Salinity effect on Cuscuta campestris Yunck. Parasitism on Arabidopsis thaliana L.

Stem holoparasitic flowering plants of the genus Cuscuta are globally distributed invasive species and agricultural pests. The present research represents the combined effect of salt stress (e.g. abiotic stress) and Cuscuta campestris infection (e.g. biotic stress) on the model host plant Arabidopsis thaliana and the response of the parasite to salinity. The response of these parasites to abiotic stress conditions including salinity is poorly studied. Arabidopsis plants were continuously irrigated with 0, 50 and 150 mM NaCl and subjected to C. campestris infection. The influence of both abiotic and biotic stresses on the major osmoprotectant L-proline and three antioxidant enzymes - catalase, superoxide dismutase and guaiacol peroxidase, was assessed in both the parasite and the host plant. All four biochemical markers were differentially affected by stress, showing that the influence of C. campestris parasitism and its interaction with salinity is mostly in the site of infection (direct response) and also in roots (indirect vertical response) rather than on non-infected leaves of infected plants (indirect horizontal response). Despite its absence of soil contact, C. campestris was also significantly affected by salinity (indirect response). The mutual adaptation of the parasite-host pair to salinity slightly altered the regular response to abiotic stress of A. thaliana, but no detrimental additive effect of biotic and abiotic stress was observed.

Aphid (Myzus persicae) feeding on the parasitic plant dodder (Cuscuta australis) activates defense responses in both the parasite and soybean host.

Dodders (Cuscuta spp.) are shoot holoparasites, whose haustoria penetrate host tissues to enable fusion between the parasite and host vascular systems, allowing Cuscuta to extract water, nutrients and other molecules from hosts. Aphids are piercing-sucking herbivores that use specialized stylets to feed on phloem sap. Aphids are known to feed on Cuscuta, but how Cuscuta and its host plant respond to aphids attacking the parasite was unknown. Phytohormone quantification, transcriptomic analysis and bioassays were performed to determine the responses of Cuscuta australis and its soybean (Glycine max) hosts to the feeding of green peach aphid (GPA; Myzus persicae) on C. australis. Decreased salicylic acid levels and 172 differentially expressed genes (DEGs) were found in GPA-attacked C. australis, and the soybean hosts exhibited increased jasmonic acid contents and 1015 DEGs, including > 100 transcription factor genes. Importantly, GPA feeding on C. australis increased the resistance of the soybean host to subsequent feeding by the leafworm Spodoptera litura and soybean aphid Aphis glycines, resulting in 21% decreased leafworm mass and 41% reduced aphid survival rate. These data strongly suggest that GPA feeding on Cuscuta induces a systemic signal, which is translocated to hosts and activates defense against herbivores.

The parasitic plant Cuscuta australis is highly insensitive to abscisic acid-induced suppression of hypocotyl elongation and seed germination.

Around 1% of angiosperms are parasitic plants. Their growth and development solely or partly depend on host plants from which they extract water, nutrients, and other molecules using a parasitic plant-specific organ, the haustorium. Strong depletion of nutrients can result in serious growth retardation and in some cases, death of the hosts. The genus Cuscuta (dodder) comprises about 200 holoparasitic species occurring on all continents. Their seedlings have no roots and cotyledons but are only string-like hypocotyls. When they contact suitable host plants, haustoria are formed and thereafter seedlings rapidly develop into vigorously growing branches without roots and leaves. This highly specialized lifestyle suggests that Cuscuta plants likely have unique physiology in development and stress responses. Using germination and seedling growth assays, we show that C. australis seeds and seedlings are highly insensitive to abscisic acid (ABA). Transcriptome analysis and protein sequence alignment with Arabidopsis, tomato, and rice homologs revealed that C. australis most likely consists of only four functional ABA receptors. Given that Cuscuta plants are no longer severely challenged by drought stress, we hypothesize that the ABA-mediated drought resistance pathway in Cuscuta spp. might have had degenerated over time during evolution.

EASTERN DODDER (CUSCUTA MONOGYNA VAHL.) SEED GERMINATION AFFECTED BY SOME HERBACEOUS DISTILLATES.

Eastern dodder (Cuscuta monogyna Vahl.) is one of the noxious parasitic weeds that infected many ornamental trees in green spaces and gardens. Our purpose is to find natural inhibitors for prevention of its seed germination. In order to reach this aim, laboratory studies were conducted by using of herbaceous distillates of Dracocephalum moldavica, Nasturtium officinalis, Malva neglecta, Mentha piperita, Mentha pulegium, Rosa damascene, Ziziphora tenuior, and Urtica dioica on seed germination of C. monogyna. Z. tenuior distillate stimulated C. monogyna seed germination, whereas others reduced this parasitic weed's seed germination. D. moldavica caused maximum inhibition on weed seed germination. Seedling growth of C. monogyna was more affected than its seed germination. All of these herbaceous distillates reduced C. monogyna seedling length so that the latter decreased from 28.2 mm in distilled water to 4.5, 3.97, 3.85, 3.67, 3.1, 2.87, 2.57, 1.9, and 1.17 in M. pulegium, M. piperita, F. officinalis, Z. tenuior, N. officinalis, M. neglecta, R. damascene, U. dioica and D. moldavica, respectively. By using these medicinal plants distillates instead of herbicides, the parasitic weed seedling length and host plant infection will reduce.

Inter-species protein trafficking endows dodder (Cuscuta pentagona) with a host-specific herbicide-tolerant trait.

· Besides photosynthates, dodder (Cuscuta spp.) acquires phloem-mobile proteins from host; however, whether this could mediate inter-species phenotype transfer was not demonstrated. Specifically, we test whether phosphinothricin acetyl transferase (PAT) that confers host plant glufosinate herbicide tolerance traffics and functions inter-specifically. · Dodder tendrils excised from hosts can grow in vitro for weeks or resume in vivo by parasitizing new hosts. The level of PAT in in vivo and in vitro dodder tendrils was quantified by enzyme-linked immunosorbent assay. The glufosinate sensitivity was examined by dipping the distal end of in vivo and in vitro tendrils, growing on or excised from LibertyLink (LL; PAT-transgenic and glufosinate tolerant) and conventional (CN; glufosinate sensitive) soybean hosts, into glufosinate solutions for 5 s. After in vitro tendrils excised from LL hosts reparasitized new CN and LL hosts, the PAT level and the glufosinate sensitivity were also examined. · When growing on LL host, dodder tolerated glufosinate and contained PAT at a level of 0.3% of that encountered in LL soybean leaf. After PAT was largely degraded in dodders, they became glufosinate sensitive. PAT mRNA was not detected by reverse transcription PCR in dodders. · In conclusion, the results indicated that PAT inter-species trafficking confers dodder glufosinate tolerance.

[Reason for dormancy of Cuscuta chinensis seed and solving method].

OBJECTIVE: To study the reason for the deep dormancy of the aged Cuscuta chinensis seed and find the solving method. METHOD: The separated and combined treatments were applied in the orthogonal designed experiments. RESULT: The aged seed had well water-absorbency; the water and ethanol extracts of the seeds showed an inhibition effect on germination capacity of the seeds. CONCLUSION: The main reason for the deep dormancy of aged C. chinensis seed is the inhibitors existed in seed. There are two methods to solve the problem. The seeds is immersed in 98% of H2SO4 for 2 min followed by 500 mg x L(-1) of GA3 treatment for 60 min, or in 100 mg x L(-1) of NaOH for 20 min followed by 500 mg x L(-1) of GA3 treatment for 120 min.

Are herbicide-resistant crops the answer to controlling Cuscuta?

BACKGROUND: Herbicide-resistant crop technology could provide new management strategies for the control of parasitic plants. Three herbicide-resistant oilseed rape (Brassica napus L.) genotypes were used to examine the response of attached Cuscuta campestris Yuncker to glyphosate, imazamox and glufosinate. Cuscata campestris was allowed to establish on all oilseed rape genotypes before herbicides were applied. RESULTS: Unattached seedlings of C. campestris, C. subinclusa Durand & Hilg. and C. gronovii Willd. were resistant to imazamox and glyphosate and sensitive to glufosinate, indicating that resistance initially discovered in C. campestris is universal to all Cuscuta species. Glufosinate applied to C. campestris attached to glufosinate-resistant oilseed rape had little impact on the parasite, while imazamox completely inhibited C. campestris growth on the imidazolinone-resistant host. The growth of C. campestris on glyphosate-resistant host was initially inhibited by glyphosate, but the parasite recovered and resumed growth within 3-4 weeks. CONCLUSION: The ability of C. campestris to recover was related to the quality of interaction between the host and parasite and to the resistance mechanism of the host. The parasite was less likely to recover when it had low compatibility with the host, indicating that parasite-resistant crops coupled with herbicide resistance could be highly effective in controlling Cuscuta.

Chemical stress induced by heliotrope (Heliotropium europaeum L.) allelochemicals and increased activity of antioxidant enzymes.

The aims of this study were to evaluate the allelopathic potential of heliotrope on some biochemical processes of dodder. The preliminary experiments revealed that the effect of aqueous extract of leaves of heliotrope is higher than its seeds and roots. So, the aqueous extract of leaves was used in remaining experiments. Leaf extracts of 5 g powder per 100 mL H2O inhibited the germination of dodder seeds up to 95% and that of radish up to 100%. While, the aqueous extract of vine leaves which is a non-allelopathic plant did not have any inhibitory effect on these seeds. Vine leaf was used as a control to show that the inhibitory effect of heliotrope is due to an inhibitory compound but not due to the concentration. The leaf extract of heliotrope at 0.0, 0.1, 1.0, 2, 3, 4 and 5 g powder per 100 mL H2O reduced the radish seedling growth from 14 cm to about 0.5 cm and that of dodder from 7.5 cm to about 0.25 cm. The effects of heliotrope allelochemicals on some physiological and biochemical processes of radish was also Investigated. The activity of auxin oxidase increased in leaves and roots of radish. Suggesting that the reduced radish growth is due to the decreased active auxin levels in its leaves and roots. The activity of alpha-amylase was reduced, so reduction of starch degradation and lack of respiratory energy is the prime reason of germination inhibition in dodder and radish seeds. The level of soluble sugars increased. This is an indication of reduction of the activity of some respiratory enzymes and reduced consumption of these sugars. Proline levels were also increased, indicating that, the chemical stress is induced by leaf extract. Finally, the activities of GPX and CAT which are antioxidant enzymes were increased, along with increased extract concentration. These finding shows that the chemical stress induced by leaf extract produces super oxide (O2*) and H2O2, which is neutralized to H2O and O2 by these enzymes.

Synthesis of phytochelatins and modulation of antioxidants in response to cadmium stress in Cuscuta reflexa--an angiospermic parasite.

Effect of cadmium on growth, antioxidative enzymes namely catalase, peroxidase, glutathione reductase, level of glutathione and phytochelatin synthesis was investigated in callus and seedlings of Cuscuta reflexa. A time, concentration and tissue dependent response of Cd was observed. Cd inhibited the growth of callus and seedlings by 50% at 300 and 500 micromol/L concentrations, respectively. Shorter exposure of low concentration of Cd led to augmentation of antioxidant activity, both in callus and seedlings, while longer exposure and high concentration of Cd led to a concentration dependent decrease in callus. Analysis of phytochelatin (PC) synthesis in callus and seedlings of C. reflexa revealed both quantitative and qualitative changes. Cd at low concentrations led to synthesis of predominantly PC4, while at higher concentrations, PC3 was the major form being synthesized. Amelioration of antioxidative systems of C. reflexa in response to Cd stress might be playing a protective role, alleviating the damaging effects of ROS, generated during Cd stress. Concomitantly, chelation and sequestering of toxic Cd ions in this parasite was mediated by synthesis of PC. The response to Cd stress shown by this holoparasitic plant was found to be similar to those of non-parasitic plants (hosts).

Glyphosate inhibits the translocation of green fluorescent protein and sucrose from a transgenic tobacco host to Cuscuta campestris Yunk.

The parasitic plant Cuscuta campestris is dependent on its host for water, assimilates and amino acids. It can be controlled by the herbicide glyphosate, which inhibits 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), resulting in shikimate accumulation. In this study, C. campestris was parasitic on transgenic tobacco plants expressing green fluorescent protein (GFP) in the phloem. Changes in [14C]sucrose and GFP accumulation in the parasite were used as indicators of the herbicide's effect on translocation between the host and parasite. Host plants were treated with glyphosate 22 days after sowing. Shikimate accumulation in the parasite 1 day after glyphosate treatment (DAGT) confirmed EPSPS inhibition in C. campestris. No damage was visible in the host plants for the first 3 DAGT, while during that same time, a significant reduction in [14C]sucrose and GFP accumulation was observed in the parasite. Thus, we propose that the parallel reduction in GFP and sucrose accumulation in C. campestris is a result of a glyphosate effect on the parasite's ability to withdraw assimilates from the host.