Cuttings of a tobacco mutant, rae, undergo cell divisions but do not initiate adventitious roots in response to exogenous auxin

Numerous studies have shown that auxin induces adventitious root initiation in stem explants from a variety of species, including tobacco. A dominant, monogenic mutation previously identified in tobacco ( Nicotiana tabacum cv. Xanthii), rac , confers tenfold auxin resistance to mesophyll-derived cell suspensions and an impaired primary root development phenotype to seedlings. Results presented here demonstrate that adventitious root formation does not occur when heterozygous and homozygous rac stem cuttings are treated in vitro with indole-3-butyric acid (IBA) concentrations ranging from 0.5 μ M to 500 μ M . Histological analysis showed that some phloem parenchyma or inner cortical parenchyma cells in wild-type stem cuttings undergo adventitious root morphogenesis when they are treated with 5 μ M IBA. The same cell types in heterozygous and homozygous rac stem cuttings undergo mitoses in response to auxin, but never form adventitious root meristems. The lack of adventitious root initiation in rac stem cuttings is phenotypically distinct from the aberrant primary root development in rac seedlings. The rac mutation appears to block an essential process for auxin induction of adventitious root initiation but not cell division in phloem parenchyma or inner cortical parenchyma cells. Comparisons of rac heterozygous and homozygous seedling primary root length and callus formation in response to auxin in stem cuttings indicate that rac copy number is correlated to the degree of expression of these two phenotypes.     

A comparison of the effects of auxin on cluster root initiation and development in Grevillea robusta Cunn. ex R. Br. (Proteaceae) and in the genus Lupinus (Leguminosae)

The effect of NAA (naphthaleneacetic acid) on the development of cluster roots in members of the Proteaceae and Leguminosae was investigated. The exogenous addition of NAA led to initiation of cluster roots in phosphate conditions normally inhibitory for their development, but initiation took place within the limits of the cluster pattern under –P conditions. There was no change in spacing within the cluster root nor between cluster roots in Grevillea robusta Cunn. ex R. Br. or in rootlet length or cluster root length. In Lupinus albus L., change in rootlet length and cluster root length was noted at 10^-10 and 10¹² M NAA. In L. albus, the length of time that roots were exposed to NAA does not appear to be important, with similar levels of cluster root initiation after 48 h and 7 days. Cluster root production in G. robusta differed from that in L. albus in terms of the concentration of NAA needed to induce initiation, and in the effects of extremely low levels of NAA on rootlet numbers and lengths. L. arboreus L. does not produce cluster roots under –P conditions. Furthermore, neither L. arboreus L., L. angustifolius L., L. luteus L. nor L. mutabilis L. were induced to produce cluster roots under –P conditions, nor under +P conditions in the presence of exogenous NAA. Thus, exogenous NAA only leads to the induction of cluster roots, at levels of P normally inhibitive of their development, in species of Lupinus that produce them under –P conditions. Auxin-induced cluster roots develop within the same constraints as those developing under –P conditions. NAA does not induce cluster roots in species of Lupinus that do not produce them under –P conditions.     

Role of cytokinin and auxin in shaping root architecture: regulating vascular differentiation, lateral root initiation, root apical dominance and root gravitropism

BACKGROUND AND AIMS: Development and architecture of plant roots are regulated by phytohormones. Cytokinin (CK), synthesized in the root cap, promotes cytokinesis, vascular cambium sensitivity, vascular differentiation and root apical dominance. Auxin (indole-3-acetic acid, IAA), produced in young shoot organs, promotes root development and induces vascular differentiation. Both IAA and CK regulate root gravitropism. The aims of this study were to analyse the hormonal mechanisms that induce the root's primary vascular system, explain how differentiating-protoxylem vessels promote lateral root initiation, propose the concept of CK-dependent root apical dominance, and visualize the CK and IAA regulation of root gravitropiosm. KEY ISSUES: The hormonal analysis and proposed mechanisms yield new insights and extend previous concepts: how the radial pattern of the root protoxylem vs. protophloem strands is induced by alternating polar streams of high IAA vs. low IAA concentrations, respectively; how differentiating-protoxylem vessel elements stimulate lateral root initiation by auxin-ethylene-auxin signalling; and how root apical dominance is regulated by the root-cap-synthesized CK, which gives priority to the primary root in competition with its own lateral roots. CONCLUSIONS: CK and IAA are key hormones that regulate root development, its vascular differentiation and root gravitropism; these two hormones, together with ethylene, regulate lateral root initiation.     

减弱UV-B辐射对烟草形态、光合及生理生化特性的影响

在地处低纬高原云南较高海拔烟区的通海县大田条件下,通过大棚覆盖不同透明膜的方式梯度减弱UV-B辐射,研究了外界环境30.39%(T1)、70.08%(T2)和75.74%(T3)强度的UV-B辐射对烟草品种K326形态、光响应特性和部分生理生化特征的影响。结果显示:减弱UV-B辐射促进了K326节间距和茎高的伸长,使叶片增大、变薄,尤以T2处理植株的叶片、茎围和节间距最大,而K326发育进程由T1至T3逐渐加快。在减弱UV-B辐射下,K326净光合速率(Pn)、最大净光合速率(Amax)和蒸腾速率(Tr)较高,但水分利用效率(WUE)、暗呼吸速率(Rd)、光补偿点(LCP)和光饱和点(LSP)较低。其中T1的Rd和LCP最低,并极大地降低了表观量子效率(AQY),T2的LSP最低而Pn最高,而T3的Amax、LCP和AQY最高。减弱UV-B辐射后烟叶叶绿素含量提高,而类黄酮、比叶重、类胡萝卜素含量和面积含水量下降,鲜重含水量和自然水分饱和亏却上升;T3处理下丙二醛含量最大,鲜重含水量和自然水分饱和亏则T2最大。结果表明,适当高强度的UV-B辐射对促进K326正常生长发育和提高其对UV-B辐射的适应性有重要作用。     

Estimation of the anatomical, stomatal and biochemical components of differences in photosynthesis and transpiration of wild-type and transgenic (expressing yeast-derived invertase targeted to the vacuole) tobacco leaves

Photosynthesis and transpiration rates of transgenic (expressing yeast-derived invertase targeted to the vacuole) tobacco (Nicotiana tabacum L.) leaves were, respectively, 50 and 70% of those of a wild type at 20°C, 350 cm³ m^?3 CO₂ concentration, 450 μmol (photons) m^?2 s^?1 of light intensity, and 70% relative air humidity. These differences could be attributed: (a) to changes in leaf anatomy and, consequently, to changes in gases diffusion between the cells' surfaces and the atmosphere; (b) to different stomatal apertures, and, for the photosynthesis rate, (c) to the altered CO₂ assimilation rate. Our objective was to estimate the relative contributions of these three sources of difference. Measurements on the wild-type and the transgenic leaf cross-sections gave values for the cell area index (CAI, cell area surface per unit of leaf area surface) of 15.91 and 13.97, respectively. The two-dimensional model 2DLEAF for leaf gas exchange was used to estimate quantitatively anatomical, stomatal and biochemical components of these differences. Transpiration rate was equal to 0.9 for the wild-type and to 0.63 mmol m^?2 s^?1 for the transgenic leaf: 24.0% of the difference (0.066 mmol m^?2 s^?1 was caused by the greater cell area surface in the wild-type leaf, and 66.0% was caused by a smaller stomatal aperture in the transgenic leaf. Photosynthetic rate was 3.10 and 1.55 μmol m^?2 s^?1 for the wild-type and transgenic leaves, respectively. Only 10.3% of this difference (0.16 μmol m^?2 s^?1) was caused by the difference in CAI, and the remaining 89.7% was caused by altered CO₂ assimilation rate.     

Effects of nitrogen and phosphorus deficiencies on levels of carbohydrates, respiratory enzymes and metabolites in seedlings of tobacco and their response to exogenous sucrose

A simple method of growing plants in agar was exploited to investigate the effect of long-term nitrogen (N) and phosphorus (P) deficiencies on respiratory metabolism and growth in shoots and roots of Nicotiana tabacum seedlings, and their interaction with exogenously supplied sucrose. Levels of hexose phosphates and 3-phosphoglyceric acid (3-PGA) were low in P-deficient shoots and roots and high in N-deficient shoots and roots. The ratio of hexose phosphates to 3-PGA and levels of fructose-2,6-bisphosphate were high in P-deficient plants and low in N-deficient plants. These data reflect differences in the way metabolism was perturbed, yet both deficiencies were associated with increased root growth relative to shoot growth, starch accumulation in the shoots, and soluble carbohydrate accumulation, especially hexoses, in the roots. Enzymes for sucrose degradation (sucrose synthase, acid and alkaline invertase) and glycolysis (phosphofructokinase, pyrophosphate-dependent phospho-fructokinase and pyruvate kinase) remained unaltered or declined in the shoots and roots. The accumulation of hexoses in roots of N- and P-deficient plants may result from maintenance of high invertase activities relative to sucrose synthase and glycolytic enzymes in the roots. The possibility that hexose accumulation may drive preferential root growth osmotically in N and P deficiencies is discussed. The addition of sucrose to roots to further investigate the interaction of carbohydrates with growth and allocation in low N and low P produced clear effects even though endogenous levels of soluble carbohydrate were already high in the nutrient-deficient plants. In complete nutrition, growth was stimulated, protein content particularly of the roots was increased and there was a preferential increase in activity of sucrose synthase in roots. At low P, enzyme activities in roots were increased, including sucrose synthase, and protein content increased, particularly in the roots, but there was no increase in growth. In N-deficient plants, exogenous sucrose led to decreased protein, Rubisco and chlorophyll content in shoots, in contrast to the other conditions, and a higher protein content and a general increase of catabolic enzyme activities and growth in the roots.     

Effect of flooding on the activities of some enzymes of activated oxygen metabolism,the levels of antioxidants,and lipid peroxidation in senescing tobacco leaves

Effects of flooding on the activities of some enzymes of activated oxygen metabolism, the levels of antioxidants, and lipid peroxidation in senescing leaves of tobacco were investigated. As judged by the decrease in chlorophyll and protein levels, flooding accelerated the senescence of tobacco leaves. Total peroxide and the lipid peroxidation product, malondialdehyde, increased in both control and flooding-treated leaves with increasing duration of the experiment. Throughout the duration of the experiment, flooded leaves had higher levels of total peroxide and malondialdehyde than did control leaves. Flooding resulted in an increase in peroxidase and ascorbate peroxidase activities and a reduction of superoxide dismutase activity in the senescing leaves. Glycolate oxidase, catalase, and glutathione reductase activities were not affected by flooding. Flooding increased the levels of total ascorbate and dehydroascorbate. Total glutathione, reduced form glutathione, or oxidized glutathione levels in flooded leaves were lower than in control leaves during the first two days of the experiment, but were higher than in control leaves at the later stage of the experiment. Our work suggests that senescence of tobacco induced by flooding may be a consequence of lipid peroxidation possibly controlled by superoxide dismutase activity. Our results also suggest that increased rates of hydrogen peroxide in leaves of flooded plants could lead to increased capacities of the scavenging system of hydrogen peroxide.Abbreviations GSH reduced form glutathione - GSSG oxidized form glutathione - GSSG reductase glutathione reductase - MDA malondialdehyde - SOD superoxide dismutase     

Transport of Rb and Sr to the ear in mature,excised shoots of wheat: Effects of temperature and stem length on Rb removal from the xylem

Wheat (Triticum aestivum L. cv. ‘Arina’) shoots grown in the field were excised post-anthesis and incubated in the laboratory for 72 h standing in 2 mM RbCl+2 mM SrCl₂. Strontium is a phloemimmobile, xylem-mobile element and indicates the distribution of the xylem sap in the plant. Rubidium is easily transported in the phloem and behaves similarly to the highly mobile K as far as the redistribution within the plant is concerned, although Rb cannot substitute physiologically or biochemically for K. The Sr contents in the ear were hardly affected by stem length or by steam-girdling (phloem-interruption). Rubidium on the other hand accumulated in the stem. A peduncle length of 5 cm was sufficient to decrease the Rb concentration in the xylem by more than 50% at 25°C. Only a minor quantity of Rb reached the ear after passing through 20 cm of stem without nodes and this transport was prevented by steam-girdling. A remarkable flux of Rb into the ear was observed in shoots with a vascular connection between the flag leaf lamina and the ear. Our results suggest that Sr was transported with the transpiration stream, while Rb was rapidly eliminated from the xylem and reached the ear via the phloem. The temperature optimum for the removal of Rb from the xylem was around 35°C. The nodes may further contribute, but are not prerequisites for this redistribution. The observed transfer processes could allow a solute specific transport via the xylem and phloem of maturing cereals and may be an important factor influencing the nutrient economy in the field.     

Characteristics of adventitious root formation in cotyledon segments of mango (Mangifera indica L. cv. Zihua): two induction patterns, histological origins and the relationship with polar auxin transport

Cotyledon segments derived from zygote embryos of mango (Mangifera indica L. cv. Zihua) were cultured on agar medium for 28 days. Depending on different pre-treatments with plant growth regulators, two distinct patterns of adventitious roots were observed. A first pattern of adventitious roots was seen at the proximal cut surface, whereas no roots were formed on the opposite, distal cut surface. The rooting ability depended on the segment length and was significantly promoted by pre-treatment of embryos with indol-3-acetic acid (IAA) or indole-3-butyric acid (IBA) for 1 h. A pre-treatment with the auxin transport inhibitor 2,3,5-triiodobenzoic acid (TIBA) completely inhibited adventitious root formation on proximal cut surfaces. A second pattern of roots was observed on abaxial surfaces of cotyledon segments when embryos were pre-treated with 2,700 μM 1-naphthalenacetic acid (NAA) for 1 h. Histological observations indicated that both patterns of adventitious roots originated from parenchymal cells, but developmental directions of the root primordia were different. A polar auxin transport assay was used to demonstrate transport of [³H] indole-3-acetic acid (IAA) in cotyledon segments from the distal to the proximal cut surface. In conclusion, we suggest that polar auxin transport plays a role in adventitious root formation at the proximal cut surface, whereas NAA levels (influx by diffusion; carrier mediated efflux) seem to control development of adventitious roots on the abaxial surface of cotyledon segments.