| 基因枪介导红叶石楠遗传转化因素分析 |
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| 引用本文： | 刘翠兰,燕丽萍,毛秀红,孙 超,夏 阳,梁慧敏.基因枪介导红叶石楠遗传转化因素分析[J].园艺学报,2013,40(11):2280-2286. |
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| 作者姓名： | 刘翠兰 燕丽萍 毛秀红 孙 超 夏 阳 梁慧敏 |
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| 作者单位： | 1 山东省林业科学研究院，济南 250014；2 江苏农林职业技术学院，江苏句容 212400；3 山东省林木遗传改良重点实验室，济南 250014 |
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| 基金项目： | 山东省农业良种工程项目（鲁农良种字〔2010〕117 号）；国家科技部成果转化项目（2006GB2C600167）；镇江市科技支撑计划项目（NY2013028）；济南市科技明星计划项目（20120126） |
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| 摘 要： | 以红叶石楠（Photinia fraseri）的胚性愈伤组织为受体材料，利用基因枪法导入外源抗寒基
因rd29A，以期建立基因枪转化红叶石楠的技术体系。结果表明，基因枪转化红叶石楠的最佳组合条件为：
胚性愈伤组织在诱导培养基MS + 6-BA 2.0 mg · L-1 + NAA 5.0 mg · L-1 + 2,4-D 0.5 mg · L-1 + 琼脂5.0
g · L-1 + 蔗糖20 g · L-1 里，用0.3 mol · L-1 甘露醇处理15 ~ 16 h；基因枪轰击时添加12.5 mol · L-1 氯化钙
50 μL + 0.1 mol · L-1 亚精胺50 μL，每枪含金粉300 μg + 质粒DNA 3.0 μg，轰击距离9 cm，轰击压力1 100 psi，
轰击次数1 次。经多次筛选获得了6 株转基因植株，转化植株经PCR 检测和Southern 分析，证实了rd29A
已经整合到红叶石楠基因组中。
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| 关 键 词： | 以红叶石楠（Photinia fraseri）的胚性愈伤组织为受体材料 利用基因枪法导入外源抗寒基 />因rd29A 以期建立基因枪转化红叶石楠的技术体系。结果表明 基因枪转化红叶石楠的最佳组合条件为 />胚性愈伤组织在诱导培养基MS + 6-BA 2.0 mg ·L-1 + NAA 5.0 mg ·L-1 + 2 4-D 0.5 mg ·L-1 + 琼脂5.0 />g ·L-1 + 蔗糖20 g ·L-1 里 用0.3 mol ·L-1 甘露醇处理15 ~ 16 h 基因枪轰击时添加12.5 mol ·L-1 氯化钙 />50 μL + 0.1 mol ·L-1 亚精胺50 μL 每枪含金粉300 μg + 质粒DNA 3.0 μg 轰击距离9 cm 轰击压力1 100 psi />轰击次数1 次。经多次筛选获得了6 株转基因植株 转化植株经PCR 检测和Southern 分析 证实了rd29A已经整合到红叶石楠基因组中。 |
Studies on the Genetic Transformation Factors of Photinia fraseri Using
Microprojectile Bombardment |
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| LIU Cui-Lan,YAN Li-Ping,MAO Xiu-Hong,SUN Chao,XIA Yang,LIANG Hui-Min.Studies on the Genetic Transformation Factors of Photinia fraseri Using
Microprojectile Bombardment[J].Acta Horticulturae Sinica,2013,40(11):2280-2286. |
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| Authors: | LIU Cui-Lan YAN Li-Ping MAO Xiu-Hong SUN Chao XIA Yang LIANG Hui-Min |
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| Institution: | 1Shandong Provincial Academy of Forestry，Ji’nan 250014，China；2Jiangsu AgricμLture and Forestry Profession
Technology College，Jurong，Jiangsu 212400，China； 3 Shandong Provincial Key Laboretory of Forest Genetic Improvement
Tree，Ji’nan 250014，China |
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| Abstract: | The study has transformed the cold resistant gene rd29A into the acceptor，the embryo
callus derived from Photinia fraseri leaf，in order to establish the transformation system by using particle
bombardment method. The results indicate that the optimal transformation scheme for Photinia fraseri leaf
callus using particle bombardment method was treating embryogenic callus tissue within 0.3 mol · L-1
mannitol for 15–16 h，in MS medium supplemented with 2.0 mg · L-1 6-benzy aminopurine，5.0 mg · L-1
napthaleneacetic acid，0.5 mg · L-1 2,4-dichlorophenoxyacetic acid，5.0 g · L-1 agar powder and 20 g · L-1
glucose. When particle bombardmenting，in addition of 12.5 mol · L-1 CaCl2 50 μL and 0.1 mol · L-1
spermidine 50 μL，and bombarding the samples once a time，at 9 cm bombardment distance，at 1 100 psi
heli μm pressure and with the dosage of gold powder 300 μg and plasmid DNA 3.0 μg per bombardment.
Six transgenic Photinia fraseri plants were obtained in the study and it was confirmed that the rd29A gene
was integrated into Photinia fraseri genomes according to the PCR and Southern analysis of plant DNA. |
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| Keywords: | Photinia fraseri rd29A microprojectile bombardment transgenic plants |
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