The Agrobacterium-mediated transient expression system with conventional binary vectors is well established in tobacco leaves,while the same system applied to tomato leaflets has relatively low expression efficiency.H...The Agrobacterium-mediated transient expression system with conventional binary vectors is well established in tobacco leaves,while the same system applied to tomato leaflets has relatively low expression efficiency.However,impacts of the leaf age,inoculation method and incubation condition after Agrobacterium infiltration on transient protein expression efficiency are seldom investigated.In this study,we optimize Agrobacterium-mediated transient expression system using conventional binary vectors to achieve the high efficiency of target gene expression in tomato leaflets.We transiently express GFP and a nucleus-localized gene SlUVI4 fused with GFP in detached 10-,20-,and 30-day-old leaflets.The cutting points of leaflets are embedded in MS medium after the Agrobacterium-mediated vacuum infiltration,and all leaflets are kept in the dark before use.The 10-and 30-day-old leaflets have more damage than 20-day-old leaflets after the infiltration.展开更多
Dissociation (Ds) transposon is one of thetransposable elements in corn. The trans-posons can be transferred into other plantswhere the transposons were not found. Oncethe transposon was inserted into target gene ofth...Dissociation (Ds) transposon is one of thetransposable elements in corn. The trans-posons can be transferred into other plantswhere the transposons were not found. Oncethe transposon was inserted into target gene ofthese plants, it could be used as a marker todistinguish and isolate the gene. The object ofthis study is to transfer Ds transposon to riceby Agrobacterium -mediated transformation.The calli of immature embryos, mature展开更多
Oat(Avena sativa L.)is a versatile,annual herbaceous species widely cultivated for both grain and forage production.It is renowned for its high biomass yield,strong resistance to abiotic stress,superior nutritional qu...Oat(Avena sativa L.)is a versatile,annual herbaceous species widely cultivated for both grain and forage production.It is renowned for its high biomass yield,strong resistance to abiotic stress,superior nutritional quality,and excellent palatability(Wu et al.,2019;Zhang et al.,2023).Despite these advantages,oat's genetic transformation system lags behind that of major crops like rice,wheat,and maize.Traditionally,oat transformation has relied on biolistic bombardment of callus tissue derived from immature embryos,mature embryos,or leaf bases(Maqbool et al.,2002).展开更多
This study was conducted to evaluate the effects of proline and glutamine on in vitro callus induction and subsequent regeneration and to develop a reproducible and highly efficient plant regeneration protocol in four...This study was conducted to evaluate the effects of proline and glutamine on in vitro callus induction and subsequent regeneration and to develop a reproducible and highly efficient plant regeneration protocol in four rice genotypes, viz. Pawana, Jaya, Indrayani and Ambemohar. Considerable variation in response to plant growth regulators and amino acid supplements used was observed in all the four genotypes. Medium supplemented with proline and glutamine was shown to be superior to medium without proline and glutamine. The best callusing from mature embryo was observed on Murashige and Skoog （MS） medium supplemented with 2.0 mg/L 2,4-dichlorophenoxyacetic acid （2,4-D）, 500 mg/L proline and 500 mg/L glutamine. Shoot induction was higher in the callus obtained from medium supplemented with 500 mg/L proline and 500 mg/L glutamine. The highest shoot regeneration frequency （83.2%） was observed on MS medium with 2.0 mg/L benzylaminopurine, 0.5 mg/L 1-naphthaleneacetic acid, 500 mg/L proline, and 500 mg/L glutamine in the callus obtained from MS medium supplemented with 2.0 mg/L 2,4-D, 500 mg/L proline and 500 mg/L glutamine. Among the four genotypes, Pawana has the highest regeneration efficiency （83.2%）, whereas the regeneration efficiency of the rest three rice genotypes was in the range of 32.0% to 72.3%. This optimized regeneration protocol can be efficiently used for Agrobacterium mediated genetic transformation in rice.展开更多
[Objectives]This study was conducted to improve the Agrobacterium-mediated genetic transformation system of Begonia wallichiana.[Methods]With sterilized tube seedling leaves as the recipient material and GFP as the re...[Objectives]This study was conducted to improve the Agrobacterium-mediated genetic transformation system of Begonia wallichiana.[Methods]With sterilized tube seedling leaves as the recipient material and GFP as the reporter gene,optimization experiments were carried out in terms of infection time and method,co-cultivation time and method,and PCR detection technology.[Results]The transformation effect was better under the conditions of shaking Agrobacterium liquid,infection time of 1-2 h,and co-cultivation on sterilized filter paper for 2 d.After co-cultivation,the recipient material was first subjected to recovery culture,and then used for Hyg gradient screening,which was conducive to obtaining resistant transformants.The designed specific PCR detection technology could quickly identify false positives in resistant regenerated plants,and the proportion of transgenic plants was 16.7%.[Conclusions]The research results provide a new technical reference for the genetic transformation of ornamental plants.展开更多
基金support of Taishan Scholar Foundation of Shandong Province(tsqn201909073,tsqn201812034)National Natural Science Foundation of China(31872951)。
文摘The Agrobacterium-mediated transient expression system with conventional binary vectors is well established in tobacco leaves,while the same system applied to tomato leaflets has relatively low expression efficiency.However,impacts of the leaf age,inoculation method and incubation condition after Agrobacterium infiltration on transient protein expression efficiency are seldom investigated.In this study,we optimize Agrobacterium-mediated transient expression system using conventional binary vectors to achieve the high efficiency of target gene expression in tomato leaflets.We transiently express GFP and a nucleus-localized gene SlUVI4 fused with GFP in detached 10-,20-,and 30-day-old leaflets.The cutting points of leaflets are embedded in MS medium after the Agrobacterium-mediated vacuum infiltration,and all leaflets are kept in the dark before use.The 10-and 30-day-old leaflets have more damage than 20-day-old leaflets after the infiltration.
文摘Dissociation (Ds) transposon is one of thetransposable elements in corn. The trans-posons can be transferred into other plantswhere the transposons were not found. Oncethe transposon was inserted into target gene ofthese plants, it could be used as a marker todistinguish and isolate the gene. The object ofthis study is to transfer Ds transposon to riceby Agrobacterium -mediated transformation.The calli of immature embryos, mature
基金funded by the National Center for Forestry and Grassland Genetic Resources (2005DKA21003).
文摘Oat(Avena sativa L.)is a versatile,annual herbaceous species widely cultivated for both grain and forage production.It is renowned for its high biomass yield,strong resistance to abiotic stress,superior nutritional quality,and excellent palatability(Wu et al.,2019;Zhang et al.,2023).Despite these advantages,oat's genetic transformation system lags behind that of major crops like rice,wheat,and maize.Traditionally,oat transformation has relied on biolistic bombardment of callus tissue derived from immature embryos,mature embryos,or leaf bases(Maqbool et al.,2002).
基金supported by grants from the Biological Breeding-National Science and Technology Major Project(2023ZD0402701 and 2024ZD04077)the National Natural Science Foundation of China(32425041)+1 种基金the National Key Research and Development Program of China(2021YFD1200701)the Joint Research on Maize Improvement of Henan(2022010202).
文摘Dear Editor,Maize(Zea mays L.)is a globally crucial crop that exhibits serious genotype dependency,which renders its genetic transformation challenging.Agrobacterium(Agrobacterium tumefaciens)-mediated transformation depends on(1)the recipient’s susceptibility to Agrobacterium infection and(2)the ability to form embryogenic callus.
基金financially supported by the Mahatma Phule Agricultural University,Rahuri,India
文摘This study was conducted to evaluate the effects of proline and glutamine on in vitro callus induction and subsequent regeneration and to develop a reproducible and highly efficient plant regeneration protocol in four rice genotypes, viz. Pawana, Jaya, Indrayani and Ambemohar. Considerable variation in response to plant growth regulators and amino acid supplements used was observed in all the four genotypes. Medium supplemented with proline and glutamine was shown to be superior to medium without proline and glutamine. The best callusing from mature embryo was observed on Murashige and Skoog （MS） medium supplemented with 2.0 mg/L 2,4-dichlorophenoxyacetic acid （2,4-D）, 500 mg/L proline and 500 mg/L glutamine. Shoot induction was higher in the callus obtained from medium supplemented with 500 mg/L proline and 500 mg/L glutamine. The highest shoot regeneration frequency （83.2%） was observed on MS medium with 2.0 mg/L benzylaminopurine, 0.5 mg/L 1-naphthaleneacetic acid, 500 mg/L proline, and 500 mg/L glutamine in the callus obtained from MS medium supplemented with 2.0 mg/L 2,4-D, 500 mg/L proline and 500 mg/L glutamine. Among the four genotypes, Pawana has the highest regeneration efficiency （83.2%）, whereas the regeneration efficiency of the rest three rice genotypes was in the range of 32.0% to 72.3%. This optimized regeneration protocol can be efficiently used for Agrobacterium mediated genetic transformation in rice.
文摘[Objectives]This study was conducted to improve the Agrobacterium-mediated genetic transformation system of Begonia wallichiana.[Methods]With sterilized tube seedling leaves as the recipient material and GFP as the reporter gene,optimization experiments were carried out in terms of infection time and method,co-cultivation time and method,and PCR detection technology.[Results]The transformation effect was better under the conditions of shaking Agrobacterium liquid,infection time of 1-2 h,and co-cultivation on sterilized filter paper for 2 d.After co-cultivation,the recipient material was first subjected to recovery culture,and then used for Hyg gradient screening,which was conducive to obtaining resistant transformants.The designed specific PCR detection technology could quickly identify false positives in resistant regenerated plants,and the proportion of transgenic plants was 16.7%.[Conclusions]The research results provide a new technical reference for the genetic transformation of ornamental plants.
