic protuberance. Cytokinins influence the development of already existed primordiums of lateral roots but do not effect the formation of new ones. Constancy of time of primordium formation and independence of their development on the state of the main root need future investigations. The authors suppose that regulation of these processes is non-hormonal.     

Ethylene Interacts with Auxin in Regulating Developmental Attenuation of Gravitropism in Flax Root

Previous research shows that gravity-sensing in flax (Linum usitatissimum) root is initiated during seed imbibition and precedes root emergence. In this study we investigated the developmental attenuation of flax root gravitropism post-germination and the involvement of ethylene. Gravity response deteriorated significantly from 3 to 11?h after root emergence, which occurred at around 19?h after imbibition (that is, from “age” 22 to 30?h). Although the root elongation rate increased from 22 to 30?h, the gravitropic curving rate declined steadily. Older roots were able to tolerate higher levels of exogenous IAA before inhibition of elongation and gravitropism occurred. The age-dependent effect of IAA on root growth and gravitropism suggests that young roots are more sensitive to auxin and respond to a smaller vertical auxin gradient than older roots upon horizontal gravistimulation. The ethylene synthesis inhibitor AVG (2-aminoethoxyvinyl glycine, 10?μM) or ethylene action inhibitor Ag⁺ (10?μM) stimulated gravitropic curvature of 30?h roots by 24 and 32%, respectively, but had no effect on 22?h roots, suggesting that as roots age, ethylene begins to play a role in root gravitropism. The auxin transport inhibitor NPA (N-naphthylphthalamic acid, 50?μM) reduced gravitropic curvature of 30?h roots by 24% but had no effect on 22?h roots. On the other hand, treating roots simultaneously with the auxin transport inhibitor and ethylene synthesis or action inhibitor stimulated gravitropic curvature of 30?h roots but not 22?h roots. Taken together, these data indicate that as roots develop, their weakened gravity response is due to decreased auxin sensitivity and possibly auxin transport regulated by ethylene.     

Boron deficiency increases putrescine levels in tobacco plants

Polyamine concentrations were determined in leaves and roots of tobacco plants (Nicotiana tabacum L.) subjected to a short-term boron deficiency. A decrease in the growth of shoots and, especially, roots was found under this mineral deficiency. Boron deficiency did not lead to a significant decrease in leaf or root ion concentrations when compared to control treatment; however, as expected, leaf boron concentration was lower in boron-deficient plants in comparison to the control. In leaves, the levels of free putrescine and spermidine were similar in both treatments. In roots, a short-term boron deficiency caused an increase in free putrescine. Moreover, boron-deficient plants had higher conjugated polyamine concentration than boron-sufficient plants, which was especially evident for conjugated putrescine in leaves. A possible link between boron and polyamine levels is proposed and discussed.     

Influence of indole-butyric acid, boron, myo-inositol, vitamin D2 and seedling age on adventitious root development in cuttings of Phaseolus aureus

Adventitious roots develop in stem cuttings of Phaseolus aureus Roxb. seedlings when treatment with indole-butyric acid (IBA) is followed by treatment with boron. Root development varies according to the age of seedlings from which cuttings are taken. Increased root number is associated with expansion of the first leaf pair but subsequently declines, whereas root growth increases with increasing seedling age. Removal of leaves furing the first 72 h of treatment impairs root initiation whereas root growth is diminished by removal of leaves at any time during the first 120 h of treatment. IBA stimulates movement of ¹⁴C-IAA out of leaves. Vitamin D₂ and myo-inositol stimulate rooting of intact cuttings provided cuttings are subsequently supplied with boron. Hypocotyls excised from cuttings pretreated with IBA develop roots in response to myo-inositol in the absence of boron. It is proposed that endogenous auxin, arising in the leaves, and myo-inositol have roles in root initiation whilst the role of boron is suggested as one of initiating or maintaining transport from the leaves.     

Effect of Main Root Decapitation on Root Branching in Flax Seedlings

Root branching patterns in intact and decapitated flax (Linum usitatissimumL.) roots were compared. The number of initiated primordia in the control and decapitated roots was similar, but decapitated roots produced an increased number of lateral roots owing to an increase in the number of primordia developed into the laterals. It is suggested that the apical meristem influences lateral root development only at the stage of root emergence from the parent root.     

Autoradiographic Examination of Meristems of Intact Boron-deficient Squash Roots Treated with Tritiated Thymidine

Intact roots of boron-sufficient squash (Cucurbita pepo L.) plants, plants entering boron deficiency, and plants recovering from boron deficiency were exposed to tritiated thymidine at the end of the treatment period to label the replicating DNA of root tip cells. Using histological sections, autoradiographs of intact root meristems were prepared. The labeling pattern in +B root tips revealed the presence of a well defined quiescent center. The ability of root tip cells to incorporate label is correlated with the total root elongation during the −B treatment period. A greater amount of total root elongation during boron deficiency and recovery reflects the fact that root tip cells have retained their ability to synthesize DNA and enter mitosis for a longer time. In roots recovering from boron deficiency, cells of the quiescent center were seen to play no part in the recovery process in roots treated for as long as 20 hours in a −B nutrient solution. They were inactive before, during, and after the −B treatment. Cessation of mitosis occurs as early as 6.5 hours after boron is withheld from the nutrient solution while DNA synthesis can occur for as long as 20 hours after withholding boron. It was concluded that boron is essential for continued DNA synthesis and mitotic activity. The absence of boron results in the cessation of mitosis and DNA synthesis within 20 hours from the time boron is withheld.     

Role of manganese in low-pH-induced root hair formation in<Emphasis Type="Italic"> Lactuca sativa</Emphasis> cv. Grand Rapids seedlings

Root hair formation is induced by low pH in lettuce (Lactuca sativa L. cv. Grand Rapids) seedlings cultured in mineral medium. The role of mineral concentrations in this phenomenon was investigated, especially for manganese. When lettuce seedlings were cultured in media that were deficient in calcium (Ca), manganese (Mn), boron (B) or molybdenum (Mo), morphological changes were induced in roots. Deficiency of other nutrients had little effect on root hair formation. Ca or B deficiency inhibited the growth of the main root and the formation of root hairs, regardless of pH. Mn or Mo deficiency increased root hair formation at pH 6 and suppressed main root growth slightly. In contrast, increasing the Mn concentration suppressed low-pH-induced root hair formation. The Mn content of roots grown at pH 4 was only about 15% of that at pH 6. In contrast, the Mo content of roots grown at low pH was about six times that of roots grown at neutral pH. These results suggest that root hair formation induced by low pH is at least partly mediated by decreased Mn uptake in root cells.     

黄瓜根尖分生组织细胞核仁超微结构类型及缺硼时的变化

硼是植物必需的一种微量元素,缺乏时植物根的生长被抑制。黄瓜根尖分生组织细胞核中核仁有四种类型。不缺硼的核仁中,丝状成分、粒状成分、丝状中心及核仁液泡能够明显地看到。缺硼的核仁中,丝状成分萎缩,变得粗短。看不见丝状中心、核仁液泡等。环形核仁中的中央大液泡变小。     

EFFECTS OF BORON DEFICIENCY ON MITOSIS AND INCORPORATION OF TRITIATED THYMIDINE INTO NUCLEI OF SUNFLOWER ROOT TIPS

Boron deprivation has multiple effects upon root growth within 6 hr after this essential micronutrient is withheld. Root elongation is inhibited and this response has been attributed to a cessation of mitosis and DNA synthesis. Our preliminary results using an autoradiographic analysis of sunflower roots labeled with [³H]-thymidine demonstrated no difference in label distribution between +/-B root tips. We found that mitosis in inhibited in -B roots but does not completely cease. Scintillation counting of whole root tips shows that boron-deficient roots up to 72 hr of treatment incorporate radioactive label at a level comparable to that of the controls. Because mitosis and presumably DNA synthesis are affected by prolonged boron deficiency, these results may be brought about by a change in membrane integrity or permeability. We propose that effects of boron deprivation on DNA synthesis and mitosis in sunflower are secondary and that primary events involve alterations in cellular membranes.     

Identification of boron transporter genes likely to be responsible for tolerance to boron toxicity in wheat and barley

Tolerance to boron (B) toxicity in cereals is known to be associated with reduced tissue accumulation of B. Genes from roots of B-tolerant cultivars of wheat and barley with high similarities to previously reported B efflux transporters from Arabidopsis and rice were cloned. Expression of these genes was strongly correlated with the ability of tolerant genotypes to lower the concentration of B in roots. The gene from barley located to chromosome 4. Backcross lines containing a B tolerance locus on chromosome 4 showed tolerance in proportion to the level of expression of the transporter gene, whereas those lacking the locus were sensitive to B and had very low levels of gene expression. The results are consistent with a widespread mechanism of tolerance to high B based on efflux of B from root cells.     

Short-term boron deprivation enhances levels of cytoskeletal proteins in maize, but not zucchini, root apices

By using indirect immunofluorescence microscopy and Western blot analysis, we have demonstrated increased levels of actin and tubulin proteins as well as an altered polymerization pattern of their cytoskeletal assemblies in maize, but not zucchini root apices, as a response to early boron deprivation. Northern blot analysis, however, did not show significant increases in the amount of steady-state mRNAs of actin and tubulin. This finding indicates that these rapid cytoskeletal responses to boron removal are very likely regulated at translational/post-translational levels. Interestingly, these increased levels of cytoskeletal proteins coincided well with a reduction in the water-extractable, but not with the cell wall-bound, fraction of boron. This implicates that free boric acid, or other more labile boron complexes, might be involved in the activation of cytoskeletal responses in maize root apices. In fact, our experimental approach revealed that maize was suffering from boron deprivation as early as zucchini. This was evidenced by its slightly reduced root elongation rate recorded within 3–5 h of boron deprivation. Importantly, however, maize roots can recover from this early inhibition indicating an effective adaptation mechanism. In contrast, zucchini roots apparently lack this boron-deprivation response pathway and suffer extensively when exposed to boron-free environment. This leads to the tentative working hypothesis for an adaptive mechanism of maize roots to boron deprivation by enhancing its cytoskeletal protein levels and altering their polymerization patterns in order to mechanically reinforce the cell periphery complex of their cells. This testable hypothesis requires further experimental verification.     

Effects of Auxins and other Factors on the Rooting of Pinus contorta Dougl. Cuttings

The effects are investigated of applications of NAA and IBAupon root number, root type and time to form first roots incuttings of Pinus contorta Dougl. Cuttings treated with NAAgenerally formed thick, fleshy, unbranched roots, while thosereceiving IBA produced a thin, finely-divided root system. Bothtypes of roots developed when the two hormones were appliedtogether. The time taken for first roots to form, and the finalnumber of rooted cuttings were also greatly increased when hormoneswere applied in the range 1–10 µg per cutting. Boricacid applied in combination with hormone treatments increasedrooting, but boron alone was ineffective. There were markeddifferences between clones in their rooting responses to bothhormone and boron treatments.     

Flax (Linum usitatissimum L.) depends on arbuscular mycorrhizal fungi for growth and P uptake at intermediate but not high soil P levels in the field

The contribution of indigenous arbuscular mycorrhizal fungi (AMF) to growth and phosphorus (P) uptake by oilseed flax (Linum usitatissimum L.) was examined in two field experiments covering soil P levels from 20–86 mg kg-1 NaHCO3-extractable P. The fumigant dazomet was applied to the soil in half of the plots to obtain control plants with reduced mycorrhiza formation. An extensive AMF colonization of up to 48% of the root length was established in untreated soil of both experiments, although P fertilization reduced colonization to 28–39% at the latest harvests. Fumigation markedly decreased or totally prevented AMF colonization throughout the experiments. Root growth responded to fumigation by increased total and specific root length. Shoot P uptake was decreased by fumigation at soil P levels lower than ca. 50 mg kg-1 whereas shoot growth was reduced by fumigation at soil P levels lower than ca. 40 mg kg-1. The effects of fumigation were ascribed to the suppression of mycorrhiza formation. The effect of the AMF increased with decreasing soil P levels. Phosphorus inflow through roots (based on shoot P uptake) was reduced more strongly by fumigation than total P uptake. The P inflow through fungal tissue in roots was estimated to 4 × 10-14 mol P cm-1 s-1. We conclude that AMF are essential to flax growth at soil P levels below ca. 40 mg P kg-1, which is representative of the conditions under which most flax is grown.     

Wheat root colonization by Azospirillum brasilense strains with different motility

Migration of associative bacteria Azospirillum brasilense in semisolid media is performed mainly by swarming (Swa⁺ phenotype), which depends on the flagellar functioning and intercellular contacts. Non-swarming mutants of A. brasilense Sp245 lacking a polar flagellum migrate in semisolid media with microcolony formation using a unrevealed mechanism (Gri⁺ phenotype). The study of wheat root colonization dynamics demonstrated that A. brasilense Sp245 Gri⁺ mutants exhibited lower capacity for wheat root adsorption. However, after “anchoring” has occurred, both A. brasilense Sp245 and its Swa-Gri⁺ mutants colonized the growing roots with virtually the same efficiency. All strains under study formed microcolonies on the surface of roots, stimulated root branching, and exhibited changes in the composition of protein antigens exposed on the bacterial cell surface. Indirect evidence was obtained for enhanced production of genus-specific protein antigens in the process of A. brasilense Sp245 adaptation to growth on plant roots.     

Is Fe deficiency rather than P deficiency the cause of cluster root formation in Casuarina species?

When subjected, directly (through nutritional deficiencies) or indirectly (through alkaline constraints leading to such deficiencies) to nutrient deficiencies, certain plants respond by developing special root structures called cluster roots. This phenomenon can be considered as an ecophysiological response to a specific nutrient deficiency enabling plants to enhance nutrient uptake. Experiments conducted on an alkaline and an acid soil showed that Casuarina glauca (Sieber ex Spreng.) produced cluster roots only in the alkaline soil and not in the acid soil. In addition, iron (Fe) and phosphorus (P) deficiencies were examined separately or together to determine their effect on cluster root formation in C. glauca seedlings grown hydroponically. Results from experiments carried out on three Casuarina species (C. glauca, C. cunninghamiana Miq. and C. equisetifolia L.) indicated that Fe is involved in cluster root formation. In nutrient media lacking P but containing Fe, no cluster roots formed while seedlings receiving P and lacking Fe developed cluster roots. When incubated on chrome-azurol S-agar on blue plates (CAS assay), a technique used routinely to detect the production of siderophores by micro-organisms, the root system of Fe-deficient plants exhibited orange halos around cluster roots, indicating production of a ferric-chelating agent. It is concluded that the capacity of cluster roots of C. glauca to chelate Fe allows the plant to grow normally on alkaline soils.     

Influence of Two Plant Species (Flax and Tomato) on the Distribution of Nitrogen Dissimilative Abilities within Fluorescent Pseudomonas spp

The distribution of nitrogen-dissimilative abilities among 317 isolates of fluorescent pseudomonads was studied. These strains were isolated from an uncultivated soil and from the rhizosphere, rhizoplane, and root tissue of two plant species (flax and tomato) cultivated on this same soil. The isolates were distributed into two species, Pseudomonas fluorescens (45.1%) and Pseudomonas putida (40.4%), plus an intermediate type (14.5%). P. fluorescens was the species with the greatest proportion of isolates in the root compartments and the greatest proportion of dissimilatory and denitrifying strains. According to their ability to dissimilate nitrogen, the isolates have been distributed into nondissimilatory and dissimilatory strains, nitrate reducers and true denitrifiers with or without N(inf2)O reductase. The proportion of dissimilatory isolates was significantly enhanced in the compartments affected by flax and tomato roots (55% in uncultivated soil and 90 and 82% in the root tissue of flax and tomato, respectively). Among these strains, the proportion of denitrifiers gradually and significantly increased in the root vicinity of tomato (44, 68, 75, and 94% in uncultivated soil, rhizosphere, rhizoplane, and root tissue, respectively) and was higher in the flax rhizoplane (66%) than in the uncultivated soil. A higher proportion of N(inf2)O reducers was also found in the root compartments. This result was particularly clear for tomato. It is hypothesized that denitrification could be a selective advantage for the denitrifiers in the root environment and that this process could contribute to modify the specific composition of the bacterial communities in the rhizosphere.