Search

Cytoplasmic male sterility (CMS) is a crucial means for the utilization of heterosis, which is of great significance for improving the yield and quality of hybrids. Currently, fertility restorationhas been extensively investigated in crops, but fertility restoration of CMS wheat with Aegilops Juvenalis cytoplasm are poorly understood.Here, a backcross population BC1F1derived from a cross between male sterile line Ju706A, its maintainer line 706B, and restorer line LK783, was used to map Rfd1locus by bulked segregant analysis and wheat 660K SNP genotyping.In which Ju706A displayedcomplete male sterility,and its fertility can be restored by LK783 with a pair of dominant genes Rfd1Rfd1. The locus was located to 2.4-Mb region on chromosome 1BS by markersAX-174254104 and AX-111201011.Combined with transcriptomic analysis and quantitative real-time PCR assay,TraesCS1B02G197400LC, the most likely candidate gene for Rfd1, encoded a pectinesterase which was localized in the cell wall, and was highly expressed in fertile anthers.The silencing of Rfd1 resulted in decreased fertility and heterogeneous expression of Rfd1 promoted pollen germination and affected the vegetative growth.It implies that Rfd1 is required for anther or pollen development and male fertility in CMS wheat with Aegilops Juvenalis cytoplasm. Further, a 7 bp deletion in Ju706A was employed to develop specific marker Xnwafu1 for molecular marker-assisted selection of restorers. Thestudy providesa new understanding for exploring the fertility restoration mechanism of CMS.

A novel waterfall aeration biofilm reactor integrated with a constructed wetland (WABR-CW) system was developed to meet the challenge of decentralized wastewater treatment with a focus on nutrient removal. In a lab-scale experiment of 70 days, the WABR-CW showed a high removal efficiency for COD (85-98 %), NH4+-N (100 %), TN (60-90 %) and TP (85 %-95 %), even when different organic loading rates (OLR) were used. The CW was responsible for improving the overall performance in view of an increased nutrient removal. The CW offers denitrification capacity when the OLR is not optimal for the WABR. Based on the lab-scale experiment, a pilotscale WABR-CW was built and tested for aquaculture wastewater treatment and reuse. A total of 63 m3 wastewater was treated of which 56.7 m3 was reused. Furthermore, the microbial structure of the WABR-CW system was investigated. A metabolic analysis highlighted the N and C metabolic pathways and functional genes distribution in the WABR-CW system. Next generation sequencing not only linked the pollutants removal performance and microbial encoding genes but also disclosed the potential ability of WABR-CW to treat more polluted and more complex wastewater. The outcomes of this study provide scale-up results and a better understanding of the functioning of the WABR-CW.

The loss of TaGAMYB function in Arabidopsis results in abnormal pollen development and leads to decreased fertility. This process may be regulated by microRNAs, which suppress the expression of fatty acid pathway genes. Development of the anthers and pollen is significantly influenced by the transcription factor GAMYB. However, our knowledge of GAMYB in wheat is limited. Here, under fertility and sterility conditions, we identified the distinct transcripts TaGAMYB-d and TaGAMYB-g in thermosensitive genic sterile wheat YanZhan 4110S and confirmed their functions. TaGAMYB-g overexpression decreased the pollen vigor and germination rates, thereby reducing fertility. TaGAMYB-d overexpression lines exhibited early flowering. Due to aberrant pollen germination, the mutant homologous TaGAMYB genes in Arabidopsis thaliana also resulted in lower fertility. Our findings indicate that TaGAMYB controls the fertility and flowering time in transgenic Arabidopsis thaliana.

Male reproductive development in higher plants experienced a series of complex biological processes, which can be regulated by Gibberellins (GA). The transcriptional factor GAMYB is a crucial component of GA signaling in anther development. However, the mechanism of GAMYB in wheat male reproduction is less understood. Here, we found that the thermo-sensitive genic male sterilitywheat line YanZhan 4110S displayed delayed tapetum programmed cell death and pollen abortive under the hot temperature stress. Combined with RNA-Sequencing data analysis, TaGAMYB associated with fertility conversion was isolated, which was located in the nucleus and highly expressed in fertility anthers. The silencing of TaGAMYB in wheat displayed fertility decline, defects in tapetum, pollen and exine formation, where the abortion characteristics were the same as YanZhan 4110S. In addition, either hot temperature or GA3 treatment in YanZhan 4110S caused the downregulation of TaGAMYB at binucleate stage and trinucleate stage, as well as fertility decrease. Further, the transcription factor TaWRKY2 significantly changed under GA3-treatment and directly interacted with the TaGAMYB promoter by W-box cis-element. Therefore, we suggested that TaGAMYB may be essential for anther development and male fertility, and GA3 activates TaGAMYB by TaWRKY2 to regulate fertility in wheat.

Major fertility restorer locus for Aegilops kotschyi cytoplasm in wheat, Rfk1 , was mapped to chromosome arm 1BS. Most likely candidate gene is TraesCS1B02G197400LC , which is predicted to encode a pectinesterase/pectinesterase inhibitor. Cytoplasmic male sterility (CMS) is widely used for heterosis and hybrid seed production in wheat. Genes related to male fertility restoration in the presence of Aegilops kotschyi cytoplasm have been reported, but the fertility restoration-associated gene loci have not been investigated systematically. In this study, a BC1F1 population derived from a backcross between KTP116A, its maintainer line TP116B, and its restorer line LK783 was employed to map fertility restoration by bulked segregant RNA-Seq (BSR-Seq). A major fertility allele restorer locus for Ae. kotschyi cytoplasm in wheat, Rfk1, was mapped to chromosome arm 1BS, and it was contributed by LK783. Morphological and cytological studies showed that male fertility restoration occurred mainly after the late uninucleate stage. Based on simple sequence repeat and single-nucleotide polymorphism genotyping, the gene locus was located between Xnwafu_6 and Xbarc137 on chromosome arm 1BS. To further isolate the specific region, six Kompetitive allele-specific polymerase chain reaction markers derived from BSR-Seq were developed to delimit Rfk1 within physical intervals of 26.0 Mb. After searching for differentially expressed genes within the candidate interval in the anthers and sequencing analysis, TraesCS1B02G197400LC was identified as a candidate gene for Rfk1 and it was predicted to encode a pectinesterase/pectinesterase inhibitor. Expression analysis also confirmed that it was specifically expressed in the anthers, and its expression level was higher in fertile lines compared with sterile lines. Thus, TraesCS1B02G197400LC was identified as the most likely candidate gene for Rfk1, thereby providing insights into the fertility restoration mechanism for K-type CMS in wheat.

Thermo-sensitive male sterility is of vital importance to heterosis, or hybrid vigor in crop production and hybrid breeding. Therefore, it is meaningful to study the function of the genes related to pollen development and male sterility, which is still not fully understand currently. Here, we conducted comparative analyses to screen fertility related genes using RNA-seq, iTRAQ, and PRM-based assay. A gene encoding expansin protein in wheat, TaEXPB5, was isolated in KTM3315A, which was in the cell wall and preferentially upregulated expression in the fertility anthers. The silencing of TaEXPB5 displayed pollen abortion, the declination or sterility of fertility. Further, cytological investigation indicated that the silencing of TaEXPB5 induced the early degradation of tapetum and abnormal development of pollen wall. These results revealed that the silencing of TaEXPB5 could eliminate the effects of temperature on male fertility, and resulting in functional loss of fertility conversion, which implied that TaEXPB5 may be essential for anther or pollen development and male fertility of KTM3315A. These findings provide a novel insight into molecular mechanism of fertility conversion for thermo-sensitive cytoplasmic male-sterility wheat, and contribute to the molecular breeding of hybrid wheat in the future.HighlightTaEXPB5 coffers to anther or pollen development and male fertility in KTM3315A, its silencing could eliminate the effects of temperature on male fertility, and resulting in functional loss of fertility conversion.

An increasing number of studies have shown that long non-coding RNAs (lncRNAs) play an important role in regulating plant fertility, however, they are less studied in wheat. Here, we analyzed lncRNA sequencing data and showed that the fixation carbon sequestration pathway was closely associated with pollen development and fertility conversion in KTM3315A, and eight differentially expressed genes under different fertility conditions were significantly regulated by TCONS_00093333 (designed as TaHTMAR) and transcription factors TaBBX25 and TaOBF1. Among them, TaBBX25 and TaOBF1 were located in the nucleus and expressed in the early stage of fertile anther development. Gene silencing experiments of TaHTMAR showed that TaHTMAR positively regulated the expression of TaBBX25 and TaOBF1 under fertile conditions, thereby reducing male fertility of KTM3315A. This study confirms the effective roles of TaHTMAR, TaBBX25, and TaOBF1 in the regulation of male fertility in wheat and provides a valuable molecular basis for hybrid wheat breeding.

Biotreated landfill leachate contains much refractory organics such as humic and fulvic acids, which can be degraded by O3. However, the low O3 mass transfer and high energy cost limit its wide application in landfill leachate treatment. Previous studies proved that packed bubble columns could enhance the O3 mass transfer and increase the synthetic humic acids wastewater degradation, but the performance of packed bubble columns in real wastewater treatment has not been investigated. Therefore, this study aims to evaluate the feasibility of application of packed bubble column in the real biotreated landfill leachates treatment and provide insights into the transformation of organic matters in leachates during ozonation. Packed bubble columns with lava rocks or metal pall rings (LBC or MBC) were applied and compared with a non-packed bubble column (BC). At an applied O3 dose of 8.35 mg/(Lwater sample min), the initial COD (400 mg/L) was only removed for 26% in BC and 32% in MBC while this was 46% in LBC, indicating LBC has the best performance. GC–MS analysis shows that raw biotreated leachate contains potential endocrine disruptors such as di(2-ethylhexyl) phthalate (DEHP). 61% of DEHP was removed in LBC and the least intermediate oxidation products from humic- and fulvic-like organics was detected in LBC. The highest O3 utilization efficiency (89%) and hydroxyl radical ( OH) exposure rate (3.0 × 10-10 M s) were observed in LBC with lowest energy consumption (EEO) for COD removal of 18 kWh/m3. The enhanced ozonation efficiency in LBC and MBC was attributed to the improved O3 mass transfer. Besides, LBC had additional adsorptive and catalytic activity that promoted the decomposition of O3 to generate OH. This study demonstrates that a packed bubble column increases removal and decreases energy use when treating landfill leachate, thus promoting the application of ozonation.

Thermo-sensitive cytoplasmic male sterility is of great significance to heterosis and hybrid seed production in wheat. Consequently, it is worthwhile to research the genes associated with male sterility. Although polygalacturonases (PGs) have been studied to play a crucial role in male reproduction of many plants, their functions in the reproductive development of wheat remain unclear. Here, TaPG (TraesCS7A02G404900) encoding a polygalacturonase was isolated from the anthers of KTM3315A, a wheat thermo-sensitive cytoplasmic male sterile with Aegilops kotschyi cytoplasm. Expression pattern analyses showed that TaPG was strongly expressed in fertile anthers and its protein was localized in the cell wall. Further verification via barley stripe mosaic virus revealed that the silencing of TaPG exhibited abnormal anthers, premature degradation of tapetum, pollen abortion, and defective pollen wall formation, resulting in the declination of fertility. Conclusively, our research suggested that TaPG contributed to the pollen development and male fertility, which will provide a novel insight into the fertility conversion of thermo-sensitive cytoplasmic male sterility in wheat.

Ozonation has been widely applied for the oxidation of contaminants in wastewater, and the disinfection of water. However, low ozone (O3) mass transfer efficiency in common ozonation reactors requires high O3 doses and causes high energy consumption. In this study, to intensify the O3 mass transfer and oxidation of humic acids (HA) solution, a lava rock packed bubble column (LBC) and a metal pall ring packed bubble column (MBC) were developed and evaluated. In comparison with non-packed bubble column (BC), both LBC and MBC enhanced the O3 mass transfer efficiency and the generation of hydroxyl radicals, thereby increasing the HA removal from an aqueous solution. At applied O3 dose of 33.3 mg/(Lcolumn h), the HA removal efficiency in BC was only 47%. When MBC and LBC were applied, it increased to 66% and 72%, respectively. Meanwhile, the O3 utilization efficiency in LBC reached 68%, which was higher than that in MBC (50%) and BC (21%). Consequently, LBC has the lowest energy consumption (EEO) for HA removal (1.4 kWh/m3), followed by MBC (1.6 kWh/m3) and BC (2.9 kWh/m3). LBC had better performance than MBC due to the adsorptive and catalytic roles of lava rock on the ozonation process. This study demonstrates the advantages of using lava rocks as packed materials in O3 bubble column over metal pall rings in intensifying O3 mass transfer and organic matters removal, which provides some insights into promoting the industrial application of O3.

Bioinformatic analysis identified the function of genes regulating wheat fertility. Barley stripe mosaic virus-induced gene silencing verified that the genes TaMut11 and TaSF3 are involved in pollen development and related to fertility conversion. Environment-sensitive genic male sterility is of vital importance to hybrid vigor in crop production and breeding. Therefore, it is meaningful to study the function of the genes related to pollen development and male sterility, which is still not fully understand currently. In this study, YanZhan 4110S, a new thermo-sensitive genic male sterility wheat line, and its near-isogenic line YanZhan 4110 were analyzed. Through comparative transcriptome basic bioinformatics and weighted gene co-expression network to further identify some hub genes, the genes TaMut11 and TaSF3 associated with pollen development and male sterility induced by high-temperature were identified in YanZhan 4110S. Further verification through barley stripe mosaic virus-induced gene silencing elucidated that the silencing of TaMut11 and TaSF3 caused pollen abortion, finally resulting in the declination of fertility. These findings provided data on the abortive mechanism in environment-sensitive genic male sterility wheat.

Background: Environment-sensitive genic male sterility is of vital importance to hybrid vigor in crop production and breeding, therefore, it is meaningful to identify and study the function of the genes related to pollen development and male sterility, which still not fully understanding currently. In this study, Yanzhan 4110S, a new thermo-sensitive genic male sterility (TGMS) wheat line, and its near isogenic line Yanzhan 4110 were carried out cytological features observation, bioinformatics analysis to investgate the abortion state and identified the genes involved in pollen development which have fertility regulation function. Barely stripe mosaic virus-induced gene silencing was used to verify the genes function.Results: Cytological analysis showed pollen abortion event of Yanzhan 4110S occur at the later uninucleate stage (Lun) under higher temperature induction (day/night temperatures of 22 °C/20 °C), when the anthers were collected and assessed for transcriptomic profiling through high-throughput sequencing. We then in-depth analyzed the differentially expressed genes (DEGs) by Gene Ontology, Kyoto Encyclopedia of Genes and Genomes, the results showed that the occurrence of Yanzhan 4110S male-sterility most likely related to metabolic pathway, including phenylpropanoid biosynthesis in the biosynthesis of other secondary metabolites, starch and sucrose metabolism in carbohydrate metabolism, carbon fixation in photosynthetic organisms as well as carbon metabolism in energy metabolism. The weighted gene co-expression network analysis in the transcriptome profiles further identified some hub genes, where the key genes involved in those pathways were intersection between the unique DEGs of Yanzhan 4110S in anther and hub genes, totally 228 genes, which were highly related to pollen development including TaMut11 and TaSF3. Moreover, further verification through barely stripe mosaic virus-induced gene silencing elucidated that the silencing of TaMut11 and TaSF3 caused pollen abortion, finally resulting in the declination of fertility. So, the genes TaMut11 and TaSF3 are related to fertility conversion of Yanzhan 4110S.Conclusion: Through comparative transcriptome bioinformatics analysis, the genes TaMut11 and TaSF3 associated with pollen development and male sterility induced by high temperature were identified in Yanzhan 4110S, and verificated by barely stripe mosaic virus-induced gene silencing. These findings provided researching the abortive mechanism in environment-sensitive genic male sterility wheat.

A large amount of disposable plastic face masks (DPFs) is produced and used during the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic, which results in an inevitable consequence of the dramatic increase of DPFs waste. However, the impact of DPFs exposure to the environment on their toxicity is rarely considered. In this study, a range of 76-276 items/L microplastics (MPs) was detected in the DPFs leachates, and fibrous (80.3%) and polypropylene (PP,89.2%) MPs were dominant. Co, Cu, Ni, Sr, Ti and Zn, were commonly detected in all leachates of the tested DPFs. Organics, such as acetophenone, 2,4-Di-tert-butylphenol, benzothiazole, bisphenol-A and phthalide, were found in the DPFs leachate, which were including organic solvents and plasticizer. Besides, we first found an emerging environmental risk substance, namely environmentally persistent free radicals (EPFRs), was generated in the DPFs leachates. The characteristic g-factors of the EPFRs was in a range of 2.003-2.004, identified as mixture of carbon- and oxygen-centered radicals. By means of in vitro toxicity assay, the DPFs leachate were confirmed to cause cytotoxicity and oxidative stress. Significantly, it is found that the formed EPFRs could contribute more toxic effects. Furthermore, when compared to N95 respirators, the tested surgical masks tend to release more MPs, leach more metals and organics, and generate more EPFRs. Surgical masks were thus showed higher risk than N95 respirators after exposure to water. This work highlights the importance of understanding the chemical complexity and possible toxicity of DPFs for their risk assessment.

LIM domain proteins were involved in organizing the cytoskeleton, adjusting the metabolism and gene expression, some of them were specific express in pollen. LIM gene family in plants were studied in sunflower, tobacco, foxtail millet, rape, rice and Arabidopsis thaliana, however, it has not been investigated in wheat to date. In the present study, we totally characterized 29 TaLIM genes through genome-wide analysis, which were divided into two categories and five subclasses according to phylogenetic analysis. RNA-Seq analysis indicated the expression patterns of TaLIM genes have specific temporal and spatial characteristics, especially TaLIM2 was highly expressed in fertility anthers. Phenotypic and cytological of BSMV: TaLIM2 showed that it had defects in the later stage of pollen development and germination, which further testified that TaLIM2 was closely related to fertility conversion. These findings will be useful for functional analysis of LIM genes in wheat fertility and contribute to hybrid wheat breeding.

Ozonation is an efficient method to remove trace organic contaminants (TrOCs) from (waste)water. However, high ozone dose requirement and energy consumption are still limiting its wider application for ozone-persistence TrOCs elimination. In this study, a packed bubble column is designed to enhance the ozone gas-liquid mass transfer and consequentially improve ozone and hydroxyl radical ( OH) exposures. An increase of 5–31% removal for five selected ozone-persistence TrOCs was observed in the packed bubble column in comparison to an unpacked bubble column under the same experimental settings. The energy consumption (EEO) for 90% TrOCs removal in the packed column was in range of 0.99–2.76 kWh/m3, which was 5–48% decrease after the use of packing materials. A kinetics-based approach based on the correlation between the concentration of target compounds, the second order reaction rate constants of these compounds with ozone and OH and the ozone and OH exposures was used to predict TrOCs removal. The predicted removal was consistent the measured removal, indicating that this prediction approach can be applied in packed system.

Four polygalacturonase gene family members were highlighted that contribute to elucidate the roles of polygalacturonase during the fertility conversion process in male-sterile wheat. Polygalacturonase (PG) belongs to a large family of hydrolases with important functions in cell separation during plant growth and development via the degradation of pectin. Specific expressed PGs in anthers may be significant for male sterility research and hybrid wheat breeding, but they have not been characterized in wheat (Triticum aestivum L.). In this study, we systematically studied the PG gene family using the latest published wheat reference genomic information. In total, 113 wheat PG genes were identified, which could be classified into six categories A–F according to their structure characteristics and phylogenetic comparisons with Arabidopsis and rice. Polyploidy and segmental duplications in wheat were proved to be mainly responsible for the expansion of the wheat PG gene family. RNA-seq showed that TaPGs have specific temporal and spatial expression characteristics, in which 12 TaPGs with spike-specific expression patterns were detected by qRT-PCR in different fertility anthers of KTM3315A, a thermo-sensitive cytoplasmic male-sterile wheat. Four of them specific upregulated (TaPG09, TaPG95, and TaPG93) or downregulated (TaPG87) at trinucleate stage of fertile anthers, and further aligning with the homologous in Arabidopsis revealed that they may undertake functions such as anther dehiscence, separation of pollen, pollen development, and pollen tube elongation, thereby inducing male fertility conversion in KTM3315A. These findings facilitate function investigations of the wheat PG gene family and provide new insights into the fertility conversion mechanism in male-sterile wheat.

Heat shock transcription factors (HSFs) play crucial roles in resisting heat stress and regulating plant development. Recently, HSFs have been shown to play roles in anther development. Thus, investigating the HSF family members and identifying their protective roles in anthers are essential for the further development of male sterile wheat breeding. In the present study, 61 wheat HSF genes (TaHsfs) were identified in the whole wheat genome and they are unequally distributed on 21 chromosomes. According to gene structure and phylogenetic analyses, the 61 TaHsfs were classified into three categories and 12 subclasses. Genome-wide duplication was identified as the main source of the expansion of the wheat HSF gene family based on 14 pairs of homeologous triplets, whereas only a very small number of TaHsfs were derived by segmental duplication and tandem duplication. Heat shock protein 90 (HSP90), HSP70, and another class of chaperone protein called htpG were identified as proteins that interact with wheat HSFs. RNA-seq analysis indicated that TaHsfs have obvious period- and tissue-specific expression patterns, and the TaHsfs in classes A and B respond to heat shock, whereas the C class TaHsfs are involved in drought regulation. qRT-PCR identified three TaHsfA2bs with differential expression in sterile and fertile anthers, and they may be candidate genes involved in anther development. This comprehensive analysis provides novel insights into TaHsfs, and it will be useful for understanding the mechanism of plant fertility conversion.

Background: Polygalacturonase (PG) belongs to a large family of hydrolases with important functions in cell separation during plant growth and development via the degradation of pectin. The specific expression of PG genes in anthers may be significant for male sterility research and hybrid wheat breeding, but it has not been characterized in wheat (Triticum aestivum L.). Results: We systematically studied the PG gene family using the latest published wheat reference genomic information. In total, 113 wheat PG genes were identified and renamed as TaPG01–113 based on their chromosomal positions. The PG genes are unequally distributed on 21 chromosomes and classified according to six categories from A–F. Analysis of the gene structures and conserved motifs demonstrated that the Class C and D TaPGs have relatively short gene sequences and a small number of introns. Class E TaPGs are the least conserved and lack conserved domain III. Polyploidy and segmental duplications in wheat were mainly responsible for the expansion of the wheat PG gene family. Predictions of cis-elements indicate that TaPGs have a wide range of functions, including the responses to light, hypothermia, anaerobic conditions, and hormonal stimulation, as well as being involved in meristematic tissue expression. RNA-seq showed that TaPGs have specific temporal and spatial expression characteristics. Twelve spike-specific TaPGs were screened using RNA-seq data and verified by qRT-PCR in the sterile and fertile anthers of thermo-sensitive male-sterile wheat. Four important candidate genes were identified as involved in the male fertility determination process. In fertile anthers, TaPG09 may be involved in the separation of pollen. TaPG87 and TaPG95 could play important roles in anther dehiscence. TaPG93 may be related to pollen development and pollen tube elongation. Conclusions: We analyzed the wheat PG gene family and identified four important TaPGs with differential expression levels in the wheat fertility conversion process. Our findings may facilitate functional investigations of the wheat PG gene family and provide new insights into the fertility conversion mechanism in male-sterile wheat.

Heat shock transcription factors (HSFs) play crucial roles in resisting heat stress and regulating plant development. Recently, HSFs have been shown to play roles in anther development. Thus, investigating the HSF family members and identifying their protective roles in anthers are essential for the further development of male sterile wheat breeding. In the present study, 61 wheat HSF genes (TaHsfs) were identified in the whole wheat genome and they are unequally distributed on 21 chromosomes. According to gene structure and phylogenetic analyses, the 61 TaHsfs were classified into three categories and 12 subclasses. Genome-wide duplication was identified as the main source of the expansion of the wheat HSF gene family based on 14 pairs of homeologous triplets, whereas only a very small number of TaHsfs were derived by segmental duplication and tandem duplication. Heat shock protein 90 (HSP90), HSP70, and another class of chaperone protein called htpG were identified as proteins that interact with wheat HSFs. RNA-seq analysis indicated that TaHsfs have obvious period- and tissue-specific expression patterns, and the TaHsfs in classes A and B respond to heat shock, whereas the C class TaHsfs are involved in drought regulation. qRT-PCR identified three TaHsfA2bs with differential expression in sterile and fertile anthers, and they may be candidate genes involved in anther development. This comprehensive analysis provides novel insights into TaHsfs, and it will be useful for understanding the mechanism of plant fertility conversion.

Background: Polygalacturonase (PG) belongs to a large family of hydrolases that undertake many important functions in cell separation during plant growth and development by degrading pectin. The specific expression of PG genes in pollen may have great significance for plant male sterile research and hybrid wheat breeding. However, it has not been reported in wheat ( Triticum aestivum L.). Results: Therefore, we systematically studied the PG gene family using the latest published wheat reference genomic information. A total of 113 PGs were identified and renamed as TaPG01 - 113 based on their position on the chromosome. They were unequally distributed on 21 chromosomes and were classified into six categories of A-F. Analysis of gene structures and conserved motifs revealed that the TaPGs of Class C and D had relatively short gene sequences and a small number of introns, and Class E TaPGs were the least conserved and all members did not have III conserved domain. Segmental duplication has been shown to be one of the major drivers of the expansion of the wheat PG gene family. The cis-element predictions indicate that wheat PGs had a wide range of functions, including response to light, hypothermia, anaerobic and hormonal stimulation, and also involved in meristematic tissue expression. In addition, twelve spike-specific expressions of TaPGs were screened using RNA-seq data, and finally three important genes were identified by expression analysis in the sterile and fertile anthers of thermo-sensitive male sterile wheat. TaPG93 was involved in the pollen development and elongation of pollen tubes, and TaPG87 and TaPG95 played important roles in the separation of pollen grains and the cracking of anthers dehiscence. Conclusions: This study, we performed a thorough analysis of the wheat PG gene family and finally obtained three TaPGs that affect wheat fertility. This will lay a solid foundation for the function exploration of wheat PG gene family and provide new enlightenment for the fertility conversion mechanism of male sterile wheat.

Background: Heat shock transcription factors (HSFs) play crucial roles in resisting heat stress and regulating plant development. Investigating the HSF family is essential for understanding the fertility conversion mechanism in thermo-sensitive male sterile wheat. Previous studies have investigated the HSF family in wheat but it is necessary to conduct more in-depth and systematic analyses based on the newly published reference genome. Results: In the present study, 61 wheat Hsf (TaHsf) genes were identified using two main strategies and renamed based on their physical locations on chromosomes. According to the gene structure and phylogenetic analyses, the 61 TaHsf genes were classified into three categories and eleven subclasses. The genes were unequally distributed on 21 chromosomes, including two pairs of tandem duplication genes and 52 TaHsf segmental duplication genes. According to the cis-elements identified, most of the TaHsfs can be activated by Ca++ and MYB, and they respond to drought, light, copper, and other stresses as well as heat shock. RNA-seq analysis indicated that the A2 class TaHsf genes exhibited persistently upregulated expression levels in the leaves/shoots, roots (except in the vegetative growth and reproductive growth stages), spikes, and grains in wheat under normal conditions. The A and B class TaHsf genes were positively regulated during the resistance to heat, whereas the C class genes were involved in drought regulation in wheat. Only the A and B class TaHsf genes were upregulated under fertile conditions in thermo-sensitive male sterile wheat. Conclusion: In this study, 61 wheat Hsf genes were identified based on the complete wheat reference genome. This comprehensive analysis provides novel insights into the TaHsf genes, including their diverse functions and involvement in metabolic pathways.

Environment-sensitive genic male sterility is a valid tool for hybrid production and hybrid breeding, but there are no previous reports of the molecular mechanism of fertility conversion. In this study, RNA-seq, phenotypic and cytological observations, and physiological indexes were applied to analyze thermo-sensitive genic male sterility line 4110S under different temperature conditions to explore the fertility transformation mechanism. In total, 3420 differentially expressed genes (DEGs) were identified comprising 2331 upregulated genes and 1089 downregulated genes. The DEGs were apparently distributed among 54 Gene Ontology functional groups. The phenylpropanoid, long-chain fatty acid, and jasmonic acid (JA) biosynthesis pathways were related to male sterility, where their downregulation blocked the synthesis of sporopollenin and JA. Phenotypic and cytological analyses showed that pollen wall defects and anther indehiscence at high temperatures induced sterility. Moreover, enzyme-linked immunosorbent assay results indicated that the abundance of JA was lower in 4110S under restrictive conditions (high temperature) than permissive conditions (low temperature). A possible regulated network of pathways associated with male sterility was suggested. These results provided insights into the molecular mechanism of fertility conversion in the thermosensitive male sterility system.

Figure S1. Breeding procedure of near-isogenic lines. (DOCX 1939 kb)

1α-Hydroxycholecalciferol (1α-OH-D3) is a vitamin D derivative. The objective of this study was to evaluate the effects of 1α-OH-D3 on the growth and the mRNA expression of vitamin D receptor (VDR) in the small intestine and kidney of chickens. A total of 240 males of one-day-old Ross 308 broilers was randomly assigned to 4 treatments with 5 replicates of 12 birds per replicate. Three levels of 1α-OH-D3 (1.25, 2.5, and 5 μg/kg) were added to a basal diet containing 0.50% calcium (Ca), 0.25% non-phytate phosphorus (NPP), and without supplemental cholecalciferol (vitamin D3). The control diet contained 1.00% Ca, 0.45% NPP, and 25 μg/kg cholecalciferol. Dietary 1α-OH-D3 levels linearly improved the average daily feed intake (ADFI), average daily gain (ADG), femur and tibia mineralization, and plasma Ca concentration, and retained Ca and total phosphorus (tP) amounts in broilers from 1 to 21 d of age (P 0.05). In addition, 1α-OH-D3 also linearly up-regulated the mRNA expression levels of VDR in the duodenum as well as those of VDR and sodium-phosphate cotransporter NaPi-IIa and NaPi-IIc in the kidney of broilers (P 0.05). However, 1α-OH-D3 did not affect the mRNA levels of 25-hydroxylase in the liver or NaPi-IIb in the duodenum (P 0.05). No differences were observed in the ADFI, ADG, bone length, plasma mineral concentration, retained tP amount, or the mRNA levels of the above genes (except for VDR in the kidney) between the birds fed the diet with 5 μg/kg 1α-OH-D3 and the birds fed the control diet (P 0.05). By contrast, the weight, ash weight, ash percentage, and Ca percentage of the bone, retained Ca amount, and the mRNA level of VDR in the kidney were lower in the birds fed the diet with 5 μg/kg 1α-OH-D3 than in the birds fed the control diet (P 0.05). These data indicate that 1α-OH-D3 up-regulates the gene expression of VDR in the small intestine and kidney at the transcriptional level, thereby improving the growth performance and bone mineralization of broiler chickens from 1 to 21 d of age.

Arsenite (As(III)) is toxic in drinking water, which becomes an environmental concern worldwide. This work was to synthesize porous hematite through the calcination of natural goethite concentrate for As(III) adsorption, including adsorption kinetics, isotherms and the influence of pH and temperature. The calcination was performed at 300 °C for 180 min, producing porous hematite with large amount of micropores. The maximum adsorption capacity of As(III) on porous hematite was achieved at pH 6.0 and 25 °C, about 14.46 mg g−1, compared with 2.965 mg g−1 on the original goethite concentrate. The improvement might be attributed to the formation of micropores and thus the increase in the surface area. Also, it was found that the adsorption was strongly pH dependent and reduced with increasing temperature. It is indicated that the low-cost porous hematite has great potential in As(III) removal from contaminated water.

This study was conducted to evaluate the relative biological value (RBV) of 1α-hydroxycholecalciferol (1α-OH-D3) to 25-hydroxycholecalciferol (25-OH-D3) in one- to 21-day-old broiler chickens fed calcium (Ca)- and phosphorus (P)-deficient diets. On the d of hatch, 450 male Ross 308 broiler chickens were weighed and randomly allotted to 9 treatments with 5 replicates of 10 birds per replicate. The basal diet contained 0.50% Ca and 0.25% non-phytate phosphorus (NPP) but was not supplemented with cholecalciferol (vitamin D3). The levels of Ca and NPP in basal diets were lower than those recommended by NRC (1994). 25-OH-D3 was fed at zero, 1.25, 2.5, 5.0, and 10.0 μg/kg, and 1α-OH-D3 was fed at 0.625, 1.25, 2.5, and 5.0 μg/kg. The RBV of 1α-OH-D3 to 25-OH-D3 based on vitamin D intake was determined by the slope ratio method. Results showed that 25-OH-D3 or 1α-OH-D3 improved the growth performance and decreased the mortality in one- to 21-day-old broilers. A linear relationship was observed between the level of 25-OH-D3 or 1α-OH-D3 and mineralization of the femur, tibia, or metatarsus. The RBV of 1α-OH-D3 to 25-OH-D3 were 234, 253, and 202% when the weight, ash weight, and Ca percentage of femur were used as criteria. The corresponding RBV of 1α-OH-D3 to 25-OH-D3 were 232 to 263% and 245 to 267%, respectively, when tibia and metatarsus mineralization were used as criteria. These data indicate that when directly feeding a hormonally active form of vitamin D as 1α-OH-D3 proportionally less is needed than when using the precursor (25-OH-D3) in diets deficient in Ca and P.

Previous studies have demonstrated that Lactobacillus has anti-inflammatory properties, but the protective functions of Lactobacillus and mechanisms of inhibition of necrotic enteritis (NE) in the intestines of chickens have not been fully clarified. In the present study, we selected a probiotic strain, Lactobacillus fermentum 1.2029, which has good adhesive ability and a high survival rate in low pH and bile salts. The objective of this study was to examine the anti-inflammatory properties of L. fermentum 1.2029 against NE in chickens. Two hundred forty 1-d-old male Arbor Acres broilers were blocked into 3 experimental groups as follows: (I) nonchallenge control group, (II) Clostridium perfringens challenge group, and (III) C. perfringens challenge + L. fermentum 1.2029 group. Lactobacillus fermentum 1.2029 (1.0 mL/d, 10(8) cfu/mL) was orally administered daily to group III during the course of the experiment, and all uninfected control chickens were inoculated accordingly with the same volume of PBS. Clostridium perfringens (0.5 mL on d 1 and 1.0 mL on d 14 to 21, 10(8) cfu/mL) was administered to chickens in group II. At 28 d, scoring of gross NE lesions was performed. Ileal segments of approximately 2 cm from 24 chickens in each experimental group were collected and fixed in 4% (wt/vol) neutral-buffered formalin solution for histological scoring. Ileal mucosa samples were also collected for mRNA analysis by real-time PCR. The results showed that L. fermentum 1.2029 reduced the severity of NE lesions in chickens. Histological scores revealed that L. fermentum 1.2029 also reduced the inflammation damage of NE in chickens. Changes in cytokines and Toll-like receptors (TLR) were determined, and L. fermentum 1.2029 was found to increase interleukin-10 levels and reduce interferon-γ and TLR2 levels in NE-infected chickens. The results showed that L. fermentum 1.2029 was able to regulate the intestinal mucosal immune response and ameliorate inflammation by changing expression levels of cytokines and TLR.

Cytoplasmic male sterility (CMS) where no functional pollen is produced has important roles in wheat breeding. The anther is a unique organ for male gametogenesis and its abnormal development can cause male sterility. However, the mechanisms and regulatory networks related to plant male sterility are poorly understood. In this study, we conducted comparative analyses using isobaric tags for relative and absolute quantification (iTRAQ) of the pollen proteins in a CMS line and its wheat maintainer. Differentially abundant proteins (DAPs) were analyzed based on Gene Ontology classifications, metabolic pathways and transcriptional regulation networks using Blast2GO. We identified 5570 proteins based on 23,277 peptides, which matched with 73,688 spectra, including proteins in key pathways such as glyceraldehyde-3-phosphate dehydrogenase, pyruvate kinase and 6-phosphofructokinase 1 in the glycolysis pathway, isocitrate dehydrogenase and citrate synthase in the tricarboxylic acid cycle and nicotinamide adenine dinucleotide (NADH)-dehydrogenase and adenosine-triphosphate (ATP) synthases in the oxidative phosphorylation pathway. These proteins may comprise a network that regulates male sterility in wheat. Quantitative real time polymerase chain reaction (qRT-PCR) analysis, ATP assays and total sugar assays validated the iTRAQ results. These DAPs could be associated with abnormal pollen grain formation and male sterility. Our findings provide insights into the molecular mechanism related to male sterility in wheat.

Adsorption is widely applied to remove arsenic from water. This paper reviewed and compared the recent progresses on the arsenic removal by adsorption using two-dimensional and three-dimensional graphene-based materials as adsorbents. Functional graphene sheet achieved the largest As(III) adsorption capacity of 138.79[Formula: see text]mg/g, while Mg-Al LDH/GO2 showed the largest As(V) adsorption capacity of 183.11[Formula: see text]mg/g. Parameters including pH, temperature, co-existing ions and loaded metal or metal oxide affected the adsorption process. The adsorption mechanisms of graphene-based materials for As(III) and As(V) could be explained by surface complexation and the electrostatic attraction, respectively. Future works are suggested to focus on regenerating of two-dimensional graphene-based adsorbents and developing the three-dimensional with large specific surface area and better adsorption performance.

Immune stress is the loss of immune homeostasis caused by external forces. The purpose of this experiment was to investigate the effects of immune stress on the growth performance, small intestinal enzymes and peristalsis rate, and mRNA expression of nutrient transporters in broiler chickens. Four hundred and thirty-two 1-d-old broilers (Cobb500) were randomly assigned to four groups for treatment; each group included nine cages with 12 birds per cage. Group 1 = no vaccine (NV); Group 2 = conventional vaccine (CV); group 3 = lipopolysaccharide (LPS)+conventional vaccine (LPS); group 4 = cyclophosphamide (CYP)+conventional vaccine (CYP). The results demonstrated that immune stress by LPS and CYP reduced body weight gain (BWG), feed intake (FI), small intestine peristalsis rate and sIgA content in small intestinal digesta (p