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1.
Cells capable of photosynthesis in the parasitic angiosperm Cuscuta reflexa Roxb. (dodder) are highly localized. Immunolocalization of ribulose-1,5 bisphosphate carboxylase-oxygenase (Rubisco) and autofluorescence of chlorophyll in transverse sections of stems showed that they were largely restricted to a band of cells adjacent to the vascular bundles, consequently, the concentrations of Rubisco and chlorophyll were low per unit area or fresh weight. When 14CO2 was supplied to stem segments of C. reflexa it preferentially accumulated in these cells adjacent to the vasculature. Although the conductance for CO2 movement to the cells containing chlorophyll and Rubisco was very low, both the light reactions and dark reactions of photosynthesis appeared to be functional. De-epoxidation of the xanthophyll-cycle pigments after exposure to high light, and the chlorophyll fluorescence parameters, photochemical quenching (qP), non-photochemical quenching (NPQ) and the quantum efficiency of photosystem II (φPSII) responded normally to changes in photon flux density, indicating functional light-driven electron transport. The response of CO2 exchange to photon flux density followed a typical hyperbolic curve, and positive rates of CO2 fixation occurred when external CO2 was increased to 5%. We propose that CO2 for carbon assimilation is derived from internally respired CO2 and that this layer of photosynthetic cells makes a positive contribution to the carbon budget of C. reflexa. Received: 23 October 1997 / Accepted: 16 December 1997  相似文献   

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Pectin degrading enzymes, hemicellulose degrading enzyme andcellulose degrading enzymes were studied in Cuscuta reflexaRoxb., its susceptible hosts, Brassica campestris L., Cocciniaindica W. & A. Datura innoxia Mill, Helianthus annuus L.,Holoptelea indica Planch, Lantana camara L., Medicago sativaL., Manihot utilissima Pohl, Petunia hybrida X Hort exvilm,Pisum sativum L., Phaseolus vulgaris L. and Solanum nigrum L.and non-susceptible plants Ipomoea batata Lam. and Solanum tuberosumL. Pectin esterase and polygalacturonase were present in higheramounts in Cuscuta parasitic on P. vulgaris and S. nigrum, whichneeded more time for haustorial establishment. Exo-l, 4-ß-D-glucosidaseactivity was found in Cuscuta but could not be detected in itshosts. Xylanase and cellulase activity of host plants increasedwhile cellobiase activity decreased as a result of infectionby the parasite. Higher pectin esterase, polygalacturonase,xylanase and exo-l, 4-ß-D-glucosidase activities inthe haustorial region of the parasite is likely to bring aboutthe lysis of the cell wall of the host plant and thus facilitatethe penetration of the parasite haustoria into the host sieveelement, which is necessary for the transport of nutrients betweenthe host and the parasite. Key words: Cell wall degrading enzymes, Cuscuta reflexa  相似文献   

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During the development of the haustorium, searching hyphae of the parasite and the host parenchyma cells are connected by plasmodesmata. Using transgenic tobacco plants expressing a GFP-labelled movement protein of the tobacco mosaic virus, it was demonstrated that the interspecific plasmodesmata are open. The transfer of substances in the phloem from host to the parasite is not selective. After simultaneous application of (3)H-sucrose and (14)C-labelled phloem-mobile amino acids, phytohormones, and xenobiotica to the host, corresponding percentages of the translocated compounds are found in the parasite. An open continuity between the host phloem and the Cuscuta phloem via the haustorium was demonstrated in CLSM pictures after application of the phloem-mobile fluorescent probes, carboxyfluorescein (CF) and hydroxypyrene trisulphonic acid (HPTS), to the host. Using a Cuscuta bridge (14)C-sucrose and the virus PVY(N) were transferred from one host plant to the another. The results of translocation experiments with labelled compounds, phloem-mobile dyes and the virus should be considered as unequivocal evidence for a symplastic transfer of phloem solutes between Cuscuta species and their compatible hosts.  相似文献   

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A recently developed empirically based modelling technique wasused to quantify uptake, flow and utilization of C and N inLupinus albus L., uninfected and parasitized by Cuscuta reflexaRoxb. plants over a 12 d period during flowering and early fruitsetting of the host. The modelling combined data on molar C:Nratios in host phloem and pressure-induced xylem sap, net incrementsof C and N in host and parasite plant parts and respiratorylosses of C. The modelling of the solute transfer between hostand Cuscuta was achieved by assuming non-specific intake fromthe xylem. The models predicted that Cuscuta derived 99.5% ofits carbon and 93.6% of its nitrogen demand from the host phloem.The overriding sink strength of the parasite diverted most ofthe basipetally translocated host assimilates and massivelycompeted with the host root and inhibited fruit setting. Carbonincorporation in Cuscuta consumed 56%, respiration 24% and secretionby extrafloral nectaries 1.8% of the current host photosynthate.Root respiration was inhibited by 59% and carbon was mobilizedfrom host root and leaves. Competition by the parasite for Nwas even more severe and Cuscuta incorporated nitrogen equalling223% of current fixation, but N2 fixation of the host was severelyrestricted to 37%. Withdrawal of N from host phloem led to severelosses of N from leaves and the root and marked decreases inN concentration. It required massive xylem-to-phloem transferof N, because the xylem as the major supply route for N wasnot exploited substantially by Cuscuta. The results are discussedin relation to likely causes for parasite-induced pathogeniceffects, suggesting that Cuscuta affected the host adverselyby depriving it mainly of its nitrogen, but that causal to incipientnitrogen deficiency and restricted N2 fixation was the superiorsink potential of Cuscuta, which prevented adequate supply ofassimilates to the nodulated root. The dominating sink potentialof Cuscuta is compared with the similarly strong sink competitionexerted by fruits at the stage of seed filling in annual plants. Key words: Cuscuta reflexa, Lupinus albus, parasitism, carbon, nitrogen, phloem, xylem, transport  相似文献   

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Nodulated Lupinus albus L. was grown on quartz sand in the greenhouseand supplied with a N-free culture solution. Half the plantswere infected with Cuscuta reflexa Roxb. at 33 DAS. An empiricallybased modelling technique was developed to quantitatively depictuptake, flow and utilization of C and N in the host plant andbetween host and parasite over a 12 d period. The modellingincorporated C: N ratios of solutes in phloem and pressure-inducedxylem sap, net increments of C and N and respiratory lossesof C. For assessing the transfer of solutes from host phloemto Cuscuta it was not possible to use the C: N ratio of phloemsap close to the site of parasite attachment, a procedure whichwould have assumed non-specific withdrawal of phloem-borne solutes,since this would have implied unimpeded mass flow from hostto parasite. The relative intake of C and N by the parasiteby specific withdrawal of nitrogenous and carbonaceous solutesfrom the phloem was obtained independently by assuming thatxylem intake occurred non-specifically. Xylem import was thusobtained (a) from transpiration and tissue water increment ofCuscuta and the concentrations of N and C in xylem sap and (b)from the Ca2+ increment of Cuscuta and the ratios Ca: N andCa: C in lupin xylem sap, assuming that Ca2+ intake occurredsolely via xylem. By subtracting net xylem import from totaluptake of C and N by Cuscuta the methods resulted in comparableratios of C: N intake from the phloem. The average ratio (53.4)was smaller than the C:N ratio in host phloem (85.6) indicatingspecific withdrawal of solutes with a distinct preference forN. Using this ratio, modelling of flows of C and N was possibleand showed that Cuscuta abstracted C and N mainly from the hostphloem, but xylem supply was nutrient-dependent and amountedto 6.4% of the N but only 0.5% of the C demand. The resultsindicated that Cuscuta exerted a very strong sink and competedefficiently with the root, the major sink of L. albus, by attracting81% of the current photosynthate and more N (223%) than wascurrently fixed. The massive demand of the parasite led to lossesparticularly of N from leaves and the root and apart from causingcarbon losses it appeared to induce a sink-dependent stimulationof photosynthesis. In contrast, nitrogen fixation in the Cuscuta-infectedlupin was inhibited to 37% of the control. Key words: Cuscuta reflexa, Lupinus albus, carbon, nitrogen, phloem, xylem, transport, parasites, modelling  相似文献   

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The parasitic angiosperm Cuscuta reflexa contains unusually high amounts of the carotenoids lutein-5,6-epoxide and 9- cis -violaxanthin. In this study the light-dependent conversions of these carotenoids in entire plant tissue and purified LHCII b was compared with that of the xanthophyll cycle carotenoid violaxanthin when plants are exposed to high irradiance followed by low irradiance. In entire tissue under high irradiance, similar conversion kinetics and stoichiometry with de-epoxidation products suggest that both lutein-5,6-epoxide and all- trans -violaxanthin are equally suitable substrates for de-epoxidase. This is not the case under low irradiance as, although epoxidation of zeaxanthin and antheraxanthin rapidly restores the violaxanthin pool, the recovery of the lutein-5,6-epoxide pool is comparatively slow and has no stoichiometric relationship with its de-epoxidation product, lutein. Light-dependent changes in the concentration of 9- cis -violaxanthin mimic violaxanthin. However, the inability to detect de-epoxidation products or to de-epoxidize 9- cis -violaxanthin in vitro suggests that it is not subject to de-epoxidation and, instead, its concentration changes may reflect the equilibrium between isomers of violaxanthin. Light exposure did not affect the composition of carotenoids bound to purified LHCII b , indicating that these bound carotenoids are not subject to de-epoxidation and do not contribute to the isomer pool equilibrium. The biosynthetic origins of lutein-5,6-epoxide and the potential role of these carotenoid cycles in photoprotection are discussed.  相似文献   

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In a study of host-parasite interrelationship and the mechanism of parasitization, ribonuclease activity was determined in Cuscuta reflexa Roxb, and in infected and control host plants of Lantana camara L. In the haustoria-bearing region of Cuscuta , the concave half of the vine bearing the haustorial site, always showed significantly higher RNase activity than the convex half, irrespective of the differing enzyme activity of the parasite tissue growing on different hosts — Brassica campestris L., Helianthus animus L., Lantana camara L., Medicago saliva L. and Solatium nigrum L. The uninfected host branch of L. camara showed the maximum specific RNase activity in the apical region which decreased toward the base, while the infected host branch showed minimum specific RNase activity in the apical region, gradually increasing towards the infected region.  相似文献   

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运用鸟枪法在Illumina测序仪上对亚洲分布的大花菟丝子(Cuscuta reflexa叶绿体基因组核苷酸全序列进行测定,并与已经发表的分布于欧洲的大花菟丝子进行了比较分析.研究结果表明亚洲分布的大花菟丝子叶绿体基因组总长度为120 972 bp,由79 499 bp的长单拷贝区,8 369 bp的短单拷贝区,以及两个16 552 bp的反向重复区组成,其长度比欧洲分布的大花菟丝子小了549 bp;基因组总GC含量为38.3%,稍高于欧洲分布的大花菟丝子.两地区的大花菟丝子叶绿体全基因组编码的功能基因完全相同,且基因排列顺序也完全一致.另外,经过进一步序列比对后发现亚洲分布的大花菟丝子与欧洲分布的存在251个插入和210个缺失现象,总插入缺失及碱基替换长度分别为7 649 bp和3 720 bp,最大的插入和缺失长度分别为426 bp和435 bp.很多插入缺失都是单碱基,但仍然存在四个长度超过200 bp的大突变,两个大的缺失发生在ycf2基因中,两个大的插入分别发生在trn F-psbE和matK-trnQ间隔区,详细的对比后发现大量的插入缺失都发生在大单拷贝区的基因间隔区,且插入缺失在反向重复区的发生频率较低.本研究首次报道不同大洲分布的同种异养植物的叶绿体全基因组比较分析,为研究这两个区域的居群多样性提供了基础资料.  相似文献   

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A time-dependent correlation of anatomical and chemical defence reactions was shown during the incompatible reaction of tomato against the phanerogamic parasite Cuscuta reflexa. Microscopical analysis of the infection sites at the tomato stem revealed the elongation of epidermal, hypodermal and collenchymatic cells beneath the parasitic prehaustorium. After 9–11 days of infection the elongated cells had collapsed forming a visible brownish plaque at the tomato stem followed by a scalariform tissue with lignified and suberized cell walls. Concomitantly, an enhanced accumulation of soluble phenolic compounds (chlorogenic acid and an unidentified hydroxycinnamic acid derivative), as well as a stimulation of peroxidases, was observed. In contrast, PAL activity was not increased. Whereas the stimulation of phenylpropanoid metabolism could also be induced by artificial wounding, the described anatomical changes were only observed during attack of Cuscuta.  相似文献   

12.
The plant parasite Cuscuta reflexa induces various responses in compatible and incompatible host plants. The visual reactions of both types of host plants including obvious morphological changes require the recognition of Cuscuta ssp. A consequently initiated signaling cascade is triggered which leads to a tolerance of the infection or, in the case of some incompatible host plants, to resistance. Calcium (Ca2+) release is the major second messenger during signal transduction. Therefore, we have studied Ca2+ spiking in tomato and tobacco during infection with C. reflexa. In our recently published study1 Ca2+ signals were monitored as bioluminescence in aequorin-expressing tomato plants after the onset of C. reflexa infestation. Signals at the attachment sites were observed from 30 to 48 h after infection. In an assay with leaf disks of aequorin-expressing tomato which were treated with different C. reflexa plant extracts it turned out that the substance that induced Ca2+ release in the host plant was closely linked to the parasite''s haustoria.Key words: cuscut, odder, calcium signaling, plant parasitismThe genera Cuscuta, also known as dodder, includes 170 parasitic species with a worldwide distribution. Members of Cuscuta ssp. belong to the 1% of angiospermic plants that live as holoparasites and depend on nutrients, water and carbohydrates from other host plants.2 Cuscuta spp. lack roots or leaves but possess specific penetrating organs, the so called haustoria, which are fully developed 5–6 days after the first contact, when an interaction between parasite and host is established.As for all dicotyledonous plants, the typical Cuscuta spp. life cycle starts with the germination of seeds. At the stage of a rootless seedling, Cuscuta ssp. has just a few days to find and successfully invade a host plant. Although Cuscuta ssp. seedlings appear to coil indiscriminately around any vertical elongated object, they seem to have an efficient “sense of smell” to recognize potential “victims” and are therefore able to infest host plants more rapidly and efficiently.3 As soon as a host is reachable, Cuscuta ssp. starts to wind around the host shoot and initiates the attachment process as well as the development of haustoria.2,4 Already at this initial phase of infection (12–48 h post attachment) the host plant senses the parasite and initiates an onset of several signals which are only partially known. Amongst the several induced genes are for example those encoding AGPs (Arabinogalactan Proteins), proteins promoting the parasite''s adherence.5 Also proteins are produced which might be important for nutrient and water uptake6 or which modify the host cell wall.7In this addendum article, we focus on signals which occur in host plants within the early infection stage prior to a vascular bundle connection and refer to our article about Ca2+ signalling in C. reflexa infected tomato plants.8 Besides phytohormones or other initial signalling molecules, such cellular calcium signals might be involved in controlling the expression of important genes for developmental or resistance related processes.In our approach, Cuscuta reflexa shoots of ∼25 cm length were wrapped around transgenic constitutively aeqourin-expressing tomato (Solanum lycopersicum) and tobacco (Nicotiana benthamiana) plants. With a highly sensitive ccd-camera we then monitored the two interacting organisms. The Ca2+-signals which are released by the host-plant could be detected as light-emission. The first cytosolic calcium signals were observed 24–48 h after the parasite attachment when the haustoria formation was already initiated. Light, indicating a cytosolic calcium influx was clearly visible directly where the parasite started to penetrate host tissue via its haustoria (Fig. 1) and often appeared several times within 1–6 h. As the light signals per recorded picture were collected for 10 min it is not clear if the duration of such cytosolic calcium influx comprises 10 ms or 10 min. An additional experiment in our study was the usage of a Cuscuta reflexa haustorium extract which was applied to aequorin expressing tomato leaf discs. Here it turned out that the Ca2+-ion influx happened steadily and slowly, because signals were only visible when summed up from 2 h recording. The finding that both boiled haustoria extract and control extract, made from Cuscuta reflexa shoots without haustoria, are inactive, suggests that a protein which is expressed during the infection process might be the direct or indirect trigger of such Ca2+-signals. These results overcome furthermore the theory that Calcium signals are induced by pressure, which might also be a step during Cuscuta ssp. infection.Open in a separate windowFigure 1Cuscuta reflexa infection induces calcium-signals in aequorin-expressing tomato. Left: Bright field; middle: light emission representing Ca2+-signals at the infection site ∼30 h post onset of the parasite; signals were monitored with a ccd camera. Right: overlay.The fact that calcium fluxes act as a second messenger in several stress responses such as cold shock, wind, touch, osmotic stress,9 phytohormone signalling pathways,10 plant—symbiotic interactions1012 or also plant pathogen interaction1315 complicates the interpretation of the signals that are induced by Cuscuta reflexa. One possibility could be that visible Ca2+-signals are part of a signalling pathway where also SA (salicylic acid) or/and JA (jasmonic acid) play an important role. Recently, Runyon et al.16 could show that tomato plants infected with Cuscuta pentagona respond with a strong induction of JA and SA 24–36 h post infections. This time frame correlates with our described calcium signals and it has been previously described that calcium fluxes might be a part of the JA- and SA-signalling cascade.The tomato—dodder interaction, however, represents an exception among dicotyledonous plants because tomato generates a hypersensitive response which is part of a successful resistance reaction.7,16 In this particular case characteristic components of C. reflexa must be sensed by its host plant. These factors indicate “non-self” for the host plant, probably following a model comparable to the MAMP concept where characteristic molecular patterns of a pathogen are recognized in host plants via pattern recognition receptors and subsequently trigger defence responses.17,18 But sensing and signalling in host plants takes place not only in the case of an “incompatible” interaction. The developmental phenomenons of a dodder—plant interaction in a “compatible” interaction are nearly a miracle. In this case, the parasite is completely tolerated and achieves the attachment and the penetration of the host plant. It interferes in developmental processes and manipulates its host to develop vascular tissues, to build up chimerical cell walls and interspecific symplastic cell connections.16,17 Finally, it is connected to the host plant and starts to withdraw nutrients and carbohydrates19,20 by mimicking endogenous sinks. Such a tolerated interaction reminds of an interaction of plants with bacterial or fungal symbionts, where also Ca2+-signals have been described and well characterized.11,12 In the case of Cuscuta ssp.—host interaction a lot of further studies have to be done to discover all important steps of signalling cascades.  相似文献   

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Protoplasts of a kanamycin-resistant (KR, nuclear genome), streptomycin-resistant (SR, chloroplast genome) and chlorophyll-deficient (A1, nuclear genome) Nicotiana tabacum (KR-SA) cell suspension cultures or X-ray-irradiated mesophyll protoplasts of kanamycin- and streptomycin-resistant green plants (KR-SR) were fused with protoplasts of a cytoplasmic male-sterile (CMS) Daucus carota L. cell suspension cultures by electrofusion. Somatic hybrid plants were selected for kanamycin resistance and the ability to produce chlorophyll. Most of the regenerated plants had a normal D. carota morphology. Callus induced from these plants possessed 23–32 chromosomes, a number lower than the combined chromosome number (66) of the parents, and were resistant to kanamycin, but they segregated for streptomycin resistance, which indicated that N. tabacum chloroplasts had been eliminated. Genomic DNA from several regenerated plants was analyzed by Southern hybridization for the presence of the neomycin phosphotransferase gene (NPTII); all of the plants analyzed were found to contain this gene. Mitochondrial (mt) DNA was analyzed by Southern hybridization of restriction endonuclease digests of mtDNA with two DNA probes, PKT5 and coxII. The results showed that the two plants analyzed possessed the mitochondria of D. carota. These results demonstrate that the regenerated plants are interfamilial somatic hybrids.  相似文献   

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Summary Fluorescence microscopy was used to visualize microtubules (Mts) and chromatin in an effort to further clarify the relationship between the generative cell (GC) and vegetative nucleus (VN) in pollen tubes of tobacco. Prominent Mt bundles are present in one or more GC extensions that can be finger-like or lamellar in form. While the VN is positioned distal to the GC in most cases, it can also straddle the cell or lie proximal to it. In all cases, however, extensions embrace, penetrate or clasp the VN. GC Mts are reorganized during the formation of the mitotic apparatus, and cell extensions are fully or partially withdrawn. By telophase in many pollen tubes, the VN shifts to a more proximal position and appears to adhere to the region of the GC containing the phragmoplast. Application of oryzalin leads to the disorganization of Mts, changes in cell shape, including the loss or alteration of cell extensions, and separation of the GC and VN in some cases. However, the position and polarity of the VN is maintained in most pollen tubes. The results indicate that GC Mts and cell extensions play a role in the association with the VN. However, the relationship appears to be controlled by other factors as well. Attention should now be directed at potential interactions involving the VN envelope, vegetative plasma membrane, GC plasma membrane and extracellular matrix.Abbreviations GC Generative cell - MGU male germ unit - Mt microtubule - VN vegetative nucleus  相似文献   

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Light is an important environment factor controlling plant growth, development, and nutritional quality and is also one of the most important factors inducing plant defence. In this study, we assayed the potential effects of light quality on the interaction between Nicotiana tabacum and cucumber mosaic virus (CMV). Our results indicated that white light‐treated N. tabacum plants displayed obvious symptoms at early stage postinoculation, while the symptoms were significantly inhibited under red light and blue light. Western blotting and quantitative real‐time PCR (qRT‐PCR) analyses showed that blue light and red light can effectively delay the replication of CMV compared with white light. The activities of various reactive oxygen species (ROS)‐scavenging enzymes and reducing substances [reduced glutathione (GSH) and ascorbic acid (ASA)] were increased under blue light and red light. In addition, hormone measurements and qRT‐PCR analyses revealed that salicylic acid (SA)‐mediated signalling pathway plays positive role in the related regulation, and cytokinin (CTK) may also participate in them. Furthermore, we found that the formation of dark green islands (DGIs) was significantly suppressed in plants under red light and blue light at 30 days postinoculation (Dpi). However, the accumulation of virus in plants under different light conditions had no notable differences at later stage of postinoculation. Taken together, these results indicated that red light and blue light could effectively delay symptom expression and replication of CMV on N. tabacum at the relatively earlier stage postinoculation.  相似文献   

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Osmotin, a plant protein, specifically binds a seven transmembrane domain receptor-like protein to exert its biological activity via a RAS2/cAMP signaling pathway. The receptor protein is encoded in the gene ORE20/PHO36 and the mammalian homolog of PHO36 is a receptor for the human hormone adiponectin (ADIPOR1). Moreover it is known that the osmotin domain I can be overlapped to the β-barrel domain of adiponectin. Therefore, these observations and some already existing structural and biological data open a window on a possible use of the osmotin or of its derivative as adiponectin agonist. We have modelled the three-dimensional structure of the adiponectin trimer (ADIPOQ), and two ADIPOR1 and PHO36 receptors. Moreover, we have also modelled the following complexes: ADIPOQ/ADIPOR1, osmotin/PHO36 and osmotin/ADIPOR1. We have then shown the structural determinants of these interactions and their physico-chemical features and analyzed the related interaction residues involved in the formation of the complexes. The stability of the modelled structures and their complexes was always evaluated and controlled by molecular dynamics. On the basis of these results a 9 residues osmotin peptide was selected and its interaction with ADIPOR1 and PHO36 was modelled and analysed in term of energetic stability by molecular dynamics. To confirm in vivo the molecular modelling data, osmotin has been purified from nicotiana tabacum seeds and its nine residues peptide synthesized. We have used cultured human synovial fibroblasts that respond to adiponectin by increasing the expression of IL-6, TNF-alpha and IL-1beta via ADIPOR1. The biological effect on fibroblasts of osmotin and its peptide derivative has been found similar to that of adiponectin confirming the results found in silico.  相似文献   

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