Factors affecting the establishment and maintenance of ‘Titan’ red raspberry root organ cultures

Root organ cultures of red raspberry (Rubus ideaus cv. Titan) were established from two cm long terminal segments of adventitious roots induced in vitro on micropropagated shoot cultures. Growth of root organ cultures was almost entirely by initiation and elongation of lateral roots rather than by terminal elongation from the apical meristem of the original root explants. Indolebutyric acid (IBA) was required for lateral root initiation and elongation. The optimal IBA concentration for lateral root initiation decreased from 0.5 to 0.1 mg/L from the first to the fifth passage in culture. Two cm root explants initiated more lateral roots per unit length than four cm explants. Liquid Anderson's medium was superior to other basal nutrient and vitamin formulations tested. Root growth in liquid culture was stimulated by aeration. Root organ cultures also grew on media solidified with gelrite or agar. A rapid transfer technique was developed for subculturing these which involved cutting and transferring 1 cm discs of roots and underlying medium. Adventitious bud formation occurred spontaneously, but sporadically in liquid cultures, and was not influenced by cytokinin, auxin (spat) light, or chilling.     

The contribution of different regions of the seminal roots of wheat to uptake of nitrate from soil

A technique was developed to determine the physiological activity of defined sections of seminal roots of wheat grown in sand. Wheat plants were grown for 2 weeks in narrow columns of N-deficient sand to which all other nutrients had been added. The columns were split longitudinally and ¹⁵N-labelled nitrate, in an agar medium, supplied to 2 cm sections of root. Shoots and roots were analysed after 24 h to determine the uptake of ¹⁵N. Three sections were examined on either the secondary or tertiary seminal root: 1 cm from the seed (basal segment), 35 cm from the seed (middle segment) and 4 cm from the root apex (apical segment). Total uptake was greatest from the basal and middle segments, declining by 50% from the apical segment. However, uptake per unit root length, including exposed sections of lateral roots, was not significantly different along the root.     

Differences in root and shoot development of tomato (Lycopersicon esculentum L.) varieties across contrasting soil environments

Summary The root and shoot growth of three tomato (Lycopersicon esculentum L.) varieties, VF-10, VF-1908 and Paste 56 were compared in either a river-washed sand or an alluvial silty loam, with or without added nutrients, to determine factors influencing root growth and development. VF-10 had consistently higher shoot weights than the other two varieties, particularly by 45 days from germination. Although the root-to-shoot ratio varied with soil treatment, no significant differences in this ratio occurred among varieties. The roots were characterised by a taproot, thickened in the uppermost 3 to 5 cm, with 86 to 190 secondary roots when harvested 30 days after germination. The variety VF-1908 had a higher proportion of fine roots than the other two varieties over all treatments. On the basis of root weight and length, VF-1908 was also more stable across the imposed environmental gradient than the varieties VF-10 or Paste 56. This arose from less variability across soil treatments in secondary branching in VF-1908; over the four soil treatments the number of secondary roots varied from 104 to 131 in VF-1908 compared to 86 to 139 in VF-10 and 95 to 190 in Paste 56. VF-1908 and Paste 56 also had less variation in tertiary branching than VF-10, which had a consistently greater number of major tertiary roots than the other two varieties over all soil treatments. With the exception of Paste 56 in the fertilized silt, shoot growth was correlated with root length.     

Periodicity in the development of the root system of young rubber trees (Hevea brasiliensis Müell. Arg.): relationship with shoot development

The growth pattern of the root system of young rubber trees (Hevea brasiliensis) was studied in relation to shoot development over a period of 3 months. Temporal and spatial variations in elongation and branching processes were examined for the different root types, by means of root observation boxes. Shoot growth was typically rhythmic. Root development was periodic and related to leaf expansion. Root elongation was depressed during leaf growth, whereas branching was enhanced. Consequently, highly branched areas with vigorous secondary roots alternated along the taproot with poorly branched areas with shorter roots. Root types were not affected to the same degree by shoot competition: during leaf expansion, taproot growth was just depressed but remained continuous, the emergence and elongation rates of secondary roots were significantly affected and the elongation rates of tertiary roots fell to zero. These results were consistent with the hypothesis that root growth is related to competition for assimilates and to the sink strength of the different root types, whereas root branching appeared to be promoted by leaf development.     

Optimization of culture conditions for in vitro rooting of argan (Argania spinosa L.)]

The root system produced of in vitro organ plantlets is of poor quality and not efficient for the transfer to out-door conditions. To overcome such problems, experimentation was undertaken where the effects of growth regulators, nitrogen, sugar, activated charcoal and coconut fiber were tested on root induction and elongation. Modified Murashige and Skoog with half strength salt was used as a basal medium. Root induction (85%) with a mean of 16 roots per explant was obtained when shoots were grown, under dark conditions for 14 days, with a combination of two auxins (IBA and NNA), added at equal concentrations (5 mg.L-1). Secondary roots, 10 cm long, were initiated in 12% of the cultures in presence of 5 g.L-1 activated charcoal. Further improvements in the growth of the primary and secondary roots were obtained when semi-solid medium was substituted with a substrate composed of coconut fibers (80 g) mixed with semi-solid medium (35 mL) and agar (2.5 g.L-1).     

Why are laterals less affected than main axes by homogeneous unfavourable physical conditions? A model-based hypothesis

When plants develop in strong soils, growth of the root system is generally depressed. However, branching and elongation of branches are often less affected than growth of the main axes, whenever the whole root system encounters even-impeded conditions. On the basis of a model simulating root growth and architecture as related to assimilate availability, we propose a simple hypothesis to explain such behaviour. In the model, growth of each root depends on its own elongation potential, which is estimated by its apical diameter. The potential elongation rate–apical diameter relationship is the same for all the roots of the system and is described by a monomolecular function. Our hypothesis is that the effect of soil strength can be simulated by introducing an impedance factor in the definition of root maximum potential elongation rate, common to the whole root system. When such impedance factor is applied, it affects more the potential of larger roots (main axes) than that of thinner roots (secondary and tertiary branches). Simulations provided in high impedance conditions led to root systems characterised by short taproots, whereas growth of secondary roots was unaffected and growth of tertiary roots was enhanced. Actual branching density was also higher, although branching rules have been unchanged. Such simulated systems where similar to that observed in strong soils. Friction laws or pore size can be involved in the larger reduction of the potential growth of main axes. Moreover, when growth of main axes is restricted, assimilate availability becomes higher for branches and that could explain that their growth could be increased in a homogeneous strong soil. This revised version was published online in June 2006 with corrections to the Cover Date.     

THE ISOLATION AND CULTURE IN VITRO OF THE QUIESCENT CENTER OF ZEA MAYS

Using 3-day-old seedling roots of Zea mays L., cv. Kelvedon 33, it was possible to remove the root cap by a simple surgical manipulation without damage to the root proper. By a further small cut, the quiescent center (QC) itself was isolated. This double-convex lens-shaped tissue piece 100 X 250 μm is composed of 1000–1500 cells representing only 0.25 mm³ in volume. The explant was demonstrated unequivocally by ³H-thymidine incorporation before excision and then by autoradiography to be composed of the specific cells usually designated the quiescent center. Using sterile techniques, the QC's were placed on nutrient agar slants and allowed to grow in culture. Of a number of nutrient media tested, only a medium supplemented with organic nitrogen components, indoleacetic acid, kinetin and inorganic nutrients plus sucrose (S2M + K -2,4-D) was effective in eliciting development. Thirty to 40 percent of the 150 isolated QC's grown on this medium formed elongated roots, up to 2 cm in length in 3–4 weeks. Roots developing on agar medium showed in their proximal portion a vascular pattern with 5–6 metaxylem elements or variations of this pattern, but as the root elongated, the vascular pattern was progressively reduced in complexity at the more distal end to a small central group of metaxylem elements. When agar-grown roots were transferred after one week in culture to a liquid nutrient medium of the same composition, the initially reduced vascular pattern evident in the proximal tissues became progressively more complex in the distal portion of the root and after 2 cm of elongation, showed an essentially normal primary vascular tissue pattern characteristic of the seedling root.     

Root elongation and branching is related to local hexose concentration in Arabidopsis thaliana seedlings

Root architecture can be profoundly affected by the carbon availability in the plant. We hypothesized that this effect could be mediated by the carbon status of root cells involved in elongation and branching processes. Arabidopsis thaliana plants were grown at several photosynthetic photon flux densities (PPFD) and were supplied with various sucrose concentrations in the root medium. Hexose and sucrose concentration was estimated in individual roots in the apical growing region of the primary root and of secondary roots as well as in the zone of primordia development. Local sugar concentration was high in fast‐growing and in highly branched roots and robust relationships between root elongation rate or branching and hexose concentration (but not sucrose) were found that were common to all situations experienced. Moreover, these relationships accounted for the plant‐to‐plant variability within a treatment as well as for the variability among individual secondary roots within a plant. These results support the view that local hexose concentration integrates changes in carbon availability from several sources and acts as a signal to induce at least part of the response of the root architecture to the environment.     

An in vitro microplant bioassay using clonal white ash to test for tall fescue allelopathy

Axillary shoots from three selected white ash (Fraxinus americana L.) clones were harvested from in vitro shoot cultures. Roots were initiated by pulsing excised shoots for eight days in the dark in MS medium supplemented with 2% sucrose, 0.7% agar, 5 M NAA, and 1 M IBA. Pulsed shoots were transferred to a root elongation medium consisting of 25% MS macrosalts, full-strength microsalts and organics, 1% sucrose, 0.7% agar and no auxins. When roots were visible (6–10 days after transfer to root elongation medium), microplants were transferred to vessels containing the same minimal medium and tall fescue (Festuca elatior var. arundinacea (Schreb.) Wimm.) leaf extracts, leaf leachates, or soil leachates from plant boxes with and without tall fescue sod. After four weeks in vitro, primary adventitious and secondary root growth was reduced by extracts obtained from 5 and 10 g ground leaves per 100 ml of medium. Leachates obtained from 5 g soaked leaves per 100 ml of medium stimulated primary root growth. Soil leachates from bare soil also stimulated primary root growth. Variation was observed among the clones for root growth when plantlets were grown in extracts or leachates from tall fescue.     

Patterns of variability in the diameter of lateral roots in the banana root system

The relative importance of root system structure, plant carbon status and soil environment in the determination of lateral root diameter remains unclear, and was investigated in this study. Banana (Musa acuminata) plants were grown at various moderate levels of soil compaction in two distinct experiments, in a field experiment (FE) and in a glasshouse experiment (GE). Radiant flux density was 5 times lower in GE. The distribution of root diameter was measured for several root branching orders. Root diameters ranged between 0.09 and 0.52 mm for secondary roots and between 0.06 and 0.27 mm for tertiary roots. A relationship was found between the diameter of the parent bearing root and the median diameter of its laterals, which appears to be valid for a wide range of species. Mean lateral root diameter increased with distance to the base of the root and decreased with branching density [number of lateral roots per unit length of bearing root (cm(-1))]. Typical symptoms of low light availability were observed in GE. In this case, lateral root diameter variability was reduced. Although primary root growth was affected by soil compaction, no effects on lateral root diameter were observed.     

Co-ordinated growth between aerial and root systems in young apple plants issued from in vitro culture

BACKGROUND AND AIMS: In several species exhibiting a rhythmic aerial growth, the existence of an alternation between root and shoot growth has been demonstrated. The present study aims to investigate the respective involvement of the emergence of new organs and their elongation in relation to this phenomenon and its possible genotypic variation in young apple plants. METHODS: Two apple varieties, X6407 (recently named 'Ariane') and X3305 ('Chantecler' x 'Baujade'), were compared. Five plants per variety, issued from in vitro culture, were observed in minirhizotrons over 4 months. For each plant, root emergence and growth were observed twice per week. Growth rates were calculated for all roots with more than two segments and the branching density was calculated on primary roots. On the aerial part, the number of leaves, leaf area and total shoot length were observed weekly. KEY RESULTS: No significant difference was observed between varieties in any of the final characteristics of aerial growth. Increase in leaf area and shoot length exhibited a 3-week rhythm in X3305 while a weaker signal was observed in Ariane. The primary root growth rate was homogeneous between the plants and likewise between the varieties, while their branching density differed significantly. Secondary roots emerged rhythmically, with a 3-week and a 2-week rhythm, respectively, in X3305 and 'Ariane'. Despite a high intra-variety variability, significant differences were observed between varieties in the secondary root life span and mean length. A synchronism between leaf emergence and primary root growth was highlighted in both varieties, while an opposition phase was observed between leaf area increments and secondary root emergence in X3305 only. CONCLUSION: A biological model of dynamics that summarizes the interactions between processes and includes the assumption of a feedback effect of lateral root emergence on leaf emergence is proposed.     

Reliable plantlet formation from embryos and seedling shoot tips of radiata pine

A method has been devised for the reliable production of plantlets from embryos and seedling shoot tips of Pinus radiata D.Don (radiata pine). Buds were induced on an agar or liquid Schenk and Hildebrandt (SH) medium containing 5.0 mg/l benzylaminopurine (BAP). Except for some abnormal buds, the buds grew into elongated shoots on an agar SH medium without cytokinin. The transfer of shoots from a SH medium to a Gresshoff and Doy (GD) medium was found to be an important pretreatment which increased the survival of the shoots when they were placed in a peat and pumice mix for root formation. Elongated shoots were induced to form roots under non-sterile conditions in a humid environment with occasional misting. An intervening 5-day treatment of shoots in an agar medium containing 2.0 mg/l indolebutyric acid (IBA) and 0.5 mg/l napthaleneacetic acid (NAA) significantly increased the percentage of shoots forming roots and the number of roots formed per shoot over control shoots placed directly in the peat:pumice mix. An enhanced level of CO₂ during root formation had no effect on the time of root formation or on the percentage of shoots forming roots. These results concerning the elongation, growth and rooting of adventitious shoots are now being applied to the development of very large numbers of plantlets starting from cotyledons from partially germinated seeds.     

Apical diameter and branching density affect lateral root elongation rates in banana

Thick roots elongate faster than thinner ones. However, within one species, the growth achieved by roots of a given diameter can be very variable, and root diameter can only be considered as a determinant of root potential elongation rate. As root elongation is highly correlated to carbon availability, it can be hypothesized that local competition for resources, expressed as the number of lateral roots per unit length (i.e. the branching density), modulates root elongation. Using novel methods in field conditions, we have estimated apical diameters, elongation rates and growth durations of nearly 3500 banana lateral roots, in a field experiment with high radiations and a shaded glasshouse experiment with low radiations. Apical diameters and branching densities were lower in the experiment with low radiation, but elongation rates were higher. In both experiments, mean elongation rates of first-order laterals and thick second-order laterals were negatively correlated with bearing root branching densities. It is hypothesized that, even though apical diameters were lower, low branching densities in the shaded glasshouse allowed enhanced lateral root elongation. In both experiments, second-order laterals elongated more slowly than first-order laterals of similar diameter. A specific effect of root order, independent of branching density and apical diameter, contributed to explain these slow second-order lateral elongation rates. Most lateral roots elongated between 9 and 21 days and growth duration was mainly correlated with root diameter.     

Phosphate Availability Alters Lateral Root Anatomy and Root Architecture of Fraxinus mandshurica Rupr. Seedlings

Plants have evolved some mechanisms to maximize the efficiency of phosphorus acquisition.Changes in root architecture are one such mechanism. When Fraxinus mandshurica Rupr. seedlings were grown under conditions of low phosphorus availability, the length of cells in the meristem zone of the lateral roots was longer, but the length of cells in the elongation and mature zones of the lateral roots was shorter,compared with seedlings grown under conditions of high phosphorus availability. The elongation rates of primary roots increased as phosphorus availability increased, but the elongation rates of the branched zones of the primary roots decreased. The number of lateral root primordia and the length of the lateral roots decreased as phosphorus availability increased. The topological index (altitude slope) decreased as phosphorus availability increased, suggesting that root architecture tended to be herringbone-like when seedlings were grown under conditions of low phosphate availability. Herringbone-like root systems exploit nutrients more efficiently, but they have higher construction costs than root systems with a branching pattern.     

根修剪对黄土旱塬冬小麦(Triticum aestivum)根系分布、根系效率及产量形成的影响

通过田间试验研究了不同时期根修剪处理对冬小麦（Triticum aestivum）根系大小与分布、根系效率、水分利用效率及产量形成的影响。设置4个根修剪处理：越冬期小剪根（WS）、越冬期大剪根（WB）,返青期小剪根（GS）、返青期大剪根（GB）,未剪根小麦作为对照（CK）。结果表明,到花期时,各根修剪处理小麦的在0～120cm总根量均显著小于对照。与对照相比各根修剪处理主要是显著地减少了上层土壤中的根量。但WS和GS两小剪根处理和对照相比在中层土壤中有较大的根量;花后各处理小麦旗叶的气孔导度和蒸腾速率均显著大于对照。这说明根修剪处理减少了小麦表层的根量,从而削弱了表土干旱信号对作物与外界气体交换的抑制作用。花期时各根修剪小麦的净光合速率均显著高于对照,而单位面积上的根呼吸速率均显著小于对照,根修剪处理提高了小麦的根系效率,使更多的光合产物用于籽粒生产,从而提高了小麦的收获指数。根修剪还提高了小麦的水分利用效率,其中WS、WB、GS处理的水分利用效率显著高于对照。但是GB处理的水分利用效率却没有显著提高。因此,本研究进一步证明了由不同年代品种得到的推测,认为在旱地农业中,通过遗传育种或采用适当农艺措施优化根系分布,既可以减少生长前期作物对水分的过度消耗,又能够削弱花后表土过度干旱对作物生长抑制作用,同时降低根系对同化产物的消耗,对作物产量及水分利用效率的提高具有积极的作用。     

The effectiveness of internal oxygen transport in a mesophyte (Pisum sativum L.)

Summary The effectiveness of oxygen movement through pea seedlings has been assessed firstly by assaying for radial oxygen loss along the roots using the cylindrical Pt electrode technique, and secondly by measuring root growth in various oxygen-free media.It was found that roots would grow in oxygen-free 3% agar to a length exceeding 20 cm, but when such plants were removed to oxygen-free 0.05% agar oxygen could not be detected in the apical segments in roots longer than 9.5 cm unless respiratory activity was curtailed by cooling.If the greater part of the root was retained in 3% agar and only the apical region exposed to 0.05% agar and assayed, oxygen loss always occurred. It was concluded that the 3% agar has a jacketing effect substantially reducing oxygen leakage from the root surface and thus allowing more oxygen to channel down to the apical regions.Root growth in unstirred air-saturated 0.05% agar matched the growth in oxygen-free 3% agar. Root growth in unstirred oxygen-free 0.05% agar was arrested at c. 9 cm.It is suggested that the effect produced by the aerated unstirred 0.05% agar is consistent with a jacketing effect mitigating oxygen loss from the root and that a growth of 9 cm in unstirred deoxygenated agar is consistent with a smaller jacketing effect due to the unstirred medium.It is proposed that the accumulation of respiratory tissue will eventually render inadequate any jacketing effect. Further aerobic development at this stage will require a supply of oxygen from the rooting medium.     

Phosphate Availability Alters Lateral Root Anatomy and Root Architecture of Fraxinus mandshurica Rupr. Seedlings

Plants have evolved some mechanisms to maximize the efficiency of phosphorus acquisition. Changes in root architecture are one such mechanism. When Fraxinus mandshurica Rupr. seedlings were grown under conditions of low phosphorus availability, the length of cells in the meristem zone of the lateral roots was longer, but the length of cells in the elongation and mature zones of the lateral roots was shorter,compared with seedlings grown under conditions of high phosphorus availability. The elongation rates of primary roots increased as phosphorus availability increased, but the elongation rates of the branched zones of the primary roots decreased. The number of lateral root primordia and the length of the lateral roots decreased as phosphorus availability increased. The topological index (altitude slope) decreased as phosphorus availability increased, suggesting that root architecture tended to be herringbone-like when seedlings were grown under conditions of low phosphate availability. Herringbone-like root systems exploit nutrients more efficiently, but they have higher construction costs than root systems with a branching pattem.     

Secondary embryonary root associated with seed germination in Garcinia intermedia (Clusiaceae) and its possible role in seedling survival

Germination tests on Garcinia intermedia (Clusiaceae) seeds showed the growth of two types of roots: additionally to the primary root, a secondary root crosses the seed lengthwise. To determine its possible role on the survival and growth of this species, 90 seedlings at least six months old (collected in Central Costa Rica) were planted in plastic bags with organic soil, and placed in a greenhouse. The seedlings were treated as follows: treatments in which the primary or secondary root was cut off, and a control group with both roots intact (30 replicates each). After three months 10 seedlings/month/treatment were extracted to measure their height, basal diameter, root length (main and secondary root), and biomass of the stem, roots and seed (without its coat). Control seedlings had the highest growth, followed by those without secondary roots. Nonetheless, more than 90% of the seedlings whose primary roots were cut off, survived after five months of the excision treatment, in part due to the capacity of this species to regenerate its radical system through the seed reserves, sprouting of a primary-like root, and/or the growth stimulus of the secondary root (60% of the total: 20% with sprouts from the primary root stump, 13.3% with a growth stimulus of the secondary root, and 26.7% with both conditions). The length of the sprouted roots was significantly different only on those plants that were extracted during the first two monthly measurements, when compared with the control (F6 = 18.6, F7 = 16.0, p < 0.01).     

Vesicular-arbuscular mycorrhizal fungi induced alteration in poplar root system morphology

Poplar was grown in a soil either inoculated with Scutellispora calospora, Glomus sp E3 or Glomus caledonium or to which a nutrient solution had been added, in order to determine effects on root morphology. Plants were harvested after 115 days. The lengths of individual roots were measured using image analysis and percentage colonisation was determined for different root orders. Colonisation did not affect plant size but induced large changes in root morphology, with lengths of individual secondary and tertiary roots increased in some cases by up to 100%. Root branching was also increased with number of laterals per unit length of colonised roots being up to 6 times greater than in non-colonised roots. These results clearly show that colonisation of roots by Vesicular-arbuscular mycorrhizal fungi can result in significant alteration to poplar root system morphology. They also suggest that the mechanisms of alteration are not entirely due to improved host plant nutrition.     

Pre-germination effects on mechanically impeded root growth of barley (Hordeum vulgare L)

The effects of conditions pre-dating germination on growth rate of impeded barley cv. Harrington roots were measured using an agar-capillary tube technique. Seedling root tips were directed into glass capillary tubes twothirds filled with agar at eight concentrations ranging from 1.6 to 9.6%, equivalent to penetrometer resistances of 25 to 1240 kPa. The rate of unrestricted root elongation (growth in air) of seed stored for 13 months (old seed), and of seed grown for a second generation without subsequent storage (new seed) was compared with growth in agar over a 24-hour interval. Root elongation rate of old and new seed was identical in the absence of resistance. At low to intermediate agar concentrations, elongation was significantly slower in roots from old, compared with new seed. At high agar concentrations root growth of old and new seed was the same. In both old and new seed, root growth through agar was greater in seed that germinated after 24, compared with 48 h. Differences in impeded root growth between old and new seed were lost in progeny of the test seed. Environmental factors that pre-date germination are an important influence on the ability of seedling roots to elongate through soil.LRS Contribution no. 3879349LRS Contribution no. 3879349