Analysis of Differentially Expressed Genes Associated with Coronatine-Induced Laticifer Differentiation in the Rubber Tree by Subtractive Hybridization Suppression

<div><p>The secondary laticifer in the secondary phloem is differentiated from the vascular cambia of the rubber tree (<i>Hevea brasiliensis</i> Muell. Arg.). The number of secondary laticifers is closely related to the rubber yield potential of <i>Hevea</i>. Pharmacological data show that jasmonic acid and its precursor linolenic acid are effective in inducing secondary laticifer differentiation in epicormic shoots of the rubber tree. In the present study, an experimental system of coronatine-induced laticifer differentiation was developed to perform SSH identification of genes with differential expression. A total of 528 positive clones were obtained by blue-white screening, of which 248 clones came from the forward SSH library while 280 clones came from the reverse SSH library. Approximately 215 of the 248 clones and 171 of the 280 clones contained cDNA inserts by colony PCR screening. A total of 286 of the 386 ESTs were detected to be differentially expressed by reverse northern blot and sequenced. Approximately 147 unigenes with an average length of 497 bp from the forward and 109 unigenes with an average length of 514 bp from the reverse SSH libraries were assembled and annotated. The unigenes were associated with the stress/defense response, plant hormone signal transduction and structure development. It is suggested that Ca²⁺ signal transduction and redox seem to be involved in differentiation, while PGA and EIF are associated with the division of cambium initials for COR-induced secondary laticifer differentiation in the rubber tree.</p></div

Reverse northern blot analysis of differentially expressed ESTs in the forward and reverse SSH libraries.

<p>Panel A, B and C, hybridization with unsubtracted cDNA probes from samples upon COR treatment for 1 day (A), 2 days (B) and 3 days (C). Panels D, E and F, hybridization with unsubtracted cDNA probes from samples upon water treatment for 1 day (D), 2 days (E) and 3 days (F). a to k, ESTs from the forward SSH library. i to t, ESTs from the reverse SSH library. The strong and corresponding weak hybridization signals were indicated with solid rings and dotted rings, respectively.</p

Diagram of <i>Hevea</i> epicormic shoots (A) and the cross-sections of bark (B-C) showing the sites of chemical application and secondary laticifer differentiation.

<p>The gray arrow in panel A indicates the site of chemical application and sampling. Panel B, treated with 20 μM coronatine (COR). Panel C, treated with water. EU, extension unit. White arrows show the primary laticifers. Black arrows show the secondary laticifers. Ca, cambium; St, sieve tube. Bars = 100 μm.</p

Expression pattern of all 10 unigenes from the forward (A) and reverse (B) SSH library by real-time PCR.

<p>Shoots were treated with 20 μM COR and water. Cambia-containing tissues were collected half an hour (0.5 h), one hour (1 h), two hours (2 h), four hours (4 h), eight hours (8 h), one day (1 d), two days (2 d) and three days (3 d) after treatments. The relative expression was normalized to the housekeeping genes of <i>HbACTIN</i> and <i>HbRH8</i>. The data were shown as averages ± SE. *, significant difference (<i>P</i> < 0.05); **, very significant difference (<i>P</i> < 0.01).</p

The GO standard analysis of 286 genes from the forward and reverse SSH libraries by BGI WEGO.

<p>The GO standard analysis of 286 genes from the forward and reverse SSH libraries by BGI WEGO.</p

Synthesis, crystal structure, and insecticidal evaluation of novel aryl pyrazole compounds containing aminophosphonate moiety

<p>A series of novel aryl pyrazole derivatives containing aminophosphonate were designed and synthesized on the basis of the commercial insecticide Fipronil. Their structures were confirmed by ¹H NMR, FT-IR, elemental analysis, and compound <b>4 m</b> was characterized by a single crystal X-ray diffraction analysis. The results of the preliminary bioassays showed that title compounds exhibited satisfactory insecticidal activities against <i>Culex pipiens</i> and <i>Musca domestica</i>. The introduction of an electron-withdrawing group to the para-position of the benzene ring was essential for high bioactivity.</p

3094904.pdf

Supplementary Informatio

Supplementary Information.docx

Supplemental document<br

A new dimeric anthraquinone from endophytic <i>Talaromyces</i> sp. YE3016

<p>A new unsymmetrical dimeric anthraquinone, 3-demethyl-3-(2-hydroxypropyl)-skyrin (<b>1</b>) was isolated from the solid-state fermentation extract of an endophytic fungal strain <i>Talaromyces</i> sp. YE 3016, together with five known compounds, skyrin (<b>2</b>), oxyskyrin (<b>3</b>), emodin (<b>4</b>), 1,3,6-trihydroxy-8-methyl-anthraquinone (<b>5</b>) and ergosterol (<b>6</b>). The structure of the new compound was elucidated on the basis of spectroscopic analysis. Compounds <b>1–3</b> exhibited moderate cytotoxic activities against MCF-7 cell line.</p

Molar Range Detection Based on Sideband Differential Absorption Spectroscopy with a Concentrated Reference

Conventional absorption spectroscopy (CAS) with a blank reference has only a slight capacity to detect high concentrations at characteristic wavelengths owing to the corresponding large molar absorption coefficient (ε) on the scale of 10³ or 10⁴ cm^–1 M^–1. To monitor concentrated analytes as high as the molar range in a plating bath and on a chemical production line, we propose a new approach using sideband differential absorption spectroscopy (SDAS). SDAS is obtained by subtracting the absorption spectra of the samples, <i>A</i>(λ,<i>C</i>_<i>x</i>), from that of a reference containing a concentrated standard analyte, <i>A</i>(λ,<i>C</i>_ref><i>C</i>_<i>x</i>), resulting in concave spectra with peaks at the sideband of conventional spectra with generally low ε values on the scale of 100 cm^–1 M^–1 or less. The negative absorbance changes linearly with the sample concentration at a certain peak wavelength, obeying Lambert–Beer’s law. In this work, SDAS was obtained and verified using inorganic and organic substances, such as chromate potassium, rhodamine B, and paracetamol