Seed plumules comprise multiple developing tissues and are key sites for above-ground plant organ morphogenesis.Here,the spatial expression of genes in developing rice seed plumules was characterized by single-cell tr...Seed plumules comprise multiple developing tissues and are key sites for above-ground plant organ morphogenesis.Here,the spatial expression of genes in developing rice seed plumules was characterized by single-cell transcriptome sequencing in Zhongjiazao 17,a popular Chinese indica rice cultivar.Of 15 cell clusters,13 were assigned to cell types using marker genes and cluster-specific genes.Marker genes of multiple cell types were expressed in several clusters,suggesting a complex developmental system.Some genes for signaling by phytohormones such as abscisic acid were highly expressed in specific clusters.Various cis-elements in the promoters of genes specifically expressed in cell clusters were calculated,and some key hormone-related motifs were frequent in certain clusters.Spatial expression patterns of genes involved in rapid seed germination,seedling growth,and development were identified.These findings enhanced our understanding of cellular diversity and specialization within plumules of rice,a monocotyledonous model crop.展开更多
Molecular marker techniques have been widely applied in the fields of genetic diversity analysis,germplasm resources identification,molecular fingerprint and genetic linkage map construction,QTL mapping and molecular ...Molecular marker techniques have been widely applied in the fields of genetic diversity analysis,germplasm resources identification,molecular fingerprint and genetic linkage map construction,QTL mapping and molecular assisted breeding.On the basis of stating the concept of molecular marker techniques based on single primer amplification reactions,this study focused on the sorting and induction of single-primer molecular marker techniques,and expounded their derivative development.Finally,the application prospect and future expectation of single-primer molecular marker techniques were described in detail.The purpose of this study was to clarify the types of molecular marker techniques based on single primer amplification reactions,so that researchers can quickly and conveniently select molecular marker techniques according to their own specific scientific research conditions.展开更多
[Objectives]To establish a non-toxic and efficient method for extracting DNA and total RNA from peanuts and laying a solid foundation for the molecular biology study of peanuts.[Methods]Based on the principle and meth...[Objectives]To establish a non-toxic and efficient method for extracting DNA and total RNA from peanuts and laying a solid foundation for the molecular biology study of peanuts.[Methods]Based on the principle and method of purifying nucleic acids by silica gel adsorption at high salt and low pH condition,a non-toxic and efficient method to extract peanut DNA and total RNA using cetyltrimethyl ammonium bromide(CTAB)extraction solution was designed.The quality and purity of nucleic acids were detected by agarose gel electrophoresis and nucleic acids protein analyzer,respectively.The quality of DNA was further verified by enzyme digestion and PCR amplification using molecular marker techniques.The quality of total RNA was further verified by reverse transcription(RT)-PCR of actin gene and cDNA-SCoT gene differential display technique.[Results]The agarose gel electrophoresis test showed that the peanut DNA extracted by a low-toxic and effective method is free of contamination and degradation.Through the detection by the nucleic acid protein analyzer,the DNA concentration,yield,A260/A280 and A260/A230 of 5 peanut varieties were 419.6-498.2 ng/μL,20.98-24.91μg/g,1.89-1.96 and 2.03-2.28,respectively.The DNA was of high quality and can be completely digested by EcoRI restriction enzymes,and also can be used for SCoT and SRAP molecular marker technology analysis.The RNA extracted from different tissues of peanuts showed no visible DNA bands by non-denaturing agarose gel electrophoresis.The separated 28S bands were brighter than 18S.The ratio of A260/A280 and A260/A230 showed that the RNA quality was good and can be used for reverse transcription,RT-PCR of actin gene and amplification of cDNA-SCoT gene differential display technique.[Conclusions]This experiment established a low-toxic and effective method for extracting DNA and total RNA from peanuts.Compared with traditional methods,this method is more time-saving and cheaper than commercial kits.The most important point is that this method does not use toxic reagents such as phenol,chloroform and isopropanol.Thus,it is expected to be widely applied in molecular biology research.展开更多
Chronic hypoxia affects stem cell function during tissue repair.Thus far,the hypoxia-associated impact on periosteal stem cells(PSCs),the main contributor to bone repair,remains unknown,and a tailored oxygen modulatio...Chronic hypoxia affects stem cell function during tissue repair.Thus far,the hypoxia-associated impact on periosteal stem cells(PSCs),the main contributor to bone repair,remains unknown,and a tailored oxygen modulation strategy for optimizing PSC function is lacking.Here,PSCs exhibit time-dependent proliferation and survival upon hypoxic exposure and a critical 48-h time-point is identified at which hypoxia transitions from beneficial to detrimental.Then,a photothermal-sensitive coaxial fiber-reinforced membrane containing oxygen and pravastatin is constructed to function as an intelligent oxygen supply system.Leveraging near-infrared light as an ON/OFF switch,the system noninvasively scales up oxygen release beginning 48 h post-implantation,counteracting prolonged hypoxia and mitigating its adverse effects on PSCs.The sustained release of pravastatin from the membrane accelerates early neovascularization both directly through its pro-angiogenic effect and indirectly by stimulating vascular endothelial growth factor secretion from PSCs,ensuring a continuous oxygen supply after exogenous oxygen exhaustion.Notably,pravastatin steers PSCs toward robust osteogenic differentiation and provides multifunctional bioactive cues for advanced bone regeneration in vivo.This time-scheduled approach to modulate oxygen supply noninvasively could be applicable beyond bone regeneration for hypoxia-related diseases and multi-tissue repair.展开更多
基金financially supported by the“STI2030-Major Project”of China(2023ZD04072)the National Key Research and Development Program of China(2021YFA1300400)+1 种基金the National Natural Science Foundation of China(32372099 and 32188102)the Young Science and Technology Talents(He Jian)in Hunan Province(2022RC1015)。
文摘Seed plumules comprise multiple developing tissues and are key sites for above-ground plant organ morphogenesis.Here,the spatial expression of genes in developing rice seed plumules was characterized by single-cell transcriptome sequencing in Zhongjiazao 17,a popular Chinese indica rice cultivar.Of 15 cell clusters,13 were assigned to cell types using marker genes and cluster-specific genes.Marker genes of multiple cell types were expressed in several clusters,suggesting a complex developmental system.Some genes for signaling by phytohormones such as abscisic acid were highly expressed in specific clusters.Various cis-elements in the promoters of genes specifically expressed in cell clusters were calculated,and some key hormone-related motifs were frequent in certain clusters.Spatial expression patterns of genes involved in rapid seed germination,seedling growth,and development were identified.These findings enhanced our understanding of cellular diversity and specialization within plumules of rice,a monocotyledonous model crop.
基金the National Natural Science Foundation of China(31960409,31960416)Guangxi Natural Science Foundation Program(2018GXNSFDA281027,2018GXNSFDA294004,2020GXNSFAA297081)Guangxi Academy of Agricultural Sciences Fund Project(GNK2017JZ13,GNK2018YM06,GNK31960409).
文摘Molecular marker techniques have been widely applied in the fields of genetic diversity analysis,germplasm resources identification,molecular fingerprint and genetic linkage map construction,QTL mapping and molecular assisted breeding.On the basis of stating the concept of molecular marker techniques based on single primer amplification reactions,this study focused on the sorting and induction of single-primer molecular marker techniques,and expounded their derivative development.Finally,the application prospect and future expectation of single-primer molecular marker techniques were described in detail.The purpose of this study was to clarify the types of molecular marker techniques based on single primer amplification reactions,so that researchers can quickly and conveniently select molecular marker techniques according to their own specific scientific research conditions.
基金Projects of National Natural Science Foundation of China(3166042831960409+2 种基金31960416)Projects of Guangxi Natural Science Foundation of China(2018GXNSFDA281027,2018GXNSFDA294004,2017GXNSFAA198032)Science and Technology Development Fund Project of Guangxi Academy of Agricultural Sciences(Gui Nong Ke 2018YM06,Gui Nong Ke 2017JZ13,31960409,Gui Nong Ke 2018YT12).
文摘[Objectives]To establish a non-toxic and efficient method for extracting DNA and total RNA from peanuts and laying a solid foundation for the molecular biology study of peanuts.[Methods]Based on the principle and method of purifying nucleic acids by silica gel adsorption at high salt and low pH condition,a non-toxic and efficient method to extract peanut DNA and total RNA using cetyltrimethyl ammonium bromide(CTAB)extraction solution was designed.The quality and purity of nucleic acids were detected by agarose gel electrophoresis and nucleic acids protein analyzer,respectively.The quality of DNA was further verified by enzyme digestion and PCR amplification using molecular marker techniques.The quality of total RNA was further verified by reverse transcription(RT)-PCR of actin gene and cDNA-SCoT gene differential display technique.[Results]The agarose gel electrophoresis test showed that the peanut DNA extracted by a low-toxic and effective method is free of contamination and degradation.Through the detection by the nucleic acid protein analyzer,the DNA concentration,yield,A260/A280 and A260/A230 of 5 peanut varieties were 419.6-498.2 ng/μL,20.98-24.91μg/g,1.89-1.96 and 2.03-2.28,respectively.The DNA was of high quality and can be completely digested by EcoRI restriction enzymes,and also can be used for SCoT and SRAP molecular marker technology analysis.The RNA extracted from different tissues of peanuts showed no visible DNA bands by non-denaturing agarose gel electrophoresis.The separated 28S bands were brighter than 18S.The ratio of A260/A280 and A260/A230 showed that the RNA quality was good and can be used for reverse transcription,RT-PCR of actin gene and amplification of cDNA-SCoT gene differential display technique.[Conclusions]This experiment established a low-toxic and effective method for extracting DNA and total RNA from peanuts.Compared with traditional methods,this method is more time-saving and cheaper than commercial kits.The most important point is that this method does not use toxic reagents such as phenol,chloroform and isopropanol.Thus,it is expected to be widely applied in molecular biology research.
基金supported by the National Natural Science Foundation of China(Grant Nos.82122043 and 82372404)the Quick Response project of AFMU(2023KXKT030).
文摘Chronic hypoxia affects stem cell function during tissue repair.Thus far,the hypoxia-associated impact on periosteal stem cells(PSCs),the main contributor to bone repair,remains unknown,and a tailored oxygen modulation strategy for optimizing PSC function is lacking.Here,PSCs exhibit time-dependent proliferation and survival upon hypoxic exposure and a critical 48-h time-point is identified at which hypoxia transitions from beneficial to detrimental.Then,a photothermal-sensitive coaxial fiber-reinforced membrane containing oxygen and pravastatin is constructed to function as an intelligent oxygen supply system.Leveraging near-infrared light as an ON/OFF switch,the system noninvasively scales up oxygen release beginning 48 h post-implantation,counteracting prolonged hypoxia and mitigating its adverse effects on PSCs.The sustained release of pravastatin from the membrane accelerates early neovascularization both directly through its pro-angiogenic effect and indirectly by stimulating vascular endothelial growth factor secretion from PSCs,ensuring a continuous oxygen supply after exogenous oxygen exhaustion.Notably,pravastatin steers PSCs toward robust osteogenic differentiation and provides multifunctional bioactive cues for advanced bone regeneration in vivo.This time-scheduled approach to modulate oxygen supply noninvasively could be applicable beyond bone regeneration for hypoxia-related diseases and multi-tissue repair.
