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红比利时杜鹃愈伤悬浮颗粒瞬时转化体系构建及植株再生 被引量:1
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作者 何凡 吴月燕 +5 位作者 谢晓鸿 杨国霞 蒋宝鑫 李东宾 鄢毅铖 贾永红 《核农学报》 CAS CSCD 北大核心 2022年第8期1546-1558,共13页
为建立红比利时杜鹃花愈伤悬浮颗粒瞬时转化体系,以嫩叶为材料,探讨了愈伤组织诱导与增殖的条件,构建并评价了愈伤悬浮颗粒瞬时转化体系,同时诱导愈伤颗粒出芽、生根,并获得组培小苗。结果表明,在2.41 g·L^(-1)木本植物基本培养基(... 为建立红比利时杜鹃花愈伤悬浮颗粒瞬时转化体系,以嫩叶为材料,探讨了愈伤组织诱导与增殖的条件,构建并评价了愈伤悬浮颗粒瞬时转化体系,同时诱导愈伤颗粒出芽、生根,并获得组培小苗。结果表明,在2.41 g·L^(-1)木本植物基本培养基(WPM)+20 g·L^(-1)蔗糖+10 g·L^(-1)麦芽糖+7.0 g·L^(-1)琼脂+0.050 mg·L^(-1)植物组培抗菌剂(PPM)为基本培养基的条件下,嫩叶愈伤组织诱导的生长调节剂最优方案为:0.3 mg·L^(-1)2,4-二氯苯氧乙酸(2,4-D)+0.3 mg·L^(-1)噻重氮苯基脲(TDZ),此时愈化率可达97.78%;愈伤组织继代增殖的生长调节剂方案为:0.61 mg·L^(-1)2,4-二氧苯氧乙酸(2,4-D)+0.65 mg·L^(-1)6-苄基腺嘌呤(6-BA)+1.37 mg·L^(-1) TDZ,此时增殖率可达167%。将继代5次后呈疏松状的愈伤颗粒接种于液体继代培养基中,大约4~12 d后愈伤颗粒进入指数生长阶段,增殖率为45%。含有GUS基因的pRI 101-AN与含有GFP基因的pCAMBIA1301-GFP分别在农杆菌GV3101介导下转化处于指数生长期的愈伤悬浮颗粒,OD_(600)为0.6的农杆菌侵染液侵染15 min后,愈伤颗粒的GUS染色效率为26.28%,愈伤颗粒的GFP荧光表达效果明显。另外,以2.41 g·L^(-1) WPM+7 g·L^(-1)琼脂+20 g·L^(-1)蔗糖为基本培养基,添加2.0 mg·L^(-1)TDZ+0.5 mg·L^(-1)2,4-D时,可诱导愈伤组织不定芽;以1/2 Read为基本培养基,添加0.5 mg·L^(-1) IBA+1 g·L^(-1)活性炭可诱导出芽的愈伤颗粒生根。本研究结果为进一步开展红比利时杜鹃花稳定遗传转化研究和转基因植株培育奠定了基础。 展开更多
关键词 红比利时杜鹃 愈伤悬浮颗粒 瞬时转化 再生
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根癌农杆菌介导的苜蓿悬浮胚性愈伤遗传转化体系的建立与优化 被引量:3
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作者 申玉华 李望丰 +4 位作者 金太成 常青 殷东旭 王德利 刘立侠 《植物研究》 CAS CSCD 北大核心 2009年第6期721-727,共7页
苜蓿基因型是限制遗传转化的关键因素之一。本实验通过对7种苜蓿胚性愈伤组织诱导,筛选出一份具有高频再生潜力的基因型公农-1号,并以该基因型为转化平台探索和建立了一套高效的苜蓿遗传转化系统。分析了影响农杆菌共培养转化苜蓿悬浮... 苜蓿基因型是限制遗传转化的关键因素之一。本实验通过对7种苜蓿胚性愈伤组织诱导,筛选出一份具有高频再生潜力的基因型公农-1号,并以该基因型为转化平台探索和建立了一套高效的苜蓿遗传转化系统。分析了影响农杆菌共培养转化苜蓿悬浮胚性愈伤的因素,优化了悬浮培养条件,建立的超声波辅助农杆菌介导苜蓿胚性愈伤的遗传转化系统为:以下胚轴诱导的愈伤组织经悬浮培养得到的胚性愈伤为转化材料,乙酰丁香酮为100μmol·L-1、超声波处理时间8s、卡那霉素筛选浓度30mg·L-1、共培养4d,接种于选择培养基上进行筛选和再生。最终,获得了大量的转基因植株,分子检测证实目的基因-角碱蓬液泡膜型Na+/H+逆向转运蛋白基因已经整合到苜蓿基因组中。 展开更多
关键词 苜蓿 悬浮胚性 NHX1基因 遗传转化
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铈对马铃薯悬浮培养的愈伤组织中花青苷积累及其生物合成基因表达的影响
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作者 卢其能 杨清 邹华文 《中国稀土学报》 CAS CSCD 北大核心 2006年第4期514-514,共1页
马铃薯中的花青苷主要为锦蔡素和天竺蔡素的衍生物。花青苷的产出率与生物量和花青苷的含量密切相关。本研究表明,0.1mmol·L^-1Ce^4+能促进细胞和愈伤组织的生长,而1mmol·L^-1Ce^4+抑制生长,2mmol·L^-1Ce^4+对细... 马铃薯中的花青苷主要为锦蔡素和天竺蔡素的衍生物。花青苷的产出率与生物量和花青苷的含量密切相关。本研究表明,0.1mmol·L^-1Ce^4+能促进细胞和愈伤组织的生长,而1mmol·L^-1Ce^4+抑制生长,2mmol·L^-1Ce^4+对细胞有害而导致细胞死亡。不同浓度牟Ce^4+能诱导正常的细胞和愈伤组织变红并积累花青苷。用0.1mmol·L^-1Ce^4+处理15d的细胞和愈伤组织花青苷的含量最高,其次是1mmol·L^-1Ce^4+处理18d的细胞和愈伤组织花青苷的含量,而最低浓度(0.01mmol·L^-1Ce^4+)和最高浓度(2mmol·L^-1Ce^4+)不利于花青苷的产生和积累。半定量RT-PCR分析表明,用0.1mmol·L^-1Ce^4+处理悬浮培养的愈伤组织能显著诱导花青苷生物合成途径中五个基因(CHS,F3H,F3′5′H,DFR和3GT)的表达,其表达量与花青量的含量呈正相关。 展开更多
关键词 花青苷 基因表达 组织悬浮培养 马铃薯(Solanum TUBEROSUM cv.Chieftain) 稀土
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非洲菊胚性愈伤组织悬浮培养方法建立及其诱变条件研究
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作者 忻雅 赵杭苹 +1 位作者 钱丽华 童建新 《杭州农业科技》 2007年第1期21-23,共3页
以紫罗兰绿心、洋红黑心、绿心黄瓣、鹅黄黑心的花托和花梗,白色黑心的叶片为外植体,在pH值为5.8、蔗糖30g/L和琼脂8g/L的条件下诱导愈伤组织,筛选出的最佳愈伤组织诱导培养基配方是MS+6B—A1.0mg/L+NAA0.2mg/L+2,4-D1.... 以紫罗兰绿心、洋红黑心、绿心黄瓣、鹅黄黑心的花托和花梗,白色黑心的叶片为外植体,在pH值为5.8、蔗糖30g/L和琼脂8g/L的条件下诱导愈伤组织,筛选出的最佳愈伤组织诱导培养基配方是MS+6B—A1.0mg/L+NAA0.2mg/L+2,4-D1.0mg/L诱导出的愈伤转到MS+6-BA2.0mg/L+NAA0.2mg/L。适合非洲菊固体和液体培养基上愈伤增长的配方是MS+6-BA2.0mg/L+NAA0.2mg/L。20Cy的Co^60辐射剂量和0.1%(V/V)的EMS浓度是较适合的诱变剂量.可以作为半致死剂量进行下一步大批量愈伤组织的诱变处理,从中筛选耐寒或花色特异等突变体。 展开更多
关键词 非洲菊 组织悬浮培养 诱变育种
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冬枣花药愈伤组织的诱导及原生质体分离 被引量:12
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作者 刘晓光 刘孟军 +3 位作者 宁强 彭艳芳 苗利军 秦子禹 《中国农学通报》 CSCD 北大核心 2009年第2期100-104,共5页
【研究目的】研究冬枣花药愈伤组织的诱导,悬浮系的建立和培养及愈伤悬浮系原生质体的分离。【方法】以冬枣花药为试材,通过选择愈伤诱导培养基和原生质体分离所用酶的浓度找到最佳诱导和分离条件。【结论】使用1/2MS基本培养基附加TDZ0... 【研究目的】研究冬枣花药愈伤组织的诱导,悬浮系的建立和培养及愈伤悬浮系原生质体的分离。【方法】以冬枣花药为试材,通过选择愈伤诱导培养基和原生质体分离所用酶的浓度找到最佳诱导和分离条件。【结论】使用1/2MS基本培养基附加TDZ0.2mg/L、NAA0.5mg/L和PVP2.0g/L,对诱导冬枣花药愈伤组织有较好效果;愈伤组织增殖采用培养基1/2MS+TDZ0.4mg/L+NAA0.2mg/L;悬浮细胞系培养采用1/2MS+TDZ0.4mg/L+NAA0.2mg/L液体培养基;冬枣花药愈伤组织悬浮系原生质体分离时以0.6M甘露醇+0.1%MES+20~25g/L纤维素酶,酶解时间为16h时得到的原生质体产量和活力较高。 展开更多
关键词 冬枣 花药 组织悬浮 原生质体 分离
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马瑟兰葡萄体细胞胚再生体系的建立 被引量:1
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作者 舒河霖 曹慧玲 +3 位作者 邵建辉 于坤淼 郭盼 马春花 《云南农业大学学报(自然科学版)》 CAS CSCD 北大核心 2023年第1期112-121,共10页
[目的]建立欧亚种葡萄马瑟兰体细胞胚再生体系,为其遗传转化体系的建立奠定基础。[方法]以马瑟兰的花蕾和未成熟种子为外植体,研究2,4-二氯苯氧乙酸、褪黑素、6-苄氨基嘌呤和噻苯隆激素不同组合(Fow1~6和Ims1~6)对其体细胞胚的诱导作用... [目的]建立欧亚种葡萄马瑟兰体细胞胚再生体系,为其遗传转化体系的建立奠定基础。[方法]以马瑟兰的花蕾和未成熟种子为外植体,研究2,4-二氯苯氧乙酸、褪黑素、6-苄氨基嘌呤和噻苯隆激素不同组合(Fow1~6和Ims1~6)对其体细胞胚的诱导作用。[结果]马瑟兰花蕾在Fow6培养基上以及未成熟种子在Ims2和Ims5培养基上愈伤诱导率最高,可达67.19%~78.00%;将其培养12~14周后的愈伤组织转移到EM6培养基上持续培养,花蕾愈伤组织未能得到体细胞胚,而来源于Ims3培养基的未成熟种子愈伤组织具有较高的体细胞胚发生率(6.77%),并可在EM6培养基上正常成苗;SCM6培养基有利于愈伤组织悬浮培养,可获得大量增殖良好的非胚性细胞团,细胞学观察结果表明愈伤细胞状态良好。[结论]以马瑟兰未成熟种子为外植体,首次建立了其体细胞胚再生体系和愈伤组织悬浮培养体系,对其遗传转化体系的建立、品质相关基因功能验证和分子育种等研究具有良好的支撑作用。 展开更多
关键词 葡萄 马瑟兰 体细胞胚发生 植株再生 愈伤悬浮
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Optimization of Inducing Conditions for Loose Callus of Pinellia ternata 被引量:3
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作者 朱艳芳 毛春娜 +3 位作者 张爱民 薛建平 李佳娣 宋运贤 《Agricultural Science & Technology》 CAS 2011年第8期1116-1117,1124,共3页
[Objective] The research aimed to establish the cell suspension culture system of Pinellia ternate.[Method] The petiole of Pinellia ternate was as the explant.The orthogonal test was used to study the influences of fo... [Objective] The research aimed to establish the cell suspension culture system of Pinellia ternate.[Method] The petiole of Pinellia ternate was as the explant.The orthogonal test was used to study the influences of four plant growth substances(2,4-D,NAA,picloram and KT)and their mixture ratios on the formation of loose callus.[Result] The induction effect of 2,4-D and picloram on the petiole callus of Pinellia ternate was the most significant.Then,the second ones were KT and NAA.The optimal medium which induced Pinellia ternate petiole to form the loose callus was MS+0.5 mg/L of 2,4-D+1.0 mg/L of NAA+1.0 mg/L of picloram+1.5 mg/L of KT.[Conclusion] The research laid the foundation for extracting the active ingredient from the cell suspension of Pinellia ternate and producing the artificial seed. 展开更多
关键词 Pinellia ternata PETIOLE Loose callus Suspension culture
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Callus Induction and Cell Suspension Culture of Eggplant(Solanum melongena L.) 被引量:1
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作者 王凤华 李光远 +2 位作者 陈双臣 蒋燕 王少先 《Agricultural Science & Technology》 CAS 2013年第9期1220-1223,1243,共5页
[Objective] The aim of this article is to establish a cell suspension culture system for eggplant. [Method] Using orthogonal design, appropriate media were screened for callus induction, subculture and cell suspension... [Objective] The aim of this article is to establish a cell suspension culture system for eggplant. [Method] Using orthogonal design, appropriate media were screened for callus induction, subculture and cell suspension culture. [Result] The appropriate medium for callus induction was MS supplemented with 0.2 mg/L 6-BA and 0.2 mg/L NAA; the appropriate subculture medium was MS supplemented with0.2 mg/L 6-BA, 0.2 mg/L NAA, and 0.1 mg/L KT; the suitable medium for cell suspension culture was liquid MS medium supplemented with 0.4 mg/L NAA and 0.2mg/L. Cell growth curve was similar to "S" shape in the above suspension medium.The cell growth included four phases, the initial phase(1-3 d), the logarithmic phase(3-7 d), the steady phase(7-8 d), and the decline phase(8-11 d). With the increasing culture time, the number of suspension cells increased, and it reached the maximum value at the 7thd, about 3.8 ×105cells/ml. Then the number of cells began to decline rapidly. The cell vigor was the highest at the initial phase. Suspension cells grew best in the liquid MS medium supplemented with NAA(0.4 mg/L)and 6-BA(0.2 mg/L). The mitotic index reached the maximum, about 4.1% in the above medium, which suggested that this medium was suitable for cell suspension culture of eggplant. [Conclusion] Cell suspension culture system of eggplant provides a significant method for eggplant biotechnology. Genetic transformation and mutants screening can be carried out with this system. 展开更多
关键词 EGGPLANT CALLUS Suspension culture Growth curve
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Study on the Browning in Cell Suspension Culture of Taxus cuspidata 被引量:1
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作者 王培忠 赵欣 张宗申 《Agricultural Science & Technology》 CAS 2012年第5期935-937,983,共4页
[Objective] This study aimed to investigate the browning of T. cuspidata cells in suspension culture and provide the guidance for the cell suspension culture of T. cuspidata. [Method] T. cuspidata callus was used as e... [Objective] This study aimed to investigate the browning of T. cuspidata cells in suspension culture and provide the guidance for the cell suspension culture of T. cuspidata. [Method] T. cuspidata callus was used as experimental materials, to explore the effect of different medium, N/P ratio, pH, shaking speed, illumination time and light intensity and other factors on browning of T. cuspidata cells in suspension culture. [Result] Non-browning callus was transferred to 2MB5 medium (pH 7.0) for illumination culture at 22℃ under light intensity of 1 500 lx with shaking speed of 90 r/min for 24 h. Results showed that the cell browning was significantly inhibited. [Conclusion] This study laid the foundation for cell suspension culture of T. cuspidata and had important significance to the large-scale industrial production of paclitaxel. 展开更多
关键词 Taxus cuspidata BROWNING CALLUS Cell suspension culture
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Optimization of Cell Line Building for Taxus chinensis var. mairei
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作者 袁梦如 沈斌 +1 位作者 孙瑜霞 沈文飚 《Agricultural Science & Technology》 CAS 2010年第7期136-137,163,共3页
[Objective] The paper aims to optimize the abduction and domestication conditions of Taxus chinensis var.Mairei callus.[Method] We compared the efficiencies of callus between different explants and investigated the mu... [Objective] The paper aims to optimize the abduction and domestication conditions of Taxus chinensis var.Mairei callus.[Method] We compared the efficiencies of callus between different explants and investigated the multiplication conditions of callus and suspended cell culture conditions with the buds,young stems and young leaves from T.chinensis var as the explants.[Results] The effect was the best with the bud as the explants; the best way for sterilizing the explants of T.chinensis var mairei was:streptomycin detergent for 2 h + suds for 3 h + 75% alcohol disinfection for 30 s + 10% sodium hypochlorite solution for 25 min + 1‰ mercury chloride for 10 min; the optimum formula of callus subculture was:B5 + 4.0 mg/L NAA + 0.5 mg/L 2,4-D + 0.2 mg/L GA + 0.5 mg/L 6-BA + 2 g/L AC.[Conclusion] This research built the high efficient regeneration system of T.chinensis var. 展开更多
关键词 Taxus chinensis var.mairei CALLUS Successive transfer culture Suspended cell culture
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Somatic embryogenesis in wild relatives of cotton (Gossypium Spp.) 被引量:9
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作者 RAO Abdul Qayyum HUSSAIN S. Sarfraz +8 位作者 SHAHZAD M. Saqib BOKHARI S. Yassir Abbas RAZA M. Hashim RAKHA Allah MAJEED A. SHAHID A. Ali SALEEM Zafar HUSNAIN Tayyab RIAZUDDIN S. 《Journal of Zhejiang University-Science B(Biomedicine & Biotechnology)》 SCIE CAS CSCD 2006年第4期291-298,共8页
Wild cotton species can contribute a valuable gene pool for agronomically desirable cultivated tetraploid cultivars. In order to exploit diploid cotton a regeneration system is required to achieve transformation based... Wild cotton species can contribute a valuable gene pool for agronomically desirable cultivated tetraploid cultivars. In order to exploit diploid cotton a regeneration system is required to achieve transformation based goals. The present studies aimed at optimizing the conditions for regeneration of local varieties as well as wild species of cotton. Different callus induction media were tested with varying concentrations of hormones in which sucrose was used as nutritional source. Different explants (hypo-cotyls, cotyledon, root) were used to check the regeneration of both local cotton plants and wild relatives using T & G medium, BAP medium, CIM medium, EMMS medium, and cell suspension medium. Different stages of embryogenicity such as early torpedo stage, late torpedo stage, heart stage, globular stage and cotyledonary stage were observed in wild relatives of cotton. The results of this study pave the way for establishing future transformation methods. 展开更多
关键词 COTTON CALLUS Somatic embryogenic Wild species Cell suspension culture
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