水稻OsAPX基因的克隆及过表达载体的构建

许惠滨; 连玲; 魏毅东; 朱永生; 谢华安; 王宗华; 张建福

doi:10.19303/j.issn.1008-0384.2013.02.003

水稻OsAPX基因的克隆及过表达载体的构建

doi: 10.19303/j.issn.1008-0384.2013.02.003

许惠滨^1,2,3,4,5,
连玲^2,3,4,5,
魏毅东^1,2,3,4,5,
朱永生^2,3,4,5,
谢华安^2,3,4,5,
王宗华¹,
张建福^2,3,4,5

1. 福建农林大学生命科学学院,福建福州 350002;
2. 农业部华南杂交水稻种质创新与分子育种重点实验室/福州(国家)水稻改良分中心/福建省作物分子育种工程实验室/福建省水稻分子育种重点实验室/福建省农业科学院水稻研究所,福建福州350003;
3. 福建省作物种质创新与分子育种省部共建国家重点实验室培育基地,福建福州350003;
4. 杂交水稻国家重点实验室华南基地,福建福州350003;
5. 水稻国家工程实验室,福建福州350003

基金项目:

国家转基因农作物新品种培育重大专项(2008ZX08001-006、2011ZX08001-006)

福建省财政专项——福建省农业科学院科技创新团队建设项目(STIF-Y04)

详细信息

作者简介:
许惠滨(1984-),女,硕士,研究方向:水稻分子生物学(E-mail:xuhuibin413@163.com);王宗华(1962-),男,博士,研究员,主要从事分子植物病理研究(E-mail:zonghuaw@163.com);张建福(1971-),男,博士,副研究员,主要从事水稻分子生物学与分子育种研究(E-mail:jianfzhang@163.com)

中图分类号: S511
计量
- 文章访问数: 221
- HTML全文浏览量: 37
- PDF下载量: 5
- 被引次数: 0
出版历程
- 收稿日期: 2012-12-12
- 刊出日期: 2013-02-18

Cloning and Construction of Over-expression Vector of OsAPX Gene

XU Hui-bin^1,2,3,4,5,
LIAN Ling^2,3,4,5,
WEI Yi-dong^1,2,3,4,5,
ZHU Yong-sheng^2,3,4,5,
XIE Hua-an^2,3,4,5,
WANG Zong-hua¹,
ZHANG Jian-fu^2,3,4,5

1. College of Life Science,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350002,China;
2. Key Laboratory of Germplasm Innovation and Molecular Breeding of Hybrid Rice for South China,Ministry of Agriculture,P.R China/Fuzhou branch,National Rice Improvement Center of China/Fujian Engineering Laboratory of Crop Molecular Breeding/ Fujian Key Laboratory of Rice Molecular Breeding/ Rice Research Institute,Fujian Academy of Agricultural Sciences,Fuzhou,Fujian.350003,China;
3. Incubator of National key Laboratory of Fujian Germplasm Innovation and Molecular Breeding Between Fujian and Ministry of Sciences & Technology,P.R.China,Fuzhou,Fujian 350003,China;
4. South China Bases of National Key Laboratory of Hybrid Rice for China,Fuzhou,Fujian 350003,China;
5. National Rice Engineering Laboratory of China,Fuzhou,Fujian 350003,China

摘要

摘要: 通过RT-PCR法从粳稻日本晴中克隆到ascorbate peroxidase(APX)基因,该基因的cDNA长为768bp,包含完整的CDS序列。将克隆到的片段连接到植物表达载体pBI121的相应位置,构建1个APX基因的过表达载体pBI121-APX。利用农杆菌EHA105介导该植物表达载体在籼稻“多系1号”和“航1号”中的遗传转化,以期成功获得转基因植株,为后期研究APX基因在水稻耐储藏方面所发挥的功能打下基础。
- 克隆 /
- APX /
- 过表达载体 /
- 遗传转化
Abstract: A full length cDNA of ascorbate peroxidase(APX) gene was cloned from Oryza sativa L.Japonica Nipponbare with RT-PCR.It was 768 bp including the CDS,and was connected to the plant expression binary vector,pBI121,to construct an over-expression vector of APX gene.The pBI121-APX vector was transformed into indica rice,Duoxi No.1 and Hang No.1,mediating by Agrobacterium tumefaciens EHA105.With this,transgenic plants could be obtained for further study on the function of APX gene in rice storability.
- clone /
- APX /
- over-expression vector /
- genetic transformation

HTML全文

参考文献(25)

[1]	BEWLEY J D.Membrane changes in seeds as related togermination and the perturbation resulting from deteriorationin storage[J].Crop Science Society of America,1984,11:27-45.
[2]	LEPRINCE O,BUITINK J,HOEKSTRA FA.Radicles andcotyledons of recalcitrant seeds of Castanea sativa Mill.exhibitcontrasting responses of respiration to drying in relation todesiccation sensitivity[J].Journal Experimental Botany,1999,338:1515-1524.
[3]	FINKEL T,HOLBROOK N J.Oxidants,oxidative stress andthe biology of ageing[J].Nature,2000,408:239-247.
[4]	KANNAN K,JAIN S K.Oxidative stress and apoptosis[J].Pathophysiology,2000,7:153-163.
[5]	HALLIWELL B,GUTTERIDGE J M C.Free radicals inbiology and mMedicine[N].Oxford University Press,2002(3).
[6]	HOEKSTRA F A,WOLKERS W F,BUITINK J,et al.Membrane stabilization in the dried state[J].ComparativeBiochemistry and Physiology,1997,117A:335-341.
[7]	HENDRY G A,FINCH-SAVAGE W E,THORPE P C,et al.Free radical processes and loss of seed viability duringdesiccation in the recalcitrant species Quercus robur L.[J].New Phytologist,1992,122:273-279.
[8]	LEPRINCE O,HENDRY G A F,MCKERSIE B D.Themechanism of desiccation tolerance in developing seeds[J].Seed Science Research,1993,3:231-246.
[9]	VERTUCCI C W,FARRANT J M.Acquisition and loss ofdesiccation tolerance[N].Seed development and germination,1995,237-271.
[10]	SMITH M T,BERJAK P.Deteriorative changes associatedwith the loss of viability of stored desiccation-Tolerant anddesiccation-sensitive seeds[N].1995,701-746.
[11]	MCDONALD M B.Seed deterioration:physiology,repair andassessment[J].Seed Science and Technology,1999,27:177-237.
[12]	BEWLEY J D,BLACK M.Seeds:physiology of developmentand germination[N].Plenum Press,1994.
[13]	BEWLEY J D.Membrane changes in seeds as related togermination and the perturbation resulting from deteriorationin storage[J].Crop Science Society of America,1986,11:27-45.
[14]	PARISH D J,LEOPOLD A C,HANNA M A.Turgorchanges with accelerated ageing of soybeans[J].CropScience,1982,22:666-669.
[15]	ROBERTS E H.Cytological,genetical and metabolic changesassociated with loss of viability[J].Viability of seeds,1972:253-306.
[16]	MAGUIRE J D.Seed quality and germination[J].Thephysiology and biochemistry of seed dormancy andgermination,1977:219-235.
[17]	BAILLY C.Active oxygen species and antioxidant in seedbiology[J].Seed Science Research,2004,14:93-107.
[18]	WILSON J O,MCDONALD J B.The lipid peroxidation modelof seed ageing[J].Seed Science and Technology,1986,14:269-300.
[19]	ALLEN R D.Dissection of oxidative stress tolerance usingtransgenic plants[J].Plant Physiology,1995,107:1049-1054.
[20]	HALLIWELL B,GUTTERIDGE J M C.Free radicals inbiology and Medicine[N],Oxford University Press,2002(3).
[21]	KWON S Y,JEONG Y J,LEE H S,et al.Enhancedtolerances of transgenic tobacco plants expressing bothsuperoxide dismutase and ascorbate peroxidase in chloroplastsagainst methyl viologen-mediated oxidative stress[J].Plant,Cell and Environment,2002,25:873-882.
[22]	YOUNG P L,KWANG-HYUN B,HAENG-SOON L,et al.Tobacco seeds simultaneously over-expressing Cu/Zn-superoxide dismutase and ascorbate peroxidase displayenhanced seed longevity and germination rates under stressconditions[J].Journal of Experimental Botany,2010,61(9):2499-2506.
[23]	BAILLY C.Active oxygen species and antioxidant in seedbiology[J].Seed Science Research,2004,14:93-107.
[24]	WILSON J D,MCDONALD J B.The lipid peroxidation modelof seed ageing[J].Seed Science and Technology,1986,14:269-300.
[25]	BAILLY C,AUDIGIER C,LADONNE F,et al.Changes inoligosaccharide content and antioxidant enzyme activities indeveloping bean seeds as related to acquisition of dryingtolerance and seed quality[J].Journal of ExperimentalBotany,2001,52:701-708.