Interactions of Cd with Zn, Cu, Mn and Fe for lettuce (Lactuca sativa L.) in hydroponic culture

Abstract. The effect and accumulation of cadmium (Cd) in lettuce grown by means of the hydroponic technique was investigated by multivariate analysis, and was found to be affected by the concentration of other trace elements. Particularly iron acted in a strongly antagonistic way against Cd. Consequently, no absolute toxicity limits for Cd can be drawn without considering other trace elements.     

Regulation of lateral root formation in lettuce (Lactuca sativa) seedling roots: Interacting effects of α-naphthaleneacetic acid and kinetin

Growth substances, α-naphthaleneacetic (NAA) and kinetin, had an important role in the regulation of lateral root (LR) formation in lettuce ( Lactuca sativa L. cv. Grand Rapids) seedling roots. NAA (10^-5 M ) was a potent stimulator of LR initiation and caused a 600% increase in the number of lateral root primordia (LRP) compared to untreated roots. NAA was required for only the first 20 h of the 72 h treatment period for maximum stimulation of LRP initiation. Kinetin (2 × 10^-5 M ) effectively prevented the spontaneous formation of LRP and inhibited the NAA-stimulated production of LRP. Kinetin inhibition was maximal during the first 20 h of NAA treatment and this effect was not overcome by subsequent supply of NAA. Also, lettuce roots were most sensitive to kinetin at 20 h of NAA treatment, when the first signs of cell division were observed in the pericycle.     

Root architecture in cultivated and wild lettuce (Lactuca spp.)

Root architecture is described for intact root systems of cultivated (Lactuca sativa L.) and wild (L. serriola L.) lettuce, grown for 5 to 6 weeks in greenhouse pot and cylinder experiments in coarse-textured soil. L. sativa cv. Salinas and a sclinas line of L. serriola attained the same biomass at 4 to 6 weeks after planting. Root biomass allocation was also similar, but root architecture differed. In the top zone along the tap root (0 to 5 cm), Salinas tended to produce more laterals, a greater total root length, and more external links (segments that originate at a branch point and end in a meristem) than wild lettuce. In the 5 to 55cm zone of the tap root, these measures were greater in the wild species. These patterns of root structure were generally corroborated by a second cylinder study with a different pair of L. sativa and L. serriola. Regressions of root structural traits were made against total root dry weight as a means to compare root architecture independently of the size of the root system. Regressions demonstrated that production of root segments differed between the two species; for example the slope for the regression of summed external link length in the top 0 to 5 cm with total root dry weight was significantly higher for Salinas, indicating that the rate of construction in the top 0 to 5cm was greater for cultivated than for wild lettuce. Yet, from 5 cm depth to the tap root tip, the rate of construction of external link length was greater in L. serriola. For many of these types of regression, r² and mean slope ± SE suggested that more variation occurred in cultivated than wild lettuce, yet genetic heterogeneity was probably low within the studied taxa. Inadvertent selection may have occurred in the breeding of cultivated lettuce varieties for increased root growth in the surface zone where water and fertilizer are applied, and for greater plasticity in construction of root segments, which might maximize the efficiency of exploitation of soil moisture and nutrients.     

莴苣大孢子发生过程中钙的分布动态

莴苣胚囊发育为蓼型,减数分裂形成的4个大孢子中只有合点端的一个大孢子可继续发育,其余3个大孢子从珠孔端依次退化.大孢子母细胞中的钙沉淀颗粒很少,减数分裂后的四分体中的钙沉淀颗粒稍有增加.以后,4个大孢子中的钙沉淀颗粒在数量上有明显差异:即将退化的大孢子中钙明显减少,而未退化大孢子细胞质中则保持有较多的细小钙沉淀颗粒.大孢子的退化是一种细胞程序死亡现象,细胞中的钙浓度降低时可能启动了大孢子细胞的程序性死亡过程,而细胞中的钙浓度高时则保持大孢子细胞的继续发育.文章首次揭示了大孢子发生过程中钙的分布特征.     

Expression analysis of polyphenol oxidase isozymes by active staining method and tissue browning of head lettuce (Lactuca sativa L.)

Browning of plant tissue is generally considered attributable to enzymatic oxidation by polyphenol oxidase (PPO). Electrophoresis followed by activity staining has been used as an effective procedure to visually detect and isolate isozymes; however, it has not been applied for examination of various PPO isozymes in lettuce. Our study demonstrated that different lettuce PPO isozymes could be detected at different pH in active staining, and multiple isozymes were detected only under alkaline conditions. As a result, we concluded that activity staining with approximately pH 8 enabled to detect various PPO isozymes in lettuce. By expression analysis of the PPO isozymes after wounding, PPO isozymes that correlated with time-course of tissue browning were detected. The wound-induced PPO may play a key role in enzymatic browning.     

Stage-specific crosstalk between light, auxin, and ethylene during low-pH-induced root hair formation in lettuce (Lactuca sativa L.) seedlings

In the light, transfer of lettuce seedlings precultured on liquid medium at pH 6.0 to fresh medium at pH 4.0 induces root hair formation. However, no root hairs form in the dark. Here, we investigated how light induces root hair formation. Randomization of the transverse cortical microtubule (CMT) arrays which occurs in root epidermal cells in the light prior to root hair initiation was not observed in the dark. However, addition of indole-3-acetic acid (IAA) or 1-aminocyclopropane-1-carboxylic acid (ACC) induced CMT randomization and root hair formation. In these cases, CMT randomization occurred in almost the same time-dependent manner as under light. However, root hair initiation was delayed for several hours in the dark. These results suggest that light promotes CMT randomization and root hair initiation via auxin and ethylene signaling but light additionally influences root hair initiation independently of these signaling mechanisms. Furthermore, addition of a microtubule-depolymerizing drug in the dark disrupted the transverse CMT arrays and initiated root hair formation; however, root hair elongation was still suppressed. Root hairs elongated when IAA or ACC was applied with the drug. These results suggest that light also promotes root hair elongation via auxin and ethylene signaling.     

Evidence of Phytochrome Mediation in the Low-pH-Induced Root Hair Formation Process in Lettuce (Lactuca sativa L. cv. Grand Rapids) Seedlings

−2 . The inductive effect of 100 Jm⁻² red light could be partially reversed by subsequent far-red light only one time. On the other hand, the inductive effect of 1,000 Jm⁻² red light was partially reversed by subsequent far-red light irradiation at least twice. These results indicate the involvement of phytochrome in this response. The inductive effect of blue light was repeatedly reversed by subsequent far-red light irradiation, suggesting that the blue-light induction was mainly mediated by phytochrome. Received 13 August 1999/ Accepted in revised form 22 December 1999     

A method for screening copper-tolerant rice (Oryza sativa L.) cultivars based on hydroponic experiments and cluster analysis

A method for screening copper (Cu)-tolerant rice cultivars was studied by combining hydroponic experiments and cluster analysis, and the classification of cultivars in Cu stress tolerance was done. In the first hydroponic experiment, seedlings of Jiahe991 and Xiushui114 were planted in nutrient solution with different Cu²⁺ concentrations from 10 to 1800 μg/L. Results indicated that the toxic threshold of Cu concentration in solution ranged from 900 to 1200 μg/L, since SPAD (Soil and Plant Analyzer Development, SPAD-502, a portable chlorophyll meter, Minolta Camera Co. Ltd., Japan) values of leaves and seedlings biomass of the treatments with ≥900 and/or 1200 µg/L were significantly lower than the control. The second experiment was conducted with 16 local rice cultivars under three Cu treatments (10, 1000, and 1500 μg/L). The 16 cultivars were well classified into tolerant, normal, and sensitive groups as a result of cluster analysis based on the relative SPAD (Soil and Plant Analyzer Development, SPAD-502, a portable chlorophyll meter, Minolta Camera Co. Ltd., Japan) value, shoot and root dry weights, root length and root dehydrogenase activity, and oxidizing capacity and shoot Cu concentration. Xiushui123, Xiushui134, Jiahe991, and Xianghu301 belonged to the tolerant group; Xiushui137 belonged to the sensitive group. The cluster analysis based on hydroponic experiments is an effective method for identifying rice cultivars that are tolerant to Cu stress. In addition, four cultivars (Xiushui123, Xiushui134, Jiahe991, and Xianghu301) are recommended in local practice.     

Low pH-Induced Root Hair Formation in Lettuce (Lactuca sativa L. cv. Grand Rapids) Seedlings: Determination of Root Hair-Forming Site

Received 20 December 1999/ Accepted in revised form 26 April 2000     

Identification of a phenylalanine ammonia-Iyase inactivating factor in harvested head lettuce (Lactuca sativa)

Exposing head lettuce ( Lactuca sativa L., crisphead or Iceberg type) leaf tissue to hormonal levels of ethylene (10 μl l⁻¹) at 5°C promotes the de novo synthesis of phenylalanine ammonia-lyase (PAL, EC 4.3.1.5) and an increase in its activity. It also promotes the appearance of the postharvest physiological disorder called russet spotting (RS). Discontinuing ethylene exposure after 4 days resulted in a rapid decline in PAL activity which was delayed by treating excised midrib leaf tissue with actinornycin D or cycloheximide at 5°C. Only cycloheximide delayed the loss of PAL activity in tissue that was transferred from 5 to 15°C. Activity of PAL from Rhodolorula glutinis was. slowly lost during incubation in buffer alone, but there was a logarithmic decline in its activity over time when it was incubated with aliquots of the resuspended 10000 g pellet from homogenized, lettuce tissue affected with RS. The in vitro loss in PAL activity was 9–fold higher in extracts from lettuce showing RS symptoms than from control lettuce, boiled samples or the buffer control. The PAL-inactivating factor isolated from lettuce affected with RS had a pH optimum around 8.0. It appears that the rapid loss in PAL activity after the discontinuation of exposure to ethylene is dependent on the de novo synthesis of a PAL-inactivating factor.     

Photoperceptive Site of the Photoinduction of Root Hairs in Lettuce (Lactuca sativa L. cv. Grand Rapids) Seedlings Under Low pH Conditions

Lactuca sativa L. cv. Grand Rapids) seedlings under acidic conditions by a phytochrome-mediated response. Microbeam irradiation of 1 mm root segments with the first (100 Jm⁻²) and second (1,000 Jm⁻²) maxima of the fluence response curve for red light induction of root hair initiation indicated that the ca. 5 mm apical portion of 12 mm long roots was the site of photoperception. The root hair-forming portion of the root was situated at a distance of 1.7 mm from the root tip at the time of red light irradiation and extended (at most 1.5 mm) towards the basal end at a later stage of development, irrespective of which portion of the root was irradiated with red light. Received 13 August 1999/ Accepted in revised form 22 December 1999     

Cloning and expression of sesquiterpene synthase genes from lettuce (Lactuca sativa L.)

Sesquiterpenoid lactones (SLs) from lettuce (Lactuca sativa L.) include constitutive components of latex such as lactucin and the induced phytoalexin, lettucenin A. A redundant primer strategy was used to recover two full length cDNA clones (LTC1 and LTC2) encoding sesquiterpene synthases from a cDNA library derived from seedlings with the red spot disorder, which accumulate phytoalexins. Recombinant enzymes produced from LTC1 and LTC2 in Escherichia coli catalysed the cyclisation of farnesyl diphosphate to germacrene A, potentially an early step in the biosynthesis of SLs. RT-PCR analysis showed LTC1 and LTC2 were expressed constitutively in roots, hypocotyls and true leaves but not in cotyledons. Expression in cotyledons was induced by challenge with the downy mildew pathogen Bremia lactucae in the disease resistant cultivar Diana. Southern hybridisation experiments showed that LTC1 and LTC2 were not part of a multigene family. The germacrene A synthases provide targets for modified expression to generate beneficial modifications to the SL profile in lettuce.     

A comparison of organic and inorganic nitrogen fertilizers: Their nitrate-N and ammonium-N release characteristics and effects on the growth response of lettuce (Lactuca sativa L. cv. Fortune)

The response of pot grown lettuce to inorganic (ammonium nitrate) and organic (dried blood and Protox) N fertilizers was determined at two temperature regimes (15°C day/10°C night and 20°C day/15°C night) and related to the NH₄–N and NO₃–N release characteristics of each material. The N release characteristics of the organic materials matched the N requirements of lettuce more closely than the inorganic fertilizer. However, was rapidly released from the protein based materials such that composts were depleted of available fertilizer N at the same time irrespective of the form supplied. The warmer temperature regimes resulted in a more rapid depletion of the fertilizers due to biological immobilization such that N recoveries in shoots, roots and leachates were reduced. Approximately 20% of the N present in Protox (a material derived from activated sewage sludge, processed to reduce the heavy metal content to minimal levels) appeared to be resistant to microbial degradation and was unavailable to the plants. Therefore, the growth response of lettuce was slightly reduced with Protox compared to the other materials at similar rates of incorporation. The organic materials did not contribute NO₃–N to the plant and small NO₃–N concentrations in petioles were derived from the water used for irrigation. However, NO₃–N levels in plants receiving inorganic ammonium nitrate were initially high but progressively declined as the fertilizer NO₃–N became depleted.     

Shoot and root physiological responses to localized zones of soil moisture in cultivated and wild lettuce (Lactuca spp.)

Cultivated crisphead lettuce (Lactuca sativa L.) has a shallower root system than its wild relative, Lactuca serriola L. The effects of localized soil water, at depth, on plant water relations, gas exchange and root distribution were examined in the two species using soil columns with the soil hydraulic-ally separated into two layers, at (0–20 cm and 20–81) cm, but permitting root growth between the layers. Three treatments were imposed on 7-week-old plants, and maintained for 4 weeks: (i) watering, both layers to field capacity; (ii) drying the upper layer while watering the lower layer to field capacity, and (iii) drying both layers. Drying only 0–20 cm of soil had no effect on leaf water status, net photosynthesis, stomatal conductance or biomass production in L. serriola compared to a well-watered control, but caused a short-term reduction (10 d) in leaf water status and photosynthesis in L. sativa that reduced final shoot production. The different responses may be explained by differences in root distribution. Just before the treatments commenced, L. serriola had 50% of total root length at 20–80 cm compared to 35% in L. sativa. Allocation of total biomass to roots in L. serriola was approximately double that in L. sativa. The wild species could provide germplasm for cultivated lettuces to extract more soil water from depth, which may improve irrigation efficiency.     

Effects of co-cropping Bidens pilosa (L.) and Tagetes minuta (L.) on bioaccumulation of Pb in Lactuca sativa (L.) growing in polluted agricultural soils

Polluted agricultural soils are a serious problem for food safety, with phytoremediation being the most favorable alternative from the environmental perspective. However, this methodology is generally time-consuming and requires the cessation of agriculture. Therefore, the purpose of this study was to evaluate two potential phytoextractor plants (the native species Bidens pilosa and Tagetes minuta) co-cropped with lettuce growing on agricultural lead-polluted soils. The concentrations of Pb, as well as of other metals, were investigated in the phytoextractors, crop species, and in soils, with the potential risk to the health of consumers being estimated. The soil parameters pH, EC, organic matter percentage and bioavailable lead showed a direct relationship with the accumulation of Pb in roots. In addition, the concentration of Pb in roots of native species was closely related to Fe (B. pilosa, r = 0.81; T. minuta r = 0.75), Cu (T. minuta, r = 0.93), Mn (B. pilosa, r = 0.89) and Zn (B. pilosa, r = 0.91; T. minuta, r = 0.91). Our results indicate that the interaction between rhizospheres increased the phytoextraction of lead, which was accompanied by an increase in the biomass of the phytoextractor species. However, the consumption of lettuce still revealed a toxicological risk from Pb in all treatments.     

Blue light control of hypocotyl elongation in etiolated seedlings of Lactuca sativa (L.) cv. Grand Rapids related to exogenous growth regulators and endogenous IAA, GA3 and abscisic acid

Results of studies on the interaction of blue light (B) and exogenous applied plant hormones (IAA, GA3 and ABA) as well as inhibitors of their synthesis on the control of hypocotyl elongation in etiolated seedlings of Lactuca sativa (L.) cv. Grand Rapids are presented, and compared with endogenous GA3, IAA and ABA levels measured by capillary gas chromatography-mass spectrometry-selected ion monitoring (GC-MS-SIM). Hypocotyl elongation of etiolated seedlings was linearly inhibited by increasing the B fluence rate between 0 and 18.3 mol m–2 s–1. Both GA3 and IAA added to the incubation medium at different concentrations were able to eliminate partially the inhibition of growth caused by 7.2 mol m–2 s–1 B. When dark (D)-grown seedlings were treated with Ca-prohexadione, a specific inhibitor of gibberellin 3-hydroxylation, they showed a growth inhibition similar to under B. Also, a suppression of growth as in B was obtained when D-grown seedlings were treated with ABA 380 M. By lowering ABA levels with fluridone (an inhibitor of ABA synthesis) a partial reversion of hypocotyl growth inhibition was obtained in B-grown seedlings. While none of the growth promoters used were able to reverse completely the growth inhibition caused by B, a proper combination of GA3, IAA and (eventually) fluridone, abolished the B effects. Correspondingly, lower levels of GA3 and IAA and a higher concentration of ABA were measured by GC-MS-SIM in B-grown hypocotyls than in D-grown ones. These results support the hypothesis that hormones are implicated in mediation of B light-dependent inhibition of hypocotyl elongation, which seems to be the result of a balance among endogenous levels of growth promoting and growth inhibiting hormones.