Subtype analysis of the HBV antigen-specific IgGs (anti-S and anti-PreS1) in sera of immunized mice before and after rAdSS1 boosting.

<p>(<b>A</b>) Sera were collected at 2 weeks after the 2^nd priming; (<b>B</b>) Sera were collected at 2 weeks after rAdSS1 boosting. Sera were diluted 1∶100. Bars indicate the average OD at 450 nm (OD₄₅₀) of each group. SS1: HBSS1. A, alum; C, CpG; P, poly(I:C). Adjs: control group with adjuvants mixture.</p

Characterisation of recombinant adenovirus rAdSS1.

<p>(<b>A</b>) Schematic representation of recombinant adenovirus viral vectors encoding HBV S and PreS1 fusion genes. ITR, inverted terminal repeat. (B) Western blot detection of SS1 fusion protein expression in HEK293 cells infected with rAdSS1 using specific rabbit anti-PreS1 polyclonal antibodies. The bands of the expressed SS1 proteins are indicated by arrowheads.</p

Total antibody-positivity rates after the 1^st and 2^nd HBSS1 priming.

<p>A, alum; C, CpG; P, poly(I:C).</p

Vaccination groups and administration strategy.

<p>A, alum; C, CpG; P, poly(I:C).</p

Table_3_Potential Root Foraging Strategy of Wheat (Triticum aestivum L.) for Potassium Heterogeneity.docx

Potassium (K) distribution is horizontally heterogeneous under the conservation agriculture approach of no-till with strip fertilization. The root foraging strategy of wheat for K heterogeneity is poorly understood. In this study, WinRHIZO, microarray, Non-invasive Micro-test Technology (NMT) and a split-root system were performed to investigate root morphology, gene expression profiling and fluxes of K+ and O2 under K heterogeneity and homogeneity conditions. The split-root system was performed as follows: C. LK (both compartments had low K), C. NK (both compartments had normal K), Sp. LK (one compartment had low K) and Sp. NK (the other compartment had normal K). The ratio of total root length and root tips in Sp. NK was significantly higher than that in C. NK, while no significant differences were found between Sp. LK and C. LK. Differential expression genes in C. LK vs. C. NK had opposite responses in Sp. LK vs. C. LK and similar responses in Sp. NK vs. C. NK. Low-K responsive genes, such as peroxidases, mitochondrion, transcription factor activity, calcium ion binding, glutathione transferase and cellular respiration genes were found to be up-regulated in Sp. NK. However, methyltransferase activity, protein amino acid phosphorylation, potassium ion transport, and protein kinase activity genes were found to be down-regulated in Sp. LK. The up-regulated gene with function in respiration tended to increase K+ uptake through improving O2 influx on the root surface in Sp. NK, while the down-regulated genes with functions of K+ and O2 transport tended to reduce K+ uptake on the root surface in Sp. LK. To summarize, wheat roots tended to perform active-foraging strategies in Sp. NK and dormant-foraging strategies in Sp. LK through the following patterns: (1) root development in Sp. NK but not in Sp. LK; (2) low-K responsive genes, such as peroxidases, mitochondrion, transcription factor activity, calcium ion binding and respiration, were up-regulated in Sp. NK but not in Sp. LK; and (3) root K+ and O2 influxes increased in Sp. NK but not in Sp. LK. Our findings may better explain the optimal root foraging strategy for wheat grown with heterogeneous K distribution in the root zone.</p

Identification of Regulatory Networks and Hub Genes Controlling Nitrogen Uptake in Tea Plants [Camellia sinensis (L.) O. Kuntze]

Nitrogen (N) uptake, as the first step of N metabolism, is a key limiting factor for plant growth. To understand the gene expression networks that control N absorption and metabolism in tea plants, we analyzed transcriptomes in the young roots of two groups of tea plants with significantly different growth rates under different N treatments (0, 0.2, and 2 mM). Using pairwise comparisons and weighted gene co-expression network analyses (WGCNA), we successfully constructed 16 co-expression modules. Among them, a specific module (turquoise) that substantially responded to the low N treatment was identified. Based on KEGG analysis, the relative genes that enriched in the “N metabolism” pathways were used to construct gene co-expression networks of N metabolism. Finally, a high-affinity ammonium (NH4+) transporter designated CsAMT1.2 was identified as a hub gene in the N metabolism network in tea plant roots and the gene expression could be highly induced by N resupply. The gene functional analysis revealed that CsAMT1.2 could make functional complementation of MEP1, MEP2, and MEP3 genes in 31019b yeast cells and improve NH4+ uptake rate in 31019b at low NH4+ level. Thus, CsAMT1.2 was a key gene controlling N uptake in tea plants and might play a vital role in promoting NH4+ uptake from the environment in tea roots. This study provided a useful foundation for improving the NUE in tea plantations

Immunisation of C57BL/6 mice.

<p>Each group (12 mice/group) was primed twice with HBSS1 together with various adjuvant combinations at weeks 0 and 4. At week 14, all immunised groups were boosted with rAdSS1. Humoral and cellular immune responses were evaluated at the indicated times.</p

ELISpot analysis of IFN-γ secretion in mouse splenocytes.

<p>(A ) Splenocytes were isolated at weeks 6 after the 2nd priming; (B) Splenocytes were isolated at weeks 16 after the rAdSS1 boost. Data are expressed as spot-forming cell (SFC) responses to S and PreS1 peptide pools, and are presented as means with SEM. Significant <i>p</i> values between vaccinated groups are shown as * <i>p</i><0.05, and ** <i>p</i><0.01. SS1: HBSS1 A, alum; C, CpG; P, poly(I:C). Adjs: control group with adjuvants mixture.Data are shown as means ± SE or ± SEMs.</p

Image_1_Potential Root Foraging Strategy of Wheat (Triticum aestivum L.) for Potassium Heterogeneity.TIF

Potassium (K) distribution is horizontally heterogeneous under the conservation agriculture approach of no-till with strip fertilization. The root foraging strategy of wheat for K heterogeneity is poorly understood. In this study, WinRHIZO, microarray, Non-invasive Micro-test Technology (NMT) and a split-root system were performed to investigate root morphology, gene expression profiling and fluxes of K+ and O2 under K heterogeneity and homogeneity conditions. The split-root system was performed as follows: C. LK (both compartments had low K), C. NK (both compartments had normal K), Sp. LK (one compartment had low K) and Sp. NK (the other compartment had normal K). The ratio of total root length and root tips in Sp. NK was significantly higher than that in C. NK, while no significant differences were found between Sp. LK and C. LK. Differential expression genes in C. LK vs. C. NK had opposite responses in Sp. LK vs. C. LK and similar responses in Sp. NK vs. C. NK. Low-K responsive genes, such as peroxidases, mitochondrion, transcription factor activity, calcium ion binding, glutathione transferase and cellular respiration genes were found to be up-regulated in Sp. NK. However, methyltransferase activity, protein amino acid phosphorylation, potassium ion transport, and protein kinase activity genes were found to be down-regulated in Sp. LK. The up-regulated gene with function in respiration tended to increase K+ uptake through improving O2 influx on the root surface in Sp. NK, while the down-regulated genes with functions of K+ and O2 transport tended to reduce K+ uptake on the root surface in Sp. LK. To summarize, wheat roots tended to perform active-foraging strategies in Sp. NK and dormant-foraging strategies in Sp. LK through the following patterns: (1) root development in Sp. NK but not in Sp. LK; (2) low-K responsive genes, such as peroxidases, mitochondrion, transcription factor activity, calcium ion binding and respiration, were up-regulated in Sp. NK but not in Sp. LK; and (3) root K+ and O2 influxes increased in Sp. NK but not in Sp. LK. Our findings may better explain the optimal root foraging strategy for wheat grown with heterogeneous K distribution in the root zone.</p

Identification of Regulatory Networks and Hub Genes Controlling Nitrogen Uptake in Tea Plants [Camellia sinensis (L.) O. Kuntze]

Nitrogen (N) uptake, as the first step of N metabolism, is a key limiting factor for plant growth. To understand the gene expression networks that control N absorption and metabolism in tea plants, we analyzed transcriptomes in the young roots of two groups of tea plants with significantly different growth rates under different N treatments (0, 0.2, and 2 mM). Using pairwise comparisons and weighted gene co-expression network analyses (WGCNA), we successfully constructed 16 co-expression modules. Among them, a specific module (turquoise) that substantially responded to the low N treatment was identified. Based on KEGG analysis, the relative genes that enriched in the “N metabolism” pathways were used to construct gene co-expression networks of N metabolism. Finally, a high-affinity ammonium (NH4+) transporter designated CsAMT1.2 was identified as a hub gene in the N metabolism network in tea plant roots and the gene expression could be highly induced by N resupply. The gene functional analysis revealed that CsAMT1.2 could make functional complementation of MEP1, MEP2, and MEP3 genes in 31019b yeast cells and improve NH4+ uptake rate in 31019b at low NH4+ level. Thus, CsAMT1.2 was a key gene controlling N uptake in tea plants and might play a vital role in promoting NH4+ uptake from the environment in tea roots. This study provided a useful foundation for improving the NUE in tea plantations