The compactness of plant chromosome and the structures of plant cell wall and cytoplasm pose a great resistance to fluorescence in situ hybridization(FISH),and consequently many new methods for improving spatial resol...The compactness of plant chromosome and the structures of plant cell wall and cytoplasm pose a great resistance to fluorescence in situ hybridization(FISH),and consequently many new methods for improving spatial resolution are being exploited to overcome these problems.However,for plants with small chromosomes like rice and Brassica,there are still many difficulties.In this article a new and effective technique for preparation of extended DNA fibers(EDFs),using a series of treatments to prophase I chromosomes of Brassica oleracea PMCs,is presented.This technique allows longitudinal extension of the chromosomes 30-107 times longer than those of their metaphase counterparts.The length of the extended DNA fibers is between 89μm and 273μm,and the space resolution is 42.8-53.0 kb.Stretching ratios were assessed in a number of FISH experiments with super-stretched chromosomes from meiotic prophase I nuclei ofB.olerecea.Through FISH to EDFs of pachytene chromosomes hybridized in situ with SRK(S-locus receptor kinase)and SPⅡ(S-locus proteinⅡ)probes,for the first time we localized the accurate positions of S-locus and quantitatively analyzed the features of S genes in B.oleracea genome to show all S genes were single-copied.In addition,the length between two linked genes was measured to be about one micron.As a result,the highest space resolution which was about 4 kb was obtained.展开更多
Fruit size is one of the most important agronomic characters,which is mainly determined by cell number and cell size.However,our current knowledge about pear is largely unknown.Through counting of pear mesocarp cells ...Fruit size is one of the most important agronomic characters,which is mainly determined by cell number and cell size.However,our current knowledge about pear is largely unknown.Through counting of pear mesocarp cells at different stages,we found that the cell number,rather than the cell size,is responsible for the differences between small- and large-fruited cultivars.Fruit weight-2.2（fw2.2） is an important quantitative trait locus（QTL） affecting fruit weight in tomato and functions as a negative regulator in carpel cell division.To get more insights into this QTL in pear fruit development,we isolated two putative homologous fw2.2 genes,which were designated as fw2.2-like（PbFWL） genes.PbFWLs encode Cys-rich proteins with the CCXXXXCPC motif and belong to the PLAC8 superfamily.In addition,results from the subcellular localization indicated that PbFWLs were localized in the plasma membrane.The expression profile of the PbFWL genes by qRT-PCR showed they expressed higher in small-sized fruit cultivar than that in large-sized fruit cultivar during the cell division period.In summary,our data suggest that these two PbFWLs might be negatively related to the cell division in pear fruit.展开更多
Amplified fragment length polymorphism （AFLP） analysis was carried out in Thatcher, near isogenic lines （NILs） canting different genes conferring resistance against wheat leaf rust, and TcLr45 × Thatcher F2 p...Amplified fragment length polymorphism （AFLP） analysis was carried out in Thatcher, near isogenic lines （NILs） canting different genes conferring resistance against wheat leaf rust, and TcLr45 × Thatcher F2 progenies were used to develop markers for Lr45 gene. Sixty AFLP primer combinations were screened and most of them provided clear amplification products, 31 primer combinations displayed polymorphism of TcLr45 in 23 NILs. Two AFLP markers closely linked to the gene Lr45 were acquired： P-AGG/M-GAG261bp, which was found closely linked to the Lr45 locus at a distance of 0.6 cM on one side, and P-ACA/M-GGT105bp, which was found at a distance of 1.3 cM on the other side. The specific hands were cloned and subsequently sequenced. The 261-bp fragment produced by P-AGG/M-GAG showed 86% similarity with the sequence of Vulgate Hort I gene; the 105-bp fragment produced by P-ACA/M-GGT showed 96% similarity with the phosphatidylserine decarboxylase gene of the Triticum monococcum. Both included an open reading frame （ORF）.展开更多
Objectives To investigate the feasibility and effect of local deliveryof c-myc antisense oligodeoxynucleotide (ASODN) by gelatin coated Platinium-Iridium stent to prevent restenosis in a normal rabbit carotid artery. ...Objectives To investigate the feasibility and effect of local deliveryof c-myc antisense oligodeoxynucleotide (ASODN) by gelatin coated Platinium-Iridium stent to prevent restenosis in a normal rabbit carotid artery. Methods Gelatin coated Platinium-Iridium stent were implanted in the right carotid arteries of 32 rabbits under vision. Animals were randomized to the control group and the treated group receiving c-myc ASODN (n=16 respectively).7, 14, 30,90 days following the stenting procedure, morphometry for caculation of neointimal area and mean neointimal thickness were performed.The expression of c-myc protein was detected by immunohistochemical methods. Results 32 stents were successfully implanted into the right carotid arteries in 32 animals.Morphometric analysis showed that neointimal area and mean neointimal thickness siginificantly increased continuously up to 12 weeks after stent implantation,and at each time point , neointimal area and mean neointimal thickness were siginificantly smaller in the treated group than control group. (P<0.001,respectively).c-myc protein expression was weak positive or negative in treated group and positive in control group. Conclusions Gelatin coated Platinium-Iridium stent mediated local delivery of c- myc ASODN is feasibility , and it can inhibit neointimal hyperplasia to prevent restenosis in a normal rabbit carotid artery.展开更多
Objectives To assess thefeasibility, efficiency and tissue distribution of local delivered c - myc antisense oligonucleotides (ASODN) by implanted gelatin coated Platinium - Iridium (Pt -Ir) stent. Methods Gelatin coa...Objectives To assess thefeasibility, efficiency and tissue distribution of local delivered c - myc antisense oligonucleotides (ASODN) by implanted gelatin coated Platinium - Iridium (Pt -Ir) stent. Methods Gelatin coated Pt - Ir stent which absorbed carboxyfluorescein - 5 - succimidyl ester (FAM) labeled c - myc ASODN were implanted in the right carotid arteries of 6 rabbits under vision. Blood samples were collected at the indicated times. The target artery, left carotid artery, heart , liver and kidney obtained at 45 minutes , 2 hours and 6 hours. The concentration of c - myc ASODN in plasma and tissues were determined by Thin Layer Fluorome-try. Tissue distribution of c - myc ASODN were assessed by fluorescence microscopy. Results At 45 min, 2 h, 6 h, the concentration of FAM labeled c -myc ASODN in target artery was 244. 39, 194. 44, 126. 94(μg/g tissues) respectively, and the delivery efficiency were 44. 4% , 35. 4% and 23. 1% respectively. At the same indicated time point, the plasma concentration was 8. 41, 5. 83, 14. 75 (μg/ml) respectively. Therefore c - myc ASODN concentrations in the target vessel were 29, 33 and 9 -fold higher than that in the plasma. There was circumferential distribution of labeled c - myc in the area of highest fluorescein coinciding with the site of medial dissecting from stent-ing, and the label was most intense in target vessel media harvested at 45 min time point and then dispersed to adventitia. Conclusions Gelatin coated Pt - Ir stent mediated local delivery of c - myc ASODN is feasible and efficient. The localization of ASODN is mainly in target vessel wall.展开更多
Structural variations(SVs)are a feature of plant genomes that has been largely unexplored despite their significant impact on plant phenotypic traits and local adaptation to abiotic and biotic stress.In this study,we ...Structural variations(SVs)are a feature of plant genomes that has been largely unexplored despite their significant impact on plant phenotypic traits and local adaptation to abiotic and biotic stress.In this study,we employed woolly grape(Vitis retordii),a species native to the tropical and subtropical regions of East Asia with both coastal and inland habitats,as a valuable model for examining the impact of SVs on local adaptation.We assembled a haplotype-resolved chromosomal reference genome for woolly grape,and conducted population genetic analyses based on whole-genome sequencing(WGS)data from coastal and inland populations.The demographic analyses revealed recent bottlenecks in all populations and asymmetric gene flow from the inland to the coastal population.In total,1,035 genes associated with plant adaptive regulation for salt stress,radiation,and environmental adaptation were detected underlying local selection by SVs and SNPs in the coastal population,of which 37.29% and 65.26% were detected by SVs and SNPs,respectively.Candidate genes such as FSD2,RGA1,and AAP8 associated with salt tolerance were found to be highly differentiated and selected during the process of local adaptation to coastal habitats in SV regions.Our study highlights the importance of SVs in local adaptation;candidate genes related to salt stress and climatic adaptation to tropical and subtropical environments are important genomic resources for future breeding programs of grapevine and its rootstocks.展开更多
基金This work was supported by the National Natural Science Foundation of China(No.60140646)the Natural Science Foundation of Chongqing(No.7914).
文摘The compactness of plant chromosome and the structures of plant cell wall and cytoplasm pose a great resistance to fluorescence in situ hybridization(FISH),and consequently many new methods for improving spatial resolution are being exploited to overcome these problems.However,for plants with small chromosomes like rice and Brassica,there are still many difficulties.In this article a new and effective technique for preparation of extended DNA fibers(EDFs),using a series of treatments to prophase I chromosomes of Brassica oleracea PMCs,is presented.This technique allows longitudinal extension of the chromosomes 30-107 times longer than those of their metaphase counterparts.The length of the extended DNA fibers is between 89μm and 273μm,and the space resolution is 42.8-53.0 kb.Stretching ratios were assessed in a number of FISH experiments with super-stretched chromosomes from meiotic prophase I nuclei ofB.olerecea.Through FISH to EDFs of pachytene chromosomes hybridized in situ with SRK(S-locus receptor kinase)and SPⅡ(S-locus proteinⅡ)probes,for the first time we localized the accurate positions of S-locus and quantitatively analyzed the features of S genes in B.oleracea genome to show all S genes were single-copied.In addition,the length between two linked genes was measured to be about one micron.As a result,the highest space resolution which was about 4 kb was obtained.
基金supported by the National High-Tech Researchand Development Program of China(2011AA10020604)the Pomology Major Projects of Xinjiang Uygur Autonomous Region of China(201130102-1)the Special Research Projects of National Forestry Industry of Public Benefit,China(201304701-1)
文摘Fruit size is one of the most important agronomic characters,which is mainly determined by cell number and cell size.However,our current knowledge about pear is largely unknown.Through counting of pear mesocarp cells at different stages,we found that the cell number,rather than the cell size,is responsible for the differences between small- and large-fruited cultivars.Fruit weight-2.2（fw2.2） is an important quantitative trait locus（QTL） affecting fruit weight in tomato and functions as a negative regulator in carpel cell division.To get more insights into this QTL in pear fruit development,we isolated two putative homologous fw2.2 genes,which were designated as fw2.2-like（PbFWL） genes.PbFWLs encode Cys-rich proteins with the CCXXXXCPC motif and belong to the PLAC8 superfamily.In addition,results from the subcellular localization indicated that PbFWLs were localized in the plasma membrane.The expression profile of the PbFWL genes by qRT-PCR showed they expressed higher in small-sized fruit cultivar than that in large-sized fruit cultivar during the cell division period.In summary,our data suggest that these two PbFWLs might be negatively related to the cell division in pear fruit.
基金Acknowledgements This work was supported by the National Natural Science Foundation of China （30170602）.
文摘Amplified fragment length polymorphism （AFLP） analysis was carried out in Thatcher, near isogenic lines （NILs） canting different genes conferring resistance against wheat leaf rust, and TcLr45 × Thatcher F2 progenies were used to develop markers for Lr45 gene. Sixty AFLP primer combinations were screened and most of them provided clear amplification products, 31 primer combinations displayed polymorphism of TcLr45 in 23 NILs. Two AFLP markers closely linked to the gene Lr45 were acquired： P-AGG/M-GAG261bp, which was found closely linked to the Lr45 locus at a distance of 0.6 cM on one side, and P-ACA/M-GGT105bp, which was found at a distance of 1.3 cM on the other side. The specific hands were cloned and subsequently sequenced. The 261-bp fragment produced by P-AGG/M-GAG showed 86% similarity with the sequence of Vulgate Hort I gene; the 105-bp fragment produced by P-ACA/M-GGT showed 96% similarity with the phosphatidylserine decarboxylase gene of the Triticum monococcum. Both included an open reading frame （ORF）.
文摘Objectives To investigate the feasibility and effect of local deliveryof c-myc antisense oligodeoxynucleotide (ASODN) by gelatin coated Platinium-Iridium stent to prevent restenosis in a normal rabbit carotid artery. Methods Gelatin coated Platinium-Iridium stent were implanted in the right carotid arteries of 32 rabbits under vision. Animals were randomized to the control group and the treated group receiving c-myc ASODN (n=16 respectively).7, 14, 30,90 days following the stenting procedure, morphometry for caculation of neointimal area and mean neointimal thickness were performed.The expression of c-myc protein was detected by immunohistochemical methods. Results 32 stents were successfully implanted into the right carotid arteries in 32 animals.Morphometric analysis showed that neointimal area and mean neointimal thickness siginificantly increased continuously up to 12 weeks after stent implantation,and at each time point , neointimal area and mean neointimal thickness were siginificantly smaller in the treated group than control group. (P<0.001,respectively).c-myc protein expression was weak positive or negative in treated group and positive in control group. Conclusions Gelatin coated Platinium-Iridium stent mediated local delivery of c- myc ASODN is feasibility , and it can inhibit neointimal hyperplasia to prevent restenosis in a normal rabbit carotid artery.
文摘Objectives To assess thefeasibility, efficiency and tissue distribution of local delivered c - myc antisense oligonucleotides (ASODN) by implanted gelatin coated Platinium - Iridium (Pt -Ir) stent. Methods Gelatin coated Pt - Ir stent which absorbed carboxyfluorescein - 5 - succimidyl ester (FAM) labeled c - myc ASODN were implanted in the right carotid arteries of 6 rabbits under vision. Blood samples were collected at the indicated times. The target artery, left carotid artery, heart , liver and kidney obtained at 45 minutes , 2 hours and 6 hours. The concentration of c - myc ASODN in plasma and tissues were determined by Thin Layer Fluorome-try. Tissue distribution of c - myc ASODN were assessed by fluorescence microscopy. Results At 45 min, 2 h, 6 h, the concentration of FAM labeled c -myc ASODN in target artery was 244. 39, 194. 44, 126. 94(μg/g tissues) respectively, and the delivery efficiency were 44. 4% , 35. 4% and 23. 1% respectively. At the same indicated time point, the plasma concentration was 8. 41, 5. 83, 14. 75 (μg/ml) respectively. Therefore c - myc ASODN concentrations in the target vessel were 29, 33 and 9 -fold higher than that in the plasma. There was circumferential distribution of labeled c - myc in the area of highest fluorescein coinciding with the site of medial dissecting from stent-ing, and the label was most intense in target vessel media harvested at 45 min time point and then dispersed to adventitia. Conclusions Gelatin coated Pt - Ir stent mediated local delivery of c - myc ASODN is feasible and efficient. The localization of ASODN is mainly in target vessel wall.
基金supported by the Science Fund Program for Distinguished Young Scholars of the National Natural Science Foundation of China(Overseas)to Yongfeng ZhouNational Natural Science Foundation of China(Nos.32300191,32372662)+1 种基金Guangxi University,Bama Institute of Integration of Industry and Education,postgraduate joint training project(Project Nos.20210020,20210039)the National Key Research and Development Program of China(grants 2023YFF1000100 and 2023YFD2200700).
文摘Structural variations(SVs)are a feature of plant genomes that has been largely unexplored despite their significant impact on plant phenotypic traits and local adaptation to abiotic and biotic stress.In this study,we employed woolly grape(Vitis retordii),a species native to the tropical and subtropical regions of East Asia with both coastal and inland habitats,as a valuable model for examining the impact of SVs on local adaptation.We assembled a haplotype-resolved chromosomal reference genome for woolly grape,and conducted population genetic analyses based on whole-genome sequencing(WGS)data from coastal and inland populations.The demographic analyses revealed recent bottlenecks in all populations and asymmetric gene flow from the inland to the coastal population.In total,1,035 genes associated with plant adaptive regulation for salt stress,radiation,and environmental adaptation were detected underlying local selection by SVs and SNPs in the coastal population,of which 37.29% and 65.26% were detected by SVs and SNPs,respectively.Candidate genes such as FSD2,RGA1,and AAP8 associated with salt tolerance were found to be highly differentiated and selected during the process of local adaptation to coastal habitats in SV regions.Our study highlights the importance of SVs in local adaptation;candidate genes related to salt stress and climatic adaptation to tropical and subtropical environments are important genomic resources for future breeding programs of grapevine and its rootstocks.
