原生质体融合构建高效降解工程菌的研究

研究了球形红假单胞菌与酿酒酵母菌的原生质体融合条件，并对融合子性能进行了测定.结果表明两种菌株分别在溶菌酶——EDTA和蜗牛酶——巯基乙醇处理后形成原生质体，两种原生质体在聚乙二醇（PEG）和Ca     

胡杨悬浮细胞原生质体游离及质膜单离子通道测定

该文以胡杨 (Populuseuphratica)悬浮细胞为材料 ,利用 1 0 %纤维素酶“onozuka”R 10、0 0 1%果胶酶Y 2 3、0 15 %离析酶R 10和 0 1%半纤维素酶的混合酶液消化细胞壁 ,得到原生质体 .并利用膜片钳细胞贴附技术分别测到质膜内向和外向单通道电流 .通道电流在不同水平上变化 ,表明并非只有一种类型的通道开放 .树木细胞原生质体的分离以及利用膜片钳测定细胞质膜离子通道的成功实践为深入研究木本植物抗盐的细胞学机制奠定了基础     

Plant regeneration from protoplasts of Nicotiana alata ‘Crimson Bedder’ for somatic hybridisation studies

Protoplasts could be isolated readily from mesophyll and suspension cells of Nicotiana alata cultivar ‘Crimson Bedder’ and could be cultured in liquid media to yield small colonies. Suspension culture protoplasts exhibited a higher plating efficiency, and colonies regenerated from mesophyll protoplasts possessed enhanced plant regeneration capacity. These protoplast systems could be used for somatic hybridisation assessment with Petunia using a simple procedure for manual isolation and culture of heterokaryons.     

siRNA对烟草原生质体中TMV RNA积累的干扰作用研究(英文)

 RNA干扰被认为是转录后基因沉默的一种机制。RNA干扰通过小干扰RNA特异性降解目标mRNA来沉默基因表达。本文以烟草花叶病毒126kD蛋白为靶蛋白,在原生质体水平上研究了小干扰RNA对病毒侵染的干扰和抑制作用。ELISA和Northern杂交的实验结果表明,共转染小干扰RNA和TMV的原生质体内检测到较低的病毒含量。在枯斑寄主上,叶片接种小干扰RNA和TMV共转染原生质体后,与对照叶片相比,仅有很少量的病斑产生。这说明,小干扰RNA能够在原生质体水平对病毒起到干扰和抑制作用。因此认为,烟草原生质体系统有利于快速和定量分析小干扰RNA介导对植物病毒的抑制作用。     

香蕉枯萎病菌TR4原生质体转化和基因敲除体系构建

正香蕉枯萎病是一种典型的维管束和土传真菌病害,也是最具毁灭性的植物病害之一~([1])。香蕉一旦感病,很难结果,而且目前尚无有效的防治措施~([1])。因此,研究植物病原菌侵染进程和致病机制,可以拓宽思路以寻求植物病害防治新途径。香蕉枯萎病是由尖孢镰刀菌古巴专化型(Fusarium oxysporum f.sp.cubense,Foc)侵染所致~([1])。对生产影响最大的Foc是侵染包括大蜜哈、     

The phenotypic characterisation of R2 generation transgenic rice plants under field and glasshouse conditions

Summary The phenotypes of seed progeny (R₂ generation) of Oryza sativa L. cv. Taipei 309, which carried the neomycin phosphotransferase II (npt II) gene, were compared with those of non-transformed, protoplast-derived plants of the same generation and non-transformed, seed-derived plants under field and glasshouse conditions. Under both conditions the transgenic plants were generally smaller, took longer to flower and had reduced fertility. Significant differences were observed between individuals within the group of transgenic plants. The npt II gene was present in most of the transgenic plants, but NPT II activity was only detected in a minority of individuals.     

Protoplast technology in relation to crop plants: Progress and problems

Summary The ready availability of isolated plant protoplasts, and the ability of these naked plant cells to fuse together, has greatly stimulated interest in the production of plant somatic hybrids. This new protoplast technology may enable interspecies hybrids to be obtained which are otherwise not possible. Many such interspecies hybrids are required amongst the crop plants, but application of this new technology to these species in presently prevented by our inability to culture cells of many of these species in vitro, and to regenerate plants reproducibly from cultured tissue.Agricultural Research Council Group, Department of Botany, University of Nottingham, U.K.     

Chromosomal instability in cell- and tissue cultures of tomato haploids and diploids

Summary The effect of the tissue culture system, the genotype and the ploidy level of the plant material used as explant source on the stability of the ploidy level of plants regenerated fromcell and tissue cultures of tomato was investigated. In addition the use of the chloroplast number in guard cells as a measure for ploidy level was evaluated. Haploids of tomato were very instable, which instability was observed already in somatic root tip and leaf cells. The number of regenerated plants that retained the original ploidy level differed significantly between the tested haploids. The plants that were regenerated from leaf explants of diploids were predominantly diploid in contrast to the plants regenerated from established callus cultures and protoplast where the majority was tetraploid.     

Protoplast technology for the breeding of top-fruit trees (Prunus, Pyrus, Malus, Rubus) and woody ornamentals

Summary Until recently, temperate fruit trees and woody ornamentals have been regarded as recalcitrant to biotechnological breeding approaches based on protoplasts. This however should no longer be the case, as procedures are now available, not only for the regeneration of complete plants from protoplasts of various tissues of such species, but also for the exploitation of protoplast technology for their genetic manipulation. This paper will examine the recent advances and state of the art in this domain, with particular attention to the use of protoplast technology as a novel tool in the breeding of rosaceous top-fruit tree species and woody ornamentals. Problems and their solutions within the context of regenerating plants from isolated protoplasts of stone (Prunus spp.), pome (Pyrus spp., Malus spp.) and small (Rubus spp.) fruits, and of several shrubby ornamental genotypes (Lonicera spp., Weigela spp., Forsythia spp., Cotoneaster spp.) will be addressed. Interspecific (Prunus spinosa + Prunus cerasifera) and intergeneric (Forsythia spp. + Syringa spp.) somatic hybridization within this group of species, as well as the use of protoplasts for host/pathogen interaction studies (Pyrus/Erwinia amylovora) will also be discussed.     

Transfer of resistance to Verticillium dahliae Kleb. from Solanum torvum S.W. into potato (Solanum tuberosum L.) by protoplast electrofusion

Summary Interspecific somatic hybrid plants were regenerated after electrofusion of mesophyll protoplasts with the objective of transferring resistance to Verticillium dahliae from Solanum torvum into potato. Early selection of the putative hybrids was based on differences in cultural behaviour of the parental and hybrid calli (particularly the ability of the latter to regenerate early) in combination with morphological markers. Four putative hybrids were recovered from hundreds of calli, probably resulting from complementation of the two parental genomes. The regenerates were tetraploids (2n=4×=48 chromosomes) and exhibited intermediate traits including leaf form, plant morphology and the presence of anthocyanin. The hybrid nature of the four selected plants was confirmed by examining isoenzyme patterns for isocitrate dehydrogenase (Idh), malate dehydrogenase (Mdh), phosphoglucoisomerase (Pgi) and 6-phosphogluconate dehydrogenase (6-Pgd). While the hybrid plants rooted readily and grew vigorously under in vitro conditions, in the greenhouse their development and growth were retarded by difficulties in rooting. When grafted on potato or S. torvum rootstocks, the hybrid plants recovered normal development and growth. Again, they exhibited intermediate morphological traits. Tests for resistance realized in vitro with medium containing 50% Verticillium wilt filtrate showed that all the somatic hybrids were resistant to the fungus filtrate.