摘要
三价铁螯合物还原酶(Ferric-chelator reductase,FCR)在植物对三价铁的吸收过程中起着关键作用,导入外源三价铁螯合物还原酶基因是改良植物耐缺铁能力的有效途径。香橙(Citrus junos Sieb.ex Tanaka)是目前表现最好的柑橘耐缺铁砧木之一,在铁胁迫时,香橙三价铁螯合物还原酶基因(CjFRO2)在根尖表皮细胞中高效表达,根系三价铁还原活性显著增强。以枳[Poncirus trifoliata(L.)Raf.]上胚轴为材料,利用根癌农杆菌介导法将CaMV 35S-CjFRO2基因表达单元导入枳基因组中,拟通过CjFRO2基因在转基因枳中的组成型表达,提高转基因枳的铁吸收能力。经过PCR检测,Southern杂交分析,获得4个单拷贝的转CjFRO2基因枳株系PF2、PF3、PF5和PF8。Real-TimePCR分析表明,CjFRO2基因在4个转基因枳株系中都能有效表达,并在其无性繁殖后代中也能够稳定表达。为培育耐缺铁优良柑橘砧木新品种奠定了基础。
Iron is an essential trace element for plants,and it is well known that iron deficiency is a significant global problem.Fruit trees are perennial plants,have been more seriously affected by iron deficiency.Breeding for tolerance to iron deficiency using conventional breeding and selection techniques is very time-consuming,laborious,and inefficient.Genetically modifying fruit trees to improve iron absorption is a complementary approach.Ferric chelate reductase（FCR） is a rate-limiting enzyme in iron uptake of plant,plays an important role in iron homeostasis.Citrus junos is characterized by its tolerance to iron deficiency in alkaline and calcareous soils,the gene of Citrus junos ferric chelate reductase（CjFRO2） show strong expression in root tip,reductase activities are greatly increased.Poncirus trifoliata is the predominant citrus rootstock in China,but it is highly sensitivity to iron deficiency.In order to enhance iron uptake ability,the gene of CjFRO2 driven by CaMV 35S promoter was introduced into the Poncirus trifoliata via Agrobacterium-mediated transformation.The primary transgenic plants were screened using PCR（polymerase chain reaction）,Southern hybridization for detection of transgenic integrations and Real-Time for investigation CjFRO2 gene expression.Four independent single copy transgenic lines were obtained and showed constitutive expression of CjFRO2,and the expression of transgenes after vegetative propagation was highly stable.
出处
《果树学报》
CAS
CSCD
北大核心
2011年第5期808-813,共6页
Journal of Fruit Science
基金
长江学者和创新团队发展计划项目(PCSIRT)
国家高科技研究发展计划项目(863)(2011AA100205)
柑桔产业技术体系(CARS-27)
农业部"引进国际先进农业科学技术"项目("948"计划)
中央高校基本科研业务费专项(XDJK2009C136)
