权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

INTRODUCE OF EFFECTIVE ANTIBIOTICS AND VACCINES ON FORAGE CROPS BY MEANS OF GENETIC ENGINEERING

通过基因工程在饲料作物上引入有效的抗生素和疫苗

基本信息

批准号：
13660270
负责人：
AKASHI Ryo
金额：
$ 2.3万
依托单位：
University of Miyazaki
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

项目摘要

We have improved our previously published technique for plant regenerationfrom seed-derived embryogenic callus of bahiagrass by keeping the callus size small through regular partition. This modifiedculture remains regenerable for at least 18 months and results in a decrease of albino plants among regenerants. Also, in transformation experiments, an increased surface area is available for gene delivery, and a greater number of cells can potentially regenerate. Using a simple self-built particle inflow gun with improved technical features, we have optimized delivery conditions and bombarded the small calli with pDB1, a construct carrying both the GUS reporter gene and a bialaphos resistance gene (bar). Numerous calli have been selected on bialaphos-supplemented medium and subjected to regeneration conditions. These calli remained resistant and continue to display GUS activity. Shoots as well as embryo-like structures formed from resistant calli under regeneration conditions and green pla … More nts were obtained.Super roots of Lotus corniculatus are a fast growing legume root culture that allows continuous root cloning, direct somatic embryogenesis and mass regeneration of plants under entirely growth regulator-free culture conditions. These features are unique to root cultures and are now stable expressed since the, culture has been isolated almost 5 years ago. In several years of experimentation, all our attempts to directly transform super roots with Agrobacterium failed for unknown reasons. Only recently, by using super root-derived callus as acceptor tissue, we were able to demonstrate transient expression of a reporter gene (GUS) in a tissuederived from super roots. GUS expression was also, visible in roots and shoots regenerated from transformed calli. The super root character is stable maintained through the callus phase. Roots derived from these calli can serve as explants for new super root cultures. This stability of the callus phase makes it unlikely that transformation was made possible through the loss of protective genetic information, such as a spontaneous A. rhizogenes infection, through callus initiation. Less

我们改进了先前发表的百喜草种子胚性愈伤组织再生技术，通过规则分化使愈伤组织保持小尺寸。这种改良的培养至少可再生18个月，并导致再生植株中白化植株的减少。此外，在转化实验中，增加的表面积可用于基因传递，并且更多的细胞可以潜在地再生。我们利用一种简单的自制颗粒流入枪，改进了技术特点，优化了输送条件，并用pDB1轰击小愈伤组织，pDB1是一个同时携带GUS报告基因和双alaphos抗性基因（bar）的结构体。在bialaphos培养基上选择了大量愈伤组织，并进行了再生条件试验。这些愈伤组织保持抗性并继续显示GUS活性。在再生条件下，抗性愈伤组织形成芽和胚样结构。莲藕超级根是豆科植物快速生长的根培养物，在完全无生长调节剂的培养条件下，可以实现根的连续克隆、直接体细胞胚发生和植株的大量再生。这些特征是根培养物所特有的，自从5年前分离出根培养物以来，这些特征现在已经稳定地表达出来了。在几年的实验中，我们所有用农杆菌直接转化超级根的尝试都失败了，原因不明。直到最近，通过使用超级根衍生的愈伤组织作为受体组织，我们才能够证明报告基因（GUS）在超级根衍生的组织中瞬间表达。GUS在愈伤组织再生的根和芽中也有表达。超根性状在愈伤组织阶段保持稳定。这些愈伤组织的根可以作为新的超级根培养的外植体。愈伤组织阶段的这种稳定性使得转化不太可能是通过失去保护性遗传信息而实现的，例如通过愈伤组织形成自发的根状芽孢杆菌感染。少