Detecção de Salmonella em tomates (Lycopersicon esculentum mill) pelos métodos fda-bam e mini vidas-biomérieux.

Resumo: Diversos surtos de Salmonella ocasionados pelo consumo de tomate contaminados com este micro-organismo têm sido relatados ultimamente, o que torna primordial a investigação sobre a presença desse patógeno nesse alimento. Métodos que permitam a avaliação rápida da presença de Salmonella em alimentos são de suma importância. O objetivo desse estudo foi comparar o método tradicional da Food and Drug Administration - Bacteriologycal Analytical Manual (FDA-BAM) com um método rápido da mini Vitek Immuno Diagnostic System Assay (Mini?Vidas-SLM)-bioMérieux, para detecção de Salmonella Brazil inoculada artificialmente na superfície de tomates. Foram analisadas 215 amostras de tomates inoculadas artificialmente com Salmonella Brazil com níveis de inóculos variando de 0,4 a 940 UFC/tomate. Os resultados obtidos mostraram que os métodos estudados apresentaram uma ótima concordância entre si, para todas as faixas de inóculo analisadas.

Resistência de acessos de Lycopersicon a Stemphylium solani e S. lycopersici.

A mancha-de-estenfílio, causada por Stemphylium solani e/ou S. lycopersici, é uma conhecida doença do tomateiro que voltou a ser importante, principalmente porque os atuais híbridos plantados no país não apresentam resistência. O controle da mancha-de-estenfílio se baseia no uso de cultivares resistentes ou outras medidas de manejo como o controle químico e a rotação de cultura. Este trabalho teve como objetivo selecionar novas fontes de resistência à doença in Lycopersicon. Entre os acessos de Lycopersicon avaliados para resistência 18 (54,5%) comportaram-se como suscetíveis, três (9,1%) comportaram-se como intermediários e 12 (36,4%) foram resistentes para as duas espécies fúngicas. Todos os acessos resistentes a S. solani também foram resistentes a S. lycopersici. Foram identificados acessos resistentes e intermediários nas espécies L. esculentum (muito provavelmente portando o gene Sm), L. peruvianum e L. hirsutum.

Tipos de tricomas em genótipos de Lycopersicon

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Influência da geometria e da temperatura na cinética de secagem de tomate (Lycopersicum esculentum)

O tomate é um fruto muito perecível por causa do seu conteúdo de umidade. A secagem é uma das práticas industriais mais utilizadas em alimentos para manter a qualidade do produto final. A pesquisa foi desenvolvida para estudo dos parâmetros de secagem de tomate (Lycopersicon esculentum Mill), cv Carmen, com relação ao tipo de corte (meio e um quarto) e à temperatura de processo (60 e 70 ºC), bem como à escolha do tempo de secagem para a obtenção de um produto com umidade de 45% (base úmida). As cinéticas de secagem foram determinadas experimentalmente por convecção forçada e ajustadas ao modelo de Page. Os resultados obtidos mostraram que a geometria de corte influenciou na taxa de secagem e no tempo de desidratação. Os tomates cortados em quatro partes e desidratados a 70 ºC alcançaram umidade de 45% em menor tempo (10 horas), quando comparados aos tomates com o corte em metades. O modelo de Page forneceu bom ajuste nas cinéticas de secagem

Suramin inhibits initiation of defense signaling by systemin, chitosan, and a β-glucan elicitor in suspension-cultured Lycopersicon peruvianum cells

Systemin-mediated defense signaling in tomato (Lycopersicon esculentum) plants is analogous to the cytokine-mediated inflammatory response in animals. Herein, we report that the initiation of defense signaling in suspension-cultured cells of Lycopersicon peruvianum by the peptide systemin, as well as by chitosan and β-glucan elicitor from Phytophtora megasperma, is inhibited by the polysulfonated naphtylurea compound suramin, a known inhibitor of cytokine and growth factor receptor interactions in animal cells. Using a radioreceptor assay, we show that suramin interfered with the binding of the systemin analog 125I-Tyr-2,Ala-15-systemin to the systemin receptor with an IC50 of 160 μM. Additionally, labeling of the systemin receptor with a photoaffinity analog of systemin was inhibited in the presence of suramin. Receptor-mediated tyrosine phosphorylation of a 48-kDa mitogen-activated protein kinase and alkalinization of the medium of suspension-cultured cells in response to systemin and carbohydrate elicitors were also inhibited by suramin. The inhibition of medium alkalinization by suramin was reversible in the presence of high concentrations of systemin and carbohydrate elicitors. Calyculin A and erythrosin B, intracellular inhibitors of phosphatases and plasma membrane proton ATPases, respectively, both induce medium alkalinization, but neither response was inhibited by suramin. The polysulfonated compound heparin did not inhibit systemin-induced medium alkalinization. NF 007, a suramin derivative, induced medium alkalinization, indicating that neither NF 007 nor heparin interact with elicitor receptors like suramin. The data indicate that cell-surface receptors in plants show some common structural features with animal cytokine and growth factor receptors that can interact with suramin to interfere with ligand binding.

Three Drought-Responsive Members of the Nonspecific Lipid-Transfer Protein Gene Family in Lycopersicon pennellii Show Different Developmental Patterns of Expression1

Genomic clones of two nonspecific lipid-transfer protein genes from a drought-tolerant wild species of tomato (Lycopersicon pennellii Corr.) were isolated using as a probe a drought- and abscisic acid (ABA)-induced cDNA clone (pLE16) from cultivated tomato (Lycopersicon esculentum Mill.). Both genes (LpLtp1 and LpLtp2) were sequenced and their corresponding mRNAs were characterized; they are both interrupted by a single intron at identical positions and predict basic proteins of 114 amino acid residues. Genomic Southern data indicated that these genes are members of a small gene family in Lycopersicon spp. The 3′-untranslated regions from LpLtp1 and LpLtp2, as well as a polymerase chain reaction-amplified 3′-untranslated region from pLE16 (cross-hybridizing to a third gene in L. pennellii, namely LpLtp3), were used as gene-specific probes to describe expression in L. pennellii through northern-blot analyses. All LpLtp genes were exclusively expressed in the aerial tissues of the plant and all were drought and ABA inducible. Each gene had a different pattern of expression in fruit, and LpLtp1 and LpLtp2, unlike LpLtp3, were both primarily developmentally regulated in leaf tissue. Putative ABA-responsive elements were found in the proximal promoter regions of LpLtp1 and LpLtp2.

De Novo Transcriptome Sequence Assembly and Analysis of RNA Silencing Genes of Nicotiana benthamiana

Background: Nicotiana benthamiana has been widely used for transient gene expression assays and as a model plant in the study of plant-microbe interactions, lipid engineering and RNA silencing pathways. Assembling the sequence of its transcriptome provides information that, in conjunction with the genome sequence, will facilitate gaining insight into the plant's capacity for high-level transient transgene expression, generation of mobile gene silencing signals, and hyper-susceptibility to viral infection. Methodology/Results: RNA-seq libraries from 9 different tissues were deep sequenced and assembled, de novo, into a representation of the transcriptome. The assembly, of16GB of sequence, yielded 237,340 contigs, clustering into 119,014 transcripts (unigenes). Between 80 and 85% of reads from all tissues could be mapped back to the full transcriptome. Approximately 63% of the unigenes exhibited a match to the Solgenomics tomato predicted proteins database. Approximately 94% of the Solgenomics N. benthamiana unigene set (16,024 sequences) matched our unigene set (119,014 sequences). Using homology searches we identified 31 homologues that are involved in RNAi-associated pathways in Arabidopsis thaliana, and show that they possess the domains characteristic of these proteins. Of these genes, the RNA dependent RNA polymerase gene, Rdr1, is transcribed but has a 72 nt insertion in exon1 that would cause premature termination of translation. Dicer-like 3 (DCL3) appears to lack both the DEAD helicase motif and second dsRNA binding motif, and DCL2 and AGO4b have unexpectedly high levels of transcription. Conclusions: The assembled and annotated representation of the transcriptome and list of RNAi-associated sequences are accessible at www.benthgenome.com alongside a draft genome assembly. These genomic resources will be very useful for further study of the developmental, metabolic and defense pathways of N. benthamiana and in understanding the mechanisms behind the features which have made it such a well-used model plant. © 2013 Nakasugi et al.

On the role of RNA silencing in the pathogenicity and evolution of viroids and viral satellites

Viroids and most viral satellites have small, noncoding, and highly structured RNA genomes. How they cause disease symptoms without encoding proteins and why they have characteristic secondary structures are two longstanding questions. Recent studies have shown that both viroids and satellites are capable of inducing RNA silencing, suggesting a possible role of this mechanism in the pathology and evolution of these subviral RNAs. Here we show that preventing RNA silencing in tobacco, using a silencing suppressor, greatly reduces the symptoms caused by the Y satellite of cucumber mosaic virus. Furthermore, tomato plants expressing hairpin RNA, derived from potato spindle tuber viroid, developed symptoms similar to those of potato spindle tuber viroid infection. These results provide evidence suggesting that viroids and satellites cause disease symptoms by directing RNA silencing against physiologically important host genes. We also show that viroid and satellite RNAs are significantly resistant to RNA silencing-mediated degradation, suggesting that RNA silencing is an important selection pressure shaping the evolution of the secondary structures of these pathogens.

The assessment of TGGE for the detection of interspecific and intergeneric DNA-marker polymorphism within Solanaceae

RFLP markers are currently the most appropriate marker system for the identification of uncharacterised polymorphism at the interspecific and intergeneric level. Given the benefits of a PCR-based marker system and the availability of sequence information for many Solanaceous cDNA clones, it is now possible to target conserved fragments, for primer development, that flank sequences possessing interspecific polymorphism. The potential outcome is the development of a suite of markers that amplify widely in Solanaceae. Temperature gradient gel electrophoresis (TGGE) is a relatively inexpensive gel-based system that is suitable for the detection of most single-base changes. TGGE can be used to screen for both known and unknown polymorphisms, and has been assessed here, for the development of PCR-based markers that are useful for the detection of interspecific variation within Solanaceae. Fifteen markers are presented where differences between Lycopersicon esculentum and L. pennellii have been detected by TGGE. The markers were assessed on a wider selection of plant species and found to be potentially useful for the identification of interspecific and intergeneric polymorphism in Solanaceous plants.

Estudio en laboratorio sobre la tasa de mortalidad, tiempo de generación y reproducción de Bemisia tabaci (Gennadius) en tomate

El estudio se llevó a cabo en el Laboratorio de Epidemiología y Manejo de Virus en la Escuela de Sanidad Vegetal de la Universidad Nacional Agraria en Managua. Pupas de Bemisia tabaci (Gennadius) fueron colectados originalmente en plantas de frijol en Estelí, para después establecer una cría de mosca blanca limpia de virus, en plantas de tomate Lycopersicon esculentum Mili., variedad VF 134. El experimento se realizó durante el período de septiembre a diciembre de 1992 bajo condiciones del laboratorio con temperatura de 26°C y con 80% de humedad relativa. El experimento consistió en colocar hembras de mosca blanca recien eclosionadas en una jaula de clip (clip cage) sobre una planta de tomate. La jaula fue retirada a las 24 horas, trasladando la hembra a una nueva planta. Se continúo transfiriendo las hembras a una nueva planta cada día hasta su muerte. Cada postura de huevos fue cuantificada y observada durante los siguientes días a fin de registrar el número de los huevos que eclosionaron a ninfas, el número de ninfas que pasaron a adultos y el tiempo transcurrido desde la postura hasta la emergencia de adultos. En base a una longevidad promedio de 9.5 días, se determinó que la tasa mortalidad diaria es de 0.10. El período de preoviposición encontrado es menor de un día. El tiempo promedio para el desarrollo de huevo a adulto fue de 19.2 días. La tasa de oviposición diaria, la sobrevivencia de huevos y la sobrevivencia de ninfas fueron 7.6 huevos por día, 0.92 y 0.74, respectivamente. Por lo tanto se estimó la tasa de reproducción de la mosca blanca en tomate de 5.2 adultos por día.

BADH基因转化番茄提高耐盐性的研究

甜菜碱是植物在盐、干旱或其它胁迫下在细胞中迅速积累的一种相容性有机小分子化合物，它在细胞中的积累与植物抗盐性的提高密切相关。甜菜碱醛脱氢酶（BADH）催化甜菜碱醛转化为甜菜碱。我们将来源于耐盐植物山菠菜（Atriplx hortensis L.）的BADH基因通过农杆菌介导法导入‘百丽春’番茄(Lycopersicon esculentum L. ‘Bailichun’)中，并获得15株转化植株，PCR、Southern和Northern检测表明，其中的6株有外源BADH基因的整合，5株中BADH基因能够正常表达，但不同植株间BADH基因的表达水平和BADH酶活力有较大差异。对叶片电导率的测定表明，转基因植株比野生型的耐盐性有较大提高。T1代分析表明，检测的两个转基因株系后代遵循孟德尔分离规律，90mmol/L NaCl胁迫下种子发芽率提高了2~4倍，幼苗的苗高、根长和须根数三个指标均明显优于对照。部分T1代植株在水培条件下能够耐受180mmol/L NaCl胁迫。 植物耐盐的另一机理就是利用液泡膜上存在的转运蛋白将细胞内的有毒离子区域化。我们将已转入编码转运蛋白基因AtNHX1的番茄品种‘Moneymaker’（L. esculentum‘Moneymaker’）株系X1OEA1通过农杆菌介导法转入山菠菜BADH基因，以期获得转双基因耐盐番茄。目前已获得转基因植株，PCR结果证明部分抗性幼苗中已整合了BADH基因，其它各项分子检测正在进行中。

不同生活型植物对机械刺激的反应

机械刺激效应（Effects of mechanical stimulation）是风生态学直接效应的主要表现形式。研究机械刺激对植物的影响，有利于将风的直接与间接效应区别开来，从而精确研究植物对风的反应格局;研究不同生活型植物对机械刺激的反应是植物力学与植物生态学的重要内容，有助于揭示植物对生境的适应机制。 本研究涉及三个实验。第一个实验探讨不同生活型植物对机械刺激和水分互作的响应格局;第二个实验研究匍匐茎草本植物蛇莓对部分机械刺激的反应;最后一个实验揭示不同水分供应条件下，番茄和紫花苜蓿对不同机械刺激频度的响应。在第一个实验中，克隆半灌木羊柴（Hedysarum laeve），一年生草本植物虫实（Corispermum mongolicum），多年生大型禾草沙鞭（Psammochloa villosa）和多年生丛生禾草黑麦草（Lolium perenne）分别接受由两个水平机械刺激（无刺激和刺激60 s d-1）和两个水平水分供应（200 ml d-1和400 ml d-1）组成的处理。在第二个实验中，匍匐茎草本蛇莓（Duchesnea indica）接受4个不同水平的机械刺激：（1）整个克隆不受机械刺激;（2）整个克隆都受机械刺激;（3）除顶端外其余克隆部分受机械刺激;（4）仅克隆顶端受机械刺激，其余部分不受机械刺激。在最后一个实验中，番茄（Lycopersicon esculentum）和紫花苜蓿（Medicago sativa）接受由三个水平机械刺激频度（0，25赫兹和50赫兹）和三个水平水分供应（50ml，150ml和250ml）组成的处理。这些实验主要回答不同生活型植物的生长和（或）机械性状如何响应机械刺激。主要结果如下： （1）在对机械刺激和水分交互效应的实验中，交互效应随物种发生变化。机械刺激和水分的交互效应对羊柴、番茄和紫花苜蓿作用不显著，但对虫实、沙鞭和黑麦草作用显著。 （2）在对机械刺激的研究中，机械刺激对植物的效应有正负之分。如机械刺激降低羊柴和沙鞭的总生物量，表明其是一种胁迫因子。但对于虫实、番茄和紫花苜蓿来说，机械刺激却能不同程度地促进植物的生长。 （3）机械刺激对虫实的机械性状没有显著影响，但对羊柴的机械性状恰好相反。此外，水分对虫实机械性状有显著影响。 （4）不同植物对机械刺激频度的敏感性存在差异。对番茄来而言，50赫兹的机械刺激对其生长具有较强的促进作用;对紫花苜蓿来说，25赫兹的机械刺激对其生长具有较强的促进作用。 （5）蛇莓对局部机械刺激具有显著反应，特别是在顶端进行机械刺激的处理中，整个克隆片段的叶柄长度缩短，根冠比发生改变，将较多的生物量分配到根。 这些结果表明：（1）不同物种对机械刺激和水分互作的反应可能与机械震动方式及物种本身有关;（2）单位植物大小所承受的机械刺激的强度及物种的生长速率是不一样的;（3）不同反应间的相互作用及相互独立可部分解释物种间的效应差异;（4）接触性形态建成的效应不能从一个物种外推到另外一个物种。 以前的研究集中探讨直立茎植物对机械刺激的响应，而对匍匐茎植物的研究极为贫乏。我们对蛇莓部分机械刺激的研究仅仅是一个初步探索。蛇莓的可塑性行为可能是一种适应性策略，因为这类植物常常生长在机械刺激频繁的开阔生境中。上述三个实验仅仅从生长和机械角度探讨了植物的适应性，而要真正揭示植物对多风生境的适应需要对不同物种进行多水平、多层面的研究，以期掌握不同生活型植物对机械刺激响应的一般格局。例如，从激素、细胞、解剖结构等方面探索其内在机制。

番茄种子萌发的高温耐性诱导及其生理效应

番茄（Lycopersicon esculentum Miller）原产于南美西部高原地带，适应原产地赤道附近高地干燥冷凉的气候特点，不耐高温多湿，种子萌发期间对高温非常敏感。本研究以佳粉17番茄种子为材料，试图寻找诱导番茄种子萌发期间高温耐性的方法，并通过研究高温耐性被诱导前后种子内部发生的生理生化变化，探索番茄种子萌发期间不耐高温以及高温耐性诱导的机理。试验结果显示： 25 ℃是种子萌发的最适温度，种子发芽率为97.5%。高温抑制种子的萌发，33 ℃和35 ℃条件下萌发率分别为58.5%和8.5%。 萌发适宜温度预吸胀、低温预吸胀、吸湿-回干预处理可提高番茄种子萌发期间的高温耐性，而水杨酸处理则没有明显效果。种子经25 ℃预吸胀30 h、0 ℃预吸胀10 h、吸湿-回干预处理后在33 ℃条件下的萌发率分别为81.5%、78.0%、90%，在35 ℃下的萌发率分别达到33.5%、42%、48.5%。经以上处理后，种子萌发速率提高，萌发高峰期提前，幼苗生长健壮，根干重增加，活力指数变大。 番茄种子萌发期间遭受高温危害时，电解质渗漏增加，相对电导率升高；脂膜过氧化作用加剧，其产物MDA的含量增加。经萌发适宜温度预吸胀、低温预吸胀、吸湿-回干等方法预处理后，高温伤害减轻，膜的完整性增强，电解质渗漏减缓，膜脂过氧化作用减弱，因而相对电导率降低，MDA含量减少。 高温抑制了抗氧化酶的活性，种子内部SOD、APX、CAT、GR等抗氧化酶活性降低。经萌发适宜温度预吸胀、低温预吸胀、吸湿-回干等方法预处理以后，抗氧化酶活性有不同程度的提高，清除细胞内超氧阴离子自由基的能力增强，因而使过氧化伤害减弱， 适宜温度预吸胀、低温预吸胀、吸湿-回干等预处理方法在保护生物膜的同时，增强抗氧化作用，抑制过氧化伤害，从而提高了番茄种子萌发的高温耐性，这是番茄种子高温耐性提高的生理机制之一。

Doenças do tomateiro cultivado em ambiente protegido.

2007

Controle de Meloidogyne javanica em tomate-salada e feijao-de-vagem com o cultivo previo de Crotalaria spectabilis.

O feijão-de-vagem é uma hortaliça severamente atacada por Meloidogyne javanica quando cultivada após o tomate-salada. Para avaliar a viabilidade de cultivo do feijão-de-vagem após o tomate-salada, cultivou-se a Crotalaria spectabilis, em faixas, por período de quatro meses, previamente ao cultivo do tomateiro. Faixas cultivadas com quiabeiro, antes do cultivo com tomate-salada, foram mantidas para comparação. Nas faixas cultivadas anteriormente com C. spectabilis, com os índices de multiplicação (IM) variando de 11 a 76 por parcela e os números de ovos do nematóide de 1.500 a 6.000 por planta, as produtividades foram de 2 a 26% maiores em cultivares de tomateiro e de 6 a 31,5% maiores em cultivares de feijão-de-vagem, em comparação com as produtividades obtidas nas faixas de quiabeiro, onde foram obtidos IM de 17 a 165 e números de ovos do nematóide que variaram de 7.200 a 19.000 por planta em cultivares de tomateiro e de feijão-de-vagem. O cultivo prévio da C. spectabilis por quatro meses é uma tecnologia viável para o controle de M. javanica, viabilizando a seqüência de cultivos com tomate-salada e feijão-de-vagem.