Tissue specific expression of leafy spurge <i>DAM</i> and <i>FT</i> genes.

<p>Gene expression in leaf, stem, shoot, crown buds, flower and root were examined at the end of a 16 hr photoperiod under non-dormancy inducing conditions. (A) <i>DAM1</i> and <i>DAM2</i> expression in different organs. (B) <i>FT2</i> and <i>FT4</i> expression in different organs. Error bars show standard deviation from three biological replicates.</p

Coordinated Expression of <i>FLOWERING LOCUS T</i> and <i>DORMANCY ASSOCIATED MADS-BOX</i>-Like Genes in Leafy Spurge

<div><p>Leafy spurge (<i>Euphorbia esula</i> L.) is a noxious perennial weed that produces underground adventitious buds, which are crucial for generating new vegetative shoots following periods of freezing temperatures or exposure to various control measures. It is also capable of flowering and producing seeds, but requires vernalization in some cases. <i>DORMANCY ASSOCIATED MADS-BOX</i> (<i>DAM</i>) genes have been proposed to play a direct role in the transition to winter-induced dormancy and maintenance through regulation of the <i>FLOWERING LOCUS T</i> (<i>FT</i>) gene, which also is likely involved in the vernalization process. To explore the regulation of <i>FT</i> and <i>DAM</i> during dormancy transitions in leafy spurge, the transcript accumulation of two previously cloned <i>DAM</i> splice variants and two different previously cloned <i>FT</i> genes was characterized. Under long-photoperiods (16 h light), both <i>DAM</i> and <i>FT</i> transcripts accumulate in a diurnal manner. Tissue specific expression patterns indicated the tissues with high <i>DAM</i> expression had low <i>FT</i> expression and vice versa. <i>DAM</i> expression is detected in leaves, stems, shoot tips, and crown buds. <i>FT</i> transcripts were detected mainly in leaves and flowers. Under dormancy inducing conditions, <i>DAM</i> and <i>FT</i> genes had an inverse expression pattern. Additionally, chromatin immunoprecipitation assays were performed using DAM-like protein specific antibodies to demonstrate that DAM or related proteins likely bind to cryptic and/or conserved CArG boxes in the promoter regions of <i>FT</i> genes isolated from endodormant crown buds. These results are consistent with the hypothesis that DAM proteins play a crucial role in leafy spurge dormancy transition and maintenance, potentially by negatively regulating the expression of <i>FT</i>.</p></div

Chromatin immunoprecipitation (CHIP) analysis of leafy spurge <i>FT2</i> promoter using a leafy spurge DAM peptide antibody from endodormant crown buds.

<p>(A) The location of primer pairs (P1 to P5) in the <i>FT2</i> promoter region used in the qRT-PCR analysis of CHIP assays. (B) The average fold enrichment analysis of immunoprecipitated DNA fragments by qRT-PCR in CHIP assay with different amounts of antibody as noted. Error bars represent the range from two separate experiments.</p

Comparison of the crown bud regrowth after 0, 4, or 7 weeks of endodormancy-inducing conditions.

<p>After the designated time points following dormancy induction or following vernalization, the aerial parts of the plants were removed, and the excised plants were grown in greenhouse for two weeks. Above chart only shows the average length of the longest shoots growing from crown buds after 0 through 7 weeks of endodormancy-inducing treatment and after twenty weeks vernalization. Error bars show range of the average of the longest stem from 7 plants for each experiment.</p

Midgut transcriptomal response of the rice leaffolder, <i>Cnaphalocrocis medinalis</i> (Guenée) to Cry1C toxin

<div><p><i>Cnaphalocrocis medinalis</i> (Guenée) is one of the important insect pests in rice field. Bt agents were recommended in the <i>C</i>. <i>medinalis</i> control and Bt rice is bred as a tactic to control this insect. However, the tolerance or resistance of insect to Bt protein is a main threat to the application of Bt protein. In order to investigate the response of <i>C</i>. <i>medinalis</i> transcriptome in defending a Cry1C toxin, high-through RNA-sequencing was carried in the <i>C</i>. <i>medinalis</i> larvae treated with and without Cry1C toxin. A total of 35,586 high-quality unigenes was annotated in the transcriptome of <i>C</i>. <i>medinalis</i> midgut. The comparative analysis identified 6,966 differently expressed unigenes (DEGs) between the two treatments. GO analysis showed that these genes involved in proteolysis and extracellular region. Among these DEGs, carboxylesterase, glutathione S-transferase and P450 were differently expressed in the treated <i>C</i>. <i>medinalis</i> midgut. Furthermore, trypsin, chymotrypsin, and carboxypeptidase were identified in DEGs, and most of them up-regulated. In addition, thirteen ABC transporters were downregulated and three upregulated in Cry1C-treated <i>C</i>. <i>medinalis</i> midgut. Based on the pathway analysis, antigen processing and presentation pathway, and chronic myeloid leukemia pathway were significant in <i>C</i>. <i>medinalis</i> treated with Cry1C toxin. These results indicated that serine protease, detoxification enzymes and ABC transporter, antigen processing and presentation pathway, and chronic myeloid leukemia pathway may involved in the response of <i>C</i>. <i>medinalis</i> to Cry1C toxin. This study provides a transcriptomal foundation for the identification and functional characterization of genes involved in the toxicity of Bt Cry protein against <i>C</i>. <i>medinalis</i>, and provides potential clues to the studies on the tolerance or resistance of an agriculturally important insect pest <i>C</i>. <i>medinalis</i> to Cry1C toxin.</p></div

Trypsin genes of <i>Cnaphalocrocis medinalis</i> midgut in response to the ingestion of Cry1C toxin.

<p>Trypsin genes of <i>Cnaphalocrocis medinalis</i> midgut in response to the ingestion of Cry1C toxin.</p

Differently expressed unigenes potentially involved in <i>Cnaphalocrocis medinalis</i> response to Cry1C toxin.

<p>Differently expressed unigenes potentially involved in <i>Cnaphalocrocis medinalis</i> response to Cry1C toxin.</p

Overview of genes annotations from <i>Cnaphalocrocis medinalis</i> midgut.

<p>(A) Species distribution of the BLASTX results. (B) GO categories of all unigenes and DEGs. (C) euKaryotic orthologous Groups (KOG) classification.</p

ABC transporters differently expressed in <i>Cnaphalocrocis medinalis</i> larvae treated with Cry1C toxin.

<p>ABC transporters differently expressed in <i>Cnaphalocrocis medinalis</i> larvae treated with Cry1C toxin.</p

Carboxypeptidase genes of <i>Cnaphalocrocis medinalis</i> midgut in response to the ingestion of Cry1C toxin.

<p>Carboxypeptidase genes of <i>Cnaphalocrocis medinalis</i> midgut in response to the ingestion of Cry1C toxin.</p