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Agrobacterium rhizogenes-mediated transformation is widely used in different species with various purposes. The development of composite plants (wild-type shoot with transgenic roots) has been a milestone for functional characterization of genes. Previously, composite plants were generated by two steps from inducing of hairy roots to growing in the growth medium. Hairy roots were induced in an induction medium and the growth of composite plants generated were in another different growth medium. The composite plants produced was subject to transplanting. Here, we describe an improved and optimized protocol for generation of composite plant achieved by one-step in cucumber, which has not been reported previously in living plants. Incubation of explants post inoculation to induce transgenic roots and the growth of rooted explants were in the same medium. The primary root of 5-day-old seedling was excised and the slant cut of residual hypocotyl with 1 cm length was inoculated with A. rhizogenes harboring the desired gene construct followed by directly planted into a pot with wet sterile vermiculite. More than 90% of the infected seedlings can produce positive transgenic root. In addition, we further used the one-step transformation protocol to analyze the function of Arabidopsis YAO promoter. The result indicated that pYAO::GUS was highly conserved expression in whole root and high activity in the root tips. Therefore, a fast, expedient, high efficient, and one-step transformation method of composite cucumber produced is established, which is suitable for promoter functional analysis and other root-related events. Key messageA highly reliable, facile and one-step generation of composite cucumber plants (with transgenic roots and wild-type shoot) protocol performed effectively in different genotypes cucumber using Agrobacterium rhizogenes-mediated transformation.Publisher's Note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

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The effect of a strengthened working memory on the detection rate through increased search efficiency is more apparent in high visual clutter. This study confirms the role of visual clutter in construction-navigated inspections, thus serving as a foundation for the optimization of inspection planning.

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Complement dependent cytotoxicity (CDC) significantly contributes to Rituximab (RTX) and Ofatumumab (OFA) efficacies in the treatment of B-cell non-Hodgkin’s lymphoma (NHL) and chronic lymphocytic leukemia (CLL). Human CD59 (hCD59) is a key complement regulatory protein that restricts the formation of the membrane attack complex and thereby inhibits CDC. hCD59 is an important determinant of the sensitivity of NHL and CLL to RTX and OFA treatment. Recently, we developed a specific and potent hCD59 inhibitor, His-tagged ILYd4, which consists of 30 amino acid sequences extending from the N-terminus of ILYd4. Our previously published results indicate that His-tagged ILYd4 can be used as a lead candidate to further develop a potential therapeutic adjuvant for RTX and OFA treatment of RTX-resistant NHL and CLL. However, these studies were conducted using ILYd4 tagged on the N-terminus with 30 additional amino acids (AA) containing 6 X His used for immobilized metal affinity chromatograph. As a further step towards the development of ILYd4-based therapeutics, we investigated the impact of the removal of this extraneous sequence on the anti-hCD59 activity. In this paper, we report the generation and characterization of tag-free ILYd4. We demonstrate that tag-free ILYd4 has over three-fold higher anti-hCD59 activities than the His-tagged ILYd4. The enhanced RTX-mediated CDC effect on B-cell malignant cells comes from tag-free ILYd4’s improved functionality and physical properties including better solubility, reduced tendency to aggregation, and greater thermal stability. Therefore, tag-free ILYd4 is a better candidate for the further development for the clinical application.

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