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Down the slippery slope: plastid genome evolution in Convolvulaceae.

Cuscuta (dodder) is the only parasitic genus found in Convolvulaceae (morning-glory family). We used long PCR approach to obtain large portions of plastid genome sequence from Cuscuta sandwichiana in order to determine the size, structure, gene content, and synteny in the plastid genome of this Cuscuta species belonging to the poorly investigated holoparasitic subgenus Grammica. These new sequences are compared with the tobacco chloroplast genome, and, where data are available, with corresponding regions from taxa in the other Cuscuta subgenera. When all known plastid genome structural rearrangements in parasitic and nonparasitic Convolvulaceae are considered in a molecular phylogenetic framework, three categories of rearrangements in Cuscuta are revealed: plesiomorphic, autapomorphic, and synapomorphic. Many of the changes in Cuscuta, previously attributed to its parasitic mode of life, are better explained either as plesiomorphic conditions within the family, i.e., conditions shared with the rest of the Convolvulaceae, or, in most cases, autapomorphies of particular Cuscuta taxa, not shared with the rest of the species in the genus. The synapomorphic rearrangements are most likely to correlate with the parasitic lifestyle, because they represent changes found in Cuscuta exclusively. However, it appears that most of the affected regions, belonging to all of these three categories, have probably no function (e.g., introns) or are of unknown function (a number of open reading frames, the function of which, if any, has yet to be discovered).

Base Sequence↗

Characterization of phenolic compounds in the Chinese herbal drug Tu-Si-Zi by liquid chromatography coupled to electrospray ionization mass spectrometry.

Phenolic compounds are the major bioactive constituents of the Chinese herbal drug Tu-Si-Zi, which is prepared from the seeds of Cuscuta chinensis. However, seeds of C. australis also are offered under the name of this drug in the herb market. In order to make a comparison of their chemical constituents, the phenolic compounds of these two Cuscuta species were analyzed by high-performance liquid chromatography/diode-array detection/electrospray ion trap tandem mass spectrometry (HPLC/DAD/ESI-MS(n)). A total of 50 compounds were observed in the methanol extracts, including 23 flavonoids, 20 lignans and 7 quinic acid derivatives. These compounds were separated on a C18 column and identified or tentatively characterized based on UV spectra and MS fragmentation behavior. In contrast to previous reports, the phenolic patterns of these two Cuscuta species were found to be very different. Kaempferol and astragalin were the predominant constituents of C. australis, while hyperoside was the major compound in C. chinensis. Most of the identified compounds, especially the acylated flavonoid glycosides, have not previously been reported from Cuscuta species. In addition, a 30 Da neutral loss observed for flavonols was investigated and could be used to differentiate flavonoid isomers such as kaempferol and luteolin. The ESI-MS fragmentation behavior of furofuran lignans was also investigated, and a characteristic pathway is proposed. The large differences observed between the phenolic constituents of C. chinensis and C. australis strongly encouraged further comparison of the bioactivities of these two species.

Chemical Fractionation↗

[Effect of wuzi yanzong pill and its disassembled prescription on mitochondrial DNA deletion, respiratory chain complexes and ATP synthesis in aged rats].

OBJECTIVE: To investigate the effect of Wuzi Yanzong Pill (WZYZP) and its disassembled prescription on mitochondrial DNA deletions, respiratory chain complexes and ATP synthesis in aged rats. METHODS: Animal experiments with polymerase chain reaction (PCR), enzyme kinetics and bioluminescence technique were conducted. RESULTS: WZYZP and its disassembled prescription of Fructus Lycii and Semen Cuscutae could reduce the mitochondrial DNA deletions and raise the activities of mitochondrial respiratory chain complexes I, IV and adenosine triphosphate (ATP) synthesis in aged rats' brain; Fructus Lycii and Semen Cuscutae could also reduce the mitochondrial DNA deletions in aged rats' heart (P < 0.05, P < 0.01). CONCLUSION: WZYZP, Fructus Lycii and Semen Cuscutae have protective effect on oxidative damage of mitochondrial DNA in aged rats.

ATP Synthetase Complexes↗

Volatile chemical cues guide host location and host selection by parasitic plants.

The importance of plant volatiles in mediating interactions between plant species is much debated. Here, we demonstrate that the parasitic plant Cuscuta pentagona (dodder) uses volatile cues for host location. Cuscuta pentagona seedlings exhibit directed growth toward nearby tomato plants (Lycopersicon esculentum) and toward extracted tomato-plant volatiles presented in the absence of other cues. Impatiens (Impatiens wallerana) and wheat plants (Triticum aestivum) also elicit directed growth. Moreover, seedlings can distinguish tomato and wheat volatiles and preferentially grow toward the former. Several individual compounds from tomato and wheat elicit directed growth by C. pentagona, whereas one compound from wheat is repellent. These findings provide compelling evidence that volatiles mediate important ecological interactions among plant species.

Cues↗

Dodder hyphae invade the host: a structural and immunocytochemical characterization.

Dodder (Cuscuta pentagona) hyphae are unique amongst the parasitic weeds for their ability to apparently grow through the walls of the host plant. Closer examination reveals, however, that the hyphae do not grow through the host but rather induce the host to form a new cell wall (or extend the existing wall) to coat the growing hypha. This chimeric wall composed of walls from two species is even traversed by plasmodesmata that connect the two cytoplasms. Compositionally, the chimeric wall is quite different from the walls of either the host or in other cells of the dodder plant, on the basis of immunocytochemical labeling. The most striking differences were in the pectins, with much stronger labeling present in the chimeric wall than in either the host or other dodder walls. Interestingly, labeling with monoclonal antibodies specific to arabinan side chains of rhamnogalacturonan I pectin fraction was highly enriched in the chimeric wall, but antibodies to galactan side chains revealed no labeling. Arabinogalactan protein antibodies labeled the plasma membrane and vesicles at the tips of the hyphae and the complementary host wall, although the JIM8-reactive epitope, associated with very lipophilic arabinogalactan proteins, was found only in dodder cells and not the host. Callose was found in the plasmodesmata and along the forming hyphal wall but was found at low levels in the host wall. The low level of host wall labeling with anticallose indicates that a typical woundlike response was not induced by the dodder. When dodder infects leaf lamina, which have more abundant intercellular spaces than petioles or shoots, the hyphae grew both intra- and extracellularly. In the latter condition, a host wall did not ensheath the parasite and there was clear degradation of the host middle lamellae by the growing hyphae, allowing the dodder to pass between cells. These data indicate that the chimeric walls formed from the growth of the host cell wall in concert with the developing hyphae are unique in composition and structure and represent an induction of a wall type in the host that is not noted in surrounding walls.

Cell Wall↗

Attachment of the parasitic weed dodder to the host.

The parasitic weed dodder (Cuscuta pentagona L.) invades a number of potential host species, but the mechanisms responsible for ensuring tight adhesion to the wide variety of host surfaces have yet to be identified. In this study, a battery of microscopy protocols is used to examine the host-parasite interface in an effort to deduce these mechanisms. As the dodder shoot approaches the host tissue, epidermal cells in the parasite shoot elongate and differentiate into secretory type trichomes. The trichome cell walls are malleable, allowing them to elongate towards the host and bend their walls to conform to the shape of the host cell surface. The presence of osmiophilic particles (probable cell-wall-loosening complexes) at far greater numbers than found in other species presages the expansion and malleable nature of the epidermal cells. In addition to the changes in cell shape, the dodder trichome cells secrete an electron-opaque cementing substance that covers the host-parasite interface. When probed with antibodies that recognize cell wall components, the cement reacted only with antibodies that recognize chiefly de-esterified pectins but not other common wall constituents. These data indicate that dodder utilizes both a cementing layer of pectin and a radically modified epidermal cell wall to secure the parasite to the perspective host.

Cell Adhesion↗

Cross-species translocation of mRNA from host plants into the parasitic plant dodder.

An intriguing new paradigm in plant biology is that systemically mobile mRNAs play a role in coordinating development. In this process, specific mRNAs are loaded into the phloem transport stream for translocation to distant tissues, where they may impact on developmental processes. However, despite its potential significance for plant growth regulation, mRNA trafficking remains poorly understood and challenging to study. Here, we show that phloem-mobile mRNAs can also traffic between widely divergent species from a host to the plant parasite lespedeza dodder (Cuscuta pentagona Engelm.). Reverse transcription-polymerase chain reaction and microarray analysis were used to detect specific tomato (Lycopersicon esculentum Mill.) transcripts in dodder grown on tomato that were not present in control dodder grown on other host species. Foreign transcripts included LeGAI, which has previously been shown to be translocated in the phloem, as well as nine other transcripts not reported to be mobile. Dodders are parasitic plants that obtain resources by drawing from the phloem of a host plant and have joint plasmodesmata with host cortical cells. Although viruses are known to move between dodder and its hosts, translocation of endogenous plant mRNA has not been reported. These results point to a potentially new level of interspecies communication, and raise questions about the ability of parasites to recognize, use, and respond to transcripts acquired from their hosts.

Arabidopsis↗

Resistance and tolerance in a host plant-holoparasitic plant interaction: genetic variation and costs.

Host organisms are believed to evolve defense mechanisms (i.e., resistance and/or tolerance) under selective pressures exerted by natural enemies. A prerequisite for the evolution of resistance and tolerance is the existence of genetic variation in these traits for natural selection to act. However, selection for resistance and/or tolerance may be constrained by negative genetic correlations with other traits that affect host fitness. We studied genetic variation in resistance and tolerance against parasitic infection and the potential fitness costs associated with these traits using a novel study system, namely the interaction between a flowering plant and a parasitic plant. In this system, parasitic infection has significant negative effects on host growth and reproduction and may thus act as a selective agent. We conducted a greenhouse experiment in which we grew host plants, Urtica dioica, that originated from a single natural population and represented 20 maternal families either uninfected or infected with the holoparasitic dodder, Cuscuta europaea. that originated from the same site. We calculated correlations among resistance, tolerance, and host performance to test for costs of resistance and tolerance. We measured resistance as parasite performance (quantitative resistance) and tolerance as the slopes of regressions relating the vegetative and reproductive biomass of host plants to damage level (measured as parasite biomass). We observed significant differences among host families in parasite resistance and in parasite tolerance in terms of reproductive biomass, a result that suggests genetic variation in these traits. Furthermore, we found differences in resistance and tolerance between female and male host plants. In addition, the correlations indicate costs of resistance in terms of host growth and reproduction and costs of tolerance in terms of host reproduction. Our results thus indicate that host tolerance and resistance can evolve as a response to infection by a parasitic plant and that costs of resistance and tolerance may be one factor maintaining genetic variation in these traits.

Analysis of Variance↗