Involvement of a vitronectin-like protein in attachment of Agrobacterium tumefaciens to carrot suspension culture cells.

Infections of dicotyledonous plants by Agrobacterium tumefaciens result in the formation of crown gall tumors. Attachment of the bacteria to plant host cells is required for tumor formation. Human vitronectin and antivitronectin antibodies both inhibited the binding of A. tumefaciens to carrot cells. Wild-type bacteria are able to bind radioactive vitronectin; nonattaching mutants showed a reduction in the ability to bind vitronectin. The binding of biotype 1 A. tumefaciens to carrot cells or to radioactive vitronectin was not affected by high ionic strength. Detergent extraction of carrot cells removed the receptor to which the bacteria bind. The extract was found to contain a vitronectin-like protein. These results suggest that A. tumefaciens utilizes a vitronectin-like protein on the plant cell surface as the receptor for its initial attachment to host cells.     

Physarum vitronectin-like protein: an Arg-Gly-Asp-dependent cell-spreading protein with a distinct NH2-terminal sequence.

A 70-kDa protein cross-reacted with anti-bovine vitronectin was isolated from slime mold Physarum polycephalum. The NH2-terminal amino acid sequence of the protein, referred to as Physarum vitronectin-like protein, did not share any homology with those of animal vitronectins. It had cell-spreading activity, which was specifically inhibited by an Arg-Gly-Asp (RGD)-containing peptide.     

A higher plant extracellular vitronectin-like adhesion protein is related to the translational elongation factor-1 alpha.

Higher plant proteins immunologically related to the animal substrate adhesion molecule vitronectin have recently been observed and implicated in a variety of biological processes, such as plasma membrane-cell wall adhesion, pollen tube extension, and bacterium-plant interaction. We provide evidence that, similar to vitronectin, one of these proteins, PVN1 (plant vitronectin-like 1), isolated from 428 mM NaCl-adapted tobacco cells binds to glass surfaces an heparin. PVN1 was isolated by glass bead affinity chromatography. Isolated PVN1 has adhesive activity based on results from a baby hamster kidney cell-spreading assay. This plant adhesion protein was detected in all tissues examined but was most abundant in roots and salt-adapted cultured cells. Immunogold labeling indicated that PVN1 is localized in the cell wall of cortical and transmitting tissue cells of pollinated mature styles. A partial amino acid sequence of PVN1 revealed no similarity with vitronectin but, instead, was nearly identical to the translational elongation factor-1 alpha (EF-1 alpha). A clone isolated by screening a tobacco cDNA expression library with anti-PVN1 encoded a protein with greater than 93% identity to sequences of EF-1 alpha from plants of numerous species. Immunological cross-reactivity between tobacco PVN1 and EF-1 alpha as well as the reaction between the EF-1 alpha antibody and the 65- and 75-kD vitronectin-like proteins of a fucoidal alga supported the conclusion that the plant extracellular adhesion protein PVN1 is related to EF-1 alpha.     

A kinematic analysis of gynoecial growth in Lilium longiflorum: surface growth patterns in all floral organs are triphasic

Sequential marking experiments combined with histological and morphological examination indicate that growth of the gynoecium in Lilium longiflorum Thunb. (Easter lily) is triphasic. Phase I (gynoecia less than 10 mm in length) is characterized by spatial and temporal variation in local relative growth rate and mitotic index throughout the entire gynoecium. In Phase II (10-100 mm), a steady growth peak is restricted to the proximal 5 to 15 mm of the style, while the ovary shows uniform growth. In Phase III (100-135 mm), the ovary ceases growth and the maximal growth zone of the gynoecium migrates from the proximal to the distal end of the style. Comparison of surface growth patterns and differentiation events in the gynoecium to those previously described in the tepal and another demonstrates the triphasic nature of growth in the whole lily flower.     

Systemic elevation of phosphatidic acid and lysophospholipid levels in wounded plants

Plants develop systemic defense responses upon exposure to pathogens or wounding by herbivores. Lipids and lipid metabolites have previously been implicated in induction of defense molecules during plant responses to physical wounding. Possible involvement of changes in lipid composition in systemic wound signal transduction was examined in leaves of seedlings of several different plant species. In the wounded tomato leaf, phosphatidic acid increased approximately fourfold within 5 min whereas lysophosphatidylcholine and lysophosphatidylethanolamine increased over twofold within 15 min of wounding. Similar changes in these lipids were observed in the neighboring non-wounded leaf. In broad bean, soybean, sunflower and pepper seedlings phosphatidic acid levels increased rapidly and systemically upon wounding. The results suggest that the role of phospholipid hydrolysis and accumulation of lipid metabolites in the early events are responsible for systemic wound signal transduction in plants. Furthermore, they indicate that the wound signal propagates outside the wounded leaf within 5 min in these plants.     

坛紫菜菌解液对植物生长和抗性指标的影响

利用琼胶降解菌处理坛紫菜粉末,降解其细胞壁多糖,释放内容物,并获得菌解液,研究不同稀释度坛紫菜菌解液对受体植物蚕豆出芽和生长,以及大豆和番茄幼苗抗性相关指标的影响.结果显示:(1)随琼胶降解菌处理时间的延长,菌解液中还原糖含量逐渐增高.(2)3.33%的菌解液对蚕豆种子萌芽促进效果最好,而2%的菌解液能增加蚕豆叶绿素含量,促进其幼苗生长.(3)3.33%的菌解液能有效提高大豆离体子叶的植保素含量,迅速增加番茄叶片表皮条的H2O2的释放量,提高苯丙氨酸解氨酶活性.研究表明,琼胶降解菌能使坛紫菜细胞壁中的营养物质释放,并产生某些激发物质,从而使坛紫菜菌解液能够促进受体植物的种子发芽和幼苗生长,并能有效诱导植物的抗性.     

Predator avoidance in phytophagous mites: response to present danger depends on alternative host quality

We studied whether volatiles released by putative host plants affect the antipredator response of an herbivorous mite, Tetranychus urticae, when the patch was invaded by Phytoseiulus persimilis. Tetranychus urticae laid a lower number of eggs on tomato leaves than on lima bean leaves, suggesting that lima bean is a preferred host food source for T. urticae. In addition, T. urticae preferred lima bean plant volatiles to tomato plant volatiles in a Y-tube olfactometer test. To investigate the antipredator response of T. urticae, we examined the migration of T. urticae from a lima bean leaf disc to a neighbouring leaf disc (either a tomato or lima bean leaf disc) when ten predators were introduced into the original lima bean disc. A Parafilm bridge allowed for migration between the leaf discs. No migrations occurred between leaf discs when there were no predators introduced to the original leaf disc. However, when predators were introduced migrations did occur. When the neighbouring leaf disc was upwind of the original disc, the migration rate of the mite from original lima bean leaf disc to a neighbouring tomato leaf disc was significantly lower than that to a neighbouring lima bean leaf disc. By contrast, when the neighbouring leaf disc was downwind of the original leaf disc, there was no difference in the migration rates between lima bean leaf discs and tomato leaf discs. The number of T. urticae killed by P. persimilis for each treatment was not different, and this clearly shows that the danger was the same in all treatments regardless of the decision made by T. urticae. From these results, we conclude that T. urticae change their antipredator response by evaluating the difference in host plant volatiles in the patch they inhabit.     

蚕豆豆类胰岛素基因的cDNA克隆及序列分析

为探明豆科植物中豆类胰岛素基因的结构特征与进化关系,在已获得大豆豆类胰岛素基因的基础上,以蚕豆种子胚根mRNA为材料,采用RT-PCR技术,克隆了蚕豆豆类胰岛素基因的cDNA序列,编码的前体多肽包括信号肽、成熟型豆类胰岛素及另一多肽的45个氨基酸残基。DNA序列分析表明,克隆片段与大豆和豌豆的同源性分别为62.5%和58.7%。在氨基酸水平上分别具有44.2%和43.6%的同源性,其中存在着高度保守的半胱氨酸位点,它们在维持豆类胰岛素的空间结构与生理功能方面,可能具有重要的作用。 Abstract:In order to elucidate the relationship between the structural features of leginsulin gene in legume plants and their phylogenetic significance,we have cloned the cDNA sequence of leginsulin gene from radicles of broad bean (Vicia faba) via RT-PCR techniques according to the leginsulin gene sequence we previously obtained from soybean (Glycine max).The cloned cDNA encoded for a precursor protein consisting of the signal peptide,mature leginsulin and an additional 45 amino acids of another polypeptide.A sequence search for homology comparison revealed the cloned leginsulin cDNA fragment shares 62.5% and 58.7% similarity to soybean and pea,respectively.The results also shown that leginsulin cDNA from broad bean presents 44.2% and 43.6% amino acid sequence homology with soybean and pea (Pisum sativum),respectively,and that there exists highly conserved cysteine sites among the leginsulin cDNAs,which may play a crucial role in maintaining the three-dimensional structure and the physiological functions of leginsulin.     

The Translocation of Antibiotics in Higher Plants: II. THE MOVEMENT OF GRISEOFULVIN IN BROAD BEAN AND TOMATO

The uptake and translocation of griseofulvin from water cultureby broad bean and tomato has been studied; observations werealso made on its decay in broad bean. In most cases griseofulvinwas determined by bioassay. Where possible the bioassay waschecked by chemical estimations and was found to be adequate. The amount of griseofulvin taken up by the broad bean was proportionalto the volume of water transpired for any single concentrationof the treating solution and the decay was exponentially relatedto the time of exposure in the tissues. The accumulation ofgriseofulvin in the tissue had, therefore, an exponential componentbut within the limits of error there was constant relationshipbetween accumulation and transpiration over the time periodsused in the trials. The rate of accumulation in tomato was alsoconstant. The amount of griseofulvin accumulated by both beanand tomato after a definite time was a linear function of theconcentration of the treating solution. There were two processes involved in the uptake of griseofulvinby the broad bean: (a) an initial rapid entry into the rootswhich was inhibited by sodium azide and dinitrophenol at concentrationswhich did not reduce transpiration; and (b) a prolonged uptakelinearly related to transpiration which was not affected bythese concentrations of the inhibitors.     

Galectin 1 inhibits incorporation of vitronectin and chondroitin sulfate B into the extracellular matrix of human vascular smooth muscle cells

Galectin-1, a beta-galactoside-binding dimeric lectin, interacts with the extracellular matrix (ECM) of smooth muscle cells (SMCs) and with particular ECM proteins. Enrichment of the ECM with galectin-1 affects adhesion and proliferation of cultured SMCs. Here we investigated whether galectin-1 (1) interacts with glycosaminoglycan (GAG) chains, (2) cross-links between ligands and facilitates the incorporation of GAGs, vitronectin and plasma fibronectin in the ECM of vascular SMCs. A recombinant galectin-1 fusion protein GalH, used in this study, formed dimers and interacted with ECM proteins. GAG chains inhibited these interactions. Among the studied GAG chains, only chondroitin sulfate B interacted with GalH in beta-galactoside-dependent manner. GalH did not bridge between ECM proteins on solid phase and [125I]-labelled ECM proteins or GAGs in solution. The ECM incorporated less vitronectin in the presence of soluble GalH. GalH-enriched ECM incorporated less vitronectin and chondroitin sulfate B. The ECM partially depleted of endogenous galectins incorporated more chondroitin sulfate B compared to untreated ECM. These results suggest that galectin-1 is likely to be involved in the ECM assembly affecting incorporation of some ECM components important for SMC behaviour.     

First Report of Tomato Yellow Leaf Curl Virus-Israel Species Infecting Tomato, Pepper and Bean in Tunisia

Tomato yellow leaf curl virus disease (TYLCVD) has been observed in Tunisia for more than 20 years. Until year 2004, only the Tomato yellow leaf curl Sardinia virus‐Sicily (TYLCSV‐[Sic]) was detected in tomato, pepper and bean crops. In the Sahel region, some tomato samples showing severe TYLCVD symptoms were collected from greenhouses in 2004 and 2005. Typing of these isolates revealed for the first time the presence of the TYLCV Israel in Tunisia. This result was confirmed by using several sets of specific primers and by sequencing. This species has also been detected on pepper and bean collected from fields in the same region. The sequencing of a tomato and a bean isolate showed that they both share more than 97% of sequence identity with the TYLCV from Dominican Republic ( AF024715 ). The TYLCV has been found in single and mixed infection with the TYLCSV‐[Sic].     

Polyphenoloxidase in higher plants: immunological detection and analysis of in vitro translation products

Antibodies to broad bean polyphenoloxidase (PPO) were used to detect and demonstrate that the PPOs found in several different plants have many similarities in common. Crude extracts from leaves of broad bean, bush bean, lettuce, mung bean, pea, soybean, spinach, tobacco, and tomato contained enzyme which cross-reacted with polyclonal anti-PPO in Ouchterlony double diffusion analysis. The results suggested that plant polyphenoloxidase from a wide range of species may contain similar antigen determinants. Poly A⁺ mRNA was isolated from leaves of each plant species and translated in vitro using a rabbit reticulocyte translation system. An in vitro synthesized product corresponding to PPO from each species was identified after specific immunoprecipitation with anti-PPO. The molecular weight of this in vitro product was similar in all plants examined and found to be approximately 45 kilodaltons. Peptide maps of the in vitro synthesized product from all plant species were similar and showed at least three peptides in common. Plant PPOs may have more structural similarities than commonly though in spite of the great variety in observed isoenzyme forms.     

Improved control capacity of the mite predator Phytoseiulus persimilis (Acari: Phytoseiidae) on tomato

The predatory mite Phytoseiulus persimilis is frequently reported to perform poorly on greenhouse tomatoes. As the predators are mass-reared on another host plant (bean), we supposed that they are poorly adapted to tomato, a plant densely packed with poisonous and sticky glandular hairs. This hypothesis was tested by comparing the control capacity of a stain of P. persimilis directly obtained from a mass rearing with the same strain after four generations on tomato. Both strains were released in a tomato crop in two identical compartments of a greenhouse and the population dynamics of prey (a tomato strain of Tetranychus urticae) and predator were recorded at weekly time intervals. It was found that the strain previously exposed to a tomato environment performed better than the unexposed strain: (1) its population increased faster; (2) the prey population declined faster; and (3) the damage to new-grown tomato leaves was considerably lower. To investigate the causes of the difference in performance between the exposed and unexposed strains, oviposition and survival rates were assessed on a diet of two-spotted spider mites on tomato leaf sections. In addition, the unexposed strain was tested on a diet of two-spotted spider mites on bean leaf sections. The difference in oviposition rates of both predator strains was small compare to the overall mean. However, the oviposition rate of the first generation of predators since transfer from bean to tomato dropped to less than half of the original value. Moreover, mortality in the first generation increased from 14% to 89%, whereas it decreased to 0% after four generations. Future research should clarify whether these changes in life history are due to selection or to physiological adaptation.     

Molecular characterisation and relative incidence of bean‐ and soybean‐infecting begomoviruses in northwestern Argentina

Recent studies identified three begomoviruses infecting soybean and bean crops in northwestern (NW) Argentina, bean golden mosaic virus (BGMV), a virus with high capsid protein identity with Sida mottle virus (SiMoV) and a possible new viral species (isolate A). Analysis of complete DNA‐A sequences confirmed that isolate A represents a new viral species for which the authors propose the name soybean blistering mosaic virus (SbBMV), whereas the SiMoV‐like virus is actually an isolate of tomato yellow spot virus (ToYSV). Molecular hybridisation‐based detection of the three begomoviruses was accomplished using a general probe obtained by mixing full‐length DNA‐A clones of the three begomoviruses and specific probes comprising part of the common region of each viral genome. These probes were used to test samples collected in NW Argentina from 2004 through 2007. Fifty‐three percent of the bean samples were infected with BGMV, 11.5% with ToYSV and 9% with SbBMV. For soybean samples, 33% were infected with SbBMV and 18% with ToYSV. BGMV was not detected in soybean. ToYSV was also detected in the wild species Abutilon theophrasti.     

增强的紫外线-B辐射对几种作物和品种生长的影响

在温室条件下,以每日2．2kJ／m^2(CK)、8．82kJ／m^2(T1)和12．6kJ／m^2(T2)紫外线-B辐射(UV-B,280～320nm)剂量研究了5个黄瓜(Cucumis satious L.)品种,7个番茄(Lycopsicon esculentum Mill)品种及大豆[Glycine max(L.)Metr．]、菜豆(Phaseolus vulgaris L.)和黄河密瓜(Cucumis melo L.)的生长反应。辐射处理25d后,测定了株高(PH)、叶重(LDW)及总生物量(TDW)、叶面积(LA)、特定叶重(SLW)、上胚轴长度(EL)和番茄品种的子叶节周长(GCN)。结果表明,种间和种内差异显著。但是大多数品种及种的反应指数为负值,并呈现强度负相关效应,说明UV—B辐射抑制了它们的生长发育,但大豆的反应指数在低剂量的处理下为正值反而促进其生长。UV—B胁迫下,大多数种类的上胚轴延伸明显受阻,特定叶重增加,叶面积和生物量减少,番茄的子叶节膨大。作物对UVB辐射的种内和种间反应是作物遗传特性上的差异和对环境的适应能力不同所致。     

Phytotoxic, clastogenic and bioaccumulation effects of the environmental endocrine disruptor bisphenol A in various crops grown hydroponically

The effects of the endocrine disruptor bisphenol A (BPA) at concentrations of 10 and 50 mg l⁻¹ were evaluated on the germination and morphology, micronuclei (MN) content in root tip cells and BPA bioaccumulation of hydroponic seedlings of broad bean (Vicia faba L.), tomato (Lycopersicon esculentum Mill.), durum wheat (Triticum durum Desf.) and lettuce (Lactuca sativa L.) after 6 and 21 days of growth. In general, BPA at any dose used did not inhibit germination and early growth (6 days) of seedlings of the species examined, with the exception of primary root length of tomato which decreased at the higher BPA dose. In contrast, an evident phytotoxicity was induced by BPA in all species after 21 days of growth with evident morphological anomalies and significant reductions of the lengths and fresh and dry weights of shoots and roots of seedlings. With respect to the nutrient medium without seedlings, BPA concentration decreased markedly during the growth period in the presence of broad bean and tomato seedlings, and limitedly in the presence of durum wheat and, especially, lettuce. Further, the presence of BPA measured in roots and shoots of broad bean and tomato after 21-day growth indicated that bioaccumulation of BPA had occurred. The number of MN in broad bean and durum wheat root tip cells increased markedly by treatment with BPA at both concentrations, thus suggesting a potential clastogenic activity of BPA in these species.     

Functional analysis of gene-silencing suppressors from tomato yellow leaf curl disease viruses

Tomato yellow leaf curl disease (TYLCD) is caused by a complex of phylogenetically related Begomovirus spp. that produce similar symptoms when they infect tomato plants but have different host ranges. In this work, we have evaluated the gene-silencing-suppression activity of C2, C4, and V2 viral proteins isolated from the four main TYLCD-causing strains in Spain in Nicotiana benthamiana. We observed varying degrees of local silencing suppression for each viral protein tested, with V2 proteins from all four viruses exhibiting the strongest suppression activity. None of the suppressors were able to avoid the spread of the systemic silencing, although most produced a delay. In order to test the silencing-suppression activity of Tomato yellow leaf curl virus (TYLCV) and Tomato yellow leaf curl Sardinia virus (TYLCSV) proteins in a shared (tomato) and nonshared (bean) host, we established novel patch assays. Using these tools, we found that viral proteins from TYLCV were able to suppress silencing in both hosts, whereas TYLCSV proteins were only effective in tomato. This is the first time that viral suppressors from a complex of disease-causing geminiviruses have been subject to a comprehensive analysis using two economically important crop hosts, as well as the established N. benthamiana plant model.     

Oviposition preference and larval performance and behavior of <Emphasis Type="Italic">Trichoplusia ni</Emphasis> (Lepidoptera: Noctuidae) on host and nonhost plants

We investigated the female oviposition preference and larval performance and behavior of Trichoplusia ni (Hubner) on cabbage, tomato, soybean (host plants), and wormseed (nonhost plant) in laboratory experiments. In addition, we investigated the role of trichomes in the oviposition of females. Females oviposited more often on cabbage than on tomato, soybean, or wormseed plants. Tomato was the second most preferred plant, followed by soybean and wormseed. Neonate larvae gained more weight on cabbage and tomato than on soybean or wormseed, while second-instar larvae grew better on cabbage than on tomato, soybean, or wormseed. The least growth of neonate larvae occurred on wormseed plants. The orientation of neonate and second-instar larvae to cabbage, tomato, soybean, and wormseed did not differ significantly. Neonate larvae settled equally on leaf discs of cabbage, tomato, soybean, and wormseed, while most second-instar larvae settled on leaf discs of cabbage in comparison with the other plants after 24 h of release. The foliar area consumed by neonate larvae was quite similar among plants, but second-instar larvae consumed more cabbage than tomato, soybean, or wormseed. Comparing different types of leaves, females oviposited more often on mature than young leaves of tomato, soybean, and wormseed. In contrast, females did not show any preference for ovipositing on young or mature leaves of cabbage. In general, we found that the density or length of nonglandular and glandular trichomes of tomato, soybean, and wormseed plants negatively affected oviposition of T. ni females.