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Agrobacterium-mediated Transformation of Kentucky Bluegrass 被引量:6
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作者 柴宝峰 梁爱华 +1 位作者 王伟 胡炜 《Acta Botanica Sinica》 CSCD 2003年第8期966-973,共8页
Embryogenic calli of Kentucky bluegrass, named Md, were induced from mature seeds and embryos, and proliferated on medium K3 containing 2,4-dichlorophenoxyacetic acid (2,4-D, 10.0 mumol/L), 6-benzylaminopurine (BAR, 0... Embryogenic calli of Kentucky bluegrass, named Md, were induced from mature seeds and embryos, and proliferated on medium K3 containing 2,4-dichlorophenoxyacetic acid (2,4-D, 10.0 mumol/L), 6-benzylaminopurine (BAR, 0.5 mumol/L) and K5 which was the K3 medium supplemented with cupric sulfa (0.5 mumol/L) under dim-light condition (20-30 mumol.m(-2).s-1, 16 h light) at 24 degreesC. Embryogenic calli were transformed with plasmids pDM805 Carring bar and gus genes, Which was mediated by an Agrobacterium strain AGL1, four transgenic lines were obtained. The important factors that affect the transformation efficiency and obtain desirable number of transgenic plants included: (1) the quality of embryogenic calli; (2) light condition and time of co-cultivation; (3) concentration of antibiotics used for suppressing the overgrowth of Agrobacterium in the course of transformed plant regeneration; (4) selection pressure, etc. The micro nutrient of cupric had significant influence on the quality of embryogenic calli. This presentation is the first successful protocol of Kentucky bluegrass transformation mediated by Agrobacterium. 展开更多
关键词 MONOCOTYLEDON CALLUS agrobacterium-mediation Poa pratensis
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Effect of Phytosulfokine-α on Agrobacterium-Mediated Transformation in Rice 被引量:3
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作者 CHEN De-xi Xu Zheng-jun MA Bing-tian LI Shi-gui 《Rice science》 SCIE 2005年第4期255-260,共6页
Phytosulfokine- α (PSK- α ), a biologically active peptide acting as a growth factor, plays a key role in cellular differentiation and proliferation. To test if PSK- α has some influence on agrobacterium-mediated... Phytosulfokine- α (PSK- α ), a biologically active peptide acting as a growth factor, plays a key role in cellular differentiation and proliferation. To test if PSK- α has some influence on agrobacterium-mediated transformation in rice, PSK-α at a series of concentrations was added into co-culture medium respectively. The results showed that PSK- α indeed affected the recovery of resistant calli and the transformation frequency of rice varieties Taipei 309 and Lijiangxintuanheigu, PSK- α at the concentration of 10 nmol/L could increase induction of resistant callus and efficiency of transformation, with a 11% and 4.9% top increase, respectively than the control. However, PSK- αat 200 nmol/L could inhibit the induction of the resistant calli. Further more, the effect of PSK-α on agrobacterium-mediated transformation is related with the concentration of 2, 4-D in selection medium. Higher induction rate of resistant calli was obtained from tissues treated with PSK- α plus 2 mg/L 2, 4-D. 展开更多
关键词 PHYTOSULFOKINE RICE agrobacterium-mediated transformation
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Efficient Regeneration and Agrobacterium-mediated Transformation of Brassica napus Cultivar Qingza No.5
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作者 Yajing XIE Aihua WU Xianjin LIU 《Agricultural Biotechnology》 CAS 2016年第3期48-53,56,共7页
Using the hypocotyl and cotyledon explants of Brassica napus L. cuhivar Qingza No. 5 as receptors, hormone combinations in bud differentiation medi- um, bud growth medium and rooting medium were optimized to establish... Using the hypocotyl and cotyledon explants of Brassica napus L. cuhivar Qingza No. 5 as receptors, hormone combinations in bud differentiation medi- um, bud growth medium and rooting medium were optimized to establish an efficient plantlet regeneration system of B. napus cuhivar Qingza No. 5. The results showed that the highest differentiation efficiency of hypocotyls of B. napus cuhivar Qingza No. 5 reached about 90%, which was three times that of cotyledons. The appropriate differentiation medium was MSB + 5 mg/L thidiazuron (TDZ) +7.5 mg/L AgNO3 + 0.1 mg/L NAA + 2 mg/L proline (L-pro) + 250 mg/L casein acid hydrolysate (CH) + 3% sucrose; the appropriate growth medium was 1/2 MSB + 1 mg/L IBA + 2 mg/L L-pro + 250 mg/L CH + 1.5% sucrose; the ap- propriate rooting medium was 1/2 MSB + 0.2 mg/L IAA + 1.5% sucrose. On this basis, a binary expression vector harboring insect resistance gene B12 was constructed and introduced into B. napus hypocotyls by Agrobacterium-mediated transformation. Positive plants were screened using hygromycin and carbenicillin. Transgenic plants were verified by PCR and GUS histochemical staining. The results showed that insect resistance gene B12 was successfully integrated into the nu- clear genome of B. napus plants and could be expressed normally. Leaves of transgenic plants with high expression levels were collected for indoor inoculation test with Plutella xylotella larvae to evaluate insect resistance of transgenic plants. 展开更多
关键词 words Brassica napus L. Efficient regeneration from hypocotyls agrobacterium-mediated transformation Insect resistance genes GUS staining Transgenicplants
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Agrobacterium-Mediated Transformation of Rice: Constraints and Possible Solutions 被引量:8
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作者 Sulaiman MOHAMMED Azman Abd SAMAD Zaidah RAHMAT 《Rice science》 SCIE CSCD 2019年第3期133-146,共14页
Genetic transformation of rice(Oryza sativa L.) by introducing beneficial traits is now a central research instrument in plant physiology and a practical tool for plant improvement. Many approaches are verified for st... Genetic transformation of rice(Oryza sativa L.) by introducing beneficial traits is now a central research instrument in plant physiology and a practical tool for plant improvement. Many approaches are verified for stable introduction of foreign genes into the plant genome. The review examined the different constraints that limit the success of rice genetic transformation via Agrobacterium-mediated approach and suggested possible solutions. Explant identification, gene transfer technique and construct to tailor the integration, transgene expression without collateral to genetic damage and transformant selection are among the technical challenges affecting the rice transformation. Despite the contests, Agrobacteriummediated transformation system has been a better option for producing transgenic rice varieties because of its exact T-DNA processing and simple integration of low copy-number transgene. This information is necessary for improving the transformation system for recalcitrant rice varieties. 展开更多
关键词 RICE agrobacterium-mediated transformation TISSUE CULTURE gene transfer T-DNA integration
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In planta Agrobacterium-Mediated Transformation of Rice 被引量:3
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作者 Kumrop RATANASUT Weerawan ROD-IN Kawee SUJIPULI 《Rice science》 SCIE CSCD 2017年第3期181-186,共6页
The floral-dip transformation, the simplest technique, is no requirement of tissue culture procedure, and can directly transfer the interest gene into plant reproductive cells. It has been successfully applied to vari... The floral-dip transformation, the simplest technique, is no requirement of tissue culture procedure, and can directly transfer the interest gene into plant reproductive cells. It has been successfully applied to various plant species. In this study, the optimal conditions of a floral-dip method for production of transgenic rice variety RD41 were explored. The simple and effective inoculation medium was composed of Murashige and Skoog(MS) medium, 5% sucrose, 44 nmol/L benzylaminopurine, and 0.075% surfactant Tween-20 with pH 5.7. The transformation efficiencies of Agrobacterium tumefaciens strains AGL1 and EHA105 were compared with the Agrobacterium density at OD_(600) = 0.8–1.0 and the co-cultivation at 25 ℃ for 48 h. A. tumefaciens strain EHA105 gave slightly higher transformation efficiency than AGL1, with statistically non-significant difference. The floral-drop transformation using the optimal floral-dip conditions showed higher transformation efficiency than the floral-dip method, but the dropped flowers turned brown and died within 2 d. Production of transgenic rice variety RD41 by the floral-dip method was achieved using A. tumefaciens strain EHA105 with the optimal conditions. Screening for the gus A gene by PCR using the gus A specific primers in the T_0 lines, there were 4 transgenic lines from 286 T_0 lines(1.4% transformation efficiency). However, histochemical glucuronidase(GUS) assay demonstrated that only three of four transgenic lines exhibited gus A expression. These results indicated that floral-dip transformation is a potential tool for production of the transgenic rice, which can be used for molecular breeding via genetic engineering in the future. 展开更多
关键词 agrobacterium-mediated transformation floral-dip floral-drop transformation efficiency in PLANTA RICE
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Factors Affecting Agrobacterium-Mediated Transformation Efficiency in Rice 被引量:7
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作者 CHENEn-hui ZHANGPing +4 位作者 ZUOShi-min LIAi-hong ZHANGYa-fang CHENZong-xiang PANXue-biao 《Rice science》 SCIE 2004年第4期181-185,共5页
Several important factors affecting the efficiency of Agrobacterium-mediated rice transformation were studied with several predominant commercial indica and japonica rice cultivars. As far as indica rice callus was co... Several important factors affecting the efficiency of Agrobacterium-mediated rice transformation were studied with several predominant commercial indica and japonica rice cultivars. As far as indica rice callus was concerned, CC medium was the best and the quality of callus was improved with the addition of 1.0 to 2.0 mg/L ABA. It decreased the percentage of browning calli and improved the callus growing state by addition of a certain amount of sorbitol to the subculture medium. NB medium was the best for callus initiation of japonica rice, but the improvement in the quality of callus of japonica subspecies was not obvious by adding ABA. During the period of subculture, to a certain degree, increasing the sucrose concentration could improve the proportion of hygromycin resistant calli. Furthermore, the transformation efficiency would be higher by applying selection pressure in the selection stage, removing selection pressure during the plantlet differentiation period and applying selection pressure again during seedling hardening period. Besides, suitable combination of plant hormones was beneficial for callus differentiation. An efficient /Agrobacterium-mediated rice transformation system had been established for several rice cultivars and a lot of transgenic rice plants had been obtained. 展开更多
关键词 RICE AGROBACTERIUM CALLUS HORMONES transformation
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The Establishment of an Agrobacterium-Mediated Transformation Platform for the Non-Embryogenic Calli of Vitis vinifera L. 被引量:3
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作者 ZHAO Feng-xia CHEN Shang-wu +3 位作者 Perl Avihai DAI Ru XU Hai-ying MA Hui-qin 《Agricultural Sciences in China》 CAS CSCD 2011年第5期686-694,共9页
Non-embryogenic calli (NEC) was inevitably and heavily produced when grape embryogenic calli (EC) was induced from explants or during the subculture of EC.A stable and highly efficient NEC transformation platform ... Non-embryogenic calli (NEC) was inevitably and heavily produced when grape embryogenic calli (EC) was induced from explants or during the subculture of EC.A stable and highly efficient NEC transformation platform is required to further sort out and verify key genes which determine/switch the identity of NEC and EC.In this research,a vector pA5 containing a chitinase signal sequence fused to gfp (green fluorescent protein) and an HDEL motive was used to target and immobilize into Agrobacterium strain EHA105 to establish a transformation platform for Vitis vinifera L.cv.Chardonnay NEC.It was determined that NEC 10 d after subculture was the best target tissue;30 min for inoculation followed by 3 d co-cultivation with the addition of 200 μmol L-1 acetosyringone (AS) was optimized as protocol.The use of bacterial densities as 1.0 at OD600 did not result in serious tissue hypersensitive reaction and it had higher efficiency.Kanamycin at 200 mg L-1 was picked for positive expression selection.The stable transformation of NEC was proved by reverse transcription-polymerase chain reaction techniques (RT-PCR) and fluorescent microscopy after three sub-cultures of the selected cell line.Highly efficient genetic transformation protocol of grape NEC was achieved and some of the optimized parameters were different from that reported for EC.This transformation platform could facilitate the verification of candidate somatic embryogenesis (SE) decisive genes,and the successfully transformed NEC with certain genes can also be used as bioreactors for the production of functional products,as NEC not only proliferates fast,but also keeps in a rather stable condition. 展开更多
关键词 Agrobacterium tumefaciens non-embryogenic calli transformation Vitis vinifera L. RT-PCR
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Study on Agrobacterium-Mediated Transformation of Pepper with Barnase and Cre Gene 被引量:4
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作者 LIU Juan-xu YU Yi-xun +2 位作者 LEI Jian-jun CHEN Guo-ju CAO Bi-hao 《Agricultural Sciences in China》 CAS CSCD 2009年第8期947-955,共9页
This study was designed to control plant fertility by cell lethal gene Barnase expressing at specific developmental stage and in specific tissue of male organ under the control of Cre/lox system, for heterosis breedin... This study was designed to control plant fertility by cell lethal gene Barnase expressing at specific developmental stage and in specific tissue of male organ under the control of Cre/lox system, for heterosis breeding of chili pepper (Capsicum annuum L.). Chili pepper inbred lines (A, D, E, and I) were transformed with Cre gene and Barnase gene situated between loxp, separately, by means of Agrobacterium co-culture. In this study, we had established a high transformation system by extensive study of affecting factors including genotype, selection of marker, and lethal dose. Cotyledon with petiole from 9-11-day-old seeding was pre-cultured on media MR[MB(MS mineral+vitamine B5)+BA(6-Benzyladenine) 5.0 mg·L^-1 +IAA(indoleacetic acid) 1.0 mg·L^-1+GA3(gibberellic acid) 1.0mg·L^-1+sucrose 3%+agar 6.5g·L^-1] for 2d. The explants were infected by Agrobacterium tumefaciens when their OD600(optical density at 600 nm)reached 0.6-0.9. After co-cultured for 4-5 d on media MC [MB+BA5.0 mg·L^-1+IAA 1.0 mg·L^-1 +GA3 1.0 mg·L^-1+sucrose 3% +agar 6.5 g·L^-1+AS (acetosyringone) 200μmol·L^-1, these cotyledons with petiole were cultured on selective differentiation medium in the media MT[MB medium supplemented with BA [5.0 mg·L^-1+ IAA 1.0 mg·L^-1+ GA3 1.0 mg·L^-1+ AgNO3 5.0 mg·L^-1+ CW (coconut water) 5% + Km (kanamycin) 65 mg·L^-1+ Cb (carbenicillin) 500 mg·L^-1+ 3% sucrose + agar 6.5 g·L^-1].The Kmr (kanamycin resistant) bud rosettes were elongated on selective elongation medium and rooted on rooting medium. PCR and Southern blotting analysis of Kmr plantlet indicated that the foreign genes had been integrated into the genome of pepper. The transgenic plants with Cre gene developed well, blossomed out, and set fruit normally. The transgenic plants with Barnase gene grew well with normal appearance of flower, but they showed different fertility from complete sterility, partial sterility to complete fertility, and similar results were obtained from in vitro pollen germination experiments. 展开更多
关键词 pepper (Capsicum annuum L.) genetic transformation male sterile TA29-Barnase Cre/lox system
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Successful Agrobacterium-mediated transformation of Populus tomentosa with apple SPDS gene 被引量:2
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作者 LIU Ting-ting PANG Xiao-ming LONG Cui ZHANG Zhi-yi 《Forestry Studies in China》 CAS 2008年第3期153-157,共5页
The problem of salinized soils has become one of the most serious constraints to agricultural and forest productivity. With the purpose of enhancing salt stress tolerance of Populus tomentosa, we transformed this tree... The problem of salinized soils has become one of the most serious constraints to agricultural and forest productivity. With the purpose of enhancing salt stress tolerance of Populus tomentosa, we transformed this tree species with spermidine synthase (SPDS) genes derived from an apple by an Agrobacterium-mediated method. Four transgenic clones were confu'med by PCR and Southern blot analysis. As well, the expression of introduced SPDS genes was analyzed by real-time quantitative PCR. 展开更多
关键词 salt tolerance spermidine synthase gene transformation Populus tomentosa
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Rapid regeneration of stable transformants in cultures of potato by improving factors influencing Agrobacterium-mediated transformation 被引量:2
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作者 Bipasha Chakravarty Gefu Wang-Pruski 《Advances in Bioscience and Biotechnology》 2010年第5期409-416,共8页
An efficient and rapid Agrobacterium tumefaciens-mediated transformation protocol was developed to generate activation-tagged mutant lines with the aim of large-scale functional analysis of the potato genome. The expl... An efficient and rapid Agrobacterium tumefaciens-mediated transformation protocol was developed to generate activation-tagged mutant lines with the aim of large-scale functional analysis of the potato genome. The explants were inoculated with an Agrobacterium strain harboring the binary plasmid pSKI074 containing four CaMV 35S enhancers in the T-DNA region which activates the downstream genes in the host plant after its integration. Various parameters investigated to increase transformation efficiency were the type and age of explant, cultivar, hormone combinations, preculture of explants, period of co-cultivation with bacteria and concentration of bacterial cultures used for transformation. Stem explants from 5 week old plantlets of cv. Bintje which had undergone phytohormone pretreatment for 4 days, inoculation with diluted bacterial concentration of OD600 = 0.2 containing acetosyringone followed by 2 days of co-cultivation and selection in media with IAA and trans-zeatin all helped in greatly improving the transformation efficiency. The total time required from infection to rooted shoots was 6-7 weeks. Initial evidence for stable integration and expression of the transgenes by PCR analysis showed that over 93% of the regenerated lines were transgenic and this was confirmed by Southern hybridization. 展开更多
关键词 BACTERIAL Concentration CULTIVAR EXPLANT POTATO transformation Preculture
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Agrobacterium-Mediated Transformation of Embryogenic Calli of Anliucheng and Regeneration of Plants Containing the Chimaeric Ribonuclease Gene
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作者 LI Dong-dong, SHI Wei and DENG Xiu-xinNational Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University , Wuhan 430070 , P. R. China 《Agricultural Sciences in China》 CAS CSCD 2003年第2期127-131,共5页
Anliucheng (Citrus sinensis Osbeck), a very seedy and widely spread acidless sweet orange cul-tivar in south of China, was transformed by the strain of Agrobacterium Tumefaciens EHA105 carrying pTA29-barnase gene, whi... Anliucheng (Citrus sinensis Osbeck), a very seedy and widely spread acidless sweet orange cul-tivar in south of China, was transformed by the strain of Agrobacterium Tumefaciens EHA105 carrying pTA29-barnase gene, which will induce pollen sterility in transgenic plants. The embryogenic calli of Anliucheng were co-cultivated with Agrobacterium tumefaciens for 3 days, and then transferred to selective medium containing 50 mg I/1 basta (a kind of herbicide) for 5 weeks. The resistant calli were recovered and regenerated 118 embryoids. A total of 13 entire plants were obtained after micro-grafted on trifoliate orange. These regenerated plants were verified by PCR amplification and confirmed by PCR-Southern blotting analysis. 展开更多
关键词 CITRUS CALLI pTA-barnase Genetic transformation agrobacterium-mediated
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A novel system for Agrobacterium-mediated transformation of wheat(Triticum aestivum L.) cells 被引量:1
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作者 XUYAO BAOJIANLI JINGFENJIA 《Cell Research》 SCIE CAS CSCD 1993年第1期48-48,50-60,共12页
A new approach for transforming the cultured cells of wheat(Triticum aestivum L. cv. Ganmai 8) was developed using Agrobacterium tumefaciens. The features of the optimum procedure were: (a) both combined synthetic sig... A new approach for transforming the cultured cells of wheat(Triticum aestivum L. cv. Ganmai 8) was developed using Agrobacterium tumefaciens. The features of the optimum procedure were: (a) both combined synthetic signal molecules and multiple natural extracts from susceptible plants were used to pretreat the primary vigorous Agrobacterium(PVA) cells for approximately 16 h; (b) the gyratory magnetic field condition was used during cocultivation; (c) the cocultivating period and selecting condition were modified; (d) the recipient cells were at exuberant metabolism and active division while infected with Agrobacterium. Both neomycin phosphotransferase and nopaline synthase assays demonstrated the expression of NPT II and NOS genes, located on the T-DNA segment of chimaeric plasmid pGV3850::1103neo, in transformed wheat cell colonies by adopting the techniques of dot blot, ndPAGE or high voltage paper electrophoresis. Integration of the foreign genes into wheat genome was confirmed by Southern blot hybridization. Moreover, a relatively rational method was described for the estimation of transformation frequencies from cultured cell levels. 展开更多
关键词 genetic engineering gyratory magnetic field natural susceptible extracts synthetic signals transformation system wheat cells.
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High-efficiency Agrobacterium-mediated transformation of chrysanthemum via vacuum infiltration of internode 被引量:1
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作者 Sujuan Xu Ze Wu +4 位作者 Jingya Zhao Fengjiao Zhang Xinqi Zhang Fadi Chen Nianjun Teng 《Ornamental Plant Research》 2022年第1期1-7,共7页
Leaf disc transformation is one of the traditional methods that are now widely used in chrysanthemum with highly economical and ornamental value in world flower production,but it depends on plant genotypes and is time... Leaf disc transformation is one of the traditional methods that are now widely used in chrysanthemum with highly economical and ornamental value in world flower production,but it depends on plant genotypes and is time consuming and complicated.In addition,the transformation success rate of this method is low,generally ranging from 0.1%to 6.25%.Therefore,a highly efficient transformation system is needed.In this study,we are the first to establish a high-efficient chrysanthemum Agrobacterium-mediated transformation system via vacuum infiltration.Chrysanthemum stem internode explants were used as research material and CmLEC1 was used as a reporter gene.After approximately 3 months of culture and selection,the positive transgenic plants were obtained.Additionally,the positive probability was about 42%.The transformation efficiency was up to 37.7%,and if the escapes were removed,it was 16%.Furthermore,stable expression of CmLEC1 in transgenic'Yuhualuoying'was confirmed by qRT-PCR analysis.These results suggest that this genetic transformation system via vacuum infiltration of chrysanthemum stem internode is highly efficient and convenient,and much better than traditional leaf disc transformation,and it will play an important role in chrysanthemum transformation and functional genetics research. 展开更多
关键词 transformation VACUUM removed
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Mutant acetolactate synthase (ALS) gene as a selectable marker for Agrobacterium-mediated transformation of soybean
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作者 陈士云 Zhang Yong 《High Technology Letters》 EI CAS 2006年第2期188-192,共5页
Soybean is one of the crops most difficult to be manipulated in vitro. Although several soybean marker genes, all the selectable markers used were from bacteria origin. To find suitable selectable marker gene from pla... Soybean is one of the crops most difficult to be manipulated in vitro. Although several soybean marker genes, all the selectable markers used were from bacteria origin. To find suitable selectable marker gene from plant origin for soybean transformation, a mutant acetolactate synthase (ALS) gene from Arabidopsis thaliana was tested for Agrobacterium-mediated soybean embryo axis transformation with the herbicide Arsenal as the selective agent. Transgenic soybean plants were obtained after the herbicide se- lection and the To transgenic lines showed resistance to the herbicide at a concentration of 100 g/ha. ALS enzyme assay of To transgenic line also showed higher activity compared to the wild type control plant. PCR analysis of the T1 transgenic lines confirmed the integration and segregation of the transgene. Taken together, our results showed that the mutant ALS gene is a suitable selectable marker for soybean transformation. 展开更多
关键词 SOYBEAN transformation AGROBACTERIUM ALS selectable marker
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Agrobacterium-mediated Transformation and Regeneration by Direct Shoot Organogenesis in Cotton (G. hirsutum)
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作者 G.BALASUBRAMANI J.AMUDHA +2 位作者 P.A.KUMAR A.B.DONGRE C.D.MAYEE 《棉花学报》 CSCD 北大核心 2002年第S1期87-87,共1页
Genotype independent transformation andregeneration of Indian cotton(Gossypiumhirsutum L.)cultivar was standardized with Bt-Cry 1A(b)gene by Agrobacterium-mediation.Apical meristem of elite G.hirsutum cultivarLRK-516 ... Genotype independent transformation andregeneration of Indian cotton(Gossypiumhirsutum L.)cultivar was standardized with Bt-Cry 1A(b)gene by Agrobacterium-mediation.Apical meristem of elite G.hirsutum cultivarLRK-516 and LRA 5166 were co-cultivated withA.tumefaciens LBA 4404 carrying synthetic Bt- 展开更多
关键词 COTTON AGROBACTERIUM COTTON ELITE CULTIVAR transformation REGENERATION carrying standardized MEDIATION
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Agrobacterium-Mediated Transformation of Cry8db Gene in Vietnam Sweet Potato Cultivar
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作者 Pham Bich Ngoc Vu Thi Lan +4 位作者 Tran Thu Trang Nguyen Hoai Thuong Le Thu Ngoc Chu Hoang Ha Le Tran Binh 《Journal of Life Sciences》 2015年第6期262-271,共10页
Sweet potato [Ipomoea batatas (L.) Lam.] is an important food crop in the world as well as in Vietnam. It is well known as a recalcitrant crop for gene transformation and tissue culture because of its genotype depen... Sweet potato [Ipomoea batatas (L.) Lam.] is an important food crop in the world as well as in Vietnam. It is well known as a recalcitrant crop for gene transformation and tissue culture because of its genotype dependent in vitro responses. In present study, Agrobacterium-mediated transformation of cry8Db from Bacillus thuringiensis into KB 1 sweet potato variety has been studied. The C58cv strain carrying a pBl 121 backbone which contained cry8Db delta-endotoxin gene regulated under 35 S CaMV prom oter, and the selection marker gene, neomycin phosphotransferase (npt11) gene, was subjected for plant transformation. Callus induced from shoot tips and leaf explants were inoculated and cocultured with A. tumefaciens. The selection occurred during callus producing and plant regenerating steps. A total of 201 transgenic putative plant lines were produced, and 21 transgenic lines were positively confirmed by PCR and finalized by Southern blot. Four putative transgenic lines confirming a single copy of the crySEIb gene were transferred into soil pots in greenhouse. Biological activity evaluation for the insecticidal capacity of these transgenic lines under controlled conditions showed that the level of infestation by sweet potato weevils (Cylasformicarius) in untransformed plants was higher than that of transgenic lines. 展开更多
关键词 Sweet potato Agrobacterium tumefaciens plant transformation cry8Db sweet potato weevil resistance cylas formicarius.
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Development of an Agrobacterium-mediated CRISPR/Cas9 gene editing system in jute(Corchorus capsularis)
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作者 Shaolian Jiang Qin Li +9 位作者 Xiangxue Meng Mengxin Huang Jiayu Yao Chuanyu Wang Pingping Fang Aifen Tao Jiantang Xu Jianmin Qi Shuangxia Jin Liwu Zhang 《The Crop Journal》 SCIE CSCD 2024年第4期1266-1270,共5页
Jute(Corchorus capsularis L.)is the second most important natural plant fiber source after cotton.However,developing an efficient gene editing system for jute remains a challenge.In this study,the transgenic hairy roo... Jute(Corchorus capsularis L.)is the second most important natural plant fiber source after cotton.However,developing an efficient gene editing system for jute remains a challenge.In this study,the transgenic hairy root system mediated by Agrobacterium rhizogenes strain K599 was developed for Meifeng 4,an elite jute variety widely cultivated in China.The transgenic hairy root system for jute was verified by subcellular localization and bimolecular fluorescence complementation(BiFC)assays.The CHLOROPLASTOS ALTERADOS 1(CcCLA1)gene,which is involved in the development of chloroplasts,was targeted for editing at two sites in Meifeng 4.Based on this hairy root transformation,the gRNA scaffold was placed under the control of cotton ubiquitin GhU6.7 and-GhU6.9 promoters,respectively,to assess the efficiency of gene editing.Results indicated the 50.0%(GhU6.7)and 38.5%(GhU6.9)editing events in the target 2 alleles(gRNA2),but no mutation was detected in the target 1 allele(gRNA1)in transgenic-positive hairy roots.CcCLA1 gene editing at gRNA2 under the control of GhU6.7 in Meifeng 4 was also carried out by Agrobacterium tumefaciens-mediated transformation.Two CcCLA1 mutants were albinic,with a gene editing efficiency of 5.3%.These findings confirm that the CRISPR/Cas9 system,incorporating promoter GhU6.7,can be used as a gene editing tool for jute. 展开更多
关键词 JUTE agrobacterium-mediated transformation Genome editing Hairy root system
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Hemorrhagic transformation in patients with large-artery atherosclerotic stroke is associated with the gut microbiota and lipopolysaccharide 被引量:1
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作者 Qin Huang Minping Wei +3 位作者 Xianjing Feng Yunfang Luo Yunhai Liu Jian Xia 《Neural Regeneration Research》 SCIE CAS CSCD 2024年第7期1532-1540,共9页
Hemorrhagic transformation is a major complication of large-artery atheroscle rotic stroke(a major ischemic stro ke subtype)that wo rsens outcomes and increases mortality.Disruption of the gut microbiota is an importa... Hemorrhagic transformation is a major complication of large-artery atheroscle rotic stroke(a major ischemic stro ke subtype)that wo rsens outcomes and increases mortality.Disruption of the gut microbiota is an important feature of stroke,and some specific bacteria and bacterial metabolites may contribute to hemorrhagic transformation pathogenesis.We aimed to investigate the relationship between the gut microbiota and hemorrhagic transformation in largearte ry atheroscle rotic stro ke.An observational retrospective study was conducted.From May 2020 to September 2021,blood and fecal samples were obtained upon admission from 32 patients with first-ever acute ischemic stroke and not undergoing intravenous thrombolysis or endovascular thrombectomy,as well as 16 healthy controls.Patients with stro ke who developed hemorrhagic transfo rmation(n=15)were compared to those who did not develop hemorrhagic transformation(n=17)and with healthy controls.The gut microbiota was assessed through 16S ribosomal ribonucleic acid sequencing.We also examined key components of the lipopolysaccharide pathway:lipopolysaccharide,lipopolysaccharide-binding protein,and soluble CD14.We observed that bacterial diversity was decreased in both the hemorrhagic transformation and non-hemorrhagic transfo rmation group compared with the healthy controls.The patients with ischemic stro ke who developed hemorrhagic transfo rmation exhibited altered gut micro biota composition,in particular an increase in the relative abundance and dive rsity of members belonging to the Enterobacteriaceae family.Plasma lipopolysaccharide and lipopolysaccharide-binding protein levels were higher in the hemorrhagic transformation group compared with the non-hemorrhagic transfo rmation group.lipopolysaccharide,lipopolysaccharide-binding protein,and soluble CD14 concentrations were associated with increased abundance of Enterobacte riaceae.Next,the role of the gut microbiota in hemorrhagic transformation was evaluated using an experimental stroke rat model.In this model,transplantation of the gut microbiota from hemorrhagic transformation rats into the recipient rats triggered higher plasma levels of lipopolysaccharide,lipopolysaccharide-binding protein,and soluble CD14.Ta ken togethe r,our findings demonstrate a noticeable change in the gut microbiota and lipopolysaccharide-related inflammatory response in stroke patients with hemorrhagic transformation.This suggests that maintaining a balanced gut microbiota may be an important factor in preventing hemorrhagic transfo rmation after stro ke. 展开更多
关键词 gut microbiota hemorrhagic transformation INFLAMMATION LIPOPOLYSACCHARIDE STROKE
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Formation and transformation of metastable LPSO building blocks clusters in Mg-Gd-Y-Zn-Zr alloys by spinodal decomposition and heterogeneous nucleation 被引量:1
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作者 Xin Zhao Zhong Yang +2 位作者 Jiachen Zhang Minxian Liang Liying Wang 《Journal of Magnesium and Alloys》 SCIE EI CAS CSCD 2024年第2期673-686,共14页
To study the formation and transformation mechanism of long-period stacked ordered(LPSO)structures,a systematic atomic scale analysis was conducted for the structural evolution of long-period stacked ordered(LPSO)stru... To study the formation and transformation mechanism of long-period stacked ordered(LPSO)structures,a systematic atomic scale analysis was conducted for the structural evolution of long-period stacked ordered(LPSO)structures in the Mg-Gd-Y-Zn-Zr alloy annealed at 300℃~500℃.Various types of metastable LPSO building block clusters were found to exist in alloy structures at different temperatures,which precipitate during the solidification and homogenization process.The stability of Zn/Y clusters is explained by the first principles of density functional theory.The LPSO structure is distinguished by the arrangement of its different Zn/Y enriched LPSO structural units,which comprises local fcc stacking sequences upon a tightly packed plane.The presence of solute atoms causes local lattice distortion,thereby enabling the rearrangement of Mg atoms in the different configurations in the local lattice,and local HCP-FCC transitions occur between Mg and Zn atoms occupying the nearest neighbor positions.This finding indicates that LPSO structures can generate necessary Schockley partial dislocations on specific slip surfaces,providing direct evidence of the transition from 18R to 14H.Growth of the LPSO,devoid of any defects and non-coherent interfaces,was observed separately from other precipitated phases.As a result,the precipitation sequence of LPSO in the solidification stage was as follows:Zn/Ycluster+Mg layers→various metastable LPSO building block clusters→18R/24R LPSO;whereas the precipitation sequence of LPSO during homogenization treatment was observed to be as follows:18R LPSO→various metastable LPSO building block clusters→14H LPSO.Of these,14H LPSO was found to be the most thermodynamically stable structure. 展开更多
关键词 LPSO Spinodal decomposition Homogenization treatment CLUSTERS Phase transformation
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Defect Detection Model Using Time Series Data Augmentation and Transformation 被引量:1
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作者 Gyu-Il Kim Hyun Yoo +1 位作者 Han-Jin Cho Kyungyong Chung 《Computers, Materials & Continua》 SCIE EI 2024年第2期1713-1730,共18页
Time-series data provide important information in many fields,and their processing and analysis have been the focus of much research.However,detecting anomalies is very difficult due to data imbalance,temporal depende... Time-series data provide important information in many fields,and their processing and analysis have been the focus of much research.However,detecting anomalies is very difficult due to data imbalance,temporal dependence,and noise.Therefore,methodologies for data augmentation and conversion of time series data into images for analysis have been studied.This paper proposes a fault detection model that uses time series data augmentation and transformation to address the problems of data imbalance,temporal dependence,and robustness to noise.The method of data augmentation is set as the addition of noise.It involves adding Gaussian noise,with the noise level set to 0.002,to maximize the generalization performance of the model.In addition,we use the Markov Transition Field(MTF)method to effectively visualize the dynamic transitions of the data while converting the time series data into images.It enables the identification of patterns in time series data and assists in capturing the sequential dependencies of the data.For anomaly detection,the PatchCore model is applied to show excellent performance,and the detected anomaly areas are represented as heat maps.It allows for the detection of anomalies,and by applying an anomaly map to the original image,it is possible to capture the areas where anomalies occur.The performance evaluation shows that both F1-score and Accuracy are high when time series data is converted to images.Additionally,when processed as images rather than as time series data,there was a significant reduction in both the size of the data and the training time.The proposed method can provide an important springboard for research in the field of anomaly detection using time series data.Besides,it helps solve problems such as analyzing complex patterns in data lightweight. 展开更多
关键词 Defect detection time series deep learning data augmentation data transformation
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