Drought has severely affected the yield and quality of commercial crops.The BRI1 family plays an important role in plant response to drought stress,and BRL3 gene plays an important role in the study of drought in Arab...Drought has severely affected the yield and quality of commercial crops.The BRI1 family plays an important role in plant response to drought stress,and BRL3 gene plays an important role in the study of drought in Arabidopsis thaliana.In this study,NtBRL3 was constructed as a vector and genetically transformed to obtain‘N.Tobacco K326’overexpression of NtBRL3.The enzyme activities of transgenic tobacco and wild-type tobacco were measured and transcriptome and metabolome analyses were performed.The results showed that the antioxidant enzymes of transgenic tobacco were more active under drought conditions,and 85 significantly differentially metabolites and 106 significantly differentially expressed genes were identified in the metabolome and transcriptome analyses,respectively.Transgenic tobacco NtBRL3ox demonstrated an excessive accumulation of droughtrelated metabolites,sugars such as sucrose and maltotetraose,and amino acids such as proline,compared with WT.We discovered drought-related differential genes in the root transcriptome,among which LOX6,RD22,WSD1,CCD8,and UGT were key genes which play an important role in plant response to drought stress.Our results demonstrate that NtBRL3 overexpression in K326 enhances drought resistance in transgenic tobacco.展开更多
Monosomic lines of Nicotiana tabacum are helpful to confirm the location of genes on specific chromosomes. In the cross N. nudicaulis and N. tabacum, hybrid seedlings express lethal symptoms, which are controlled by t...Monosomic lines of Nicotiana tabacum are helpful to confirm the location of genes on specific chromosomes. In the cross N. nudicaulis and N. tabacum, hybrid seedlings express lethal symptoms, which are controlled by the S subgenome of N. tabacum. To identify the responsible chromosome, we needed to produce chromosome lacking lines (CLLs) of N. tabacum L. “Red Russian” and use them to cross with N. nudicaulis. From a cross of (N. tabacum × N. tomentosiformis) × N. tabacum, 380 BC1 individuals were obtained. Using a Haplo-Q line (a monosomic line lacking the single linkage group 11) and N. tabacum, we found that qPCR is a simple and reliable screening method for CLLs of N. tabacum. The marker PT30342 is located on linkage group 11, and the -Ct value (Ct Actin - Ct PT30342) was 2.0 for a disomic line and was 1.097 for a Haplo-Q line. By the use of flow cytometry, qPCR and chromosome counting together as a screening method, we identified 6 CLLs lacking 2 to 6 chromosomes. Compared with conventional methods, our method is a rapid technique for making and screening CLLs ofthe S or S/T subgenome of N. tabacum. Further, these CLLs will be useful to identify the location of two or more factors on chromosomes controlling a variety of genetic problems affecting breeding. Here, we only made CLLs of the S or S/T subgenome of N. tabacum. We will use the method established in this study to produce CLLs of the T subgenome of N. tabacum, and gather a full set of CLLs of N. tabacum. qPCR could also be applied to the identification of chromosome aberrations in other plants.展开更多
Investigations into the potential application of nanoparticles acting as nanofungicides in sustainable agriculture are rapidly expanding due to the high antimicrobial properties of these compounds,which do not risk in...Investigations into the potential application of nanoparticles acting as nanofungicides in sustainable agriculture are rapidly expanding due to the high antimicrobial properties of these compounds,which do not risk inducing pathogen resistance to fungicides.A detailed understanding of the impact of copper oxide nanoparticles(CuO NPs)on soil-borne phytopathogenic fungi is yet to be obtained.This study aimed to explore the in vitro antifungal activity and control efficacy of CuO NPs applied via irrigation with respect to tobacco black shank(TBS)disease caused by Phytophthora nicotianae.The results revealed that CuO NPs greatly interfered with the reproductive growth process of this fungus,repressing hyphal growth,spore germination and sporangium production.Additionally,morphological damage,intracellular ROS accumulation and increased SOD enzyme activity in hyphae were the antifungicidal mechanisms of these NPs.In pot experiments,treatment with CuO NPs at 100 mg L^(–1)significantly suppressed TBS development,compared with the effect on control plants,and the control efficacy reached 33.69%without inducing phytotoxicity.Exposure to CuO NPs significantly activated a series of defense enzymes,and resistance genes in tobacco can further explain the mechanisms by which CuO NPs suppressed fungal infection.The Cu content in both the leaves and roots of P.nicotianae-infested plants increased by 50.03 and 27.25%,respectively,after treatment with 100 mg L^(–1)CuO NPs,compared with that of healthy plants.In particular,a higher Cu content was observed in infected roots than in leaves.Therefore,this study showed the potential of CuO NPs applied as nanofungicides and as nanoinducers of fungus resistance genes for the management of TBS through inhibition of pathogen infection and stimulation of plant defenses.展开更多
Vacuolar ATPase is the key enzyme in the aspect of plant stress,which contains 13 subunits.In this study,VHA-c which was cloned from the salt-tolerant plants Iris lacteal was transformed into tobacco,and then treated ...Vacuolar ATPase is the key enzyme in the aspect of plant stress,which contains 13 subunits.In this study,VHA-c which was cloned from the salt-tolerant plants Iris lacteal was transformed into tobacco,and then treated the transgenic tobacco with 0.7% NaCl for 7 days.Detected the MDA content,the SOD activity and the POD activity.Moreover,the transgenic plants displayed obviously better growth than the normal plants under salt stress.These results suggested that VHA-c could confer the stress tolerance through enhancing the POD and the SOD activities,and by protecting membranes from damage by decreasing lipid peroxidation under salt stress.展开更多
The relationships between chemical components and quality indexes were studied in the tobacco leaves with different aromatic styles. A total of 16 chemical components, 4 quality indexes, and 6 smoking quality indexes ...The relationships between chemical components and quality indexes were studied in the tobacco leaves with different aromatic styles. A total of 16 chemical components, 4 quality indexes, and 6 smoking quality indexes from 366 tobacco leaf samples with 4 different types of aroma from Guizhou Province, China, were subjected to principal component analysis and stepwise regression analysis. The tobacco leaves with different types of aroma showed remarkable difference in the contents of chemical components, quality indexes, and smoking quality indexes. The first principal factors (carbohydrates and nitrogen-containing compounds) of the chemical composition of the leaf were similar among different types of aroma, which showed that the quality of the leaf was mainly influenced by carbohydrates and nitrogen-containing compounds and their ratios. The factors for the second through the seventh principal components varied largely among various aromatic types, suggesting the contribution of other chemical components to the leaf quality. In addition, the smoking quality of four different aromatic leaves showed significant correlation with the different chemical components. The quality of tobacco leaves with different types of aroma was influenced by multiple factors, especially ecological conditions and culture techniques, which may provide guidance for directive cultivation of high-quality tobacco leaves.展开更多
The constitutive promoter of cauliflower mosaic virus 35S (CaMV 35S) is a polar unidirectional promoter and is widely used in plant genetic engineering. In the present study, the unidirectional CaMV 35S promoter has b...The constitutive promoter of cauliflower mosaic virus 35S (CaMV 35S) is a polar unidirectional promoter and is widely used in plant genetic engineering. In the present study, the unidirectional CaMV 35S promoter has been modified to a bi-directional promoter by fusing its minimal promoter element to the 5′end of CaMV 35S promoter in the opposite orientation. To qualitatively and quantitatively esti-mate its bi-directional transcriptional function and activity, two visible reporter genes, gusA (β-glucuronidase, GUS) and gfp (green fluo-rescent protein, GFP), were fused to the two ends of the promoter in bi-orientations ending with NOS terminator sequences, respectively. Stable expression of gusA and gfp genes in transgenic tobacco (Nicotiana tabacum L.) was visulized by histochemically staining for GUS and fluorescence microscopic observation under UV for GFP in transgenic plants. The expression of two reporter genes showed that the constructed bi-directional promoter did have the bi-directional transcriptional function in both expected orientations. The quantitative estimation of GUS and GFP were determined on a HITACHI F1000 Fluorescence Spectrophotometer with various wavelengths of excita-tion and emission. The GUS activity varied from 8 to 250 pmol 4-MU / min / mg protein and the GFP content varied from 0.9 to 1.8 μg / mg protein in various lines of transgenic tobacco plants. Higher GUS activity generally coupled with lower GFP content, and vice versa.展开更多
Plant genotypic difference of pot assiu m-enr ich ment capab ility and p ot assi um (K ) d ist rib ut ion at root-soil ioterface of different plant genotypes were studied by using seven plant species and eight varieti...Plant genotypic difference of pot assiu m-enr ich ment capab ility and p ot assi um (K ) d ist rib ut ion at root-soil ioterface of different plant genotypes were studied by using seven plant species and eight varieties oftobacco (Nicotiana tabacum L.). The results indicated that K enrichment capability was: Ethiopian guizotia(Guizotia abyssinica Cass.)>feather cockscomb (Celosia arpentea L.)>alligator alternanthera (Alternantheraphiloxeroides (Mart.) Griseb.)> tobacco>sesbania (Sesbania cannabina (Retz.) Pers.)>wheat (Thticumaestivum L.)>broadbean (Vicia faba L.). Ethiopian guizotia showed very high K-enrichment capability atdifferent soil K levels, and the K content in its dry matter was over 110 mg kg-1 when soil K was fullysupplied, and about 60 mg kg-1 when no K fertilizer was applied. For alligator alternanthera, the capabilityto accumulate K was closely related with its growth medium. When it was grown on soils, both the K contentand K uptake rate of the plant were similar to those of tobacco. Evident K dep1etion was observed in therhizosphere of all plant species, and the depletion rate was related to the capability of K enrichment of plant.展开更多
基金supported by the Science and Technology Project of Guizhou Tobacco Company(2021XM04)the Creation of“Tobacco T-DNA Activation Insertion Mutant Library and Screening of Important Trait Mutants”Project of Guizhou University Talent Introduction(Guizhou University Hezi[2013]50).
文摘Drought has severely affected the yield and quality of commercial crops.The BRI1 family plays an important role in plant response to drought stress,and BRL3 gene plays an important role in the study of drought in Arabidopsis thaliana.In this study,NtBRL3 was constructed as a vector and genetically transformed to obtain‘N.Tobacco K326’overexpression of NtBRL3.The enzyme activities of transgenic tobacco and wild-type tobacco were measured and transcriptome and metabolome analyses were performed.The results showed that the antioxidant enzymes of transgenic tobacco were more active under drought conditions,and 85 significantly differentially metabolites and 106 significantly differentially expressed genes were identified in the metabolome and transcriptome analyses,respectively.Transgenic tobacco NtBRL3ox demonstrated an excessive accumulation of droughtrelated metabolites,sugars such as sucrose and maltotetraose,and amino acids such as proline,compared with WT.We discovered drought-related differential genes in the root transcriptome,among which LOX6,RD22,WSD1,CCD8,and UGT were key genes which play an important role in plant response to drought stress.Our results demonstrate that NtBRL3 overexpression in K326 enhances drought resistance in transgenic tobacco.
文摘Monosomic lines of Nicotiana tabacum are helpful to confirm the location of genes on specific chromosomes. In the cross N. nudicaulis and N. tabacum, hybrid seedlings express lethal symptoms, which are controlled by the S subgenome of N. tabacum. To identify the responsible chromosome, we needed to produce chromosome lacking lines (CLLs) of N. tabacum L. “Red Russian” and use them to cross with N. nudicaulis. From a cross of (N. tabacum × N. tomentosiformis) × N. tabacum, 380 BC1 individuals were obtained. Using a Haplo-Q line (a monosomic line lacking the single linkage group 11) and N. tabacum, we found that qPCR is a simple and reliable screening method for CLLs of N. tabacum. The marker PT30342 is located on linkage group 11, and the -Ct value (Ct Actin - Ct PT30342) was 2.0 for a disomic line and was 1.097 for a Haplo-Q line. By the use of flow cytometry, qPCR and chromosome counting together as a screening method, we identified 6 CLLs lacking 2 to 6 chromosomes. Compared with conventional methods, our method is a rapid technique for making and screening CLLs ofthe S or S/T subgenome of N. tabacum. Further, these CLLs will be useful to identify the location of two or more factors on chromosomes controlling a variety of genetic problems affecting breeding. Here, we only made CLLs of the S or S/T subgenome of N. tabacum. We will use the method established in this study to produce CLLs of the T subgenome of N. tabacum, and gather a full set of CLLs of N. tabacum. qPCR could also be applied to the identification of chromosome aberrations in other plants.
基金financial support by the National Natural Science Foundation of China(32001934)the Key Science and Technology Project of Sichuan Tobacco Company,China(SCYC202114)。
文摘Investigations into the potential application of nanoparticles acting as nanofungicides in sustainable agriculture are rapidly expanding due to the high antimicrobial properties of these compounds,which do not risk inducing pathogen resistance to fungicides.A detailed understanding of the impact of copper oxide nanoparticles(CuO NPs)on soil-borne phytopathogenic fungi is yet to be obtained.This study aimed to explore the in vitro antifungal activity and control efficacy of CuO NPs applied via irrigation with respect to tobacco black shank(TBS)disease caused by Phytophthora nicotianae.The results revealed that CuO NPs greatly interfered with the reproductive growth process of this fungus,repressing hyphal growth,spore germination and sporangium production.Additionally,morphological damage,intracellular ROS accumulation and increased SOD enzyme activity in hyphae were the antifungicidal mechanisms of these NPs.In pot experiments,treatment with CuO NPs at 100 mg L^(–1)significantly suppressed TBS development,compared with the effect on control plants,and the control efficacy reached 33.69%without inducing phytotoxicity.Exposure to CuO NPs significantly activated a series of defense enzymes,and resistance genes in tobacco can further explain the mechanisms by which CuO NPs suppressed fungal infection.The Cu content in both the leaves and roots of P.nicotianae-infested plants increased by 50.03 and 27.25%,respectively,after treatment with 100 mg L^(–1)CuO NPs,compared with that of healthy plants.In particular,a higher Cu content was observed in infected roots than in leaves.Therefore,this study showed the potential of CuO NPs applied as nanofungicides and as nanoinducers of fungus resistance genes for the management of TBS through inhibition of pathogen infection and stimulation of plant defenses.
基金Supported by the Post-doctoral Research Start-up Funding of Heilongjiang Province (LBH-210144)the Post-doctoral Funds Project of Heilongjiang Province (520-415037)
文摘Vacuolar ATPase is the key enzyme in the aspect of plant stress,which contains 13 subunits.In this study,VHA-c which was cloned from the salt-tolerant plants Iris lacteal was transformed into tobacco,and then treated the transgenic tobacco with 0.7% NaCl for 7 days.Detected the MDA content,the SOD activity and the POD activity.Moreover,the transgenic plants displayed obviously better growth than the normal plants under salt stress.These results suggested that VHA-c could confer the stress tolerance through enhancing the POD and the SOD activities,and by protecting membranes from damage by decreasing lipid peroxidation under salt stress.
文摘The relationships between chemical components and quality indexes were studied in the tobacco leaves with different aromatic styles. A total of 16 chemical components, 4 quality indexes, and 6 smoking quality indexes from 366 tobacco leaf samples with 4 different types of aroma from Guizhou Province, China, were subjected to principal component analysis and stepwise regression analysis. The tobacco leaves with different types of aroma showed remarkable difference in the contents of chemical components, quality indexes, and smoking quality indexes. The first principal factors (carbohydrates and nitrogen-containing compounds) of the chemical composition of the leaf were similar among different types of aroma, which showed that the quality of the leaf was mainly influenced by carbohydrates and nitrogen-containing compounds and their ratios. The factors for the second through the seventh principal components varied largely among various aromatic types, suggesting the contribution of other chemical components to the leaf quality. In addition, the smoking quality of four different aromatic leaves showed significant correlation with the different chemical components. The quality of tobacco leaves with different types of aroma was influenced by multiple factors, especially ecological conditions and culture techniques, which may provide guidance for directive cultivation of high-quality tobacco leaves.
文摘The constitutive promoter of cauliflower mosaic virus 35S (CaMV 35S) is a polar unidirectional promoter and is widely used in plant genetic engineering. In the present study, the unidirectional CaMV 35S promoter has been modified to a bi-directional promoter by fusing its minimal promoter element to the 5′end of CaMV 35S promoter in the opposite orientation. To qualitatively and quantitatively esti-mate its bi-directional transcriptional function and activity, two visible reporter genes, gusA (β-glucuronidase, GUS) and gfp (green fluo-rescent protein, GFP), were fused to the two ends of the promoter in bi-orientations ending with NOS terminator sequences, respectively. Stable expression of gusA and gfp genes in transgenic tobacco (Nicotiana tabacum L.) was visulized by histochemically staining for GUS and fluorescence microscopic observation under UV for GFP in transgenic plants. The expression of two reporter genes showed that the constructed bi-directional promoter did have the bi-directional transcriptional function in both expected orientations. The quantitative estimation of GUS and GFP were determined on a HITACHI F1000 Fluorescence Spectrophotometer with various wavelengths of excita-tion and emission. The GUS activity varied from 8 to 250 pmol 4-MU / min / mg protein and the GFP content varied from 0.9 to 1.8 μg / mg protein in various lines of transgenic tobacco plants. Higher GUS activity generally coupled with lower GFP content, and vice versa.
文摘Plant genotypic difference of pot assiu m-enr ich ment capab ility and p ot assi um (K ) d ist rib ut ion at root-soil ioterface of different plant genotypes were studied by using seven plant species and eight varieties oftobacco (Nicotiana tabacum L.). The results indicated that K enrichment capability was: Ethiopian guizotia(Guizotia abyssinica Cass.)>feather cockscomb (Celosia arpentea L.)>alligator alternanthera (Alternantheraphiloxeroides (Mart.) Griseb.)> tobacco>sesbania (Sesbania cannabina (Retz.) Pers.)>wheat (Thticumaestivum L.)>broadbean (Vicia faba L.). Ethiopian guizotia showed very high K-enrichment capability atdifferent soil K levels, and the K content in its dry matter was over 110 mg kg-1 when soil K was fullysupplied, and about 60 mg kg-1 when no K fertilizer was applied. For alligator alternanthera, the capabilityto accumulate K was closely related with its growth medium. When it was grown on soils, both the K contentand K uptake rate of the plant were similar to those of tobacco. Evident K dep1etion was observed in therhizosphere of all plant species, and the depletion rate was related to the capability of K enrichment of plant.
