Hydrangea bretschneideri Dipp is a highly popular ornamental plant for garden decoration.Genetic engineering technology has been successfully used in many plant species,but it is limited in Hydrangea.Here we establish...Hydrangea bretschneideri Dipp is a highly popular ornamental plant for garden decoration.Genetic engineering technology has been successfully used in many plant species,but it is limited in Hydrangea.Here we established an efficient regeneration system by using stem segments as explants for the first time.In our study,the plant growth regulators(PGRs)were evaluated at the different regeneration processes,including axillary shoots regeneration and root induction.We found that the optimal concentration for axillary buds’induction was 2.0 mgL^(-1)6-BA and 0.5 mgL^(-1)1 IAA,its highest induction rate was 70%.Moreover,the highest axillary shoots proliferation coefficient was 10.7 on the Murashige and Skoog(MS)medium with 2.0 mgL^(-1)6-benzyladenine(BA),0.2 mgL^(-1)indole-3-butyric acid(IBA),and 1.0 mgL^(-1)gibberellin A3(GA3).The highest frequency of root induction was 80.0±0.06%by culturing the elongated shoots in 1/2 MS medium containing 0.1 mgL^(-1)IBA.In summary,our study will provide an effective technology for large-scale propagation and important pathway for promoting the popularization and application of Hydrangea bretschneideri Dipp.展开更多
Genetic transformation with mature material as the explants could shorten the transgenic period and avoid seed dependence compared with genetic transformation using the epicotyl seedling stem segments as the receptor....Genetic transformation with mature material as the explants could shorten the transgenic period and avoid seed dependence compared with genetic transformation using the epicotyl seedling stem segments as the receptor. Here, we constructed an Agrobacterium tumefaciensmediated transformation for generation of marker-free transgenic plants from navel orange(Citrus sinensis Osbeck) mature stems using a CreloxP recombination system. To efficiently recover the regenerated buds from mature tissues, five recovery methods were compared: in vitro micrografting of 0.1-0.5(1-2 weeks), > 0.5 cm(3-4 weeks) and > 1 cm long lignified bud and in vitro micrografting of explants with a bud and rooting regenerated bud. The data showed that in vitro micrografting of > 1 cm long regenerated bud with expanded leaves after one month of continuous culture for lignification was the optimal solution for plant recovery from mature tissues. Transgenic plants without selectable marker genes were created from navel orange(Citrus sinensis Osbeck) tissue using a transformation vector PLI-35SPR1aCB containing a Cre/loxP system recombination together with genes encoding the selectable marker isopentenyl transferase(IPT) and an anti-bacterial peptide(PR1aCB).Using IPT positive selection, the transformation efficiency determined by PCR was 0.9%, and in total, 20 transgenic plants were obtained.Southern blotting confirmed further their transgenicity. PCR and sequencing analysis demonstrated that both the Cre and IPT genes had been successfully removed from the transgenic plants(deletion efficiency 100%). Over all, using Cre/loxP system recombination together with the IPT positive selection, marker-free transgenic plants can be recovered efficiently from mature tissues of navel orange(Citrus sinensis Osbeck), which provides a potential method for production of transgenic plants from citrus mature tissue.展开更多
An efficient in vitro regeneration system by direct organogenesis from mature nodal and internodal stem segments ofNewhall navel orange (Citrus sinensis L. Osbeck) was developed. Illuminating conditions together with...An efficient in vitro regeneration system by direct organogenesis from mature nodal and internodal stem segments ofNewhall navel orange (Citrus sinensis L. Osbeck) was developed. Illuminating conditions together with plant growthregulators affected the adventitious bud regeneration frequency and efficiency. The initial 15 d darkness inoculation isbeneficial for the adventitious bud regeneration. The highest regeneration frequency (85.2%) and bud formationefficiency (3.7 per responsive internodal stem segment) were obtained in the media supplemented with 1.0 mg L-1 BAPand 0.5 mg L-1 NAA. ABA at 0.2 mg L-1 positively affected the bud formation efficiency, which amounted to 8.5 buds perinternodal segment in the presence of BAP at 1.0 mg L-1. The adventitious shoots successfully rooted and weretransferred to the soil.展开更多
[ Objective] This study aimed to investigate the differences in morphological characteristics of Dendrobium officinale plantlets propagated from different explants. [ Method] Randomly 1 000 D. offtcinale plantlets pro...[ Objective] This study aimed to investigate the differences in morphological characteristics of Dendrobium officinale plantlets propagated from different explants. [ Method] Randomly 1 000 D. offtcinale plantlets propagated via different regeneration pathways were selected for morphological investigation and classification. [ Result] D. officinale plantlets propagated from stem segment explants exhibited highly consistent morphological characteristics, while those propagated from seed explants exhibited a variety of morphological characteristics. [ Conclusion] Therefore, using seed explants for regeneration can effectively broaden the germplasms resources of D. officinale.展开更多
[Objectives]The purpose was to establish an induction system for friable callus of Hedera nepalensis var.sinensis with different parts.[Methods]By screening the most suitable explant and adjusting the hormone ratio of...[Objectives]The purpose was to establish an induction system for friable callus of Hedera nepalensis var.sinensis with different parts.[Methods]By screening the most suitable explant and adjusting the hormone ratio of medium,friable calli of H.nepalensis var.sinensis were induced.[Results]The calli could be induced from leaves,petioles and stem segments,but the ideal explant was stem segments,with induction rate reaching 98%.The optimal medium for callus proliferation was MS+0.5 mg/L KT+1.0 mg/L 2,4-D+30.0 g/L sucrose.After 3-4 generations of subculture on MS+0.5 mg/L BA+1.0 mg/L 2,4-D+30.0 g/L sucrose,favorable friable calli of H.nepalensis var.sinensis were obtained.[Conclusions]The friable calli induced in this experiment can lay a foundation for in-vitro regeneration and cellular secondary metabolite production of H.nepalensis var.sinensis.展开更多
基金This work is supported by the Grassland Talent Project:The Innovation Team of New Varieties Breeding at the Economic and Ecological Shrub,and the Evaluation of the Economic and Ecological Shrub Resources and New Variety Breeding in Inner Mongolia(No.201702077)。
文摘Hydrangea bretschneideri Dipp is a highly popular ornamental plant for garden decoration.Genetic engineering technology has been successfully used in many plant species,but it is limited in Hydrangea.Here we established an efficient regeneration system by using stem segments as explants for the first time.In our study,the plant growth regulators(PGRs)were evaluated at the different regeneration processes,including axillary shoots regeneration and root induction.We found that the optimal concentration for axillary buds’induction was 2.0 mgL^(-1)6-BA and 0.5 mgL^(-1)1 IAA,its highest induction rate was 70%.Moreover,the highest axillary shoots proliferation coefficient was 10.7 on the Murashige and Skoog(MS)medium with 2.0 mgL^(-1)6-benzyladenine(BA),0.2 mgL^(-1)indole-3-butyric acid(IBA),and 1.0 mgL^(-1)gibberellin A3(GA3).The highest frequency of root induction was 80.0±0.06%by culturing the elongated shoots in 1/2 MS medium containing 0.1 mgL^(-1)IBA.In summary,our study will provide an effective technology for large-scale propagation and important pathway for promoting the popularization and application of Hydrangea bretschneideri Dipp.
基金supported by the Fundamental Research Funds for the Central Universities (Grant No. XDJK 2018B016)the National Natural Sciences Foundation of China (Grant No. 31972393)+1 种基金he earmarked fund for China Agriculture Research System (Grant No. CARS-26)the Natural Science Foundation of Chongqing (Grant No. cstc2020jcyj-msxmX1064)。
文摘Genetic transformation with mature material as the explants could shorten the transgenic period and avoid seed dependence compared with genetic transformation using the epicotyl seedling stem segments as the receptor. Here, we constructed an Agrobacterium tumefaciensmediated transformation for generation of marker-free transgenic plants from navel orange(Citrus sinensis Osbeck) mature stems using a CreloxP recombination system. To efficiently recover the regenerated buds from mature tissues, five recovery methods were compared: in vitro micrografting of 0.1-0.5(1-2 weeks), > 0.5 cm(3-4 weeks) and > 1 cm long lignified bud and in vitro micrografting of explants with a bud and rooting regenerated bud. The data showed that in vitro micrografting of > 1 cm long regenerated bud with expanded leaves after one month of continuous culture for lignification was the optimal solution for plant recovery from mature tissues. Transgenic plants without selectable marker genes were created from navel orange(Citrus sinensis Osbeck) tissue using a transformation vector PLI-35SPR1aCB containing a Cre/loxP system recombination together with genes encoding the selectable marker isopentenyl transferase(IPT) and an anti-bacterial peptide(PR1aCB).Using IPT positive selection, the transformation efficiency determined by PCR was 0.9%, and in total, 20 transgenic plants were obtained.Southern blotting confirmed further their transgenicity. PCR and sequencing analysis demonstrated that both the Cre and IPT genes had been successfully removed from the transgenic plants(deletion efficiency 100%). Over all, using Cre/loxP system recombination together with the IPT positive selection, marker-free transgenic plants can be recovered efficiently from mature tissues of navel orange(Citrus sinensis Osbeck), which provides a potential method for production of transgenic plants from citrus mature tissue.
基金The authors acknowledge the financial support by the National Natural Science Foundation of China(002002)
文摘An efficient in vitro regeneration system by direct organogenesis from mature nodal and internodal stem segments ofNewhall navel orange (Citrus sinensis L. Osbeck) was developed. Illuminating conditions together with plant growthregulators affected the adventitious bud regeneration frequency and efficiency. The initial 15 d darkness inoculation isbeneficial for the adventitious bud regeneration. The highest regeneration frequency (85.2%) and bud formationefficiency (3.7 per responsive internodal stem segment) were obtained in the media supplemented with 1.0 mg L-1 BAPand 0.5 mg L-1 NAA. ABA at 0.2 mg L-1 positively affected the bud formation efficiency, which amounted to 8.5 buds perinternodal segment in the presence of BAP at 1.0 mg L-1. The adventitious shoots successfully rooted and weretransferred to the soil.
基金Supported by of Spark Program of Guangdong Department of Science and Technology"Development of Cultivation Techniques of Dendrobium officinale Seedlings"(2013B020503062)Special Innovation Fund for Small and Medium Enterprise in Maoming City"Research and Demonstration of Alpine Organic Efficient Cultivation Technology of Precious Chinese Herb Dendrobium officinale"(2012B01088)
文摘[ Objective] This study aimed to investigate the differences in morphological characteristics of Dendrobium officinale plantlets propagated from different explants. [ Method] Randomly 1 000 D. offtcinale plantlets propagated via different regeneration pathways were selected for morphological investigation and classification. [ Result] D. officinale plantlets propagated from stem segment explants exhibited highly consistent morphological characteristics, while those propagated from seed explants exhibited a variety of morphological characteristics. [ Conclusion] Therefore, using seed explants for regeneration can effectively broaden the germplasms resources of D. officinale.
基金Tianjin Science and Technology Plan Project(18ZXBFNC00370)Industrial Innovation and Entrepreneurship Team Project of Hebei Province(199A2905H)Fund of Central Government for Guiding Science and Technology Development in Hebei Province(206Z6303G).
文摘[Objectives]The purpose was to establish an induction system for friable callus of Hedera nepalensis var.sinensis with different parts.[Methods]By screening the most suitable explant and adjusting the hormone ratio of medium,friable calli of H.nepalensis var.sinensis were induced.[Results]The calli could be induced from leaves,petioles and stem segments,but the ideal explant was stem segments,with induction rate reaching 98%.The optimal medium for callus proliferation was MS+0.5 mg/L KT+1.0 mg/L 2,4-D+30.0 g/L sucrose.After 3-4 generations of subculture on MS+0.5 mg/L BA+1.0 mg/L 2,4-D+30.0 g/L sucrose,favorable friable calli of H.nepalensis var.sinensis were obtained.[Conclusions]The friable calli induced in this experiment can lay a foundation for in-vitro regeneration and cellular secondary metabolite production of H.nepalensis var.sinensis.