The resistance of lettuce (Lactuca sativa L.) toNasonovia ribisnigri: Bionomics ofN. ribisnigri on near isogenic lettuce lines

The intrinsic rate of increase (r_m-values), mean relative growth rates and mortality ofNasonovia ribisnigri (Mosley) (Homoptera, Aphididae) on different lines of lettuce (Lactuca sativa L.) were determined. Near isogenic resistant and susceptible lines, plus their ancestors, were used. Bionomics ofN. ribisnigri on the resistant lines (NrNr) with the dominantNasonovia resistance gene (Nr-gene) differed clearly from the susceptible lines (nrnr). Mortality was high, no larvae reached adulthood, and no reproduction nor honeydew production was seen on the resistant lines. Transfer of aphids to susceptible plants after a period of 2 days on the resistant lines showed no signs of intoxication of aphids. Apparently there is no feeding on the resistant lines. It is not clear whether the aphids can not reach the phloem or do not accept it on the resistant line.     

Transactivation of Ds elements in plants of lettuce (Lactuca sativa)

The maize transposable element, Activator (Ac), is being used to develop a transposon mutagenesis system in lettuce, Lactuca sativa. In this paper, we describe somatic and germinal transactivation of Ds by chimeric transposase genes in whole plants. Constructs containing either the Ds element or the Ac transposase open reading frame (ORF) were introduced into lettue. The Ds element was located between either the 35S or the Nos promoter and a chimeric spectinomycin resistance gene (which included a transit peptide), preventing expression of spectinomycin resistance. The genomic coding region of the Ac transposase was expressed from the 35S promoter. Crosses were made between 104 independent R₁ plants containing Ds and three independent R₁ plants expressing transposase. The excision of Ds in F₁ progenies was monitored using a phenotypic assay on spectinomycin-containing medium. Green sectors in one-third of the F₁ families indicated transactivation of Ds by the transposase at different developmental stages and at different frequencies in lettuce plants. Excision was confirmed using PCR and by Southern analysis. The lack of green sectors in the majority of F₁ families suggests that the majority of T-DNA insertion sites are not conducive to excision. In subsequent experiments, the F₁ plants containing both Ds and the transposase were grown to maturity and the F₂ seeds screened on medium containing spectinomycin. Somatic excision was again observed in several F₂ progeny; however, evidence for germinal excision was observed in only one F₂ family.     

Analysis of wheat resistance to the cereal aphidSitobion avenae using electrical penetration graphs and flow charts combined with correspondence analysis

The behaviour ofSitobion avenae (F.), was compared on resistant wheat lines ofTriticum monococcum (L.) and a susceptible variety ofTriticum aestivum (L.). Firstly, stylet penetration activities were monitored with the Electrical Penetration Graph (EPG) technique and subsequently analysed using flow charts combined with correspondence analysis. Plant resistance was shown to be associated with repeated penetrations without access to either the xylem or the phloem, and with numerous failures in starting a sustained sap ingestion (as represented by pattern E2). Access to sieve elements of the phloem did not seem to be much affected on resistant plants but it took the aphid three times as long to produce a sap ingestion pattern when maintained on the resistant lineT. monococcum n^o 44 (Tm44) as compared with aphids maintained on susceptible plants. As a result the total time spent in ingesting from sieve elements was reduced by 72% on Tm44. Secondly, direct observations of freely-moving apterous adults were performed. Aphids did not discriminate between resistant and susceptible wheat during the first 30 min of access to test leaves, but only 4 out of 25 aphids were still probing after eight hours on resistant Tm44. The relevance of these results to possible location of the resistance factor(s) are discussed. Although detection of plant resistance before sieve elements are reached can not be rigorously excluded, the factors involved inT. monococcum resistance toS. avenae undoubtedly occur within the phloem vessels.     

FRET hybridization probes for the rapid detection of disease resistance alleles in plants: detection of corky root resistance in lettuce

We describe the development of a non-electrophoresis PCR-based assay for allele discrimination at a disease resistance locus. The assay is based on the emission of light by fluorescence resonance energy transfer (FRET) upon annealing of two hybridization probes. The analysis of melting curve profiles of the probes and templates allowed the detection of single nucleotide polymorphisms. The assay was applied to the detection of alleles at the cor locus in lettuce (Lactuca sativa) that confers recessive resistance to corky root disease. Probes and primers for the assay were designed after the characterization of a single nucleotide polymorphism between alleles of PCR products amplified using a linked marker. That polymorphism was validated in a collection of lettuce varieties representing different genetic backgrounds. The FRET hybridization probes approach provided fast and accurate genotyping of breeding material directly in a one-tube reaction. The absence of electrophoresis makes this approach suitable for applications that require automation and high-throughput genotyping analyses such as marker-assisted selection programs.     

Response of lettuce to pre- and post-transplant phosphorus and pre-transplant inoculation with a VA-mycorrhizal fungus

Pre-transplant inoculation of lettuce (Lactuca sativa L.) seedlings with the vesicular-arbuscular mycorrhizal fungusGlomus aggregatum (Smith and Schenck emend. Koske) increased P uptake and dry matter yields after transplanting into soil when the concentration of P in the soil solution was 0.02 mg L^–1 but had little affect in soil with 0.30 mg L^–1 solution P. Tissue P concentrations and dry matter yields after transplanting were increased by supplying adequate P prior to transplanting. Adequate levels of pre-transplant P appeared to be more important than maximum mycorrhizal infection of transplants for promoting post-transplant growth of the fast maturing lettuce crop.Journal Series No. 0000 of the Hawaii Institute of Tropical Agriculture and Human Resources.     

Molecular analysis of irradiation-induced and spontaneous deletion mutants at a disease resistance locus inLactuca sativa

The major cluster of disease resistance genes in lettuce (Lactuca sativa) contains at least nine downy mildew resistance genes (Dm) spanning a genetic distance of 20 cM and a physical distance of at least 6 Mb. Nine molecular markers that were genetically tightly linked toDm3 were used to analyze nine independent deletion mutants and construct a map of the region surroundingDm3. This analysis identified a linear order of deletion breakpoints and markers along the chromosome. There was no evidence for chromosomal rearrangements associated with the deletions. The region is not highly recombinagenic and the deletion breakpoints provided greater genetic resolution than meiotic recombinants. The region contains a mixture of high- and low-copy-number sequences; no single-copy sequences were detected. Three markers hybridized to low-copy-number families of sequences that are duplicated predominantly close toDm3. This was not true for sequences related to the triose-phosphate isomerase gene; these had been shown previously to be linked toDm3, as well as to two independent clusters ofDm genes, and elsewhere in the genome. Two spontaneous mutants ofDm3 were identified; several markers flankingDm3 are absent in one of these two mutants. The stability of theDm3 region was also studied by analyzing the genotypes of diverse related cultivars. The 1.5 Mb region surroundingDm3 has remained stable through many generations of breeding with and without selection forDm3 activity.     

Stable transformation of lettuce cultivar South Bay from cotyledon explants

Transgenic plants of lettuce cultivar (cv.) South Bay were produced by using Agrobacterium tumefaciens vectors containing the -glucuronidase (GUS) reporter gene and the NPT II gene for kanamycin resistance as a selectable marker. High frequency of transformation, based on kanamycin resistance and assays for GUS expression, was obtained with 24 to 72-h-old cotyledon explants cocultivated for 48 h with Agrobacterium tumefaciens. After the cocultivation period, the explants were placed in selection medium containing 50 or 100 mg l^–1 of kanamycin, 100 mg l^–1 cefotaxime and 500 mg l^–1 carbenicillin for 10 days. Surviving explants were transferred every 14 days on shoot elongation medium. Progenies of R₀ plants demonstrated linked monogenic segregation for kanamycin resistance and GUS activity.Florida Agricultural Experiment Station Journal Series R-02231. This research was partially supported by CNPq/RHAE (Brazil).     

Variability among lettuce cultivars grown at two levels of available phosphorus

Fifteen lettuce cultivars representing three different morphological types were grown in a sand alumina system under conditions of low (deficient) and high (sufficient) P supply. An efficient plant was defined as one that produced a large shoot fresh weight under low P conditions. Cultivars within their respective groups varied significantly for some traits that appeared to be important in determining adaptation to P. This led to the conclusion that accumulation of P in shoot tissue or the total plant was the main difference between efficient and inefficient cultivars. Accumlation of P seemed to be due to a greater absorption capability of roots or greater root mass (weight), depending on the different lettuce groups. Differences in internal use of P did not contribute to differences in shoot fresh weight.The butterhead cultivars were the least efficient plants when grown under low P. Compared to the other groups, plants has lower translocation efficiency and a greater root: shoot ratio. Never-the-less, butterhead cultivars as efficient as the best cultivars of other groups were found. There were no differences between Brazilian and American cultivars for any of the traits analysed, probably due to the fact that in both countries vegetables are bred under high fertility levels and grown with heavy applications of fertilizers. The results of this study demonstrate that there were genotypic variability and/or genotype×environment interaction effects for shoot weight (yield) among the lettuce cultivars grown under low P conditions imposed in the sand-alumina system.     

Infrequent transposition of Ac in lettuce,Lactuca sativa

The maize transposable element Activator (Ac) is being used to develop a transposon mutagenesis system in lettuce, Lactuca sativa. Two constructs containing the complete Ac from the waxy-m7 locus of maize were introduced into lettuce and monitored for activity using Southern analysis and PCR amplification of the excision site. No transposition of Ac was detected in over 32 transgenic R₁ plants, although these constructs were known to provide frequent transposition in other species. Also, no transposition was observed in later generations. In subsequent experiments, transposition was detected in lettuce calli using constructs that allowed selection for excision events. In these constructs, the neomycin phosphotransferase II gene was interrupted by either Ac or Ds. Excision was detected as the ability of callus to grow on kanamycin. Synthesis of the transposase from the cDNA of Ac expressed from the T-DNA 2 promoter resulted in more frequent excision of Ds than was observed with the wild-type Ac. No excision was observed with Ds in the absence of the transposase. The excision events were confirmed by amplification of the excision site by PCR followed by DNA sequencing. Excision and reintegration were also confirmed by Southern analysis. Ac/Ds is therefore capable of transposition in at least calli of lettuce.     

Growth effects of nagilactone E on cultured cells of lettuce

Nagilactone E, a norditerpene dilactone isolated from a gymnosperm, Podocarpus nagi (Podocarpaceae), was able to stimulate the growth of cultured cells of Lactuca sativa cv. Grand Rapids at 0.1 g ml^-1 but not at 0.01 or 1 g ml^-1. Cell wet weight/unit time and cell number/unit wet weight were increased when they were recorded at the end of the 14-day incubation period. Also, the population of cells treated with that concentration of nagilactone E had a higher percentage of cells with shorter cell length compared to the control (untreated).     

A study of factors which regulate the membrane appression of lettuce thylakoids in relation to irradiance

Factors that may influence the extent of thylakoid membrane appression have been examined using lettuce (Lactuca sativa cv. Celtuce) grown under different irradiances. Electron microscopy and salt-induced chlorophyll fluorescence suggest that the percentage of membrane appression is increased in plants grown in low light (20 Wm^–2) compared with those grown in high light (150 Wm^–2). In high light plants surface charge, as measured by 9-aminoacridine, was found to be twice that measured in low light plants. There was a similar difference in ATPase activity of CF₁ and in light saturated photophosphorylation. The chlorophyll content of LHC-2 as a proportion of the total chlorophyll was greatest in thylakoids of low light plants. Measurement of non-cyclic photophosphorylation rates suggested that membrane appression has a stimulatory role in the photophosphorylation process. The importance of these inter-related factors for the mechanism of thylakoid appression is discussed.Abbreviations PS photosystem - chl chlorophyll - LHC-2 light harvesting chlorophyll-protein complex serving PS 2 - CF₁ coupling factor 1 - NADP nicotinamide-adenine dinucleotide phosphate     

纳米二氧化钛对生菜的生长效应分析

为了阐明纳米二氧化钛颗粒(TiO₂NPs)对生菜(Lactuca sativa)生长的影响，采用自行设计的水培装置探究不同浓度TiO₂NPs (300~1 200 mg/L)下，生菜生长和生理生化指标的变化。结果表明，300 mg/L TiO₂NPs能促进生菜幼苗的根长、茎长、叶表面积、鲜重和干重；随着TiO₂ NPs浓度增大，生菜的生长指标呈现下降趋势，但仍优于对照组。生菜体内的抗氧化酶(SOD、POD)在低TiO₂ NPs浓度(300 mg/L)时，活性明显下降；随着TiO₂ NPs浓度增大，这两种抗氧化酶活性逐渐增强。因此，生菜对TiO₂NPs胁迫具有浓度依赖性，表现为“低促高抑”，且能够通过抗氧化酶系统来减轻TiO₂NPs伤害。     

Vesicular-arbuscular mycorrhizal infection in lettuce (Lactuca sativa) in relation to calcium supply

Summary Infection of lettuce roots (Lactuca sativa L.) by the vesicular-arbuscular mycorrhizal fungiGlomus caledonium andGlomus mosseae was dependent on the amount of calcium (supplied as CaCl₂·2H₂O or CaSO₄·2H₂O) in the nutrient solution; those plants growing at low calcium concentrations being poorly infected.     

罗马直立生菜生长发育及营养指标对臭氧浓度升高和持续熏蒸的响应

环境臭氧(O₃)已成为影响植物生长发育的重要生态因子。为探究地面O₃污染对蔬菜形态学特征及营养指标的影响,选罗马直立生菜(Lactuca sativa var. roman)为实验材料,采用开顶式气室开展熏蒸实验。实验设置4个O₃熏蒸浓度(NF:未过滤的环境空气;NF40:环境空气+40 nmol/mol;NF80:环境空气+80 nmol/mol;NF120:环境空气+120 nmol/mol),每个处理设置3个重复组,分析评价O₃污染对植物造成的可见伤害、生产量、叶片解剖学特征以及食用部位营养指标的影响。研究表明:(1)O₃熏蒸对生菜叶片产生不可逆的可见伤害,叶片出现浅黄色斑点和棕色斑点,且随着熏蒸时间延长,叶片出现黄化,大面积的坏死斑块,衰老加速。(2)高浓度O₃胁迫显著降低了生长阶段的株高(P<0.05)。与NF组相比,NF40、NF80、NF120组分别使生物量下降5.90%、14.99%、39.21%。(3)随着O₃熏蒸浓度升高,气孔密度增加,气孔开度减小。叶片厚度、海绵组织厚度、栅栏组织厚度与O₃暴露剂量AOT40呈显著负相关关系(P<0.05)。(4)高浓度O₃暴露使蔬菜中Ca、Na、Fe、Zn、Mg等元素含量显著降低,脂肪和蛋白质含量增加,生菜的营养指标发生改变。研究表明,罗马直立生菜对环境O₃污染敏感,其生长发育及营养指标在O₃胁迫条件下发生明显变化。目前,关于O₃污染对蔬菜形态学特征影响的研究较少,研究系统探讨蔬菜的叶片厚度、栅栏组织、海绵组织、气孔密度及开度等形态学指标在臭氧污染条件下的变化。蔬菜的品质是关系到"三农"问题的重要方面,研究探讨了臭氧污染对蔬菜的产量及营养指标的影响,可为O₃污染条件下蔬菜的生产提供科学参考。     

Effect of hypobaric conditions on ethylene evolution and growth of lettuce and wheat

Elevated levels of ethylene occur in enclosed crop production systems and in spaceflight environments, leading to adverse plant growth and sterility. There are engineering advantages in growing plants at hypobaric (reduced atmospheric pressure) conditions in biomass production for extraterrestrial base or spaceflight environments. Objectives of this research were to characterize the influence of hypobaria on growth and ethylene evolution of lettuce (Lactuca sativa) and wheat (Triticum aestivum). Plants were grown under variable total gas pressures [from 30 to 101 kPa (ambient)]. In one study, lettuce and wheat were direct seeded, germinated and grown in the same chambers for 28 d at 50 or 101 kPa. Hypobaria increased plant growth and did not alter germination rate. During a 10-day study, 28-day-old lettuce and 40-day-old wheat seedlings were transplanted together in the same low and ambient pressure chambers; ethylene accumulated in the chambers, but the rate of production by both lettuce and wheat was reduced more than 65% under 30 kPa compared with ambient pressure (101 kPa). Low O2 concentrations [partial pressure of O2 (pO2) = 6.2 kPa] inhibited ethylene production by lettuce under both low (30 kPa) and ambient pressure, whereas ethylene production by wheat was inhibited at low pressure but not low O2 concentration. There was a negative linear correlation between increasing ethylene concentration and decreasing chlorophyll content of lettuce and wheat. Lettuce had higher production of ethylene and showed greater sensitivity to ethylene than wheat. The hypobaric effect on reduced ethylene production was greater than that of just hypoxia (low oxygen).     

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.     

基于EPG的麦长管蚜、麦二叉蚜和禾谷缢管蚜取食行为比较

麦长管蚜Sitobion avenae (Fabricius),麦二叉蚜Schizaphis graminum (Rondani)和禾谷缢管蚜Rhopalosiphum padi (Linnaeus)虽然都取食小麦,但其取食部位、传毒能力均有差异。利用EPG研究了3种麦蚜在同一寄主植物上的取食行为,结果显示:(1)禾谷缢管蚜在取食过程中G波出现的时间最早,G波总的持续时间和每次刺探G波的平均持续时间都明显长于麦长管蚜和麦二叉蚜。麦长管蚜第1次E1波的时间、E1波总的持续时间和每次刺探E1波的平均持续时间都明显比麦二叉蚜和禾谷缢管蚜要长。麦二叉蚜E2波总的持续时间和每次刺探E2波的平均持续时间都明显长于麦长管蚜和禾谷缢管蚜。表明3种蚜虫各自拥有不同取食策略。禾谷缢管蚜比其它两种蚜虫需要更多的水分和无机盐,麦长管蚜通过分泌大量唾液增大对食物的利用率,麦二叉蚜通过增加韧皮部取食时间以满足对营养的需要。(2)刺探过程中麦二叉蚜pd波出现的次数为(51.97±7.82)次,高于麦长管蚜的(44.73±4.52)次和禾谷缢管蚜的(32.99±4.22)次。麦二叉蚜pd波Ⅱ-2和Ⅱ-3亚波的时间最长,禾谷缢管蚜最短,三者之间差异达显著水平。表明与其它两种麦蚜相比,麦二叉蚜在取食过程中口针与细胞间的接触频率更高,与细胞内获毒和传毒相关的Ⅱ-3波和Ⅱ-2波持续时间更长,增加了获毒和传毒的几率,是其传毒能力最强的机理。     

致病性葡萄球菌cfr基因介导的多重耐药机制研究进展

葡萄球菌是临床常见致病菌及食源性致病菌,可在食品原料加工、包装及运输过程中污染食品,引起人体多种严重感染,其耐药性的不断增强对公共卫生安全产生了重大的威胁。葡萄球菌中cfr (chloramphenicol-florfenicol resistance)基因编码的甲基转移酶,可引起细菌核糖体RNA的甲基化,从而阻碍或减弱多种化学结构不同的抗生素与肽基转移酶活性中心(peptidyl transferase center,PTC)的结合,导致葡萄球菌多重耐药表型的出现。噁唑烷酮类药物−利奈唑胺是继万古霉素后治疗耐药革兰氏阳性菌所致感染的最后一道防线,cfr基因的出现大大加速了利奈唑胺耐药性的传播。cfr基因广泛分布于多种致病性葡萄球菌中,cfr基因与各类型可转移元件(质粒、转座子和整合相关元件等)密切关联的遗传环境是其广泛传播的结构基础。在cfr基因水平传播的过程中,食源性致病葡萄球菌作为中间者扮演着重要的角色。本文就近年来国内外对致病性葡萄球菌中cfr基因的分布状况、耐药机制、遗传环境、传播机制等进行综述,以期为防控致病性葡萄球菌的传播提供参考,以遏制多重耐药菌的进一步传播。     

Acyrthosiphon pisum performance in response to the sugar and amino acid composition of artificial diets, and its relation to lucerne varietal resistance

In order to understand the mechanisms of resistance of two lucerne cultivars (susceptible Resistador and resistant Lahontan clones) to a French biotype of Acyrthosiphon pisum Harris, several biological parameters of this aphid were investigated on sap-copied holidic diets differing either by their amino acid or sucrose concentrations or by the relative proportions of some amino acids.We recorded the quantitative influence of nutrient levels and amino acid profiles on adult survival and reproduction, as well as on larval growth and development. The differences in A. pisum performances on artificial diets may partly by explained by variable ingestion rates, but also by differences in metabolic efficiency of the ingested nutrients, which greatly depends on amino acid composition.Although holidic diet experiments partially confirm the biological assays of resistance on plants, they also point out a lack in our understanding of the factors present in the sap which are involved in phagostimulation in vivo, thus preventing a complete interpretation of observed resistance.

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Résumé Sur des milieux holidiques se différenciant par leur concentration en acides aminés ou en saccharose ou par leur spectre en acides aminés et copiant la sève de deux variétés de luzerne (sensible et résistante) nous avons étudié différents paramètres biologiques d'un biotype d'Acyrthosiphon pisum Harris afin de mieux comprendre les mécanismes de résistance à ce puceron.Que ce soit pour le reproduction et la survie des adultes ou pour la croissance et le développment larvaire, nous montrons l'influence spécifique de la concentration en saccharose et en acides aminés ainsi que celle du spectre en acides aminés des régimes artificiels. Ces différences de performances de A. pisum peuvent en partie être expliquées par les différences d'ingestion. Néanmoins pour des rapport saccharose/acides aminés identiques et pour des niveaux d'ingestion égaux, il existe selon les milieux de grandes différences de taux de reproduction et donc d'efficacités métaboliques.Les milieux copiant la sève de variétés sensible et résistante permettent jusqu'à présent d'expliquer une partie des observations faites sur végétal, mais démontrent que l'ignorance de certains facteurs phagostimulants présents dans la plante empêche une interprétation complète des effets observés.