Dynamic nitrification and denitrification processes are affected by changes in soil redox conditions,and they play a vital role in regulating soil N_(2)O emissions in rice-based cultivation.It is imperative to underst...Dynamic nitrification and denitrification processes are affected by changes in soil redox conditions,and they play a vital role in regulating soil N_(2)O emissions in rice-based cultivation.It is imperative to understand the influences of different upland crop planting systems on soil N_(2)O emissions.In this study,we focused on two representative rotation systems in Central China:rapeseed–rice(RR)and wheat–rice(WR).We examined the biotic and abiotic processes underlying the impacts of these upland plantings on soil N_(2)O emissions.The results revealed that during the rapeseed-cultivated seasons in the RR rotation system,the average N_(2)O emissions were 1.24±0.20 and 0.81±0.11 kg N ha^(–1)for the first and second seasons,respectively.These values were comparable to the N_(2)O emissions observed during the first and second wheat-cultivated seasons in the WR rotation system(0.98±0.25 and 0.70±0.04 kg N ha^(–1),respectively).This suggests that upland cultivation has minimal impacts on soil N_(2)O emissions in the two rotation systems.Strong positive correlations were found between N_(2)O fluxes and soil ammonium(NH_(4)^(+)),nitrate(NO_(3)^(–)),microbial biomass nitrogen(MBN),and the ratio of soil dissolved organic carbon(DOC)to NO_(3)^(–)in both RR and WR rotation systems.Moreover,the presence of the AOA-amoA and nirK genes were positively associated with soil N_(2)O fluxes in the RR and WR systems,respectively.This implies that these genes may have different potential roles in facilitating microbial N_(2)O production in various upland plantation models.By using a structural equation model,we found that soil moisture,mineral N,MBN,and the AOA-amoA gene accounted for over 50%of the effects on N_(2)O emissions in the RR rotation system.In the WR rotation system,soil moisture,mineral N,MBN,and the AOA-amoA and nirK genes had a combined impact of over 70%on N_(2)O emissions.These findings demonstrate the interactive effects of functional genes and soil factors,including soil physical characteristics,available carbon and nitrogen,and their ratio,on soil N_(2)O emissions during upland cultivation seasons under rice-upland rotations.展开更多
The nuclear factor Y(NF-Y) gene family is a class of transcription factors that are widely distributed in eukaryotes and are involved in various biological processes. However, the NF-Y gene family members in watermelo...The nuclear factor Y(NF-Y) gene family is a class of transcription factors that are widely distributed in eukaryotes and are involved in various biological processes. However, the NF-Y gene family members in watermelon, a valued and nutritious fruit, remain largely unknown and their functions have not been characterized. In the present study, 22 ClNF-Y genes in watermelon, 29 CsNF-Y genes in cucumber, and 24CmNF-Y genes in melon were identified based on the whole-genome investigation and their protein properties, gene location, gene structure, motif composition, conserved domain, and evolutionary relationship were investigated. ClNF-YB9 from watermelon and its homologs in cucumber and melon were expressed specifically in seeds. Its expression remained low in the early stages of watermelon seed development,increased at 20 days after pollination(DAP), and peaked at 45–50 DAP. Moreover, the knockout mutant Clnf-yb9 exhibited abnormal leafy cotyledon phenotype, implying its critical role during seed formation.Finally, protein interaction assays showed that ClNF-YB9 interacts with all ClNF-YCs and the ClNF-YB9-YC4 heterodimer was able to recruit a ClNF-YA7 subunit to assemble a complete NF-Y complex, which may function in seed development. This study revealed the structure and evolutionary relationships of the NF-Y gene family in Cucurbitaceae and the novel function of ClNF-YB9 in regulating seed development in watermelon.展开更多
Many economically important crops and vegetables belonging to the cruciferous family are heavily endangered by clubroot disease caused by Plasmodiophora brassicae infection.Breeding of clubroot resistant cultivars bas...Many economically important crops and vegetables belonging to the cruciferous family are heavily endangered by clubroot disease caused by Plasmodiophora brassicae infection.Breeding of clubroot resistant cultivars based on mapping and cloning of resistant genes is commonly regarded as the most cost-effective and efficient way to fight against this disease.The traditional way of R gene functional validation requires stable transformation that is both time-and labor-consuming.In this study,a rapid and efficient hairy-root transgenic protocol mediated by Agrobacterium rhizogenes was developed.The transformation positive rate was over 80%in Brassica napus showed by GUS reporter gene and this transformation only took 1/6 of the time compared with stable transformation.The system was applicable to different B.napus varieties and other cruciferous crops including Brassica rapa and Brassica oleracea.In particular,two known CR genes,CRA3.7.1 and CRA8.2.4 were used respectively,as example to show that the system works well for CR gene study combined with subsequent P.brassicae infection in B.napus.Most importantly,it works both in over-expression that led to disease resistance,as well as in RNAi which led to disease susceptible phenotype.Therefore,this system can be used in batch-wise identification of CR genes,and also offered the possibility of manipulating key genes within the P.brassicae genome that could improve our knowledge on host-pathogen interaction.展开更多
Rice is a model plant for genomic study of grass species. Functional identification and definition of rice genes becomes the object of its functional genomics research. WRKY gene superfamily, one of the transcription ...Rice is a model plant for genomic study of grass species. Functional identification and definition of rice genes becomes the object of its functional genomics research. WRKY gene superfamily, one of the transcription factor gene families, was recently suggested to play important roles in plant development and stress response. In rice, the results of analyses of expression pattern and ectopic overexpressor lines also support this viewpoint, and the evidences implicate rice WRKY proteins in transcriptional reprogramming during biotic or abiotic stresses, senescence, sugar metabolites, and morphological architecture. In this paper, we review the advance in study of rice WRKY gene family and also propose unified nomenclature for rice WRKY factors to eliminate confusion.展开更多
It is very important in the field of bioinformatics to apply computer to perform the function annotation for new sequenced bio-sequences. Based on GO database and BLAST program, a novel method for the function annotat...It is very important in the field of bioinformatics to apply computer to perform the function annotation for new sequenced bio-sequences. Based on GO database and BLAST program, a novel method for the function annotation of new biological sequences is presented by using the variable-precision rough set theory. The proposed method is applied to the real data in GO database to examine its effectiveness. Numerical results show that the proposed method has better precision, recall-rate and harmonic mean value compared with existing methods.展开更多
Functional dyspepsia(FD) is a constellation of functional upper abdominal complaints with poorly elucidated pathophysiology. However, there is increasing evidence that susceptibility to FD is influenced by hereditary ...Functional dyspepsia(FD) is a constellation of functional upper abdominal complaints with poorly elucidated pathophysiology. However, there is increasing evidence that susceptibility to FD is influenced by hereditary factors. Genetic association studies in FD have examined genotypes related to gastrointestinal motility or sensation, as well as those related to inflammation or immune response. G-protein b3 subunit gene polymorphisms were first reported as being associated with FD. Thereafter, several gene polymorphisms including serotonin transporter promoter, interlukin-17 F, migration inhibitory factor, cholecystocynine-1 intron 1, cyclooxygenase-1, catechol-o-methyltransferase, transient receptor potential vanilloid 1 receptor, regulated upon activation normal T cell expressed and secreted, p22 PHOX, Toll like receptor 2, SCN10 A, CD14 and adrenoreceptors have been investigated in relation to FD; however, the results are contradictory. Several limitations underscore the value of current studies. Among others, inconsistencies in the definitions of FD and controls, subject composition differences regarding FD subtypes, inadequate samples, geographical and ethnical differences, as well as unadjusted environmental factors. Further well-designed studies are necessary to determine how targeted genes polymorphisms, influence the clinical manifestations and potentially the therapeutic response in FD.展开更多
A field experiment involving cry1Ab transgenic rice(GM) and its parental non-cry1Ab rice(M) has been on-going since 2014. The diversity of the bacterial communities and the abundance of the microbial functional genes ...A field experiment involving cry1Ab transgenic rice(GM) and its parental non-cry1Ab rice(M) has been on-going since 2014. The diversity of the bacterial communities and the abundance of the microbial functional genes which drive the conversion of nitrogen in paddy soil were analyzed during the growth period of rice in the fifth year of the experiment, using 16 S rRNAbased Illumina Mi Seq and real-time PCR on the amoA, nirS and nirK genes. The results showed no differences in the alpha diversity indexes of the bacterial communities, including Chao1, Shannon and Simpson, between the fields cultivated with line GM and cultivar M at any of the growth stages of rice. However, the bacterial communities in the paddy soil with line GM were separated from those of paddy soil with cultivar M at each of the growth stages of rice, based on the unweighted Uni Frac NMDS or PCoA. In addition, the analyses of ADONIS and ANOSIM, based on the unweighted Uni Frac distance, indicated that the above separations between line GM and cultivar M were statistically significant(P<0.05) during the growth season of rice. The increases in the relative abundances of Acidobacteria or Bacteroidetes, in the paddy soils with line GM or cultivar M, respectively, led to the differences in the bacterial communities between them. At the same time, functional gene prediction based on Illumina Mi Seq data suggested that the abundance of many functional genes increased in the paddy soil with line GM at the maturity stage of rice, such as genes related to the metabolism of starch, amino acids and nitrogen. Otherwise, the copies of bacterial amo A gene, archaeal amo A gene and denitrifying bacterial nir K gene significantly increased(P<0.05 or 0.01) in the paddy soil with line GM. In summary, the release of cry1Ab transgenic rice had effects on either the composition of bacterial communities or the abundance of microbial functional genes in the paddy soil.展开更多
Field resistances of nine accessions of common wild rice (Oryza rufipogon Griff.) and one rice variety (IR24) were evaluated by using nine strains of bacterial blight pathogen (Xanthomonas oryzae pv. oryzae) fro...Field resistances of nine accessions of common wild rice (Oryza rufipogon Griff.) and one rice variety (IR24) were evaluated by using nine strains of bacterial blight pathogen (Xanthomonas oryzae pv. oryzae) from the Philippines. IR24 was highly susceptible to all the strains, and six common wild rice accessions resisted all the nine strains, with a resistance frequency of 67%. The accessions Yulin and Wanning were only susceptible to PXO280 and PXO71, respectively. The accession Gaozhou was susceptible to the three strains PXO79, PXO99 and PXO339, whereas resistant to the other six strains. It could be concluded that there is at least one resistance gene in each common wild rice accession. The functional markers of the genes xa5, xa13, Xa21 and Xa27 were used to detect the presence of these resistance genes in the nine tested wild rice accessions, and it was found that four wild rice accessions contained heterozygous xa13. Among the nine common wild rice accessions, five were homozygous for Xa27 and three homozygous for xa27, and the accession Laibin contained neither xa27 nor Xa27. In addition, there were no xa5 and Xa21 in all of these accessions.展开更多
AIM: To investigate the influence of heme oxygenase-1 (HO-1) gene transfer on the viability and function of cultured rat islets in vitro. METHODS: Islets were isolated from the pancreata of Sprague-Dawley rats by intr...AIM: To investigate the influence of heme oxygenase-1 (HO-1) gene transfer on the viability and function of cultured rat islets in vitro. METHODS: Islets were isolated from the pancreata of Sprague-Dawley rats by intraductal collagenase diges- tion, and purified by discontinuous Ficoll density gradient centrifugation. Purified rat islets were transfected with adenoviral vectors containing human HO-1 gene (Ad- HO-1) or enhanced green fluorescent protein gene (Ad- EGFP), and then cultured for seven days. Transfection was confirmed by fluorescence microscopy and Western blot. Islet viability was evaluated by acridine orange/ propidium iodide fluorescent staining. Glucose-stimulated insulin release was detected using insulin radioimmuno- assay kits and was used to assess the function of islets. Stimulation index (SI) was calculated by dividing the insulin release upon high glucose stimulation by the insulin release upon low glucose stimulation. RESULTS: After seven days culture, the viability of cultured rat islets decreased significantly (92% ± 6% vs 52% ± 13%, P < 0.05), and glucose-stimulated insulin release also decreased significantly (6.47 ± 0.55 mIU/ L/30IEQ vs 4.57 ± 0.40 mIU/L/30IEQ, 14.93 ± 1.17 mIU/L/30IEQ vs 9.63 ± 0.71 mIU/L/30IEQ, P < 0.05). Transfection of rat islets with adenoviral vectors at an MOI of 20 was efficient, and did not impair islet function. At 7 d post-transfection, the viability of Ad-HO-1 transfected islets was higher than that of control islets(71% ± 15% vs 52% ± 13%, P < 0.05). There was no significant difference in insulin release upon low glucose stimulation (2.8 mmol/L) among Ad-HO-1 transfected group, Ad-EGFP transfected group, and control group (P > 0.05), while when stimulated by high glucose (16.7 mmol/L) solution, insulin release in Ad-HO-1 transfected group was significantly higher than that in Ad-EGFP transfected group and control group, respectively (12.50 ± 2.17 mIU/L/30IEQ vs 8.87 ± 0.65 mIU/L/30IEQ; 12.50 ± 2.17 mIU/L/30IEQ vs 9.63 ± 0.71 mIU/L/30IEQ, P < 0.05). The SI of Ad-HO-1 transfected group was also significantly higher than that of Ad-EGFP transfected group and control group, respectively (2.21 ± 0.02 vs 2.08 ± 0.05; 2.21 ± 0.02 vs 2.11 ± 0.03, P < 0.05). CONCLUSION: The viability and function of rat islets decrease over time in in vitro culture, and heme oxygenase-1 gene transfer could improve the viability and function of cultured rat islets.展开更多
The present research was designed to study the association of polymorphism of natural resistance-associated macrophage proteinl (Nrampl) with some immune function and the production performance in Large White pig. T...The present research was designed to study the association of polymorphism of natural resistance-associated macrophage proteinl (Nrampl) with some immune function and the production performance in Large White pig. The PCR-RFLP technique was applied to analyze the correlation between the polymorphisms of Nrampl gene and immune function [value of Polymorphonuclear Leukocytes (PMN) obtained by Nitroblue Tetrazolium (NBT) Reduction and effect of Cytotoxin in Monocyte] and production performance in 165 Large White pigs. The results showed that there was one Nde I restriction locus in Large White pig, and both values of PMN by NBT Reduction and effect of Cytotoxin in Monocyte in genotype BB were higher than those in genotype AB (P〈0.05). Simultaneously, the weight of 180-day-old pigs with genotype BB was higher than that with genotype AB (P〈0.05). The results indicated that there was a significant correlation between different genotypes of Nrampl gene and Immune function and production performance, and it can be regarded as a candidate gene of disease resistance. All these results provide valuable reference to further studies of pig disease resistance.展开更多
This paper reported firstly successful cloning of lycopene ε-cyclase (lbLCYe) gene from sweetpotato, lpomoea batatas (L.) Lam. Using rapid amplification of cDNA ends (RACE), lbLCYe gene was cloned from sweetpot...This paper reported firstly successful cloning of lycopene ε-cyclase (lbLCYe) gene from sweetpotato, lpomoea batatas (L.) Lam. Using rapid amplification of cDNA ends (RACE), lbLCYe gene was cloned from sweetpotato cv. Nongdafu 14 with high carotenoid content. The 1 805 bp cDNA sequence oflbLCYe gene contained a 1236 bp open reading frame (ORF) encoding a 411 amino acids polypeptide with a molecular weight of 47 kDa and an isoelectric point (pI) of 6.95. IbLCYe protein contained one potential lycopene ε-cyclase domain and one potential FAD (flavinadenine dinucleotide)/NAD(P) (nicotinamide adenine dinucleotide phosphate)-binding domain, indicating that this protein shares the typical characteristics of LCYe proteins. The gDNA oflbLCYe gene was 4 029 bp and deduced to contain 5 introns and 6 exons. Real-time quantitative PCR analysis revealed that the expression level of IbLCYe gene was significantly higher in the storage roots of Nongdafu 14 than those in the leaves and stems. Transgenic tobacco (cv. Wisconsin 38) expressing [bLCYe gene accumulated significantly more ^-carotene compared to the untransformed control plants. These results showed that lbLCYe gene has an important function for the accumulation of carotenoids of sweetpotato.展开更多
High concentrations of ammonium nitrogen released from tannery sludge during storage in open air may cause nitrogen pollution to soil and groundwater.To study the transformation mechanism of NH_(4)^(+)-N by nitrifying...High concentrations of ammonium nitrogen released from tannery sludge during storage in open air may cause nitrogen pollution to soil and groundwater.To study the transformation mechanism of NH_(4)^(+)-N by nitrifying functional bacteria in tannery sludge contaminated soils,a series of contaminated soil culture experiments were conducted in this study.The contents of ammonium nitrogen(as NH_(4)^(+)-N),nitrite nitrogen(as NO_(2)^(−)-N)and nitrate nitrogen(as NO_(3)^(−)-N)were analyzed during the culture period under different conditions of pollution load,soil particle and redox environment.Sigmodial equation was used to interpret the change of NO_(3)^(−)-N with time in contaminated soils.The abundance variations of nitrifying functional genes(amoA and nxrA)were also detected using the real-time quantitative fluorescence PCR method.The results show that the nitrification of NH_(4)^(+)-N was aggravated in the contaminated silt soil and fine sand under the condition of lower pollution load,finer particle size and more oxidizing environment.The sigmodial equation well fitted the dynamic accumulation curve of the NO_(3)^(−)-N content in the tannery sludge contaminated soils.The Cr(III)content increased with increasing pollution load,which inhibited the reproduction and activity of nitrifying bacteria in the soils,especially in coarse-grained soil.The accumulation of NO_(2)^(−)-N contents became more obvious with the increase of pollution load in the fine sand,and only 41.5%of the NH_(4)^(+)-N was transformed to NO_(3)^(−)-N.The redox environment was the main factor affecting nitrification process in the soil.Compared to the aerobic soil environment,the transformation of NH_(4)^(+)-N was significantly inhibited under anaerobic incubation condition,and the NO_(3)^(−)-N contents decreased by 37.2%,61.9%and 91.9%under low,medium and high pollution loads,respectively.Nitrification was stronger in the silt soil since its copy number of amoA and nxrA genes was two times larger than that of fine sand.Moreover,the copy numbers of amoA and nxrA genes in the silt soil under the aerobic environment were 2.7 times and 2.2 times larger than those in the anaerobic environment.The abundance changes of the amoA and nxrA functional genes have a positive correlation with the nitrification intensity in the tannery sludge-contaminated soil.展开更多
Biological desilication process is an effective way to remove silicate from rutile so that high purity rutile could be obtained. However, little is known about the molecular mechanism of this process. In this work, a ...Biological desilication process is an effective way to remove silicate from rutile so that high purity rutile could be obtained. However, little is known about the molecular mechanism of this process. In this work, a newly developed rutile bio-desilication reactor was applied to enrich rutile from rough rutile concentrate obtained from Nanzhao rutile mine and a comprehensive high through-put functional gene array(Geo Chip 4.0) was used to analyze the functional gene diversity, structure and metabolic potential of microbial communities in the biological desilication reactor. The results show that TiO2 grade of the rutile concentrate could increase from 78.21% to above 90% and the recovery rate could reach to 96% or more in 8-12 d. The results also show that almost all the key functional genes involved in the geochemical cycling process, totally 4324 and 4983 functional microorganism genes, are detected in the liquid and ore surface, respectively. There are totally 712 and 831 functional genes involved in nitrogen cycling for liquid and ore surface samples, respectively. The relative abundance of functional genes involved in the phosphorus and sulfur cycling is higher in the ore surface than liquid. These results indicate that nitrogen, phosphorus and sulfur cycling are also present in the desiliconization process of rutile. Acetogenesis genes are detected in the liquid and ore surface, which indicates that the desiliconizing process mainly depends on the function of acetic acid and other organic acids. Four silicon transporting genes are also detected in the sample, which proves that the bacteria have the potential to transfer silicon in the molecule level. It is shown that bio-desilication is an effective and environmental-friendly way for enrichment of rough rutile concentrate and presents an overview of functional diversity and structure of desilication microbial communities, which also provides insights into our understanding of metabolic potential in biological desilication reactor ecosystems.展开更多
Understanding the sequence diversity of rice blast resistance genes is important for breeding new resistant rice cultivars against the rice blast fungusMagnaporthe oryzae. In this study, we selected 24 rice cultivars ...Understanding the sequence diversity of rice blast resistance genes is important for breeding new resistant rice cultivars against the rice blast fungusMagnaporthe oryzae. In this study, we selected 24 rice cultivars with different genetic back-grounds to study the alelic diversity of rice blast resistance genesPiz-t, Pitaand Pik. For Piz-t, a total of 17 alelic types were found within the 24 cultivars. Blast inoculations showed that most of the mutations can affect the function of the resistance gene. For Pita, except for the difference at the 918th amino acid, a majority of the 21 mutations were detected among the cultivars. Inoculations with blast isolates carryingAvr-Pita revealed that cultivars with mutations in other sites except for the 918th amino acid did not affect the function of thePita gene. ForPik, a total of six alelic types were found within the 24 cultivars, but ifve of them lost the function of the resistance gene. In addition, we found thatPiz-t, Pita and Pik were expressed constitutively in the 24 rice cultivars and the expression level was not related to resistance. Our results have provided the sequence diversity information of the resistance genesPiz-t, Pita and Pik among the popular rice cultivars grown in the northeast region of China. Keywords:resistance gene, avirulence gene, aleles, function, genetic evolution zae(M. oryzae), is one of the most destructive diseases in rice production worldwide. Over the years, comprehensive studies on rice blast resistance have been conducted (Silue et al. 1992). The resistance in newly cultivated rice cultivars to M. oryzae can be lost quickly due to the high level of instability in the genome of the fungus (Bonmanet al. 1992). Previous studies show that cultivars with durable and broad-spectrum resistance againstM. oryzae carry multiple major resistance (R) and minor resistance genes (Liuet al. 2014). An effective way to control rice blast disease is, therefore, to breed rice cultivars with multiple R and QTL genes. To date, over 83 rice blast R genes have been identiifed, and are distributed on 11 rice chromosomes except Received 22 May, 2015 Accepted 26 October, 2015 WANG Yan, E-mail: 8806wy@163.com; Correspondence LIU Zhi-heng, Tel: +86-24-23738857, E-mail: lzhh1954@163.com; ZHENG Wen-jing, Tel: +86-24-31021081, E-mail: zwj27@126. com *These authors contributed equaly to this study. ? 2016, CAAS. Al rights reserved. Published by Elsevier Ltd. doi: 10.1016/S2095-3119(15)61207-2 1. Introduction Rice blast disease, caused by the fungusMagnaporthe ory-展开更多
Obtaining transgenic plants is a common method for analyzing gene function. Unfortunately, stable genetic transformation is difficult to achieve, especially for plants(e.g., soybean), which are recalcitrant to genet...Obtaining transgenic plants is a common method for analyzing gene function. Unfortunately, stable genetic transformation is difficult to achieve, especially for plants(e.g., soybean), which are recalcitrant to genetic transformation. Transient expression systems, such as Arabidopsis protoplast, Nicotiana leaves, and onion bulb leaves are widely used for gene functional studies. A simple method for obtaining transgenic soybean callus tissues was reported recently. We extend this system with simplified culture conditions to gene functional studies, including promoter analysis, expression and subcellular localization of the target protein, and protein-protein interaction. We also evaluate the plasticity of this system with soybean varieties, different vector constructs, and various Agrobacterium strains. The results indicated that the callus transformation system is efficient and adaptable for gene functional investigation in soybean genotype-, vector-, and Agrobacterium strain-independent modes. We demonstrated an easy set-up and practical homologous strategy for soybean gene functional studies.展开更多
[Objectives]This study was conducted to determine the functions of eight ORF genes of porcine circovirus-like virus P1.[Methods]The double-copy tandem molecular cloning of porcine circovirus-like virus P1 genome was u...[Objectives]This study was conducted to determine the functions of eight ORF genes of porcine circovirus-like virus P1.[Methods]The double-copy tandem molecular cloning of porcine circovirus-like virus P1 genome was used to construct molecular clones with eight ORFs deleted by DNA site-directed mutagenesis technology.After transfected into PK15 cells for a certain period of time,RNA were extracted and was used to verify whether the eight ORFs were deleted or not and used for gene microarry analysis.The GO functions and KEGG pathway enrichment of differentially expressed genes were analyzed.[Results]P1 ORF1 is mainly involved in the biological processes of defense response to virus,signal transduction,regulation of Rab GTPase activity,and lipid metabolic process,and involved in the molecular functions of protein phosphatase inhibitor activity,phosphatidylinositol phospholipase C activity,2 iron,2 sulfur cluster binding,phosphoric diester hydrolase activity,and Rab GTPase activator activity,and in the KEGG pathways of secretion of digestive gland and nervous system development.P1 ORF2 is mainly involved in the biological processes of positive regulation of leukocyte chemotaxis,positive regulation of cell proliferation,positive regulation of cell migration,defense response to virus,regulation of cell growth,and involved in the molecular functions of insulin-like growth factor binding,and chemokine activity,and in the KEGG pathways of cytosolic DNA-sensing pathway,RIG-I-like receptor signaling pathway,toll-like receptor signaling pathway,chemokine signaling pathway,and cytokines,cytokine-cytokine receptor interaction.The biological processes,molecular functions and related pathways involving P1 ORF3 and ORF5 are basically similar to those of ORF2.P1 ORF8 is mainly involved in the biological processes of purine ribonucleotide biosynthetic process,amino acid transport,defense response to virus,amino acid transmembrane transport,and involved in molecular functions of N6-(1,2-dicarboxyethyl)AMP AMP-lyase(fumarate-forming)activity,iron-sulfur cluster binding,amino acid transmembrane transporter activity.[Conclusions]The analysis of the ORF functions of P1 virus lays a foundation for the study of its pathogenicity and pathogenesis.展开更多
As a major raw material for the textile industry and the most important fiber crop in the world,cotton is of great significance in Chinese economy.The development of cotton fiber can be divided
基金the National Key Research and Development Program of China(2017YFD0800102)the Hubei Provincial Key Research and Development Program,China(2021BCA156)。
文摘Dynamic nitrification and denitrification processes are affected by changes in soil redox conditions,and they play a vital role in regulating soil N_(2)O emissions in rice-based cultivation.It is imperative to understand the influences of different upland crop planting systems on soil N_(2)O emissions.In this study,we focused on two representative rotation systems in Central China:rapeseed–rice(RR)and wheat–rice(WR).We examined the biotic and abiotic processes underlying the impacts of these upland plantings on soil N_(2)O emissions.The results revealed that during the rapeseed-cultivated seasons in the RR rotation system,the average N_(2)O emissions were 1.24±0.20 and 0.81±0.11 kg N ha^(–1)for the first and second seasons,respectively.These values were comparable to the N_(2)O emissions observed during the first and second wheat-cultivated seasons in the WR rotation system(0.98±0.25 and 0.70±0.04 kg N ha^(–1),respectively).This suggests that upland cultivation has minimal impacts on soil N_(2)O emissions in the two rotation systems.Strong positive correlations were found between N_(2)O fluxes and soil ammonium(NH_(4)^(+)),nitrate(NO_(3)^(–)),microbial biomass nitrogen(MBN),and the ratio of soil dissolved organic carbon(DOC)to NO_(3)^(–)in both RR and WR rotation systems.Moreover,the presence of the AOA-amoA and nirK genes were positively associated with soil N_(2)O fluxes in the RR and WR systems,respectively.This implies that these genes may have different potential roles in facilitating microbial N_(2)O production in various upland plantation models.By using a structural equation model,we found that soil moisture,mineral N,MBN,and the AOA-amoA gene accounted for over 50%of the effects on N_(2)O emissions in the RR rotation system.In the WR rotation system,soil moisture,mineral N,MBN,and the AOA-amoA and nirK genes had a combined impact of over 70%on N_(2)O emissions.These findings demonstrate the interactive effects of functional genes and soil factors,including soil physical characteristics,available carbon and nitrogen,and their ratio,on soil N_(2)O emissions during upland cultivation seasons under rice-upland rotations.
基金supported by the National Youth Talent Program (A279021801)Key-Area R&D Program of Guangdong province (2022B0202060001)+4 种基金Key R&D Program of Shaanxi Province (2023-YBNY-008)the Science and Technology Innovation Team of Shaanxi (2021TD-32)the Natural Science Foundation of Shaanxi Province (2021JM-089, 2022JM-112 and 2022JQ-162)the Key R&D Project from Yangling Seed Industry Innovation Center (K3031322016)the Fundamental Research Fund from Northwest A&F University (2452022111)。
文摘The nuclear factor Y(NF-Y) gene family is a class of transcription factors that are widely distributed in eukaryotes and are involved in various biological processes. However, the NF-Y gene family members in watermelon, a valued and nutritious fruit, remain largely unknown and their functions have not been characterized. In the present study, 22 ClNF-Y genes in watermelon, 29 CsNF-Y genes in cucumber, and 24CmNF-Y genes in melon were identified based on the whole-genome investigation and their protein properties, gene location, gene structure, motif composition, conserved domain, and evolutionary relationship were investigated. ClNF-YB9 from watermelon and its homologs in cucumber and melon were expressed specifically in seeds. Its expression remained low in the early stages of watermelon seed development,increased at 20 days after pollination(DAP), and peaked at 45–50 DAP. Moreover, the knockout mutant Clnf-yb9 exhibited abnormal leafy cotyledon phenotype, implying its critical role during seed formation.Finally, protein interaction assays showed that ClNF-YB9 interacts with all ClNF-YCs and the ClNF-YB9-YC4 heterodimer was able to recruit a ClNF-YA7 subunit to assemble a complete NF-Y complex, which may function in seed development. This study revealed the structure and evolutionary relationships of the NF-Y gene family in Cucurbitaceae and the novel function of ClNF-YB9 in regulating seed development in watermelon.
基金supported by grants from the Wuhan Science and Technology Major Project on Key techniques of biological breeding and Breeding of new varieties(Grant No.2022021302024851)the special project for sustainable development agenda of innovation demonstration zone(Grant No.202204AC100001-A04)the National Key R&D Program of China(Grant No.2022YFD1200400)。
文摘Many economically important crops and vegetables belonging to the cruciferous family are heavily endangered by clubroot disease caused by Plasmodiophora brassicae infection.Breeding of clubroot resistant cultivars based on mapping and cloning of resistant genes is commonly regarded as the most cost-effective and efficient way to fight against this disease.The traditional way of R gene functional validation requires stable transformation that is both time-and labor-consuming.In this study,a rapid and efficient hairy-root transgenic protocol mediated by Agrobacterium rhizogenes was developed.The transformation positive rate was over 80%in Brassica napus showed by GUS reporter gene and this transformation only took 1/6 of the time compared with stable transformation.The system was applicable to different B.napus varieties and other cruciferous crops including Brassica rapa and Brassica oleracea.In particular,two known CR genes,CRA3.7.1 and CRA8.2.4 were used respectively,as example to show that the system works well for CR gene study combined with subsequent P.brassicae infection in B.napus.Most importantly,it works both in over-expression that led to disease resistance,as well as in RNAi which led to disease susceptible phenotype.Therefore,this system can be used in batch-wise identification of CR genes,and also offered the possibility of manipulating key genes within the P.brassicae genome that could improve our knowledge on host-pathogen interaction.
基金supported by the Science Foundation of the Chinese Academy of Sciences (Grant No.KSCX2-YW-N-007)the National Natural Science Foundation of China (Grant No.30370803)+1 种基金the Ministry of Science and Technology of China (Grant No.2005DKA21006)‘Hundred Talents’ Program of the Chinese Academy of Sciences
文摘Rice is a model plant for genomic study of grass species. Functional identification and definition of rice genes becomes the object of its functional genomics research. WRKY gene superfamily, one of the transcription factor gene families, was recently suggested to play important roles in plant development and stress response. In rice, the results of analyses of expression pattern and ectopic overexpressor lines also support this viewpoint, and the evidences implicate rice WRKY proteins in transcriptional reprogramming during biotic or abiotic stresses, senescence, sugar metabolites, and morphological architecture. In this paper, we review the advance in study of rice WRKY gene family and also propose unified nomenclature for rice WRKY factors to eliminate confusion.
基金the support of the National Natural Science Foundation of China under Grant No.60673023,60433020,10501017,3040016the European Commission for TH/Asia Link/010 under Grant No.111084.
文摘It is very important in the field of bioinformatics to apply computer to perform the function annotation for new sequenced bio-sequences. Based on GO database and BLAST program, a novel method for the function annotation of new biological sequences is presented by using the variable-precision rough set theory. The proposed method is applied to the real data in GO database to examine its effectiveness. Numerical results show that the proposed method has better precision, recall-rate and harmonic mean value compared with existing methods.
基金This study was supported by the Oklahoma Applied Research Support (OARS), Oklahoma Center for the Advancement of Science and Technology (OCAST), the State of Oklahoma through the Project AR062-034, and the United States Department of Energy under the Genomics: GTL program through the Virtual Institute of Microbial Stress and Survival (VIMSShttp://vimss.lbl.gov), Environmental Remediation Science Program (ERSP), Office of Biological and Environmental Research, Office of Science.
文摘Functional dyspepsia(FD) is a constellation of functional upper abdominal complaints with poorly elucidated pathophysiology. However, there is increasing evidence that susceptibility to FD is influenced by hereditary factors. Genetic association studies in FD have examined genotypes related to gastrointestinal motility or sensation, as well as those related to inflammation or immune response. G-protein b3 subunit gene polymorphisms were first reported as being associated with FD. Thereafter, several gene polymorphisms including serotonin transporter promoter, interlukin-17 F, migration inhibitory factor, cholecystocynine-1 intron 1, cyclooxygenase-1, catechol-o-methyltransferase, transient receptor potential vanilloid 1 receptor, regulated upon activation normal T cell expressed and secreted, p22 PHOX, Toll like receptor 2, SCN10 A, CD14 and adrenoreceptors have been investigated in relation to FD; however, the results are contradictory. Several limitations underscore the value of current studies. Among others, inconsistencies in the definitions of FD and controls, subject composition differences regarding FD subtypes, inadequate samples, geographical and ethnical differences, as well as unadjusted environmental factors. Further well-designed studies are necessary to determine how targeted genes polymorphisms, influence the clinical manifestations and potentially the therapeutic response in FD.
基金the National Science and Technology Major Project of the Ministry of Science and Technology of China (2016ZX08001-001)。
文摘A field experiment involving cry1Ab transgenic rice(GM) and its parental non-cry1Ab rice(M) has been on-going since 2014. The diversity of the bacterial communities and the abundance of the microbial functional genes which drive the conversion of nitrogen in paddy soil were analyzed during the growth period of rice in the fifth year of the experiment, using 16 S rRNAbased Illumina Mi Seq and real-time PCR on the amoA, nirS and nirK genes. The results showed no differences in the alpha diversity indexes of the bacterial communities, including Chao1, Shannon and Simpson, between the fields cultivated with line GM and cultivar M at any of the growth stages of rice. However, the bacterial communities in the paddy soil with line GM were separated from those of paddy soil with cultivar M at each of the growth stages of rice, based on the unweighted Uni Frac NMDS or PCoA. In addition, the analyses of ADONIS and ANOSIM, based on the unweighted Uni Frac distance, indicated that the above separations between line GM and cultivar M were statistically significant(P<0.05) during the growth season of rice. The increases in the relative abundances of Acidobacteria or Bacteroidetes, in the paddy soils with line GM or cultivar M, respectively, led to the differences in the bacterial communities between them. At the same time, functional gene prediction based on Illumina Mi Seq data suggested that the abundance of many functional genes increased in the paddy soil with line GM at the maturity stage of rice, such as genes related to the metabolism of starch, amino acids and nitrogen. Otherwise, the copies of bacterial amo A gene, archaeal amo A gene and denitrifying bacterial nir K gene significantly increased(P<0.05 or 0.01) in the paddy soil with line GM. In summary, the release of cry1Ab transgenic rice had effects on either the composition of bacterial communities or the abundance of microbial functional genes in the paddy soil.
基金supported by the Project of the National Ministry of Science and Technology,China (Grant No.2006AA10Z1C8)the Knowledge Innovative Program of the Chinese Academy of Sciences (Grant Nos.KSCX-YW-N-009-02 and KSCX1-YW-03)+1 种基金the National Basic Research Program of China (Grant No.2009CB126004)the Natural Science Foundation of Hainan Province,China (Grant No.309019)
文摘Field resistances of nine accessions of common wild rice (Oryza rufipogon Griff.) and one rice variety (IR24) were evaluated by using nine strains of bacterial blight pathogen (Xanthomonas oryzae pv. oryzae) from the Philippines. IR24 was highly susceptible to all the strains, and six common wild rice accessions resisted all the nine strains, with a resistance frequency of 67%. The accessions Yulin and Wanning were only susceptible to PXO280 and PXO71, respectively. The accession Gaozhou was susceptible to the three strains PXO79, PXO99 and PXO339, whereas resistant to the other six strains. It could be concluded that there is at least one resistance gene in each common wild rice accession. The functional markers of the genes xa5, xa13, Xa21 and Xa27 were used to detect the presence of these resistance genes in the nine tested wild rice accessions, and it was found that four wild rice accessions contained heterozygous xa13. Among the nine common wild rice accessions, five were homozygous for Xa27 and three homozygous for xa27, and the accession Laibin contained neither xa27 nor Xa27. In addition, there were no xa5 and Xa21 in all of these accessions.
基金Supported by the National Natural Science Foundation of China, No. 30571759Social Development Foundation of Shanghai, No. 200253
文摘AIM: To investigate the influence of heme oxygenase-1 (HO-1) gene transfer on the viability and function of cultured rat islets in vitro. METHODS: Islets were isolated from the pancreata of Sprague-Dawley rats by intraductal collagenase diges- tion, and purified by discontinuous Ficoll density gradient centrifugation. Purified rat islets were transfected with adenoviral vectors containing human HO-1 gene (Ad- HO-1) or enhanced green fluorescent protein gene (Ad- EGFP), and then cultured for seven days. Transfection was confirmed by fluorescence microscopy and Western blot. Islet viability was evaluated by acridine orange/ propidium iodide fluorescent staining. Glucose-stimulated insulin release was detected using insulin radioimmuno- assay kits and was used to assess the function of islets. Stimulation index (SI) was calculated by dividing the insulin release upon high glucose stimulation by the insulin release upon low glucose stimulation. RESULTS: After seven days culture, the viability of cultured rat islets decreased significantly (92% ± 6% vs 52% ± 13%, P < 0.05), and glucose-stimulated insulin release also decreased significantly (6.47 ± 0.55 mIU/ L/30IEQ vs 4.57 ± 0.40 mIU/L/30IEQ, 14.93 ± 1.17 mIU/L/30IEQ vs 9.63 ± 0.71 mIU/L/30IEQ, P < 0.05). Transfection of rat islets with adenoviral vectors at an MOI of 20 was efficient, and did not impair islet function. At 7 d post-transfection, the viability of Ad-HO-1 transfected islets was higher than that of control islets(71% ± 15% vs 52% ± 13%, P < 0.05). There was no significant difference in insulin release upon low glucose stimulation (2.8 mmol/L) among Ad-HO-1 transfected group, Ad-EGFP transfected group, and control group (P > 0.05), while when stimulated by high glucose (16.7 mmol/L) solution, insulin release in Ad-HO-1 transfected group was significantly higher than that in Ad-EGFP transfected group and control group, respectively (12.50 ± 2.17 mIU/L/30IEQ vs 8.87 ± 0.65 mIU/L/30IEQ; 12.50 ± 2.17 mIU/L/30IEQ vs 9.63 ± 0.71 mIU/L/30IEQ, P < 0.05). The SI of Ad-HO-1 transfected group was also significantly higher than that of Ad-EGFP transfected group and control group, respectively (2.21 ± 0.02 vs 2.08 ± 0.05; 2.21 ± 0.02 vs 2.11 ± 0.03, P < 0.05). CONCLUSION: The viability and function of rat islets decrease over time in in vitro culture, and heme oxygenase-1 gene transfer could improve the viability and function of cultured rat islets.
文摘The present research was designed to study the association of polymorphism of natural resistance-associated macrophage proteinl (Nrampl) with some immune function and the production performance in Large White pig. The PCR-RFLP technique was applied to analyze the correlation between the polymorphisms of Nrampl gene and immune function [value of Polymorphonuclear Leukocytes (PMN) obtained by Nitroblue Tetrazolium (NBT) Reduction and effect of Cytotoxin in Monocyte] and production performance in 165 Large White pigs. The results showed that there was one Nde I restriction locus in Large White pig, and both values of PMN by NBT Reduction and effect of Cytotoxin in Monocyte in genotype BB were higher than those in genotype AB (P〈0.05). Simultaneously, the weight of 180-day-old pigs with genotype BB was higher than that with genotype AB (P〈0.05). The results indicated that there was a significant correlation between different genotypes of Nrampl gene and Immune function and production performance, and it can be regarded as a candidate gene of disease resistance. All these results provide valuable reference to further studies of pig disease resistance.
基金supported by the National Transgenic Crops Program, China (Grant No. 2008ZX08001-006)the Research Funds for Public Benefit in Ministry of Agriculture, China (Grant No. 200803056)+1 种基金the Key Support Program of Jiangsu Science and Technology, China (Grant No. BE2008354)the Self-directed Innovation Fund of Agricultural Science and Technology in Jiangsu Province, China (Grant No. CX [08]603)
基金supported by the China Agriculture Research System (Sweetpotato)the National High-Tech Research and Development Project of China(2011AA100607 and 2012AA101204)
文摘This paper reported firstly successful cloning of lycopene ε-cyclase (lbLCYe) gene from sweetpotato, lpomoea batatas (L.) Lam. Using rapid amplification of cDNA ends (RACE), lbLCYe gene was cloned from sweetpotato cv. Nongdafu 14 with high carotenoid content. The 1 805 bp cDNA sequence oflbLCYe gene contained a 1236 bp open reading frame (ORF) encoding a 411 amino acids polypeptide with a molecular weight of 47 kDa and an isoelectric point (pI) of 6.95. IbLCYe protein contained one potential lycopene ε-cyclase domain and one potential FAD (flavinadenine dinucleotide)/NAD(P) (nicotinamide adenine dinucleotide phosphate)-binding domain, indicating that this protein shares the typical characteristics of LCYe proteins. The gDNA oflbLCYe gene was 4 029 bp and deduced to contain 5 introns and 6 exons. Real-time quantitative PCR analysis revealed that the expression level of IbLCYe gene was significantly higher in the storage roots of Nongdafu 14 than those in the leaves and stems. Transgenic tobacco (cv. Wisconsin 38) expressing [bLCYe gene accumulated significantly more ^-carotene compared to the untransformed control plants. These results showed that lbLCYe gene has an important function for the accumulation of carotenoids of sweetpotato.
基金supported jointly by Natural Science Foundation of Hebei Province(D2020504003)National Key Research and Development Program of China(No.2019YFC1805300).
文摘High concentrations of ammonium nitrogen released from tannery sludge during storage in open air may cause nitrogen pollution to soil and groundwater.To study the transformation mechanism of NH_(4)^(+)-N by nitrifying functional bacteria in tannery sludge contaminated soils,a series of contaminated soil culture experiments were conducted in this study.The contents of ammonium nitrogen(as NH_(4)^(+)-N),nitrite nitrogen(as NO_(2)^(−)-N)and nitrate nitrogen(as NO_(3)^(−)-N)were analyzed during the culture period under different conditions of pollution load,soil particle and redox environment.Sigmodial equation was used to interpret the change of NO_(3)^(−)-N with time in contaminated soils.The abundance variations of nitrifying functional genes(amoA and nxrA)were also detected using the real-time quantitative fluorescence PCR method.The results show that the nitrification of NH_(4)^(+)-N was aggravated in the contaminated silt soil and fine sand under the condition of lower pollution load,finer particle size and more oxidizing environment.The sigmodial equation well fitted the dynamic accumulation curve of the NO_(3)^(−)-N content in the tannery sludge contaminated soils.The Cr(III)content increased with increasing pollution load,which inhibited the reproduction and activity of nitrifying bacteria in the soils,especially in coarse-grained soil.The accumulation of NO_(2)^(−)-N contents became more obvious with the increase of pollution load in the fine sand,and only 41.5%of the NH_(4)^(+)-N was transformed to NO_(3)^(−)-N.The redox environment was the main factor affecting nitrification process in the soil.Compared to the aerobic soil environment,the transformation of NH_(4)^(+)-N was significantly inhibited under anaerobic incubation condition,and the NO_(3)^(−)-N contents decreased by 37.2%,61.9%and 91.9%under low,medium and high pollution loads,respectively.Nitrification was stronger in the silt soil since its copy number of amoA and nxrA genes was two times larger than that of fine sand.Moreover,the copy numbers of amoA and nxrA genes in the silt soil under the aerobic environment were 2.7 times and 2.2 times larger than those in the anaerobic environment.The abundance changes of the amoA and nxrA functional genes have a positive correlation with the nitrification intensity in the tannery sludge-contaminated soil.
基金Project(2011-622-40) supported by the Mineral Exploration Foundation of Henan Province,ChinaProject(51104189) supported by the National Natural Science Foundation of ChinaProject(2013M531814) supported by the Postdoctoral Science Foundation of China
文摘Biological desilication process is an effective way to remove silicate from rutile so that high purity rutile could be obtained. However, little is known about the molecular mechanism of this process. In this work, a newly developed rutile bio-desilication reactor was applied to enrich rutile from rough rutile concentrate obtained from Nanzhao rutile mine and a comprehensive high through-put functional gene array(Geo Chip 4.0) was used to analyze the functional gene diversity, structure and metabolic potential of microbial communities in the biological desilication reactor. The results show that TiO2 grade of the rutile concentrate could increase from 78.21% to above 90% and the recovery rate could reach to 96% or more in 8-12 d. The results also show that almost all the key functional genes involved in the geochemical cycling process, totally 4324 and 4983 functional microorganism genes, are detected in the liquid and ore surface, respectively. There are totally 712 and 831 functional genes involved in nitrogen cycling for liquid and ore surface samples, respectively. The relative abundance of functional genes involved in the phosphorus and sulfur cycling is higher in the ore surface than liquid. These results indicate that nitrogen, phosphorus and sulfur cycling are also present in the desiliconization process of rutile. Acetogenesis genes are detected in the liquid and ore surface, which indicates that the desiliconizing process mainly depends on the function of acetic acid and other organic acids. Four silicon transporting genes are also detected in the sample, which proves that the bacteria have the potential to transfer silicon in the molecule level. It is shown that bio-desilication is an effective and environmental-friendly way for enrichment of rough rutile concentrate and presents an overview of functional diversity and structure of desilication microbial communities, which also provides insights into our understanding of metabolic potential in biological desilication reactor ecosystems.
基金supported by the National Natural Science Foundation of China (31571993)the Natural Science Foundation of Liaoning Province of China (2014027027 and 2013020074)the Doctoral Fund of Liaoning Province of China (20131053)
文摘Understanding the sequence diversity of rice blast resistance genes is important for breeding new resistant rice cultivars against the rice blast fungusMagnaporthe oryzae. In this study, we selected 24 rice cultivars with different genetic back-grounds to study the alelic diversity of rice blast resistance genesPiz-t, Pitaand Pik. For Piz-t, a total of 17 alelic types were found within the 24 cultivars. Blast inoculations showed that most of the mutations can affect the function of the resistance gene. For Pita, except for the difference at the 918th amino acid, a majority of the 21 mutations were detected among the cultivars. Inoculations with blast isolates carryingAvr-Pita revealed that cultivars with mutations in other sites except for the 918th amino acid did not affect the function of thePita gene. ForPik, a total of six alelic types were found within the 24 cultivars, but ifve of them lost the function of the resistance gene. In addition, we found thatPiz-t, Pita and Pik were expressed constitutively in the 24 rice cultivars and the expression level was not related to resistance. Our results have provided the sequence diversity information of the resistance genesPiz-t, Pita and Pik among the popular rice cultivars grown in the northeast region of China. Keywords:resistance gene, avirulence gene, aleles, function, genetic evolution zae(M. oryzae), is one of the most destructive diseases in rice production worldwide. Over the years, comprehensive studies on rice blast resistance have been conducted (Silue et al. 1992). The resistance in newly cultivated rice cultivars to M. oryzae can be lost quickly due to the high level of instability in the genome of the fungus (Bonmanet al. 1992). Previous studies show that cultivars with durable and broad-spectrum resistance againstM. oryzae carry multiple major resistance (R) and minor resistance genes (Liuet al. 2014). An effective way to control rice blast disease is, therefore, to breed rice cultivars with multiple R and QTL genes. To date, over 83 rice blast R genes have been identiifed, and are distributed on 11 rice chromosomes except Received 22 May, 2015 Accepted 26 October, 2015 WANG Yan, E-mail: 8806wy@163.com; Correspondence LIU Zhi-heng, Tel: +86-24-23738857, E-mail: lzhh1954@163.com; ZHENG Wen-jing, Tel: +86-24-31021081, E-mail: zwj27@126. com *These authors contributed equaly to this study. ? 2016, CAAS. Al rights reserved. Published by Elsevier Ltd. doi: 10.1016/S2095-3119(15)61207-2 1. Introduction Rice blast disease, caused by the fungusMagnaporthe ory-
基金supported by the Transgenic Programs,China(2014ZX0800930B and 2016ZX08009-001)the National Natural Science Found of China(31371703)
文摘Obtaining transgenic plants is a common method for analyzing gene function. Unfortunately, stable genetic transformation is difficult to achieve, especially for plants(e.g., soybean), which are recalcitrant to genetic transformation. Transient expression systems, such as Arabidopsis protoplast, Nicotiana leaves, and onion bulb leaves are widely used for gene functional studies. A simple method for obtaining transgenic soybean callus tissues was reported recently. We extend this system with simplified culture conditions to gene functional studies, including promoter analysis, expression and subcellular localization of the target protein, and protein-protein interaction. We also evaluate the plasticity of this system with soybean varieties, different vector constructs, and various Agrobacterium strains. The results indicated that the callus transformation system is efficient and adaptable for gene functional investigation in soybean genotype-, vector-, and Agrobacterium strain-independent modes. We demonstrated an easy set-up and practical homologous strategy for soybean gene functional studies.
基金National Natural Science Foundation of China(30972184,31272574).
文摘[Objectives]This study was conducted to determine the functions of eight ORF genes of porcine circovirus-like virus P1.[Methods]The double-copy tandem molecular cloning of porcine circovirus-like virus P1 genome was used to construct molecular clones with eight ORFs deleted by DNA site-directed mutagenesis technology.After transfected into PK15 cells for a certain period of time,RNA were extracted and was used to verify whether the eight ORFs were deleted or not and used for gene microarry analysis.The GO functions and KEGG pathway enrichment of differentially expressed genes were analyzed.[Results]P1 ORF1 is mainly involved in the biological processes of defense response to virus,signal transduction,regulation of Rab GTPase activity,and lipid metabolic process,and involved in the molecular functions of protein phosphatase inhibitor activity,phosphatidylinositol phospholipase C activity,2 iron,2 sulfur cluster binding,phosphoric diester hydrolase activity,and Rab GTPase activator activity,and in the KEGG pathways of secretion of digestive gland and nervous system development.P1 ORF2 is mainly involved in the biological processes of positive regulation of leukocyte chemotaxis,positive regulation of cell proliferation,positive regulation of cell migration,defense response to virus,regulation of cell growth,and involved in the molecular functions of insulin-like growth factor binding,and chemokine activity,and in the KEGG pathways of cytosolic DNA-sensing pathway,RIG-I-like receptor signaling pathway,toll-like receptor signaling pathway,chemokine signaling pathway,and cytokines,cytokine-cytokine receptor interaction.The biological processes,molecular functions and related pathways involving P1 ORF3 and ORF5 are basically similar to those of ORF2.P1 ORF8 is mainly involved in the biological processes of purine ribonucleotide biosynthetic process,amino acid transport,defense response to virus,amino acid transmembrane transport,and involved in molecular functions of N6-(1,2-dicarboxyethyl)AMP AMP-lyase(fumarate-forming)activity,iron-sulfur cluster binding,amino acid transmembrane transporter activity.[Conclusions]The analysis of the ORF functions of P1 virus lays a foundation for the study of its pathogenicity and pathogenesis.
基金This work was funded by grants fromthe National Basic Research and Development Program(2004CB117304)the Hi-tech Research and Development Program of China (2007AA10Z115)
文摘As a major raw material for the textile industry and the most important fiber crop in the world,cotton is of great significance in Chinese economy.The development of cotton fiber can be divided