Functional Characterization of the Photosynthetic Machinery in Smicronix Galls on the Parasitic Plant Cuscuta campestris by JIP-Test.

Members of the genus Cuscuta are generally considered to be non-photosynthetic, stem-holoparasitic flowering plants. Under certain circumstances, at least some members of the genus are capable of limited photosynthesis. The galls of the Smicronyx weevils formed on Cuscuta campestris are particularly rich in chlorophylls compared to the stem of the parasitic plant. In the present study, we aimed to characterize the photosynthetic activity in the inner and outer gall cortices in comparison to the non-photosynthetic stems and a reference plant (Arabidopsis thaliana). The recorded prompt chlorophyll fluorescence transients were analyzed using JIP test. Detailed analysis of the chlorophyll fluorescence confirmed the presence of actively functioning photosynthetic machinery, especially in the inner cortex of the galls. This photosynthesis, induced by the insect larvae, did not reach the levels of the photosynthetic activity in Arabidopsis thaliana plants. Thylakoid protein complexes were identified by separation with two-dimensional Blue Native/SDS PAGE. It appeared that some of the complexes presented in A. thaliana are missing in C. campestris. We hypothesize that the insect-triggered transition from non-photosynthetic to photosynthetic tissue in the gall is driven by the increased requirements for nutrients related to the larval nutrition.

Autoinduction as Means for Optimization of the Heterologous Expression of Recombinant Single-Chain Fv (scFv) Antibodies.

The monoclonal antibodies and the recombinant antibody fragments are widely used in the biotechnology studies and in medicine as a powerful therapeutic and diagnostic tool. The most commonly used recombinant antibody fragments are single-chain fragment variable (scFv) because of their small size and minimal immunogenicity while still retaining high-affinity antigen binding. A wide range of expression systems such as bacterial and eukaryotic cell systems enable the sufficient production of scFv antibodies. However, their stable expression in soluble form and correct protein folding are often insufficient. In the present study, we present the autoinduction as a key element of the optimized scheme for heterologous expression of human monoclonal scFv antibodies (clones A1 and A12) in Escherichia coli HB2151, which resulted in two-fold increase of the total protein yield in 24 h.

Salinity Effect on Germination and Further Development of Parasitic Cuscuta spp. and Related Non-Parasitic Vines.

Plants are continuously subjected to the unfavorable impact of abiotic stress factors, of which soil salinity is among the most adverse. Although away from direct soil contact throughout most of their lifecycle, stem parasitic plants of the genus Cuscuta, family Convolvulaceae are also affected by salinity. The present study aimed to assess salt stress impact on germination and early establishment of three Cuscuta species, in comparison to related nonparasitic vines of the same family. It was found, that Cuscuta spp. are highly sensitive to NaCl concentration within the range of 200 mM. Germination was delayed in time and reduced by nearly 70%, accompanied by decrease in further seedling growth, ability to infect host plants and growth rate of established parasites. The nonparasitic vines showed similar sensitivity to salinity at germination level, but appeared to adapt better after the stress factor was removed. However, the negative effect of salinity did not fully prevent some of the Cuscuta species from infecting hosts, probably a beneficial characteristic at a species level, allowing the parasite to successfully thrive under the scarce host availability under saline conditions.