A mixed-model QTL analysis for salt tolerance in seedlings of crop-wild hybrids of lettuce

Cultivated lettuce is more sensitive to salinity stress than its wild progenitor species potentially due to differences in root architecture and/or differential uptake and accumulation of sodium. We have identified quantitative trait locis (QTLs) associated with salt-induced changes in root system architecture (RSA) and ion accumulation using a recombinant inbred line population derived from a cross between cultivated lettuce (Lactuca sativa ‘Salinas’) and wild lettuce (L. serriola). Components of RSA were quantified by replicated measurements of seedling growth on vertical agar plates containing different concentrations of NaCl in a controlled growth chamber environment. Accumulation of sodium and potassium ions was measured in replicates of greenhouse-grown plants watered with 100 mM NaCl water. A total of 14 QTLs were identified using multi-trait linkage analysis, including three major QTLs associated with general root development, root growth in salt stress condition, and ion accumulation. The three major QTLs, qRC9.1, qRS2.1, and qLS7.2, were linked with markers E35/M59-F-425, LE9050, and LE1053, respectively. This study provides regions of lettuce genome contributing to salt-induced changes in RSA and ion accumulation. Future fine-mapping of major QTLs will identify candidate genes underlying salt stress tolerance in cultivated lettuce.     

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.     

Identification and comparison of natural rubber from two Lactuca species

Renewed interest in the identification of alternative sources of natural rubber to Hevea brasiliensis has focused on the Compositae family. In our search for Compositae models for rubber synthesis, we extracted latex from stems of two lettuce species: Lactuca serriola, prickly lettuce, and Lactuca sativa cv. Salinas, crisphead lettuce. Both species contained cis-1,4-polyisoprene rubber in the dichloromethane-soluble portions of their latex, and sesquiterpene lactones in their acetone-soluble portions. The rubber from both species and their progeny had molecular weights in excess of 1,000,000g/mol, and polydispersity values of 1.1. Rubber transferase activity was detected across a range of farnesyl diphosphate initiator concentrations, with decreased activity as initiator concentrations exceeded putative saturation. These results add lettuce to the short list of plant species that produce high molecular weight rubber in their latex. Due to the genomic and agronomic resources available in lettuce species, they provide the opportunity for further dissection of natural rubber biosynthesis in plants.     

Chromosomal study of lettuce and its allied species (Lactuca spp., Asteraceae) by means of karyotype analysis and fluorescence in situ hybridization

In this study, in addition to the karyotype analysis, the chromosomal distributions of 5 S and 18 S rDNAs, and the Arabidopsis-type (T3AG3) telomeric sequences were detected by means of fluorescence in situ hybridization (FISH) to promote the information of chromosomal organization and evolution in the cultivated lettuce and its wild relatives, L. sativa, L. serriola, L. saligna and L. virosa. The karyotype analysis revealed the dissimilarity between L. virosa and the remaining species. In all four Lactuca species studied, one 5 S rDNA and two 18 S rDNA loci were detected. The simultaneous FISH of 5 S and 18 S rDNAs revealed that both rDNA loci of L. sativa, L. serriola and L. saligna were identical, however, that of L. virosa was different from the other species. These analyses indicate the closer relationships between L. sativa/L. serriola and L. saligna rather than L. virosa. Arabidopsis-type telomeric sequences were detected at both ends of their chromatids of all chromosomes not in the other regions. This observation suggests the lack of telomere-mediated chromosomal rearrangements among the Lactuca chromosomes.     

Development and application of a 6.5 million feature affymetrix genechip® for massively parallel discovery of single position polymorphisms in lettuce (Lactuca spp.)

ABSTRACT: BACKGROUND: High-resolution genetic maps are needed in many crops to help characterize the genetic diversity that determines agriculturally important traits. Hybridization to microarrays to detect single feature polymorphisms is a powerful technique for marker discovery and genotyping because of its highly parallel nature. However, microarrays designed for gene expression analysis rarely provide sufficient gene coverage for optimal detection of nucleotide polymorphisms, which limits utility in species with low rates of polymorphism such as lettuce (Lactuca sativa). RESULTS: We developed a 6.5 million feature Affymetrix GeneChip? for efficient polymorphism discovery and genotyping, as well as for analysis of gene expression in lettuce. Probes on the microarray were designed from 26,809 unigenes from cultivated lettuce and an additional 8,819 unigenes from four related species (L. serriola, L. saligna, L. virosa and L. perennis). Where possible, probes were tiled with a 2 bp stagger, alternating on each DNA strand; providing an average of 187 probes covering approximately 600 bp for each of over 35,000 unigenes; resulting in up to 13 fold redundancy in coverage per nucleotide. We developed protocols for hybridization of genomic DNA to the GeneChip? and refined custom algorithms that utilized coverage from multiple, high quality probes to detect single position polymorphisms in 2 bp sliding windows across each unigene. This allowed us to detect greater than 18,000 polymorphisms between the parental lines of our core mapping population, as well as numerous polymorphisms between cultivated lettuce and wild species in the lettuce genepool. Using marker data from our diversity panel comprised of 52 accessions from the five species listed above, we were able to separate accessions by species using both phylogenetic and principal component analyses. Additionally, we estimated the diversity between different types of cultivated lettuce and distinguished morphological types. CONCLUSION: By hybridizing genomic DNA to a custom oligonucleotide array designed for maximum gene coverage, we were able to identify polymorphisms using two approaches for pair-wise.     

Lettuce, a shallow-rooted crop, and Lactuca serriola, its wild progenitor, differ at QTL determining root architecture and deep soil water exploitation

Wild plant species are often adapted to more stressful environments than their cultivated relatives. Roots are critical in exploiting soil resources that enable plants to withstand environmental stresses, but they are difficult to study. Cultivated lettuce (Lactuca sativa L.) and wild L. serriola L. differ greatly in both shoot and root characteristics. Approximately 100 F_2:3 families derived from an interspecific cross were evaluated in greenhouse and field experiments. In the greenhouse, root traits (taproot length, number of laterals emerging from the taproot, and biomass) and shoot biomass were measured 4 weeks after planting. In the field, plants were grown for 9 weeks (close to harvest maturity of the cultivated parent); mild drought stress was induced by withholding water for 1 week, and gravimetric moisture of soil was then determined for five depth increments between 0–100 cm. The families were genotyped using codominantly scored AFLP markers distributed throughout the genome. Composite interval mapping was used to analyze marker-trait associations. Quantitative trait loci were identified for differences between wild and cultivated lettuce for root architectural traits and water acquisition. Thirteen QTL were detected that each accounted for 28–83% of the phenotypic variation. The loci for taproot length (i.e., cm taproot length g^–1 plant biomass) and the ability to extract water from deep in the soil profile co-localized in the genome. These coincident loci were identified in separate experiments. The wild L. serriola is therefore a potential source of agriculturally important alleles to optimize resource acquisition by cultivated lettuce, thereby minimizing water and fertilizer inputs and ultimately enhancing water quality. Received: 25 February 2000 / Accepted: 31 March 2000     

QTLs for shelf life in lettuce co-locate with those for leaf biophysical properties but not with those for leaf developmental traits

Developmental and biophysical leaf characteristics that influence post-harvest shelf life in lettuce, an important leafy crop, have been examined. The traits were studied using 60 informative F9 recombinant inbed lines (RILs) derived from a cross between cultivated lettuce (Lactuca sativa cv. Salinas) and wild lettuce (L. serriola acc. UC96US23). Quantitative trait loci (QTLs) for shelf life co-located most closely with those for leaf biophysical properties such as plasticity, elasticity, and breakstrength, suggesting that these are appropriate targets for molecular breeding for improved shelf life. Significant correlations were found between shelf life and leaf size, leaf weight, leaf chlorophyll content, leaf stomatal index, and epidermal cell number per leaf, indicating that these pre-harvest leaf development traits confer post-harvest properties. By studying the population in two contrasting environments in northern and southern Europe, the genotype by environment interaction effects of the QTLs relevant to leaf development and shelf life were assessed. In total, 107 QTLs, distributed on all nine linkage groups, were detected from the 29 traits. Only five QTLs were common in both environments. Several areas where many QTLs co-located (hotspots) on the genome were identified, with relatively little overlap between developmental hotspots and those relating to shelf life. However, QTLs for leaf biophysical properties (breakstrength, plasticity, and elasticity) and cell area correlated well with shelf life, confirming that the ideal ideotype lettuce should have small cells with strong cell walls. The identification of QTLs for leaf development, strength, and longevity will lead to a better understanding of processability at a genetic and cellular level, and allow the improvement of salad leaf quality through marker-assisted breeding.     

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.     

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.     

The involvement of the root apex and cytokinins in the control of lateral root emergence in lettuce seedlings

Removal of the apical 3 mm of the primary root of hydroponically-grown lettuce seedlings 3 or 5 days after sowing, prevented further elongation of the root and increased both the number and total length of lateral roots. The length of the lateral zone, i.e. the distance from the base of the parent root to the lateral nearest the tip, except on one occasion, remained the same as the control in both 3 and 5 day treatments, until the length of the decapitated root (which had ceased elongating) became limiting.Zeatin applied via the roots, at a concentration range from 3 × 10^–10 M to 10^–8 M reduced tap root extension growth at all concentrations. Lateral root emergence was enhanced by low zeatin concentrations and retarded by higher ones. In general, the lateral zone length was the same in cytokinin-treated plants as in untreated controls.     

Clonal strategies of a woody weed: Melia azedarach

Melia azedarach L., a weedy tree that typically reproduces by seeds, may exhibit clonal growth following disturbance (e.g. fire, herbivory, animal injury). A dynamic and holistic study (sensu Hallé et al., 1978) was carried out in order to determine the architecture and the vegetative growth strategies, in plants collected from undisturbed and disturbed areas in the ‘El Palmar’ National Park. The architectural and morphological observations can be summarized as follows: (a) Tree architecture is determined by a monopodial trunk that produces tiers of orthotropic monopodial branches. (b) The root system consists of a tap root with woody primary lateral roots that exhibit plagiotropic growth and are typically found within the first few centimeters (ca. 4 cm) of the soil profile. (c) Root buds were produced in all (fire treated and control) plagiotropic root cuts when incubated under controlled conditions. Root suckers developed from the differentiation of parenchymatous cells produced by meristematic activity in the cambial zone. The ecological implications of vegetative reproduction on the invasive capacity of this species are discussed. This revised version was published online in June 2006 with corrections to the Cover Date.     

Field water relations of a compass plant, Lactuca serriola L.

Abstract Field water relations of Lactuca serriola serriola and L. serriola integrifolia were examined. Leaf conductance to water vapour was high early in the morning and declined rapidly during the midday hours. Leaf water potentials decreased to their minima early in the morning and remained low all day. Afternoon recovery of leaf conductance occurred occasionally. Leaf conductance was shown to have a linear response to vapour concentration difference. No differences were seen between L. serriola serriola and L. serriola integrifolia. The pattern of diurnal gas-exchange activity appeared to be complemented by the pattern of intercepted solar irradiance which results from the compass plant leaf orientation observed in L. serriola.     

Experimental evaluation of an efflux-influx model of C exudation by individual apical root segments

The aim of this study was to evaluate if a model describing the efflux and the influx of C through the root surface could be fitted to experimental short-term kinetics of carbon (C) exudation by individual apical root segments in maize (Zea mays L.). The efflux of C was set constant or modelled by a power function of the distance from the apex to simulate the greater release of C around the root tip commonly reported in the literature. The influx was proportional to the C concentration in the external solution to simulate the active re-uptake of exudates by the root. Plants were exposed to full light or to shade to manipulate C allocation to roots. The model with a constant efflux gave satisfactory fits to the kinetics of exudation (average R(2)=0.66). The average gross efflux was then 2.1 mug C cm(-2) root surface h(-1). The model was improved if exudation was set more intense towards the root apex (average R(2)=0.74). The estimated gross efflux decreased then from 5.2 mug C cm(-2) h(-1) at the apex to 1.8 mug C cm(-2) h(-1) for the region located 5-25 cm from the root tip. The decrease in net exudation of individual roots due to the shading of plants was weak, which may indicate that the import of C by the primary roots studied was not reduced significantly. By describing the exudation of an apical root segment of variable length and diameter, the model is a first step in linking exudation to root system architecture models and to whole plant functioning.     

小麦和玉米根系取样位置优化确定及根系分布模拟

为了确定小麦(Triticum aestivum)、玉米(Zea mays)根系的最优取样位置和更准确地模拟根长密度在土壤剖面的分布, 在冬小麦和夏玉米的灌浆后期, 采用根钻法取样, 比较了不同取样位置对根系分布的影响; 采用Gerwitz和Page模型对根长密度的分布进行了模拟。结果表明, 冬小麦行间和行上取样在0-10 cm土层根长密度差异显著, 在10 cm以下土层差异减少。在确定根长密度分布的取样中, 在0-20 cm土层应考虑根长密度分布的空间差异, 即行上密度大于行间密度; 而在20-100 cm土层, 需要考虑行间根长密度大于行上的空间差异; 在1 m以下土层两个位置的差异逐渐消失, 可不考虑空间差异。夏玉米根长密度在上层土壤表现出距离植株不同位置差异显著的特征。植株位置(株上)、距植株10 cm和距植株20 cm位置根长密度在土壤中的分布特征是: 0-10 cm土层3个位置根长密度差异在50%以上, 根长密度大小是株上>距植株10 cm>距植株20 cm; 而在10-30 cm层次, 根长密度表现为距植株10 cm>株上>距植株20 cm, 30-50 cm土层株上位置的根长密度最小, 50 cm以下各位置根长密度差异不明显。对于玉米根系取样, 50 cm以上土层需要考虑根长密度的空间差异, 50 cm以下土层可不考虑。采用Gerwitz和Page模型, 结合华北平原机械化耕作下形成的土壤犁底层变厚及其犁底层容重增加对根系分布的影响, 在模型中加入土壤容重参数订正可以使模型更准确地模拟根长密度在土壤剖面的分布。     

Phylogenetic relationships among Lactuca (Asteraceae) species and related genera based on ITS-1 DNA sequences

Internal transcribed spacer (ITS-1) sequences from 97 accessions representing 23 species of Lactuca and related genera were determined and used to evaluate species relationships of Lactuca sensu lato (s.l.). The ITS-1 phylogenies, calculated using PAUP and PHYLIP, correspond better to the classification of Feráková than to other classifications evaluated, although the inclusion of sect. Lactuca subsect. Cyanicae is not supported. Therefore, exclusion of subsect. Cyanicae from Lactuca sensu Feráková is proposed. The amended genus contains the entire gene pool (sensu Harlan and De Wet) of cultivated lettuce (Lactuca sativa). The position of the species in the amended classification corresponds to their position in the lettuce gene pool. In the ITS-1 phylogenies, a clade with L. sativa, L. serriola, L. dregeana, L. altaica, and L. aculeata represents the primary gene pool. L. virosa and L. saligna, branching off closest to this clade, encompass the secondary gene pool. L. virosa is possibly of hybrid origin. The primary and secondary gene pool species are classified in sect. Lactuca subsect. Lactuca. The species L. quercina, L. viminea, L. sibirica, and L. tatarica, branching off next, represent the tertiary gene pool. They are classified in Lactuca sect. Lactucopsis, sect. Phaenixopus, and sect. Mulgedium, respectively. L. perennis and L. tenerrima, classified in sect. Lactuca subsect. Cyanicae, form clades with species from related genera and are not part of the lettuce gene pool.     

Recent amplification of triose phosphate isomerase related sequences in lettuce.

A random cDNA clone was identified as distinguishing near-isogenic lines for downy mildew resistance in lettuce. The clone detected multiple restriction fragments in genomic Southern blots of lettuce. Restriction fragment length polymorphisms (RFLPs) detected by this clone mapped to separate clusters of resistance genes; therefore, these sequences were studied in a greater detail. Sequence analysis indicated that the cDNA encoded the glycolytic enzyme triose phosphate isomerase (TPI). The lettuce clone shares 85% sequence similarity at the amino acid level with TPI from maize. TPI-related sequences were mapped in lettuce using three crosses. Ten loci were distributed in six linkage groups. Possible mechanisms of amplification and dispersion were investigated. Retrotransposition was excluded, since intron five is retained in all TPI-related genomic sequences. Large scale chromosomal rearrangements were not involved, as RFLP markers flanking TPI loci were not duplicated. A high level of genomic variability was detected by the TPI clone; 37 different restriction fragments were detected in Southern hybridizations to 64 populations of lettuce including 47 cultivars of Lactuca sativa and five wild species. Species distantly related to L. sativa had few TPI loci, indicating that their amplification and dispersion were recent and had occurred after the emergence of the L. serriola complex.     

Photosynthetic characteristics of Lactuca serriola L.

Abstract. Gas exchange responses of intact leaves of Lactuca serriola serriola and L. serriola integrifolia were examined. The light, temperature and intercellular CO₂ dependence of photosynthesis of these two forms of L. serriola were indistinguishable. Dependence of photosynthesis on leaf temperature differed only slightly between plants grown at approximately 10/3°C and those grown at 33/25°C with the thermal optimum shifting 4°C. Transpiration was shown to increase exponentially with leaf temperature. The gas exchange characteristics of L. serriola are discussed in relation to water relations, carbon gain and compass leaf orientation under field conditions.     

Promotive effect of capillarol and related compounds on root growth

The promotion of root growth by capillarol [methyl 3-(3-methylbut-2-enoyl)-4-hydroxycinnamate] and related phenolic compounds were studied in relation to structure-activity relationships. Concentrations above 5 × 10⁻⁵ M capillarol stimulated the root growth of rice ( Oryza sativa L. cv. Tanginbozu and cv. Nihonbare) seedlings to about 180% of the control value at 5 × 10⁻⁴ M . Capillarol had no promotive and hardly any inhibitory effect on the growth of the second leaf sheath. Capillarol promoted the root growth also in seedlings of lettuce ( Lactuca sativa L. cv. Grand Rapids) to ca 150% of the control value at 5 × 10⁻⁴ M . The free acid form of capillarol (capillaric acid) was about as effective as capillarol. Para -hydroxy- but not m -methoxy- substituted cinnamic acid, phenylpyruvic acid, phenylacetic acid and amino-hydrocinnamic acid could stimulate root growth, but p -hydroxybenzoic acid was inactive. It is concluded that the important structural requirements for high root growth-promoting activity of phenolic compounds are the hydroxyl group substitution at the C-4 position of the benzene ring, and the propanoic or propenoic acid side chain at the C-1 position. A possible mode of the action of capillarol on root growth-promoting activity is also discussed.     

检疫性杂草毒莴苣的光合特征及其入侵地群落学生态调查

毒莴苣是我国东南沿海地区的新入侵杂草,也是国家进境植物的检疫对象.调查发现,毒莴苣植株高大,易在入侵地形成群落优势种;常见的伴生杂草有小飞蓬、野塘蒿、鬼针草、裂叶月见草、裂叶牵牛、狗尾草、野胡萝卜、苍耳、一年蓬、山莴苣、葎草、龙葵和钻形紫菀等.应用LCA 4光合蒸腾测定系统对毒莴苣进行净光合速率测定,结果表明：该种实测净光合速率高达21.22±0.45 μmol CO₂·m^-2·s^-1,比入侵性杂草一年蓬、野塘蒿稍低,比藜、北美车前、山莴苣等高,是一种高光效植物;根据毒莴苣的光合-响应曲线,该外来入侵种的理论光补偿点为37.58 μmol·m^-2·s^-1, 光饱和点为1 480 μmol·m^-2·s^-1,理论最大净光合速率20.81 μmol CO₂·m^-2·s^-1;毒莴苣的光合作用具有午休现象,是由于高光照和高温导致气孔阻力增加、气孔关闭,影响了植株对外的气体交换;影响净光合速率的主要因素是气孔导度、叶面光合有效辐射和叶片的蒸腾.     

Molecular diversity at the major cluster of disease resistance genes in cultivated and wild Lactuca spp.

Diversity was analyzed in wild and cultivated Lactuca germplasm using molecular markers derived from resistance genes of the NBS-LRR type. Three molecular markers, one microsatellite marker and two SCAR markers that amplified LRR-encoding regions, were developed from sequences of resistance gene homologs at the main resistance gene cluster in lettuce. Variation for these markers were assessed in germplasm including accessions of cultivated lettuce, Lactuca sativa L. and three wild Lactuca spp., L. serriola L., L. saligna and L. virosa L. Diversity was also studied within and between natural populations of L. serriola from Israel and California; the former is close to the center of diversity for Lactuca spp. while the latter is an area of more recent colonization. Large numbers of haplotypes were detected indicating the presence of numerous resistance genes in wild species. The diversity in haplotypes provided evidence for gene duplication and unequal crossing-over during the evolution of this cluster of resistance genes. However, there was no evidence for duplications and deletions within the LRR-encoding regions studied. The three markers were highly correlated with resistance phenotypes in L. sativa. They were able to discriminate between accessions that had previously been shown to be resistant to all known isolates of Bremia lactucae. Therefore, these markers will be highly informative for the establishment of core collections and marker-aided selection. A hierarchical analysis of the population structure of L. serriola showed that countries, as well as locations, were significantly differentiated. These differences may reflect local founder effects and/or divergent selection. Received: 7 March 1999 / Accepted: 25 March 1999