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Enemies atpeace:Recentprogressin Agrobacterium-mediated cereal transformation
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作者 Shaoshuai Liu Ke Wang +5 位作者 Shuaifeng Geng Moammar Hossain Xingguo Ye Aili Li Long Mao Karl-Heinz Kogel 《The Crop Journal》 SCIE CSCD 2024年第2期321-329,共9页
Agrobacterium tumefaciens mediated plant transformation is a versatile tool for plant genetic engineering following its discovery nearly half a century ago.Numerous modifications were made in its application to increa... Agrobacterium tumefaciens mediated plant transformation is a versatile tool for plant genetic engineering following its discovery nearly half a century ago.Numerous modifications were made in its application to increase efficiency,especially in the recalcitrant major cereals plants.Recent breakthroughs in transformation efficiency continue its role as a mainstream technique in CRISPR/Cas-based genome editing and gene stacking.These modifications led to higher transformation frequency and lower but more stable transgene copies with the capability to revolutionize modern agriculture.In this review,we provide a brief overview of the history of Agrobacterium-mediated plant transformation and focus on the most recent progress to improve the system in both the Agrobacterium and the host recipient.A promising future for transformation in biotechnology and agriculture is predicted. 展开更多
关键词 agrobacterium tumefaciens Cereal species Genome editing Genetic engineering Plant breeding
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根癌农杆菌介导的紫花苜蓿遗传转化体系的优化
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作者 王之杰 李明序 张万军 《草地学报》 CAS CSCD 北大核心 2024年第6期1665-1671,共7页
为进一步提高根癌农杆菌介导的紫花苜蓿遗传转化效率,本试验以紫花苜蓿(Medicago sativa L.)品种‘中苜1号’为试验材料,通过筛选高效再生的植株,以筛选出的植株叶片为外植体,并通过调节培养基激素配比,在农杆菌侵染过程中添加谷氨酰胺... 为进一步提高根癌农杆菌介导的紫花苜蓿遗传转化效率,本试验以紫花苜蓿(Medicago sativa L.)品种‘中苜1号’为试验材料,通过筛选高效再生的植株,以筛选出的植株叶片为外植体,并通过调节培养基激素配比,在农杆菌侵染过程中添加谷氨酰胺、进行冷处理等措施优化苜蓿由农杆菌介导的遗传转化体系。试验结果表明:转化过程中以嫩叶作为外植体,菌液和重悬液中添加150μmol·L^(-1)乙酰丁香酮,侵染过程中使用300μmol·L^(-1)的谷氨酰胺将瞬时转化效率从32%提高至92%,转化效率提高到31%。研究紫花苜蓿的遗传转化体系可为紫花苜蓿高效遗传改良及基因功能研究提供了技术支持。 展开更多
关键词 紫花苜蓿 根癌农杆菌 遗传转化 体系优化
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石油生物催化脱硫菌Agrobacterium tumefaciens UP3的分离筛选 被引量:9
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作者 史德青 赵金生 +2 位作者 侯影飞 杨金荣 孔瑛 《微生物学报》 CAS CSCD 北大核心 2004年第2期248-250,共3页
从胜利油田被原油污染的土壤中筛选到一株能有效降解模型化合物二苯并噻吩 (DBT)的菌株。根据常规的形态分析、生理生化性状及 16SrDNA序列分析 ,将其鉴定为根癌土壤杆菌 (AgrobacteriumtumefaciensUP3)。该菌不能以十二烷、十六烷、液... 从胜利油田被原油污染的土壤中筛选到一株能有效降解模型化合物二苯并噻吩 (DBT)的菌株。根据常规的形态分析、生理生化性状及 16SrDNA序列分析 ,将其鉴定为根癌土壤杆菌 (AgrobacteriumtumefaciensUP3)。该菌不能以十二烷、十六烷、液体石蜡和萘作为唯一碳源和能源生长 ,具有工业应用的潜力。对该菌株DBT降解能力的初步研究表明 ,5 4h内可将 5 0 0mg L的DBT降解至 15 0mg L。对降解产物的分析表明 ,根癌土壤杆菌降解DBT的途径与Kodama路线及 4_S路线不同。 展开更多
关键词 二苯并噻吩 DBT 菌株 根癌土壤杆菌 土壤 石油 生物催化脱硫 筛选 降解
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根癌农杆菌介导彩色马蹄莲遗传转化体系的建立及优化
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作者 李菲 曹永琼 +3 位作者 王纲 郭彦兵 李紫薇 吴红芝 《南方农业学报》 CAS CSCD 北大核心 2024年第6期1692-1699,共8页
【目的】建立及优化根癌农杆菌介导的彩色马蹄莲(Zantedeschia hybrida)遗传转化体系,为提高彩色马蹄莲遗传转化效率及培育彩色马蹄莲抗性品种提供理论参考依据。【方法】将不同外植体(茎基、叶、不定芽)和不同浓度噻苯隆(TDZ)(0.001、0... 【目的】建立及优化根癌农杆菌介导的彩色马蹄莲(Zantedeschia hybrida)遗传转化体系,为提高彩色马蹄莲遗传转化效率及培育彩色马蹄莲抗性品种提供理论参考依据。【方法】将不同外植体(茎基、叶、不定芽)和不同浓度噻苯隆(TDZ)(0.001、0.002和0.005 g/L)进行正交试验筛选外植体和丛生芽增殖培养基;液氮冻融法将pCAM BIA2301和pBI121质粒转入农杆菌LBA4404感受态细胞。以β-葡萄糖苷酸酶(GUS)基因作为农杆菌介导转化测定的报告基因,对彩色马蹄莲的丛生芽进行遗传转化,阳性苗经PCR扩增验证转化结果。通过不同硫酸卡那霉素(Kan)浓度(50、100和150 mg/L)、侵染时间(25、30和40 min)及共培养时间(2和3 d)3因素正交试验筛选最佳转化条件。【结果】外植体为不定芽的丛生芽诱导率极显著高于茎基和叶(P<0.01),当不定芽为外植体、TDZ浓度为0.002 mg/L时,丛生芽增殖倍数(5.12倍)显著高于0.001和0.005 mg/L TDZ处理(P<0.05),筛选出M6培养基+0.002 mg/L TDZ+30 g/L蔗糖+6.25 g/L琼脂为丛生芽增殖培养基;影响阳性频率因素排序为:侵染时间>Kan浓度>共培养时间。当Kan浓度为50 mg/L、侵染时间25 min及共培养时间为3 d为最优侵染条件,此时,GUS+丛生芽出芽频率和PCR阳性频率最高,分别为54.0%和15.27%。【结论】成功优化彩色马蹄莲遗传转化体系,最佳外植体为不定芽,转化体系最佳组合为Kan浓度为50 mg/L、侵染时间为25 min及共培养时间为3 d。 展开更多
关键词 彩色马蹄莲 根癌农杆菌 遗传转化 优化
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农杆菌介导的黄瓜遗传转化体系优化研究
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作者 陈梅 陈露倩 +2 位作者 陈思 杜长霞 樊怀福 《四川农业大学学报》 CSCD 北大核心 2024年第3期540-545,571,共7页
【目的】筛选适用于农杆菌介导的黄瓜遗传转化体系,为通过生物技术进行黄瓜新品种培育奠定基础。【方法】以华北型黄瓜品种‘9930’子叶为试材,探讨卡那霉素浓度(kanamycin, Kan)、预培养时间、预培养基添加乙酰丁香酮(acetosyringone, ... 【目的】筛选适用于农杆菌介导的黄瓜遗传转化体系,为通过生物技术进行黄瓜新品种培育奠定基础。【方法】以华北型黄瓜品种‘9930’子叶为试材,探讨卡那霉素浓度(kanamycin, Kan)、预培养时间、预培养基添加乙酰丁香酮(acetosyringone, AS)、分化培养基pH及生根培养基中添加不同外源物质对黄瓜遗传转化的影响。【结果】不定芽分化、生根阶段的Kan临界浓度为150 mg/L;不进行预培养的黄瓜不定芽诱导率最高,为77.17%;当分化培养基pH为5.6时不定芽诱导率最高,为75.26%;外植体在添加0.5 mg/L的IBA生根培养基上生根率达100%,根长也显著增加。【结论】在以黄瓜品种‘9930’子叶节为外植体的遗传转化体系中,150 mg/L Kan可作为不定芽分化和生根阶段的筛选选择压;不进行预培养更有利于其抗性芽诱导;适合不定芽诱导的分化培养基pH为5.6;0.5 mg/L的IBA为诱导外植体生根最佳激素浓度。 展开更多
关键词 根癌农杆菌 黄瓜 遗传转化
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等离子体活化水对根癌土壤杆菌的抑制作用及其对樱花根癌病的防治
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作者 潘越 司红波 +1 位作者 潘宇伟 孙永莲 《北方园艺》 CAS 北大核心 2024年第9期54-61,共8页
以樱花幼苗为试材,使用低温等离子体活化水冷杀菌技术(PAW)对樱花幼苗及其病原菌进行处理,研究了PAW对樱花苗木的抗病作用,以期为PAW在樱花根癌病防治中的应用提供参考依据。结果表明:等离子体活化水能有效地抑制樱花根癌病病菌Agrobact... 以樱花幼苗为试材,使用低温等离子体活化水冷杀菌技术(PAW)对樱花幼苗及其病原菌进行处理,研究了PAW对樱花苗木的抗病作用,以期为PAW在樱花根癌病防治中的应用提供参考依据。结果表明:等离子体活化水能有效地抑制樱花根癌病病菌Agrobacterium tumefaciens的生长,其可以对Agrobacterium tumefaciens的细胞结构造成破坏。CK组樱花幼苗的根瘤发生率为100%,根瘤的平均直径为18.8 mm,处理后的幼苗发病率为56.7%,根瘤的平均直径为8.4 mm。此外,PAW可以提高樱花根系多酚氧化酶(PPO)和苯丙氨酸解氨酶(PAL)及超氧化物歧化酶(SOD)活性的表达,激活樱花植株的抗性。综上,PAW具有一定的生物防治潜力,可以防御樱花根癌病。 展开更多
关键词 樱花 根癌病 agrobacterium tumefaciens 等离子体活化水
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Genetic Transformation of Aloe barbadensis Miller by Agrobacterium tumefaciens 被引量:1
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作者 何聪芬 张佳星 +4 位作者 陈杰 叶兴国 杜丽璞 董银卯 赵华 《Journal of Genetics and Genomics》 SCIE CAS CSCD 北大核心 2007年第12期1053-1060,共8页
Despite the importance of aloe in cosmetic and pharmaceutical industries, improvement of aloe (Aloe barbadensis Miller) by genetic engineering was seldom reported previously. In this study, regeneration and transfor... Despite the importance of aloe in cosmetic and pharmaceutical industries, improvement of aloe (Aloe barbadensis Miller) by genetic engineering was seldom reported previously. In this study, regeneration and transformation conditions, including explant selection and surface sterilization, use of different Agrobacterium strains, and co-culture processing, are optimized. The use of 20.0% sodium hypochloride (25 rain) for sterilization was less detrimental to the health of explant than 0.1% mercuric chloride (10 min). Regeneration frequency from stems was much higher than that from leaves or sheaths. Explants were infected by Agrobacterium (30 rain) in liquid co-cultural medium, and this was followed by three days co-culture on sterile filter papers with light for 10 h per day at 24℃. Histochemical data demonstrated that the transient expression of GUS gene in the stem explants of aloe infected with Agrobacterium strains EHAI05 and C58CI was 80.0% and 30.0%, respectively, suggesting the higher sensitivity of the explants to EHAI05 than to C58C1. Infected tissues were selected using G418 (10.0-25.0 mg/L) to generate transformants. Sixty-seven G418 resistant plantlets were generated from the infected explants. Southern blotting, PCR, and ELISA analyses indicated that the alien gene were successfully transferred into aloe and was expressed in the transgenic plants. This newly established transformation system could be used for the genetic improvement of aloe. 展开更多
关键词 ALOE agrobacterium tumefaciens TRANSFORMATION molecular test ELISA analysis
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耐盐芦苇高频遗传转化体系的建立
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作者 王美玲 韩建玮 +2 位作者 陈方军 尹海波 张侠 《烟台大学学报(自然科学与工程版)》 2024年第1期69-73,共5页
以耐盐芦苇(Phragmites australis)种子愈伤组织为材料,以根瘤农杆菌EHA105为媒介建立高效的芦苇遗传转化体系。结果表明:在潮霉素浓度为50 mg/L、6-BA浓度为0.8 mg/L时不定芽分化率达到最高。将沉默载体pFGC1008-GRPi通过根瘤农杆菌EHA... 以耐盐芦苇(Phragmites australis)种子愈伤组织为材料,以根瘤农杆菌EHA105为媒介建立高效的芦苇遗传转化体系。结果表明:在潮霉素浓度为50 mg/L、6-BA浓度为0.8 mg/L时不定芽分化率达到最高。将沉默载体pFGC1008-GRPi通过根瘤农杆菌EHA105在芦苇愈伤组织中进行遗传转化,共获得8个抗性株系,转化率为3.9%。实时荧光定量PCR检测显示6个株系实现了基因沉默,表明该转化体系高效可用。 展开更多
关键词 芦苇 根瘤农杆菌 遗传转化
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异养硝化好氧反硝化菌株Agrobacterium tumefaciens LAD9羟胺氧化酶的分离纯化 被引量:2
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作者 陈倩 马涛 王婷 《生物技术通报》 CAS CSCD 北大核心 2014年第7期69-73,共5页
羟氨氧化酶(Hydroxylamine oxidase,HAO)的作用方式直接决定了异养硝化好氧反硝化细菌的代谢途径,分离得到纯度较高的HAO也就成为研究这类细菌脱氮机制的重要环节。以异养硝化好氧反硝化细菌Agrobacterium tumefaciens LAD9为代表,建立... 羟氨氧化酶(Hydroxylamine oxidase,HAO)的作用方式直接决定了异养硝化好氧反硝化细菌的代谢途径,分离得到纯度较高的HAO也就成为研究这类细菌脱氮机制的重要环节。以异养硝化好氧反硝化细菌Agrobacterium tumefaciens LAD9为代表,建立了该菌株HAO的分离纯化方法:首先采用渗透压休克法提取细胞周质液,然后采用DEAE Sepharose CL-6B离子交换层析和Sephacryl S-100凝胶过滤层析对细胞周质液进行分离纯化。结果表明,经过离子交换层析可得到分子量分别为55.3、35.7和19.2kD的杂蛋白,进一步经过凝胶过滤层析即可得到电泳纯的HAO,纯化倍数为5.79,产率为39.71%。对其酶学性质的初步研究表明,该菌株HAO的分子量为18.8 kD,能够将羟胺氧化为亚硝酸盐氮,且Fe2+的加入可显著增强其酶活。 展开更多
关键词 脱氮 羟胺氧化酶 分离纯化 agrobacterium TUMEFACIENS LAD9 异养硝化好氧反硝化
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石油生物脱硫菌Agrobacterium tumefaciens UP-3 被引量:2
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作者 侯影飞 孔瑛 +2 位作者 杨金荣 辛伟 张建辉 《微生物学通报》 CAS CSCD 北大核心 2005年第1期52-56,共5页
对能降解二苯并噻吩(DBT)的根癌土壤杆菌AgrobacteriumtumefaciensUP3菌株进行了固定化研究,以聚乙烯醇(PVA)和海藻酸钠(SA)混合物为包埋法固定化载体,固定化最佳操作条件为4℃交联,PVA和SA混合物总浓度7%,两者最佳浓度比为6,细胞浓度为... 对能降解二苯并噻吩(DBT)的根癌土壤杆菌AgrobacteriumtumefaciensUP3菌株进行了固定化研究,以聚乙烯醇(PVA)和海藻酸钠(SA)混合物为包埋法固定化载体,固定化最佳操作条件为4℃交联,PVA和SA混合物总浓度7%,两者最佳浓度比为6,细胞浓度为0.05g/mL。当DBT加入量为2.7mmol/L时,UP-3的静息细胞最高脱硫率为13%,而固定化细胞的脱硫效率超过了60%;固定化细胞的最佳使用条件为降解5d,温度28℃~32℃。 展开更多
关键词 石油生物脱硫 二苯并噻吩 固定化细胞 根癌土壤杆菌UP-3
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一株脱有机硫菌Agrobacterium tumefaciens W3的分离鉴定及特性的研究 被引量:3
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作者 邱乐泉 杨琳 钟卫鸿 《浙江工业大学学报》 CAS 北大核心 2009年第5期520-524,共5页
从杭州炼油厂的土壤中筛选到的能降解二苯并噻吩(Dibenzothiophene,DBT)的菌株,并进行了显微形态、生理生化分析和16S rDNA序列分析,将其鉴定为Agrobacteriumtume faciens W3;对该菌株的降解产物分析表明,其降解DBT的终产物为2-羟基... 从杭州炼油厂的土壤中筛选到的能降解二苯并噻吩(Dibenzothiophene,DBT)的菌株,并进行了显微形态、生理生化分析和16S rDNA序列分析,将其鉴定为Agrobacteriumtume faciens W3;对该菌株的降解产物分析表明,其降解DBT的终产物为2-羟基联苯(2-hydroxybiphenyl,2-HBP),符合4-S脱硫途径;对该菌株的部分脱硫特性进行分析表明,5 g/L葡萄糖,3 g/L NH4Cl,0.15 mmol/L DBT,初始pH 6.9-7.5为其适宜的培养条件,培养时间72-96 h可达到最大生长,脱硫效果可达83%. 展开更多
关键词 生物脱硫 二苯并噻吩 根癌土壤杆菌W3
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利用绿色荧光蛋白优化辣椒遗传转化体系
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作者 董依萍 刘画 +3 位作者 刘丹 周迎佳 李峰 邓颖天 《广东农业科学》 CAS 2024年第3期103-113,共11页
【目的】农杆菌介导的辣椒遗传转化较为困难,至今仍未建立高效转化体系。绿色荧光蛋白(GFP)基因是植物遗传转化中常用的报告基因,以GFP为报告基因,优化农杆菌介导的辣椒遗传转化体系。【方法】利用GFP表达系统,统计3个辣椒品种(‘HP’‘... 【目的】农杆菌介导的辣椒遗传转化较为困难,至今仍未建立高效转化体系。绿色荧光蛋白(GFP)基因是植物遗传转化中常用的报告基因,以GFP为报告基因,优化农杆菌介导的辣椒遗传转化体系。【方法】利用GFP表达系统,统计3个辣椒品种(‘HP’‘8214’和‘L55’)中3种不同外植体(子叶、下胚轴和Flamingo-bill外植体)的不定芽分化率、不定根分化率与荧光阳性率,探究农杆菌侵染浓度、侵染时间、预培养时间和共培养时间等因素对不定芽、不定根分化率及荧光阳性率的影响。【结果】3个辣椒品种的Flamingo-bill外植体不定芽分化率均显著高于下胚轴与子叶外植体,其中‘L55’Flamingo-bill外植体的不定芽分化率最高、达77.59%,故选用‘L55’Flamingo-bill外植体进行后续研究。4种农杆菌不同侵染浓度和时间组合下,‘L55’Flamingo-bill外植体均可产生不定芽、不定根及表达GFP的愈伤组织,当农杆菌侵染浓度为OD_(600)=0.05,侵染时间为30 min时,不定芽分化率和荧光阳性率最高,分别达48.39%、4.84%。在6种不同预培养与共培养时间组合处理下,Flamingo-bill外植体也均产生不定芽和不定根,预培养1 d、共培养1~2 d处理下的不定芽分化率和荧光阳性率最高,分别达48.44%、12.50%。最后对表达GFP荧光的不定根与愈伤组织进行PCR检测,结果显示GFP、Kan和Cas9基因在荧光阳性的组织中均被检测到,表明农杆菌介导的T-DNA插入成功且转化稳定。【结论】辣椒的外植体类型、农杆菌侵染浓度、侵染时间、预培养和共培养时间均对辣椒遗传转化效率有影响。以GFP为报告基因,筛选合适的转化条件可提高农杆菌介导的辣椒遗传转化效率。 展开更多
关键词 辣椒 Flamingo-bill外植体 绿色荧光蛋白 遗传转化 农杆菌 组织培养
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Optimization of Genetic Transformation System of Tobacco K326 Mediated by Agrobacterium 被引量:2
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作者 赵勤 《Agricultural Science & Technology》 CAS 2011年第1期62-64,共3页
[Objective]The aim was to optimize genetic transformation system in tobacco K326 mediated by Agrobacterium.[Method]The leaf of tobacco aseptic seedling was taken as explants to study the optimization of Agrobacterium-... [Objective]The aim was to optimize genetic transformation system in tobacco K326 mediated by Agrobacterium.[Method]The leaf of tobacco aseptic seedling was taken as explants to study the optimization of Agrobacterium-mediated genetic transformation system.[Result] The highest transformation efficiency was obtained when the explants were pre-cultured in the medium of MS + 2 mg/L 6-BA + 0.2 mg/L IAA for 2 d,and then infected with Agrobacterium GV3101(OD600 =0.6) for 5 min.The PCR detection proved that npt II gene had been integrated into the regenerated tobacco plants.[Conclusion]A highly efficient genetic transformation system of tobacco leaf mediated by Agrobacterium was established. 展开更多
关键词 agrobacterium tumefaciens TOBACCO Genetic transformation
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Establishing VIGS and CRISPR/Cas9 techniques to verify RsPDS function in radish
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作者 Jiali Ying Yan Wang +10 位作者 Liang Xu Tiaojiao Qin Kai Xia Peng Zhang Yinbo Ma Keyun Zhang Lun Wang Junhui Dong Lianxue Fan Yuelin Zhu Liwang Liu 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2024年第5期1557-1567,共11页
Virus-induced gene silencing(VIGS)and clustered regularly interspaced short palindromic repeats/CRISPR-associated protein(CRISPR/Cas)systems are effective technologies for rapid and accurate gene function verification... Virus-induced gene silencing(VIGS)and clustered regularly interspaced short palindromic repeats/CRISPR-associated protein(CRISPR/Cas)systems are effective technologies for rapid and accurate gene function verification in modern plant biotechnology.However,the investigation of gene silencing and editing in radish remains limited.In this study,a bleaching phenotype was generated through the knockdown of RsPDS using tobacco rattle virus(TRV)-and turnip yellow mosaic virus(TYMV)-mediated gene silencing vectors.The TYMV-mediated gene silencing efficiency was higher than the TRV-based VIGS system in radish.The expression level of RsPDS was significantly inhibited using VIGS in'NAU-067'radish leaves.The rootless seedlings of‘NAU-067'were infected with Agrobacterium rhizogenes using the 2300GN-Ubi-RsPDS-Cas9 vector with two target sequences.Nine adventitious roots were blue with GUs staining,and four of these adventitious roots were edited at target sequence 1 of the RsPDS gene as indicated by Sanger sequencing.Furthermore,albino lines were generated with A.tumefaciens-mediated transformation of radish cotyledons.Five base substitutions and three base deletions occurred at target sequence 2 in Line 1,and three base insertions and three base substitutions occurred at target sequence 1 in Line 2.This study shows that VIGS and CRISPR/Cas9 techniques can be employed to precisely verify the biological functions of genes in radish,which will facilitate the genetic improvement of vital horticultural traits in radish breeding programs. 展开更多
关键词 Raphanus sativus L. VIGS CRISPR/Cas9 agrobacterium rhizogenes A.tumefaciens RsPDS
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Study on Agrobacterium tumefaciens-mediated Transformation of Brassica campestris L. with Fusion Gene Ycoil-bFGF
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作者 徐岩 肖艳双 +4 位作者 杜金霞 汪洪 郑伟 李营 庞实锋 《Agricultural Science & Technology》 CAS 2009年第4期31-36,共6页
[ Objective] The study is to generate pharmaceutical protein via plant transgenic technique. [Methed] Using the cotyledons with petiole as transformation receptor, the fusion gene of rapeseed oil-body gene and bFGF wa... [ Objective] The study is to generate pharmaceutical protein via plant transgenic technique. [Methed] Using the cotyledons with petiole as transformation receptor, the fusion gene of rapeseed oil-body gene and bFGF was introduced into the rapeseed ( Brassica campestris L. ) by Agrobacterium tumefaciens-mediated transformation; meanwhile regeneration conditions of rapeseed were also optimized, and the regenerated resistant plantlets were detected by PCR and Southern blot. [ Result] This fusion gene had been integrated into rapeseed genome successfully, and the optimized conditions of transformation and regeneration were as follows: explants pre-culture for 2 d, co-culture for 3 d, bacteria solution OD600 for 0.3 and infection time for 5 min. [ Conclusion] The results laid a solid foundation for extraction, isolation and purification of protein in transgenic plant seeds. 展开更多
关键词 Basic fibroblast growth factor (bFGF) Plant bioreactor Oil-body system agrobacterium tumefaciens Brassica campestris L.
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Optimization of Agrobacterium Mediated Genetic Transformation System of Tomato Meifen No.1
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作者 李立芹 《Agricultural Science & Technology》 CAS 2010年第11期92-94,共3页
[Objective] The aim of this study was to carry out study on the optimization of Agrobacterium mediated genetic transformation system of tomato Meifen No.1.[Method] The cotyledon of tomato cultivar Meifen No.1 was used... [Objective] The aim of this study was to carry out study on the optimization of Agrobacterium mediated genetic transformation system of tomato Meifen No.1.[Method] The cotyledon of tomato cultivar Meifen No.1 was used as the explant,and the Agrobacterium mediation method was used to optimize its genetic transformation efficiency so as to establish the efficient Agrobacterium mediated genetic transformation system of tomato cotyledon.[Result] The highest transformation efficiency was obtained when the explants were cultivated for 2 d on MS + 2.0 mg/L 6-BA+ 0.5 mg/L IAA medium and then infected with Agrobacterium EHA105(OD = 0.4)for 5 min;it was proved by PCR analysis that the target nptII gene had been integrated into the genome of regenerated plants.[Conclusion] The result in this study had provided basis for the transfer of valuable genes into tomato Meifen No.1. 展开更多
关键词 agrobacterium tumefaciens TOMATO Transgenic transformation
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根癌农杆菌(Agrobacterium tumefaciens)对毛白杨五种防御酶系统的影响 被引量:2
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作者 王召元 袁玉欣 +3 位作者 杜克久 王颖 侯西成 王艳芬 《河北农业大学学报》 CAS CSCD 北大核心 2007年第4期31-35,共5页
用野生根癌农杆菌接种毛白杨后,定期测定叶片内5种酶活性的动态变化,旨在揭示毛白杨对根癌农杆菌侵染后的生理响应,阐明杨树根癌病的病害生理。毛白杨接种病原菌后,SOD、POD、PPO前期活性高于对照,有1个酶活高峰,后期下降接近对照;CAT出... 用野生根癌农杆菌接种毛白杨后,定期测定叶片内5种酶活性的动态变化,旨在揭示毛白杨对根癌农杆菌侵染后的生理响应,阐明杨树根癌病的病害生理。毛白杨接种病原菌后,SOD、POD、PPO前期活性高于对照,有1个酶活高峰,后期下降接近对照;CAT出现2个酶活高峰,25 d虽有下降,但一直高于对照;PAL活性变化幅度比较小,基本保持在对照水平,与对照相比差异不显著。研究结果表明:SOD和CAT对根癌农杆菌最敏感,POD和PPO次之,PAL敏感性最差。 展开更多
关键词 毛白杨 根癌农杆菌 超氧化物歧化酶 过氧化物酶 过氧化氢酶 多酚氧化酶 苯丙氨酸解氨酶
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根癌农杆菌(Agrobacterium tumefaciens)介导培育富含叶黄素的基因工程小球藻(Chlorella vulgaris)的研究 被引量:2
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作者 马瑞娟 林祥志 《海洋与湖沼》 CAS CSCD 北大核心 2013年第1期159-164,共6页
采用基因克隆技术构建双元载体pCAMBIA2301-idi,通过电转转入农杆菌LBA4404中。利用根癌农杆菌介导的转化方法,将pCAMBIA2301-idi质粒的T-DNA区转入小球藻,以G418抗性基因(NPTⅡ)作为筛选标记,筛选出阳性转化子。通过PCR扩增表明idi基因... 采用基因克隆技术构建双元载体pCAMBIA2301-idi,通过电转转入农杆菌LBA4404中。利用根癌农杆菌介导的转化方法,将pCAMBIA2301-idi质粒的T-DNA区转入小球藻,以G418抗性基因(NPTⅡ)作为筛选标记,筛选出阳性转化子。通过PCR扩增表明idi基因和NPTⅡ基因已经整合到小球藻基因组中。测定转化子的生物量,结果表明大部分转化子的生物量与野生型相似。测定转化子小球藻干粉的叶黄素含量,发现转化子叶黄素含量最高达到0.84mg/g,与野生型相比提高了30.95%。进一步分析藻液中叶黄素的产量,发现转化子的叶黄素产量最高达到1.98mg/L,比野生型提高了36.77%。 展开更多
关键词 小球藻 农杆菌 遗传转化 idi基因 叶黄素
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Improvement of Plant Regeneration from Immature Embryos of Wheat Infected by Agrobacterium tumefaciens 被引量:10
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作者 TAO Li-li YIN Gui-xiang DU Li-pu SHI Zheng-yuan SHE Mao-yun XU Hui-jun YE Xing-guo 《Agricultural Sciences in China》 CAS CSCD 2011年第3期317-326,共10页
Wheat, one of the most important food crops, has been extensively studied with respects to plant regeneration and transformation employing the immature embryos as recipient tissues. However, the transformed tissues of... Wheat, one of the most important food crops, has been extensively studied with respects to plant regeneration and transformation employing the immature embryos as recipient tissues. However, the transformed tissues often become severely necrotic after co-cultivation with Agrobacterium, which is one of the major obstacles in gene delivery. In this study, wheat varieties CB037, Kenong 199, Xinchun 9, Lunxuan 987, and Shi 4185 showed desirable culture potential or high infection ability in Agrobacterium-mediated transformation. Similarly, optimal regeneration conditions were determined by testing their ability to inhibit the cell necrosis and cell death phenotype. Two auxins of 2,4-dichlorophenoxyacetic acid (2,4-D) and 3,6-dichloro-o-anisic acid (dicamba) were evaluated for highly significant effect on both callus and plantlet production, although they were genotype-dependent in wheat. Substitution of 2,4-D by dicamba enhanced the growth and regeneration ability of callus from the immature embryos of most genotypes tested. The callus growth state couldn’t be modified by adding antioxidants such as ascorbic acid, cysteine, and silver nitrate or organic additives such as thiamine HCl and asparagine to the media. On the contrary, the best tissue statement and plant regeneration was achieved by employing the media containing the simplest MS (Murashige and Skoog) components and dicamba without organic components and vitamins. Thereby, our results are thought to inhibit wheat cell necrosis effectively and suggested to be used for more wheat genotypes. 展开更多
关键词 WHEAT immature embryos agrobacterium tumefaciens TRANSFORMATION GENOTYPES plant regeneration
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Meropenem as an Alternative Antibiotic Agent for Suppression of Agrobacterium in Genetic Transformation of Orchid 被引量:9
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作者 CAO Ying Niimi Yoshiyuki HU Shang-lian 《Agricultural Sciences in China》 CAS CSCD 2006年第11期839-846,共8页
A case of Meropenem as a novel antibacterial agent to suppress and eliminate Agrobacterium tumefaciens in the Agrobacterium-mediated transformation of orchid protocorm-like bodies (PLBs) has been reported in this ar... A case of Meropenem as a novel antibacterial agent to suppress and eliminate Agrobacterium tumefaciens in the Agrobacterium-mediated transformation of orchid protocorm-like bodies (PLBs) has been reported in this article. The in vitro activities of meropenem and four comparator antibacterial agents against three Agrobacterium tumefaciens strains, LBA4404, EHA101, and GV3101, were assessed. In addition, the effect of meropenem on the growth of Dendrobium phalaenopsis PLBs was determined. Compared with other commonly used antibiotics (including ampicillin, carbenicillin, cefotaxime, and cefoperazone), meropenem showed the highest activity in suppressing all tested A. tumefaciens strains (minimum inhibitory concentration [MIC] 〈 0.5 mg L^-1, which is equal to minimum bactericidal concentration [MBC]). Meropenem, at all tested concentrations, except for 10 mg L^-1 concentration, had little negative effect on the growth of orchid tissues. The A. tumefaciens strain EHA101 in genetic transformation with vector plG121Hm in infected PLBs of the orchid was visually undetectable after a two-month subculture in 1/2 MS medium with 50 mg L^-1 meropenem and 25 mg L^-1 hygromacin. The expression and incorporation of the transgenes were confirmed by GUS histochemical assay and PCR analysis. Meropenem may be an alternative antibiotic for the effective suppression of A. tumefaciens in genetic transformation. 展开更多
关键词 ANTIBIOTICS MEROPENEM agrobacterium tumefaciens antibacterial activity genetic transformation protocormlike bodies (PLBs)
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