发根农杆菌Ri质粒rol基因研究进展及在林木改良上的应用

Ri质粒上携带的诱根基因的表达不仅能导致植物被感染部位形成大量的毛根,而且由毛根容易获得再生的转化植株,这些植株可表现出许多能稳定遗传的表型变异,在植物遗传改良中有着广阔的应用前景。20世纪80年代以来,国内外学者对发根农杆菌Ri质粒及其rol基因进行了广泛深入的研究。本文重点综述了农杆碱型发根农杆菌Ri质粒的结构与功能,rol基因的位点与特征,Ri质粒的转化与rol基因的表达对植物生长发育的影响及在林木遗传改良上应用等方面的研究现状,并讨论了Ri质粒rol基因在林木遗传改良应用上存在的问题与应用前景。     

发根农杆菌Ri质粒rol基因研究进展及在林木改良上的应用

Ri质粒上携带的诱根基因的表达不仅能导致植物被感染部位形成大量的毛根,而且由毛根容易获得再生的转化植株,这些植株可表现出许多能稳定遗传的表型变异,在植物遗传改良中有着广阔的应用前景。20世纪80年代以来,国内外学者对发根农杆菌Ri质粒及其rol基因进行了广泛深入的研究。本文重点综述了农杆碱型发根农杆菌Ri质粒的结构与功能,rol基因的位点与特征,Ri质粒的转化与rol基因的表达对植物生长发育的影响及在林木遗传改良上应用等方面的研究现状,并讨论了Ri质粒rol基因在林木遗传改良应用上存在的问题与应用前景。     

Ri质粒诱导的植物发根培养系及其应用

阐述了具Ri质粒的发根农杆菌的生物学特性,介绍了毛状根的诱导、筛选及增殖培养的具体方法,提出了农杆菌转化植物细胞的影响因素,对国内外利用植物发根培养系进行次生代谢物质生产方面的研究进行了综述,并对Ri质粒诱导的发根培养系的应用作了展望。     

Ri质粒诱导的植物发根培养系及其应用

本文阐述了具Ri质粒的发根在杆菌的生物学特性,介绍了毛状根的诱导、筛选及增殖培养的具体方法,提出了农杆菌转化植物细胞的影响因素,对国内外利用植物发根培养系进行次生代谢物质生产方面的研究进行了综述,并对Ri质粒诱导的发根培养系的应用做了展望。     

Ri质粒在植物科学中应用的新进展

本文综述了发根农杆菌Ri质粒在植物学理论研究、有用植物次生代谢物生产、植物品种改良及植物栽培四个方面应用的最新进展。     

Ri质粒介导的毛状根体系建立及其在植物次生代谢产物合成中的研究进展

发根农杆菌Ri质粒可诱导植物产生毛状根体系,该体系具有遗传性状稳定且增殖速度快的特点,可用于药用植物次生代谢产物的生产研究,为利用生物反应器技术进行药用植物有效成分工业化水平的发酵培养开辟了新途径。本文主要综述了发根农杆菌Ri质粒介导的植物毛状根体系遗传转化机理,并对毛状根体系在药用植物次生代谢产物生产中的研究现状进行了深入分析,为从基因水平上调控植物次生代谢产物的合成提供新思路。     

Ri质粒基因转化植物细胞的机理

发根农杆菌(Agrobacteriumrhizogenes)含有一个约250Kb的大质粒,与Ti质粒转化植物细胞产生冠瘿瘤(crowngall)相似,它也可以侵染双子叶植物受伤部位的细胞并产生大量不定根(所谓发根或毛状根,(hairyroot),因此,这个大质粒被称为Ri质粒(根诱导质粒,Root-inducingplasmid)。     

用土壤杆菌(Agrobacterium)Ti质粒转化单子叶植物

<正> 迄今,植物直接遗传操作方面所取得的进展主要在双子叶植物,而单子叶植物包括主要的谷类作物尚不是容易进行遗传工程的对象。然而,最近的两篇论文发表后,看来,单子叶植物有可能在某一天同样会成为遗传工程的研究课题。利用Ti质粒和Ri质粒尤其是土壤杆菌的Ti质粒可以将外来基因插入对土壤杆菌易感的     

发根农杆菌研究进展

ReviewofStudiesonObtainingTransgenicPlantsbyAgrobacteriumrhizogenesMediatedGeneTransferSystemZhouYanqingZhanggenfaYuanBaojun(DepartmentofBiology,HenanNormalUniversity,Xinxiang453002)②苑保军现在河南省周口地区农业科学技术研究所工作．在植物基因工程中,农杆菌质粒介导的基因转移系统[37]是比较完善与有效的基因转移方法。目前,在根癌农杆菌Ti质粒的结构、功能及其被改造为载体系统与应用等方面均已取得很大进展的情况下,与之同属于根瘤菌科的发根农杆菌及其所携带的Ri质粒开始被广泛研究。本文就发根农杆菌Ri…     

农杆菌T—DNA介导的植物转基因的分子机制

前言 根瘤农杆菌(Agrobacterium tumefaciens)与发根农根菌(A.rhizogenes)同属根瘤菌科,是革兰氏阴性植物病原菌。前者感染植物引起冠瘿病(Crop gall tumour),冠瘿病用含有抗生素的培养基培养与除菌。可无限增殖;后者感染植物诱发出许多不定根,把不定根培养在上述培养基上也可迅速生长,多次分枝成毛状称毛状根(hairy root)。农杆菌感染机理很复杂。其感染作用起始于受伤的植物细胞分泌的大量化学物质时期。受伤植物细胞分泌的酚分子诱导农杆菌怀有的Ti质粒或Ri质粒上的基因活化,尔后农杆菌紧附植物细胞并把Ti质粒或Ri质粒上一部分DNA(称作T—DNA,tranfer DNA)转移到植物染色体上。T.DNA共价整合后编译合成的新的低分量的代谢物,称为冠瘿碱。     

T-DNA fragments of hairy root plasmid pRi8196 are distantly related to octopine and nopaline Ti plasmid T-DNA

Agrobacterium Ti (tumor-inducing) and Ri (root-inducing) plasmids transform dicot plant cells by insertion of a specific plasmid sector called T-DNA (transferred DNA) into host plant nuclear DNA. The mannopine-type Ri plasmid pRi8196 contains four BamHI fragments that encompass core T-DNA. We report Southern hybridization studies that show that these four fragments have no strong homology to octopine-, nopaline-, or agropine-type Ti plasmids. We detected and mapped very weak homology regions, most of which are assignable to opine synthase or opine catabolic functions on the Ti plasmid. We found no homology between Ri T-DNA and the region of Ti T-DNA that encodes tumor morphology functions.     

Transfer of the octopine T-DNA segment to plant cells mediated by different types of Agrobacterium tumor- or root-inducing plasmids: generality of virulence systems.

Genetic complementation studies demonstrated that the transfer to plant cells of the octopine T-DNA, entirely present as the only part of the tumor-inducing (Ti) plasmid on the plasmid pAL1050, was effected by the virulence systems from related plasmids, viz. the nopaline Ti plasmid pTiC58, the limited host range plasmid pTiAg57, and the root-inducing (Ri) plasmid pRi1855. Rhizobium symbiosis plasmids were not capable of effecting the introduction of pAL1050 into plant cells.     

The role of endogenous auxin in root initiation

This paper is the second part of a review which considers evidence for the involvement of auxin in root initiation. Part II examines the research being carried out with transformed plant tissues. Agrobacterium rhizogenes causes abundant root initiation at the site of inoculation. Ri plasmid T-DNA contains several genes which encode enzymes involved in the biosynthesis and metabolism of indole-3-acetic acid. Transfer of various fragments of the Ri plasmid has also been reported to confer increased sensitivity to auxin upon plant cells. Controlled expression of these genes in the plant genome potentially offer an insight for developmental plant physiologists into the role of plant growth substances in the process of root initiation. The importance of absolute levels of IAA in the stimulation of root initiation is discussed.     

Binary plant vector based on Ri plasmid and part of T-DNA of the Ti plasmid

A binary vector system inA. tumefaciens for the introduction of foreign genes into the plant genome was developed. The first component is the Ri plasmid and the second component is a small vector plasmid, replicating inAgrobacterium, which carries the 25 bp terminal sequence of the Ti plasmid, theNos gene as the selectable marker and neighbouring sequences of the Ti plasmid. Functions necessary for integration are providedin trans by the virulence region of the Ri plasmid. Transformed cells are selected on the basis of hairy root tumor proliferation and agropine synthesis. They also showNos activity coded by the gene on the small part of Ti T-DNA.     

A comparison of virulence determinants in an octopine Ti plasmid, a nopaline Ti plasmid, and an Ri plasmid by complementation analysis of Agrobacterium tumefaciens mutants

Transposon-insertion mutants with vir^? Ti plasmids were characterized and then used in complementation experiments. One of the mutants (LBA 1517) had a mutation in a newly discovered vir locus called virF. The virF mutation led to a strongly diminished virulence on tomato and tobacco, but not on certain other plant species. Also a mutant (LBA 1505) was isolated with a mutation somewhere in the bacterial genome but outside the octopine Ti plasmid that caused a restriction in host range for tumor induction. Introduction of a nopaline Ti plasmid or an Ri plasmid into LBA 1505 did not restore normal virulence, showing that the vir gene affected in LBA 1505 determines a factor which is essential for normal tumor induction both by different types of Ti plasmids and by the Ri plasmid. The introduction of R primes containing part or all of the octopine Ti plasmid virulence region led to a restoration of virulence in strains with a vir^? nopaline Ti plasmid. Also the transfer of an Ri plasmid to a large number of different vir^? octopine or nopaline Ti plasmid mutants rendered these strains virulent. These results indicate that the octopine Ti plasmid, the nopaline Ti plasmid, and the Ri plasmid each have a similar virulence system which can mediate the transfer of T-DNA to plant cells from different types of Ti or Ri plasmids. In complementation experiments between vir^? octopine Ti plasmid mutations and vir^? nopaline Ti plasmid mutations it was found that equivalent functions are determined by the areas of DNA homology in the virulence regions of these two types of Ti plasmids. The previously defined octopine Ti plasmid virC locus appeared to consist of two different loci. One of these loci was found to be in a region of the octopine Ti plasmid which does not share DNA homology with the nopaline Ti plasmid, and was therefore called virO (octopine Ti plasmid specific). For the other locus the name virC was retained. Whereas mutations in the virC locus were avirulent on all plant species tested, mutations in virO were avirulent on tomato and pea, but virulent on sunflower and Nicotiana rustica. VirO^? mutants produced rooty tumors on Kalanchoë tubiflora.     

A critical review on use of Agrobacterium rhizogenes and their associated binary vectors for plant transformation

Agrobacterium rhizogenes, along with A. tumefaciens, has been used to affect genetic transformation in plants for many years. Detailed studies conducted in the past have uncovered the basic mechanism of foreign gene transfer and the implication of Ri/Ti plasmids in this process. A number of reviews exist describing the usage of binary vectors with A. tumefaciens, but no comprehensive account of the numerous binary vectors employed with A. rhizogenes and their successful applications has been published till date. In this review, we recollect a brief history of development of Ri-plasmid/Ri-T-DNA based binary vectors systems and their successful implementation with A. rhizogenes for different applications. The modification of native Ri plasmid to introduce foreign genes followed by development of binary vector using Ri plasmid and how it facilitated rapid and feasible genetic manipulation, earlier impossible with native Ri plasmid, have been discussed. An important milestone was the development of inducible plant expressing promoter systems which made expression of toxic genes in plant systems possible. The successful application of binary vectors in conjunction with A. rhizogenes in gene silencing and genome editing studies which are relatively newer developments, demonstrating the amenability and adaptability of hairy roots systems to make possible studying previously intractable research areas have been summarized in the present review.     

Genetic transformation of Medicago truncatula using Agrobacterium with genetically modified Ri and disarmed Ti plasmids

Summary Fertile transgenic plants of the annual pasture legume Medicago truncatula were obtained by Agrobacterium-mediated transformation, utilising a disarmed Ti plasmid and a binary vector containing the kanamycin resistance gene under the control of the cauliflower mosaic virus 35S promoter. Factors contributing to the result included an improved plant regeneration protocol and the use of explants from a plant identified as possessing high regeneration capability from tissue culture. Genes present on the T-DNA of the Ri plasmid had a negative effect on somatic embryogenesis. Only tissue inoculated with Agrobacterium strains containing a disarmed Ti plasmid lacking the T-DNA region or a Ri plasmid with an inactivated rol A gene regenerated transgenic plants. Fertile transgenic plants were only obtained with disarmed A. tumefaciens, and the introduced NPT II gene was transmitted to R₁ progeny.Abbreviations BAP 6-benzylaminopurine - NAA 1-naphthaleneacetic acid - NPT neomycin phosphotransferase