Expression of antisense chalcone synthase RNA in transgenic hybrid walnut microcuttings. Effect on flavonoid content and rooting ability

Walnut somatic embryos (Juglans nigra × Juglans regia) were transformed with a vector containing a neomycin phosphotransferase II, a -glucuronidase and an antisense chalcone synthase (chs) gene. This antisense construct included a 400 bp cDNA fragment of a walnut chs gene under the control of the duplicated CaMV-35S promoter. Molecular, biochemical and biological characterizations were performed both on transformed embryos propagated by secondary somatic embryogenesis and on microshoots developed by in vitro culture of embryonic epicotyls from somatic embryos. Thirteen transformed lines with the vector containing the antisense chs gene, one line with only the gus and nptII genes and one untransformed line were maintained in tissue culture. Six of the antisense lines were shown to be flavonoid-deficient. They exhibited a strongly reduced expression of chs genes, very low chalcone synthase activity and no detectable amounts of quercitrin, myricitrin, flavane-3-ols and proanthocyanidins in stems. Rooting tests showed that decreased flavonoid content in stems of antisense chs transformed lines was associated with enhanced adventitious root formation. Free auxin and conjugated auxin contents were determined during the latter phase of the micropropagation, and no variations were detected between control and antisense chs transformed lines. The in vitro plants developed a large basal callus and apical necrosis upon auxinic induction and the transformed lines highly deficient in flavonoids were more sensitive to exogenous application of indolebutyric acid (IBA).     

Etiolation of stock plants affects adventitious root formation and hormone content of pea stem cuttings

Stock pea plants (Pisum sativum L.) were etiolated fully or partially at the third internode that acted as the cutting base. The etiolation started the fifth day after sowing and lasted till cutting preparation. Cuttings derived from partially etiolated plants rooted more than non-etiolated ones while fully etiolated ones rooted more only after treatment with 1% sucrose solution for 4 days. Endogenous IAA in the base of etiolated cuttings was higher during the first 24 h after cutting preparation than in the control. Z/ZR did not show significant differences while iAde/iAdo was higher in the control. Ethylene was increased 24 h after cutting preparation and the increase was greater from partially etiolated cuttings. The results showed that besides IAA and cytokinins, which played a role in the rooting of cuttings, sucrose influenced rooting in the case of fully etiolated stock plants.     

Low activity of amine-oxidases and accumulation of conjugated polyamines in disfavour of organogenic programs in Chrysanthemum leaf disc explants

Foliar discs (8 mm diameter) from expanding leaves of the middle part of vegetative shoots of Chrysanthemum morifolium Ramat raised in vitro were induced to form directly on specific media in vitro either roots or vegetative buds, or callus. The budding programme, on its specific medium, was deviated to callus formation by the addition of 2 mM β-OH-E (β-OH-ethyldrazine, an inhibitor of diamine oxidase). Conversely vegetative buds instead of callus were formed on the callus medium in the presence of 2 mM DFMO (difluoromethylornithine, an inhibitor of ornithine decarboxylase). Callus formation was characterized by high accumulation of free and particularly conjugated polyamines (PA), very low or undetectable activities of diamine- and polyamine oxidases, and transglutaminase. DFMO-deviation of callus initiation in favour of bud formation lowered the accumulation of PA and increased the activity of amine-oxidases. The high catabolism of PA in the organogenic (rooting, budding) programs was questioned as to its role in developmental processes. This revised version was published online in June 2006 with corrections to the Cover Date.     

Development of a sterile cultivation system for rooting of shoot tip cultures (red raspberries) in duroplast foam

In order to improve the availability of oxygen for shoot tip cultures during the course of rooting and to overcome some further agar-related problems a sterile foam cultivation system was developed. Three commercially available foam substrates have been tested, but only a wide pore phenol resin foam proved to be useful in the rooting of red raspberry shoots (Rubus idaeus cv. ‘Gigant’). In comparison to root growth in agarous medium, the newly developed foam cultivation system improved the ramification of the root system, increased the number of root hairs significantly and reduced the friability (lignification) of roots. Also shoot length increased significantly. From the growth experiments it can be concluded that the chemical rather than physical properties of the foam substrate are growth limiting. Still some efforts must be made to improve and standardize the properties of the foam substrate for routine use in mass propagation of organ cultures in vitro.     

彩色马蹄莲的组织培养

以彩色马蹄莲芽眼为外植体,在添加不同激素配比的MS培养基上进行培养.结果表明:(1)较适宜诱导愈伤组织的激素组合是1.5 mg/L Zt+0.1 mg/L NAA、3.0 mg/L 6-BA+0.1 mg/L NAA或0.5 mg/L 2,4-D+0.1 mg/L NAA,出愈率分别为94.12%、100%和86.67%;较适宜诱导不定芽的激素组合是2.0 mg/L 6-BA+0.1 mg/L NAA,芽丛诱导率可达87.3%;芽增殖较适宜的激素组合是2.0 mg/L 6-BA+0.1 mg/L NAA或2.0 mg/L KT+1.0 mg/L NAA,而且在继代培养中,交替使用这两种激素组合,繁殖系数可达4倍以上;生根培养基以1/2 MS+0.3 mg/L NAA+0.2 mg/L IAA较为适宜,生根率达95%以上.(2)彩色马蹄莲小块茎形成的主要影响因素是培养基中的蔗糖浓度,最适宜蔗糖浓度为10%;10 mg/L多效唑和8 mg/L NAA对小块茎的形成有明显的促进作用,而分裂素则表现出较强的抑制作用.     

In vitro screening of potato against water-stress mediated through sorbitol and polyethylene glycol

With the objective to develop a practical and effective method of screening potato for drought tolerance, shoot and root growth in microtuber-derived plantlets was studied in vitro in three genotypes with known root mass production under field conditions. Different levels of water-stress were induced using five concentrations of either sorbitol or polyethylene glycol (PEG) in MS medium. Water potential of various media ranged from −0.80 MPa to −2.05 MPa. Water-stress in culture adversely affected plantlet growth, and genotypes differed for their responses. Genotype IWA-1 was less affected than IWA-3 and IWA-5. At the same level of water potential, sorbitol had lower adverse effect than PEG; the latter being sticky. Genotype × sorbitol and genotype × PEG interactions were significant. At 0.2 M sorbitol and 0.003 M PEG, IWA-1 had significantly more roots with higher total root length, root volume, as well as root-dry weight than those of IWA-3 and IWA-5, whereas the latter two genotypes were at par for all these characters. This pattern was similar to the reported pattern of these genotypes for root-dry weight under field conditions. It is concluded that in vitro screening of potato under specific and limited water-stress conditions may provide a system for effectively differentiating the genotypes for their expected root mass production under field conditions.     

An efficient in vitro shoot regeneration from leaf petiolar explants and ex vitro rooting of Bixa orellana L.- A dye yielding plant

Physiology and Molecular Biology of Plants - Bixa orellana L. (Bixaceae) is a multipurpose tree grown for the production of commercially important dyes. In the present study, an efficient,...     

向日葵离体再生体系的建立

为了建立高效的向日葵离体再生体系,从基因差异、外植体取材、生长素和细胞分裂素浓度、附加物的添加等方面出发,对向日葵愈伤诱导、分化、生根等过程进行了系统优化。结果表明：杂交材料相对于自交材料更容易实现再生;最佳外植体是生长4 d的子叶;最佳愈伤诱导培养基是MS培养基 (MS) +2.0 mg/L 6-苄基腺嘌呤 (6-BA)+0.5 mg/L奈乙酸 (NAA)+1.0 mg/L激动素 (KT),诱导率最高可达100％;最佳分化培养基是MS+0.2 mg/L 6-BA+0.5 mg/L NAA+0.3 mg/L KT+0.3 mg/L硝酸银 (AgNO3)+0.2 g/L活性炭 (AC),芽分化率可达71％;最佳生根培养基是1/2 MS+0.6 mg/L吲哆丁酸 (IBA),生根率最高为77％。方差分析表明,材料基因型、外植体生长时间、激素、AgNO3、AC对向日葵再生呈现显著性影响。     

欧洲云杉的扦插基质选择和穗条效应研究

以欧洲云杉6年生母树为采穗母株,系统研究了不同插穗条件对生根和幼苗生长的响应,同时观测了网袋容器基质配比对扦插幼苗生长的影响.结果表明:扦插后20 d愈伤组织开始形成,37 d时根尖开始出现,50～65 d是生根的高峰期和根系发育期,65 d时扦插苗根系基本形成.不同基质对欧洲云杉插穗生根率影响达到极显著水平,1份泥炭...     

杜鹃花属植物扦插繁殖研究进展

我国有着丰富的杜鹃花资源,但有关其繁殖应用的研究还具有一定的局限。种子育苗耗时长,组培育苗成本和技术要求高,都不适于杜鹃花属植物的大面积生产。扦插繁殖快,还可保持母本的优良性状。从插条的选择,准备,插条的生根激素处理,扦插基质的选择,外界环境条件对扦插成活率的影响及扦插后的养护管理等六个方面对杜鹃的扦插繁殖技术的研究进行综述,以期推进杜鹃花属植物,尤其是中国野生杜鹃的引种驯化和大面积的推广应用。