Plant-specific GRAS transcription factors regulate various biological processes in plant growth, development and stress responses. However, this important gene family was not fully characterized in pepper (Capsicum annuum L.), an economically important vegetable crop. Here, a total of 50 CaGRAS members were identified in pepper genome and renamed by their respective chromosomal distribution. Genomic organization revealed that most CaGRAS genes (84%) have no intron. Phylogenetic analysis divided pepper CaGRAS members into 10 subfamilies, with each having distinct conserved domains and functions. For the expansion of the GRAS genes in pepper, segmental duplication contributed more than tandem duplication did. Gene expression analysis in various tissues demonstrated that most of CaGRAS genes exhibited a tissue- and development stage-specific expression pattern, uncovering their potential functions in pepper growth and development. Moreover, 21 CaGRAS genes were differentially expressed under cold, drought, salt and gibberellin acid (GA) treatments, indicating that they may implicated in plant response to abiotic stress. Notably, GA responsive cis-elements were detected in the promoter regions of the majority of CaGRAS genes, suggesting that CaGRAS may involve in signal cross-talking. The first comprehensive analysis of GRAS gene family in pepper genome by this study provide insights into understanding the GRAS-mediated regulation network, benefiting the genetic improvements in pepper and some other relative plants.

Pyrolysis of dried sewage sludge is regarded as an effective treatment method as well as a promising technology for energy/fuel production. In this work, thermochemical conversion of dried sewage sludge in a continuous reactor at different pyrolysis temperatures (400–800 °C) and solid residence times (6–46 min) was conducted. The pyrolysis products obtained using different pyrolysis conditions were extensively investigated. It is indicated that high pyrolysis temperature (>700 °C) and long solid residence time (>23 min) could enhance secondary reactions and decrease the yield of bio-oil. The maximum yield of bio-oil of 16.69% was achieved at reaction temperature of 700 °C and solid residence time of 23 min. The FTIR and GC-MS analyses of the bio-oil obtained at optimum condition indicated that it contained large amounts of phenols and esters. H2 and CO2 were the main components of pyrolysis gas, with a total amount that exceeded 52.18%. The characteristics of the char, including elemental composition, functional groups, and combustion property were investigated by XRF, FTIR, and TG. FTIR analysis of the char showed that the main functional groups are M–X, C–O, and C–H. The volatile content in the char decreased with an increase of pyrolysis temperature, while it increased with an increase of solid residence time. Heavy metals distribution in pyrolysis products of dried sewage sludge were investigated by inductively coupled plasma atomic emission spectroscopy. The results showed that most of the heavy metals remained in char. In addition, the potential ecological risk assessment of heavy metals was assessed.

Palmitoleic acid (16:1Δ 9 ) is one kind of ω-7 fatty acids (ω-7 FAs) widely used in food, nutraceutical and industry. However, such high-valued ω-7 FA only has a trace level in mature seeds of cotton and other common oil crops. We found that palmitoleic acid (>10.58 Mol%) was specially enriched in developing cotton endosperm which is disappeared in its mature seed. The present study was conducted to investigate the mechanism underlying high accumulation of palmitoleic acid in developing endosperm but not in embryo of upland cotton ( Gossypium hirsutum L.) seed. Of 17 stearoyl-ACP Δ 9 desaturases (SAD) gene family members identified in upland cotton, six GhSADs may specifically work in the desaturation of palmitic acid (16:0-ACP) to produce palmitoleic acid (16:1Δ 9 -ACP), which were revealed by examining the key amino acids in the catalytic center and their cis -elements. Gene expression analysis showed that spatial patterns of these GhSADs were different in developing ovules, with GhA-SAD6 and GhD-SAD8 preferentially expressed in developing endosperms. Functional analysis by transient expression in Nicotiana benthamiana leaves and genetic complementary assay using yeast mutant BY4389 strain unable to synthesize unsaturated fatty acids demonstrated that GhA-SAD6 and GhD-SAD8 have strong substrate specificity for 16:0-ACP. In contrast, GhA-SAD5 and GhA-SAD7 exhibited high specific activity on 18:0-ACP. Taken together, these data evidence that GhA-SAD6 and GhD-SAD8 are responsible for making palmitoleic acid in developing cotton endosperms, and provide endogenous gene targets for genetic modification to enrich ω-7 FAs in cotton seed oil required for sustainable production of functionality-valued products.

The plant-specific WRINKLED1 (WRI1) is a member of the AP2/EREBP class of transcription factors that positively regulate oil biosynthesis in plant tissues. Limited information is available for the role of WRI1 in oil biosynthesis in castor bean (Ricinus connunis L.), an important industrial oil crop. Here, we report the identification of two alternatively spliced transcripts of RcWRI1, designated as RcWRI1-A and RcWRI1-B. The open reading frames of RcWRI1-A (1341 bp) and RcWRI1-B (1332 bp) differ by a stretch of 9 bp, such that the predicted RcWRI1-B lacks the three amino acid residues “VYL” that are present in RcWRI1-A. The RcWRI1-A transcript is present in flowers, leaves, pericarps and developing seeds, while the RcWRI1-B mRNA is only detectable in developing seeds. When the two isoforms were individually introduced into an Arabidopsis wri1-1 loss-of-function mutant, total fatty acid content was almost restored to the wild-type level, and the percentage of the wrinkled seeds was largely reduced in the transgenic lines relative to the wri1-1 mutant line. Transient expression of each RcWRI1 splice isoform in N. benthamiana leaves upregulated the expression of the WRI1 target genes, and consequently increased the oil content by 4.3–4.9 fold when compared with the controls, and RcWRI1-B appeared to be more active than RcWRI1-A. Both RcWRI1-A and RcWRI1-B can be used as a key transcriptional regulator to enhance fatty acid and oil biosynthesis in leafy biomass.

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