In this paper, we estimate the Fekete-Szego functional with k-th root transform for the inverse of certain classes of analytic univalent functions using quasi-subordination.
Agrobacterium tumefaciens-mediated transformation has been widely adopted for plant genetic engineering and the study of gene function(Krenek et al.,2015).This method is prevalent in the genetic transformation of herb...Agrobacterium tumefaciens-mediated transformation has been widely adopted for plant genetic engineering and the study of gene function(Krenek et al.,2015).This method is prevalent in the genetic transformation of herbaceous plants,with notable applications in species such as Arabidopsis(Yin et al.,2024),soybean(Zhang et al.,2024),rice(Zhang et al.,2020),and Chinese cabbage(Li et al.,2021).However,its application in fruit trees is limited.This is primarily due to their long growth cycles and lack of rapid,efficient,and stable transgenic systems,which severely hinders foundational research involving plant genetic transformation(Mei et al.,2024).Furthermore,for subtropical fruit trees,the presence of recalcitrant seeds adds an extra layer of difficulty to genetic transformation(Umarani et al.,2015),as most methods rely on seed germination as a basis for transformation.展开更多
The CRISPR/Cas9 genome-editing system serves as a pivotal tool for enhancing crop genetics.Within this system,single guide RNAs(sgRNAs)are instrumental in the precise cleavage of DNA double strands.However,the efficie...The CRISPR/Cas9 genome-editing system serves as a pivotal tool for enhancing crop genetics.Within this system,single guide RNAs(sgRNAs)are instrumental in the precise cleavage of DNA double strands.However,the efficiency of gene editing varies among sgRNAs,emphasizing the need to meticulously select target sites,especially in the context of Citrullus lanatus,a species notorious for its challenging and inefficient generation of transgenic plants through stable transformation.This study employed an Agrobacterium rhizogenes-mediated hairy root method to assess effective target sites for gene editing of ClCIPK17 across various Citrullus species.Hairy roots were successfully induced in different plant tissues at diverse growth stages of Citrullus lanatus,Citrullus mucosospermus,and Citrullus amarus.Employing a vector with two sgRNAs(sgRNA1 and sgRNA5)positioned within conserved regions of exon 1 and exon 5 of ClCIPK17 in the CRISPR/Cas9 system,targeted mutations were detected in 90.9%of accessions across the four Citrullus species.Notably,73.94%of all examined hairy roots exhibited mutations at the sgRNA1 site,while the sgRNA5 site showed no mutations.Among the 31 different mutation types identified at the sgRNA1 site,base deletion was the most prevalent.Using the sgRNA1 site of ClCIPK17,stable transgenic watermelon buds were obtained from explants,and the targeted mutations of the sgRNA1 site were confirmed.These findings underscore the viability of the hairy root transformation system in assessing the editing efficiency of sgRNA targets in diverse Citrullus species.展开更多
Soybean is one of the world's most important oil and protein crops. Efficient transformation is a key factor for the improvement of soybean by genetic modification. We describe an optimized protocol for the Agroba...Soybean is one of the world's most important oil and protein crops. Efficient transformation is a key factor for the improvement of soybean by genetic modification. We describe an optimized protocol for the Agrobacterium rhizogenes-mediated transformation of soybean and the induction of hairy root development in vitro. Cotyledons with 0.5-cm hypocotyls were cut from 5-day-old seedlings and used as explants. After infection and co-cultivation,hairy roots were produced in induction culture medium after 10–12 days. Using this method, 90%–99% of the infected explants of five different cultivars produced hairy roots within one month. Observations using reporter constructs showed that 30%–60% of the hairy roots induced were transformed. Based on high transformation efficiency and short transformation period, this method represents an efficient and rapid platform for study of soybean gene function.展开更多
Summary： To study the efficacy and the mechanism of Colquhoumia root （ Tripterygium hypoglaucure （Le,vL） Hutch） in the treatment of mesangial proliferation glomerulonephritis （MsPGN）, SD rats were injected with...Summary： To study the efficacy and the mechanism of Colquhoumia root （ Tripterygium hypoglaucure （Le,vL） Hutch） in the treatment of mesangial proliferation glomerulonephritis （MsPGN）, SD rats were injected with anti-thymoeyte serum （ATS） to make MsPGN model （anti-Thyl model）. The rats were then divided into 3 groups： normal control group, anti-Thyl model group and treatment group. Histopathologieal （HE, PAS）, immunohistoehemieal, RT-PCR technique and computer imaging analysis system were used to evaluate mesangial matrix production, the expression of TGF-β protein and mRNA in the tissues of kidney. Our result showed that proteinuria and the ratio of extraeellular matrix/glomerular capillaries area （ECM/CA） were increased significantly in model group. The expression of both TGF-β protein and mRNA in glomeruli was much higher in model group than in control group （P〈0.01）. After the treatment with Colquhoumia root, proteinuria, ECM/CA and the expression of both TGF-β1 protein and mRNA in glomeruli were significantly decreased in treatment group as compared with those in model group. It is concluded that Colquhoumia root is effective in reducing proteinuria and mesangial matrix proliferation in MsPGN and it may achieve these effects by inhibiting the expressions of TGF-β1 protein and mRNA of mesangial cells.展开更多
Non-heading Chinese cabbage, a variety of Brassica campestris, is an important vegetable crop in the Yangtze River Basin of China. However,the immaturity of its stable transformation system and its low transformation ...Non-heading Chinese cabbage, a variety of Brassica campestris, is an important vegetable crop in the Yangtze River Basin of China. However,the immaturity of its stable transformation system and its low transformation efficiency limit gene function research on non-heading Chinese cabbage. Agrobacterium rhizogenes-mediated(ARM) transgenic technology is a rapid and effective transformation method that has not yet been established for non-heading Chinese cabbage plants. Here, we optimized conventional ARM approaches(one-step and two-step transformation methods) suitable for living non-heading Chinese cabbage plants in nonsterile environments. Transgenic roots in composite non-heading Chinese cabbage plants were identified using phenotypic detection, fluorescence observation, and PCR analysis. The transformation efficiency of a two-step method on four five-day-old non-heading Chinese cabbage seedlings(Suzhouqing, Huangmeigui, Wuyueman, and Sijiu Caixin) was 43.33%-51.09%, whereas using the stout hypocotyl resulted in a transformation efficiency of 54.88% for the 30-day-old Sijiu Caixin.The one-step method outperformed the two-step method;the transformation efficiency of different varieties was above 60%, and both methods can be used to obtain transgenic roots for functional studies within one month. Finally, optimized ARM transformation methods can easily,quickly, and effectively produce composite non-heading Chinese cabbage plants with transgenic roots, providing a reliable foundation for gene function research and non-heading Chinese cabbage genetic improvement breeding.展开更多
Genetic transformation experiments of the different explants from Citrus grandis cv. Changshou Shatian You infected with Agrobacterium rhizogenes were carried out in darkness or in light. The optimizing culture system...Genetic transformation experiments of the different explants from Citrus grandis cv. Changshou Shatian You infected with Agrobacterium rhizogenes were carried out in darkness or in light. The optimizing culture system of Ri T-DNA transformed roots for C. grandis cv. Changshou Shatian You was constructed as follows： After the ventral wounded striations on the single activation cotyledon were inoculated by A. rhizogenes A4 （logarithmic period）, they were cocultured at （25 ±2）℃ in darkness for 25-30 days; some transformed roots were generated from wounded striations of most cotyledons. The genetically transformed ratio is （83 ± 11）%. Axenic Ri T-DNA transformed roots （hairy roots） were harvested after five subcultures. Explants were activated on MT medium. The MS medium was used for subculture of transformed roots. Mass Ri T-DNA transformed roots in which the hormone was produced independently were harvested from this optimizing culture system. White, fresh Ri T-DNA transformed roots were （1.14 ±0.07） cm long, （0.73 ±0.04） mm wide, and the growth direction of transformed roots was negative geotropism.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
A rapid, efficient and high-performance transformation protocol employing Agrobacterium rhizogenes was developed for the common bean Phaseolus vulgaris. In this study, we examined competencies of various protocols to ...A rapid, efficient and high-performance transformation protocol employing Agrobacterium rhizogenes was developed for the common bean Phaseolus vulgaris. In this study, we examined competencies of various protocols to induce and explants that respond to hairy root transformation in bean plants. Utilizing young seedlings with severed radicles/hypocotyls, we developed a highly efficient procedure for achieving hairy root transformation frequencies as high as 100% as visualized by GUS reporter gene expression system. Transgenic hairy roots in these young composite plants were susceptible to nodulation by rhizobia, and form an excellent system for high throughput genomic analysis to study root biology and endosymbiosis in common bean.展开更多
Let n≥2 and let L be a second-order elliptic operator of divergence form with coefficients consisting of both an elliptic symmetric part and a BMO anti-symmetric part in ℝ^(n).In this article,we consider the weighted...Let n≥2 and let L be a second-order elliptic operator of divergence form with coefficients consisting of both an elliptic symmetric part and a BMO anti-symmetric part in ℝ^(n).In this article,we consider the weighted Kato square root problem for L.More precisely,we prove that the square root L^(1/2)satisfies the weighted L^(p)estimates||L^(1/2)(f)||L_(ω)^p(R^(n))≤C||■f||L_(ω)^p(R^(n);R^(n))for any p∈(1,∞)andω∈Ap(ℝ^(n))(the class of Muckenhoupt weights),and that||■f||L_(ω)^p(R^(n);R^(n))≤C||L^(1/2)(f)||L_(ω)^p(R^(n))for any p∈(1,2+ε)andω∈Ap(ℝ^(n))∩RH_(2+ε/p),(R^(n))(the class of reverse Hölder weights),whereε∈(0,∞)is a constant depending only on n and the operator L,and where(2+ε/p)'denotes the Hölder conjugate exponent of 2+ε/p.Moreover,for any given q∈(2,∞),we give a sufficient condition to obtain that||■f||L_(ω)^p(R^(n);R^(n))≤C||L^(1/2)(f)||L_(ω)^p(R^(n))for any p∈(1,q)andω∈A_(p)(R^(n))∩pRH_(q/p),(R^(n)).As an application,we prove that when the coefficient matrix A that appears in L satisfies the small BMO condition,the Riesz transform∇L^(−1/2)is bounded on L_(ω)^(p)(ℝ^(n))for any given p∈(1,∞)andω∈Ap(ℝ^(n)).Furthermore,applications to the weighted L^(2)-regularity problem with the Dirichlet or the Neumann boundary condition are also given.展开更多
基金Supported by the National Natural Science Foundation of China (Grant No. 11561001)the Natural Science Foundation of Inner Mongolia of China (Grant No. 2018MS01026)+4 种基金the Natural Science Foundation of Anhui Provincial Department of Education (Grant Nos. KJ2018A0833KJ2020A0993KJ2020ZD74)Provincial Quality Engineering Project of Anhui Colleges and Universities (Grant No. 2018mooc608)the Key Cultivated Project at School Level of the National Science Fund of Guangzhou Civil Aviation College (Grant No. 18X0428)。
文摘In this paper, we estimate the Fekete-Szego functional with k-th root transform for the inverse of certain classes of analytic univalent functions using quasi-subordination.
基金funded by the Key-Area Research and Development Program of Guangdong Province(Grant No.2022B0202070002)the Guangxi Science and Technology Major Program(Grant No.GuikeAA23023007-2)+1 种基金the Guangdong Province Modern Agricultural Industry Technology System Innovation Team Construction Project(2024CXTD19)Guangdong Basic and Applied Basic Research Foundation(Grant No.2023A1515010303)。
文摘Agrobacterium tumefaciens-mediated transformation has been widely adopted for plant genetic engineering and the study of gene function(Krenek et al.,2015).This method is prevalent in the genetic transformation of herbaceous plants,with notable applications in species such as Arabidopsis(Yin et al.,2024),soybean(Zhang et al.,2024),rice(Zhang et al.,2020),and Chinese cabbage(Li et al.,2021).However,its application in fruit trees is limited.This is primarily due to their long growth cycles and lack of rapid,efficient,and stable transgenic systems,which severely hinders foundational research involving plant genetic transformation(Mei et al.,2024).Furthermore,for subtropical fruit trees,the presence of recalcitrant seeds adds an extra layer of difficulty to genetic transformation(Umarani et al.,2015),as most methods rely on seed germination as a basis for transformation.
基金supported by the Basic Research Funds of Chinese Academy of Agricultural Sciences(Grant No.1610192023201)the National Key Research and Development Program of China(Grant No.SQ2023YFE0201239)+3 种基金China Agriculture Research System of MOF and MARA(Grant No.CARS-25-03)the Major Science and Technology Projects of Henan Province(Grant No.221100110400)the Funding of Joint Research on Agricultural Variety Improvement of Henan Province(Grant No.2022010503)the Agricultural Science and Technology Innovation Program(Grant No.CAAS-ASTIP-2021-ZFRI)。
文摘The CRISPR/Cas9 genome-editing system serves as a pivotal tool for enhancing crop genetics.Within this system,single guide RNAs(sgRNAs)are instrumental in the precise cleavage of DNA double strands.However,the efficiency of gene editing varies among sgRNAs,emphasizing the need to meticulously select target sites,especially in the context of Citrullus lanatus,a species notorious for its challenging and inefficient generation of transgenic plants through stable transformation.This study employed an Agrobacterium rhizogenes-mediated hairy root method to assess effective target sites for gene editing of ClCIPK17 across various Citrullus species.Hairy roots were successfully induced in different plant tissues at diverse growth stages of Citrullus lanatus,Citrullus mucosospermus,and Citrullus amarus.Employing a vector with two sgRNAs(sgRNA1 and sgRNA5)positioned within conserved regions of exon 1 and exon 5 of ClCIPK17 in the CRISPR/Cas9 system,targeted mutations were detected in 90.9%of accessions across the four Citrullus species.Notably,73.94%of all examined hairy roots exhibited mutations at the sgRNA1 site,while the sgRNA5 site showed no mutations.Among the 31 different mutation types identified at the sgRNA1 site,base deletion was the most prevalent.Using the sgRNA1 site of ClCIPK17,stable transgenic watermelon buds were obtained from explants,and the targeted mutations of the sgRNA1 site were confirmed.These findings underscore the viability of the hairy root transformation system in assessing the editing efficiency of sgRNA targets in diverse Citrullus species.
基金supported by the Major Science and Technology Projects of China (2016ZX08010-004)the Ministry of Science and Technology of China (2016YFD0100504)the CAAS (Chinese Academy of Agriculture Sciences) Innovation Project
文摘Soybean is one of the world's most important oil and protein crops. Efficient transformation is a key factor for the improvement of soybean by genetic modification. We describe an optimized protocol for the Agrobacterium rhizogenes-mediated transformation of soybean and the induction of hairy root development in vitro. Cotyledons with 0.5-cm hypocotyls were cut from 5-day-old seedlings and used as explants. After infection and co-cultivation,hairy roots were produced in induction culture medium after 10–12 days. Using this method, 90%–99% of the infected explants of five different cultivars produced hairy roots within one month. Observations using reporter constructs showed that 30%–60% of the hairy roots induced were transformed. Based on high transformation efficiency and short transformation period, this method represents an efficient and rapid platform for study of soybean gene function.
文摘Summary： To study the efficacy and the mechanism of Colquhoumia root （ Tripterygium hypoglaucure （Le,vL） Hutch） in the treatment of mesangial proliferation glomerulonephritis （MsPGN）, SD rats were injected with anti-thymoeyte serum （ATS） to make MsPGN model （anti-Thyl model）. The rats were then divided into 3 groups： normal control group, anti-Thyl model group and treatment group. Histopathologieal （HE, PAS）, immunohistoehemieal, RT-PCR technique and computer imaging analysis system were used to evaluate mesangial matrix production, the expression of TGF-β protein and mRNA in the tissues of kidney. Our result showed that proteinuria and the ratio of extraeellular matrix/glomerular capillaries area （ECM/CA） were increased significantly in model group. The expression of both TGF-β protein and mRNA in glomeruli was much higher in model group than in control group （P〈0.01）. After the treatment with Colquhoumia root, proteinuria, ECM/CA and the expression of both TGF-β1 protein and mRNA in glomeruli were significantly decreased in treatment group as compared with those in model group. It is concluded that Colquhoumia root is effective in reducing proteinuria and mesangial matrix proliferation in MsPGN and it may achieve these effects by inhibiting the expressions of TGF-β1 protein and mRNA of mesangial cells.
基金funded by National Natural Science Foundation of China (Grant No.32072575)Postgraduate Research & Practice Innovation Program of Jiangsu Province (Grant No.KYCX20_0588)National Vegetable Industry Technology System (Grant No.CARS-23-A16)。
文摘Non-heading Chinese cabbage, a variety of Brassica campestris, is an important vegetable crop in the Yangtze River Basin of China. However,the immaturity of its stable transformation system and its low transformation efficiency limit gene function research on non-heading Chinese cabbage. Agrobacterium rhizogenes-mediated(ARM) transgenic technology is a rapid and effective transformation method that has not yet been established for non-heading Chinese cabbage plants. Here, we optimized conventional ARM approaches(one-step and two-step transformation methods) suitable for living non-heading Chinese cabbage plants in nonsterile environments. Transgenic roots in composite non-heading Chinese cabbage plants were identified using phenotypic detection, fluorescence observation, and PCR analysis. The transformation efficiency of a two-step method on four five-day-old non-heading Chinese cabbage seedlings(Suzhouqing, Huangmeigui, Wuyueman, and Sijiu Caixin) was 43.33%-51.09%, whereas using the stout hypocotyl resulted in a transformation efficiency of 54.88% for the 30-day-old Sijiu Caixin.The one-step method outperformed the two-step method;the transformation efficiency of different varieties was above 60%, and both methods can be used to obtain transgenic roots for functional studies within one month. Finally, optimized ARM transformation methods can easily,quickly, and effectively produce composite non-heading Chinese cabbage plants with transgenic roots, providing a reliable foundation for gene function research and non-heading Chinese cabbage genetic improvement breeding.
基金The research was supported by the Science Fund of China Post Ph.D （2003034492）Science Foundation of Chongqing Educational Committee （040216）, China.
文摘Genetic transformation experiments of the different explants from Citrus grandis cv. Changshou Shatian You infected with Agrobacterium rhizogenes were carried out in darkness or in light. The optimizing culture system of Ri T-DNA transformed roots for C. grandis cv. Changshou Shatian You was constructed as follows： After the ventral wounded striations on the single activation cotyledon were inoculated by A. rhizogenes A4 （logarithmic period）, they were cocultured at （25 ±2）℃ in darkness for 25-30 days; some transformed roots were generated from wounded striations of most cotyledons. The genetically transformed ratio is （83 ± 11）%. Axenic Ri T-DNA transformed roots （hairy roots） were harvested after five subcultures. Explants were activated on MT medium. The MS medium was used for subculture of transformed roots. Mass Ri T-DNA transformed roots in which the hormone was produced independently were harvested from this optimizing culture system. White, fresh Ri T-DNA transformed roots were （1.14 ±0.07） cm long, （0.73 ±0.04） mm wide, and the growth direction of transformed roots was negative geotropism.
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Centro de Ciencias de Genomicas,Universidad Autonoma de Mexico,Cuernavaca,Morelos,Mexico for financial and lab support
文摘A rapid, efficient and high-performance transformation protocol employing Agrobacterium rhizogenes was developed for the common bean Phaseolus vulgaris. In this study, we examined competencies of various protocols to induce and explants that respond to hairy root transformation in bean plants. Utilizing young seedlings with severed radicles/hypocotyls, we developed a highly efficient procedure for achieving hairy root transformation frequencies as high as 100% as visualized by GUS reporter gene expression system. Transgenic hairy roots in these young composite plants were susceptible to nodulation by rhizobia, and form an excellent system for high throughput genomic analysis to study root biology and endosymbiosis in common bean.
基金supported by the Key Project of Gansu Provincial National Science Foundation(23JRRA1022)the National Natural Science Foundation of China(12071431)+1 种基金the Fundamental Research Funds for the Central Universities(lzujbky-2021-ey18)the Innovative Groups of Basic Research in Gansu Province(22JR5RA391).
文摘Let n≥2 and let L be a second-order elliptic operator of divergence form with coefficients consisting of both an elliptic symmetric part and a BMO anti-symmetric part in ℝ^(n).In this article,we consider the weighted Kato square root problem for L.More precisely,we prove that the square root L^(1/2)satisfies the weighted L^(p)estimates||L^(1/2)(f)||L_(ω)^p(R^(n))≤C||■f||L_(ω)^p(R^(n);R^(n))for any p∈(1,∞)andω∈Ap(ℝ^(n))(the class of Muckenhoupt weights),and that||■f||L_(ω)^p(R^(n);R^(n))≤C||L^(1/2)(f)||L_(ω)^p(R^(n))for any p∈(1,2+ε)andω∈Ap(ℝ^(n))∩RH_(2+ε/p),(R^(n))(the class of reverse Hölder weights),whereε∈(0,∞)is a constant depending only on n and the operator L,and where(2+ε/p)'denotes the Hölder conjugate exponent of 2+ε/p.Moreover,for any given q∈(2,∞),we give a sufficient condition to obtain that||■f||L_(ω)^p(R^(n);R^(n))≤C||L^(1/2)(f)||L_(ω)^p(R^(n))for any p∈(1,q)andω∈A_(p)(R^(n))∩pRH_(q/p),(R^(n)).As an application,we prove that when the coefficient matrix A that appears in L satisfies the small BMO condition,the Riesz transform∇L^(−1/2)is bounded on L_(ω)^(p)(ℝ^(n))for any given p∈(1,∞)andω∈Ap(ℝ^(n)).Furthermore,applications to the weighted L^(2)-regularity problem with the Dirichlet or the Neumann boundary condition are also given.
