Taxol-induced apoptotic cell death in suspension cultures of Taxus cuspidata

Taxol caused apoptotic cell death of Taxus cuspidata in suspension cultures. Typical morphological and biochemical changes of apoptosis were observed by microscopy and total DNA agarose gel electrophoresis. Taxus cuspidata responded to the added Taxol by increasing the biosynthesis of Taxol. The percentage of apoptotic cells in total cells increased with the concentration of added Taxol. With Taxol added at 10 mg l^–1, the maximum concentration of Taxol produced was 23 mg l^–1, 3 times higher than that of the control culture.     

Cryopreservation of hairy root cultures of Vinca minor (L.) by encapsulation-dehydration

Hairy root cultures of Vinca minor and Ajuga reptans var. atropurpurea could be cryopreserved when the roots were precultured and encapsulated in 2% (w/v) alginate beads with 0.3 M sucrose and 0.5 M glycerol and dehydrated until the bead weight reached 25% of the initial weight before cooling in liquid nitrogen. Preculture and encapsulation of the roots with abscisic acid was effective in increasing the survival rates. For V. minor root tips moreover a sufficiently high survival rate of more than 70% was attained by eliminating glycerol from the preculture medium and dehydration of beads until 23% of the initial weight was reached instead of 25%.     

Extracellular production of a glycolipid biosurfactant, mannosylerythritol lipid, from Candida antarctica

Candida antarctica (sp. SY16) required avegetable oil as the carbon source to produce a biosurfactant, mannosylerythritol lipid (MEL-SY16). Biosurfactant production was 31 g l^–1 after 7 days in a batch culture and was not growth associated. In a two-stage culture, glycerol and oleic acid were used as an initial and a feeding carbon source, respectively, and 41 g l^–1 biosurfactant was produced after 8 days.     

Variation in potato microplant morphology in vitro and DNA methylation

Heterotrophic and autotrophic culture in agar and in polyurethane foam, the latter used as an alternative tissue support to agar, resulted in potato microplants with different in vitro morphologies. The microplants were visually characterised in terms of their relative developmental maturity, by comparing the respective leaf shapes in vitro with ontogenetic differences in leaf shape in glasshouse-grown potato plants. Cytosine methylation in the DNA of microplants of the different morphologies was determined using a method based on the AFLP technique but employing methylation-sensitive restriction enzymes (MSAP analysis) to test the hypothesis that DNA methylation could be used to characterise differences in microplant development in vitro. In three of the four treatments there was a good correlation between the visual assessment of relative morphological maturity and DNA base methylation levels. In these microplants there was increased DNA methylation in the leaves with mature leaf morphology represented by a decreased number of restriction fragments. The fourth in vitro morphology had the most juvenile leaf shape but did not have the predicted level of DNA methylation, having a relatively low number of restriction fragments. Subtraction analysis was used to discriminate the fragments that were unique to the juvenile and mature in vivo leaf morphologies. Comparison of the fragment patterns from the microplants with the latter reference profiles, confirmed the relationship with the total DNA methylation as detected by MSAP analysis, that is, the number of common fragments with the juvenile or mature in vivo leaf profiles, respectively. However, none of the fragment profiles, while sharing some common bands at random, was identical to any other; or to that of either the juvenile or mature in vivo leaf. The anomalous relationship of the microplants with most juvenile leaf shape and highest DNA methylation was confirmed. The measurement of DNA methylation in in vitro plants is discussed in the context of the development of a method to assess the quality of microplants produced by different in vitro protocols.     

Evaluation of a new culture method for mass-propagation of a photoperiod-temperature sensitive genic male sterile rice strain N19S

A newly established culture method was evaluated for application in mass-propagation of a photoperiod-temperature sensitive genic male sterile rice strain. It was found that the new culture method was very efficient for the regeneration of plants from callus of this strain, and a high concentration of 6-benzyladenine necessary for highly efficient plant regeneration culture did not seem to have negative effects on some agronomic traits investigated of the regenerated plants and their progenies. By the new culture method, a large number of plants could be regenerated in a relatively short time (approximately 8×10¹² plants within 300 days) from one naked seed-explant. Because of this, prominent drift of the PTGMS rice strain, which accumulates gradually during the course of propagation of the seeds through generations, in the response to photoperiod-temperature conditions could be avoided when this in vitro culture protocol is employed instead of seed propagation. Differences in rates of pollen fertility and seed setting were found between regenerated plants and seed-grown plants, and between their respective first and second progenies. These differences are also of important application value in two-line rice production: Higher pollen fertility of the regenerated plants can be applied with higher efficiency for the propagation of the strain seeds because they have much higher seed setting by self-fertilization; lower pollen fertility of the regenerated plant progenies will result in much lower seed setting by self-fertilization and is, therefore, conducive to the production of hybrid seeds of higher purity by natural crossing with the other line in the two-line system.     

Influence of carbon source and concentration on the in vitro development of olive zygotic embryos and explants raised from them

The influence of sucrose or mannitol on in vitro zygotic embryo germination, seedling development and explant propagation of olive tree (Olea europaea L.) was compared. Embryos germinated without sucrose in the medium but for adequate development of the seedlings to yield viable plants, a carbohydrate supply was necessary; both sucrose and mannitol were equally suitable for this purpose. However, when explants obtained from in vitro germinated embryos were cultured with mannitol or sucrose, then the polyalcohol promoted significantly more growth than sucrose by increasing shoot length, pairs of leaves formed, and breaking apical dominance. This improved the in vitro culture of olive plant material, thus allowing new olive clonal lines to be obtained in shorter times. This will assist in future breeding experiments with the species.     

In vitro plantlet regeneration from cotyledons of the tree-legume Leucaena leucocephala

Two plant regeneration methods applicable to Leucaenaleucocephala were developed. In the first method, involvingorganogenesis via callus formation, cotyledon, hypocotyl and root segments wereinitiated on MS medium containing different concentrations ofN⁶-benzyladenine (BA), 2,4-dichlorophenoxyacetic acid (2,4-D), andnaphthaleneacetic acid (NAA). Both compact (type I) and friable (type II) calliwere obtained from the cotyledon and hypocotyl explants treated with differentconcentrations of the growth regulators. Shoots were generated only from thefriable calli formed from the cotyledon explants. The calli formed from thehypocotyl explants did not generate shoots and the root explants died withoutforming callus. Cotyledon explants from 3–4 day old seedlings showedmaximum callus induction compared to those from older seedlings. In a secondmethod involving direct organogenesis, excised cotyledons were cultured on 1/2MS medium containing 10–35 mg l^–1N⁶-benzyladenine (BA) for 7–14 days. Transfer of thecotyledonsto regeneration medium containing low BA resulted in callus formation andsubsequent shoot regeneration from the base of the excised cotyledon explants,with up to 100% frequency. Regenerated shoots rooted best on a basal mediumcontaining no growth regulators.     

家猫的胚胎工程

家猫是惟一一种没有被列为珍稀或濒危的猫科动物。通过家猫的胚胎工程研究,对保护其它濒危猫科物种有重要的借鉴意义。本文描述了家猫的一般生殖特点,着床前的胚胎在体内的发育概况;综述了近年来对家猫的超数排卵,卵母细胞的体外成熟,体外受精,胚胎的体外培养,胚胎移植,冷冻保存和胚胎克隆等方面的研究进展。     

酵母菌在红葡萄酒酒精发酵串罐中稳定性研究

在生产条件下,对酵母菌在红葡萄酒酒精发酵串罐过程中的稳定性进行了研究。结果表明,在近1个月的时间内（相当于酵母菌细胞无性繁殖了200代）,串罐过程中的酵母菌细胞不仅能保持初始酵母菌的发酵活性和优良特性的稳定性,而且由于葡萄汁的选择作用,串罐用的酵母菌细胞的发酵活性比初始酵母菌的活性更强,因而其酒精发酵的启动和速度都更快。