Preparation of Chloroplast DNA from Barley and Lettuce and Comparison of Restriction Fragments

Chloroplast DNA from three barley cultivars and from one lettuce cultivar was prepared from chloroplasts isolated by Conventional differential centrifugation. Barley chloroplast DNA size was sensibly lower (130 kpb) than lettuce chloroplast DNA (150 kpb). Chloroplast DNAs from the three barley cultivars showed similar restriction fragment patterns after digestion with: BamHI, EcoRI or HindIII. The lettuce chloroplast DNA restriction pattern was very different from the barley chloroplast DNA restriction pattern.     

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.     

Identification in lettuce seedlings of a catecholamine active in synergistically enhancing the gibberellin effect on lettuce hypocotyl elongation

The occurrence of catecholamines in lettuce seedlings was examined by bioassay and gas chromatography-mass spectrometry (GC-MS), since synthetic catecholamines can synergistically enhance the stimulating effect of gibberellic acid (GA₃) on hypocotyl elongation of decotylized lettuce seedlings. The catecholamine fraction on TLC obtained from lettuce seedlings synergistically enhanced the GA₃ effect on hypocotyl elongation. The analysis of the catecholamine fraction from lettuce seedlings by GC-MS demonstrated the occurrence of dopamine.     

Cinnamaldehyde Inhibits Phenylalanine Ammonia-Lyase and Enzymatic Browning of Cut Lettuce

Stored cut lettuce gradually turns brown on the cut section after several days of storage, because cutting induces phenylalanine ammonia-lyase (PAL) activity, the biosynthesis of polyphenol is promoted, and the polyphenols are oxidized by polyphenol oxidase. In this study, we screened for inhibitors of PAL derived from fermented broths of microbes and from foods and found that a cinnamon extract definitely inhibited PLA of cut lettuce. An active component was isolated by chromatographic procedures and was identified as trans-cinnamaldehyde. Browning of cut lettuce immersed in a solution containing trans-cinnamaldehyde was definitely repressed.     

Fragment length distributions and collision probabilities for AFLP markers

AFLP is a DNA fingerprinting technique frequently used in plant and animal sciences. A drawback of the technique is the occurrence of multiple DNA fragments of the same length in a single AFLP lane, which we name a collision. In this article we quantify the problem. The well-known birthday problem plays a role. Calculation of collision probabilities requires a fragment length distribution (fld). We discuss three ways to estimate the fld: based on theoretical considerations, on in-silico determination using DNA sequence data from Arabidopsis thaliana, or on direct estimation from AFLP data. In the latter case we use a generalized linear model with monotone smoothing of the fragment length probabilities. Collision probabilities are calculated from two perspectives, assuming known fragment counts and assuming known band counts. We compare results for a number of fld's, ranging from uniform to highly skewed. The conclusion is that collisions occur often, with higher probabilities for higher numbers of bands, for more skewed distributions, and, to a lesser extent, for smaller scoring ranges. For a typical plant genome an AFLP with 19 bands is likely to contain the first collision. Practical implications of collisions are discussed. AFLP examples from lettuce and chicory are used for illustration.     

Effect of a plant-derived smoke extract, N6-benzyladenine and gibberellic acid on the thermodormancy of lettuce seeds

The effects of a plant-derived smoke extract, BA and GA₃ on the thermodormancy of Grand Rapids lettuce seeds were studied. Thermodormant lettuce seeds treated either with BA, GA₃ or smoke extract alone did not germinate. Combinations of BA with smoke extract and BA with GA₃ were most effective in overcoming induced thermodormancy. GA₃ plus smoke did not break the induced thermodormancy. The effects of the different treatments on germination were concentration dependent. BA was most effective at 10^–5 to 10^–3 M in combination with smoke dilutions 1:5,000 to 1:1,000,000 in overcoming thermodormancy.Abbreviations BA N⁶-benzyladenine; - GA₃ gibberellic acid; - SM smoke extract     

First Record of Bremia lactucae Infecting Sonchus oleraceus and Sonchus asper in Brazil and its Infectivity to Lettuce

Bremia lactucae is recorded for the first time causing downy mildew on common sowthistle (Sonchus oleraceus) and spiny sowthistle (Sonchus asper) in Brazil. The disease and etiological agent are described. Pathogencity of sporangia obtained from S. oleraceus was tested on 12 species belonging to the Asteraceae, already recorded in the literature as hosts of B. lactucae, and four commonly cultivated varieties of lettuce. All four cultivars of lettuce, S. oleraceus and S. asper showed symptoms of the disease and sporulation of the pathogen 5 days after inoculation. It has been observed that the disease occurs on Sonchus spp. throughout the year in Viçosa (state of Minas Gerais), being more common on these two hosts than on lettuce. Both weeds are very common invaders of vegetable growing areas in Brazil. This indicates that those two hosts for the fungus may be important inoculum reservoirs for disease occurring in lettuce, highlighting the need for control of these weeds as part of the procedures aimed at controlling this disease. These results are also in agreement with the views that B. lactucae is split into host‐specific infra‐specific taxa. Lettuce and sowthistles are regarded as closely related, belonging to the same subfamily and tribe in the Asteraceae (Subfamily Cichorioideae: Tribe Lactuceae).     

Direct isolation of vacuoles from leaf tissue of lettuce (Lactuca sativa) retaining protoplasts within the leaves

A procedure is described in which vacuoles are isolated from leaf tissue of lettuce ( Lactuca sativa L.). After incubation in an enzyme solution, the vacuoles are directly extracted from the leaf tissue by osmotic shock using a phosphate buffer. In this method no protoplasts are released from the leaf tissue. This procedure avoids the problems of separating vacuoles from protoplasts with similar density. To evaluate the purity of the vacuoles, the activity of glucan synthetase 11 (EC 2.4.1.34), NAD(P) H-cytochrome c reductase (EC 1.6.99.3) and malate dehydrogenase (EC 1.1.1.37) was measured. To measure vanadate- and nitrate-sensitive ATPase activity (EC 3.6.1.8) vesicles were prepared from the vacuoles and ATP-dependent vesicle acidification was measured as acridine orange quenching. Nitrate inhibited the quenching, while addition of vanadate had no effect. It was concluded that the vacuoles were not contaminated with plasma membranes. To evaluate the viability of the vacuoles [¹⁴C]-malate uptake was measured. The vacuoles showed a constant rate of [¹⁴C]-malate uptake during 45 min. This rate was maximal at pH 6.8.     

Crossing experiments of lettuce cultivars and species (Lactuca sect.Lactuca,Compositae)

The degree of relationships withinLactuca sativa and three wild relativesL. serriola, L. saligna, andL. virosa was studied by observing the performance, vigour and fertility of the F 1 hybrids obtained from crosses made in and between the four species. The crosses ofL. saligna ×L. virosa and the reciprocal crosses produced no hybrids.L. saligna andL. virosa are the least related of the four species.L. sativa ×L. serriola and the reciprocal crosses were successful and produced fertile hybrids These two species are genetically very closely related.L. saligna is known to produce, as a female parent, hybrids withL. sativa andL. serriola. Now the reciprocal cross was successful for the first time, so the unability to obtain hybrids in the past was based on the choice of accessions and not caused by unilateral incompatibility.L. virosa ×L. sativa and the reciprocal combination produced hybrids. The combinationL. serriola ×L. virosa produced hybrids with very limited fertility. In contrast to earlier reports (sterile hybrids) one combination of the reciprocal cross too produced hybrids with very limited fertility.—Some of theL. saligna ×L. sativa (and reciprocal) hybrids were found to look strikingly likeL. serriola. This adds evidence for the descent ofL. serriola andL. sativa:L. saligna also made part of the ancestral complex of the cultivated lettuce.     

The effects of placing small amounts of phosphate fertilizer close to the seed on growth and nutrient concentrations of lettuce

Summary Two glasshouse experiments are described in which the effects of applying starter phosphate fertilizer, 1 cm beneath the seeds, on early growth and nutrient concentrations of lettuce (Lactuca sativa L. cv. Avondefiance) in well fertilized soil were determined. In Experiment 1 various rates of starter P in the form of NH₄H₂PO₄ were applied to soil containing a range of rates of incorporated triple superphosphate. Although there was little response of lettuce dry weight to the incorporated triple superphosphate there was a large response (about 65% increase after 36 days) to the starter. N and P concentrations within the plants were increased by the starter treatments whereas K concentration was reduced. The per cent P in the plants at 36 days from sowing could account for 60% of the variation in plant dry weight. In Experiment 2 the starter P was added as either the Ca, Na or K salt, with or without added (NH₄)₂SO₄. Adding the starter P without ammonum increased the P concentration of the plants by an average of 12% and the dry weight by an average of 39% at 30 days from sowing. The addition of ammonium ions increased plant concentrations of P, Mg and N but decreased plant K concentration. The effect of the ammonium ions on growth depended on the form of phosphate supplied as the starter. This variation in effect of ammonium ions was attributed to the effects of other starter ions on the relative concentration of ammonium in the soil solution.