Methods and Techniques to Enhance the Production of Secondary Metabolites in Hairy Roots

Hairy roots culture is a new means to gain the plant secondary metabolites. How to enhance the production of secondary metabolites in hairy roots becomes a new hotspot recently. The authors summarize recent trends and results about the increase of the production of secondary metabolites in medicinal plant hairy roots. The change of culture conditions and the application of biotechnologies can enhance the production of secondary metabolites in hairy roots.     

不同保存方法对光皮桦总DNA提取效果的影响

光皮桦是一种顽拗类植物,其鲜叶组织中富含酚类、糖类及其他次生代谢物。这些物质很容易使鲜叶发生褐变,从而严重影响植物基因组DNA的提取质量。本文以新鲜嫩叶为对照,比较了-20℃冷藏、硅胶干燥、改进的饱和NaCl-CTAB溶液保存和核分离缓冲液固定4种鲜叶保存方法对提取光皮桦基因组总DNA效果的影响,寻找最佳的光皮桦嫩叶保存方法。并以提取的核酸得率、纯度、片段分布情况、是否含有PCR反应抑制剂等指标来评价其保存效果。结果表明:-20℃冷藏的样品未能提到DNA;新鲜样品、改进的饱和NaCl-CTAB溶液保存和核分离缓冲液保存的样品均提到纯度较好的DNA,大小在48Kb左右,得率为400!g/g鲜叶左右,并获得条带清晰、多态性好的PCR扩增图谱;用硅胶干燥保存的样品也能提到DNA,但得率低,为51.2!g/g鲜叶,且未获得条带完整、清晰的PCR扩增图谱。     

Effect of in vitro propagation methods on field performance of two strawberry cultivars

Summary Strawberry cultivars, Redcoat and Veestar, propagated by meristem culture (MC), callus culture (CC) and direct shoot regeneration (DR) from leaf disks were compared for their vegetative and reproductive characters with standard runner (SR) propagated plants under field conditions. In the planting year, in vitro propagated plants of both cultivars had the same number of leaves as SR plants, but in vitro propagated Redcoat produced fewer stolons per plant than SR plants. However, in the following year, in vitro propagated mother plants of both cultivars had more leaves and higher runner production than SR mother plants. Flowering and fruiting behaviour of Veestar was not appreciably influenced by in vitro propagation methods. However, in vitro propagated plants of Redcoat flowered earlier and produced more flowers and fruits than SR plants, but still maintained normal berry weight. Among in vitro propagated plants, DR plants of Redcoat were the earliest to flower, whereas MC plants produced more flowers and fruits. The field performance of the first daughter plants derived from the in vitro propagated plants was consistent with their respective mother plants. Leaf shape of both cultivars was not altered by in vitro propagation. Phenotypic abnormalities were mainly confined to occurrence of yellow leaf variants in MC and CC plants and occasional appearance of plants with irregular flowering and growth habit among CC plants.NRCC No. 38004     

The Experimental and Commercial Release of Transgenic Crop Plants

With advances in recombinant DNA methods and transformation procedures, it is possible to transfer genes into crop plants from unrelated plants, microbes and animals. Many of the modifications being carried out, or envisaged, are for disease and pest resistance, product quality and tolerance to environmental stress, but there are additional opportunities to modify crops to give specialized products for industrial or pharmaceutical use. Some of the characteristics of transgenic plants are considered, including: transgene copy number, position, expression, stability, pleiotropy, selectable marker genes and somaclonal variation. There have been several hundreds of field trials with transgenic plants, and the first transgenic varieties are likely to be approved for commercial production in 1993. Before releasing transgenic plants, it is necessary to carry out a risk assessment to determine whether the transgenic variety will behave differently from a conventionally bred variety. Assessment procedures are being harmonized internationally by various organizations. There is a growing commitment to apply these genetic modification methods to crops in developing countries, as genes relevant to their crops and environments become available.     

Comparison of selection indices to screen lowland rice for drought resistance

Summary Although many selection indices have been used to screen rices (Oryza sativa L.) for drought resistance, there has been little comparison of the relative merits of these indices. Research was conducted to compare drought resistance as estimated from grain yields, canopy-temperature-based stress indices, visual scoring, and uprooting force for 30 rice genotypes grown in the field with a puddled Maahas clay (Typic Tropaquept) and to evaluate traits related to drought resistance from nonstressed plants grown in the field and in aeroponic culture. Water deficit was imposed in the field by withholding irrigation from 45 to 75 days after transplanting compared to a continuously flooded control. Grain yields in the stress treatment were most strongly correlated with visual assessment of drought stress symptoms according to a standard evaluation system (r = 0.66). Canopy-temperature-based indices were also significantly correlated with grain yields of the stress treatment (r from –0.55 to –0.63). No trait of aeroponically grown plants was correlated with traits of stressed plants in the field. We conclude that visual scoring of stressed plants was the best method of screening for drought resistance, but if controlled water deficit cannot be imposed, then drought resistance may be estimated by measuring both uprooting force and grain yield.     

Heterosis: its genetic basis and its utilisation in plant breeding

Summary To discuss the utilisation of heterosis in plant breeding it is necessary to consider its genetic basis. In cross fertilized plants in which the phenomenon of heterosis is very important, such as maize, it appears that a great part can be explained by the mutation load. However in both autogamous or allogamous plants it is difficult to exclude a role of marginal overdominance which could justify hybrid variety production.To know in a relatively short time whether it will be better to develop lines or hybrids it is necessary to know the parameters (mean and variance) of the distribution of all varieties of a given type which can be derived from the breeding population. A numerical application allows the determination of the situations where the best single crosses will be better than the best lines. With some approximate values of the necessary parameters, the case of grain yield in maize and in wheat is briefly discussed.To develop lines or hybrids, a general strategy is given. Its main axis is population improvement which has to be adapted to the type of varieties to develop. It is shown that according to the situation the type of varieties to develop can change with time: single crosses can be more justified at the beginning and lines can be justified later if all heterosis were fixable. To develop hybrids the best scheme is reciprocal recurrent selection with half-sib or full-sib progenies followed by pedigree reciprocal selection initiated by full-sib progeny tests. With this strategy there is no problem of the prediction of hybrid performances. However it seems possible to develop some predictors of heterosis or of specific combining ability between two lines using some criteria of genetic distances. Preliminary results with distances computed from the behaviour of lines in a two-tester top cross design appears very stimulating.Communication given at the206th meeting of teh Genetical Society of Grain Britain     

Resistance in Cucumis sativus L. to Tetranychus urticae Koch. 2. Designing a reliable laboratory test for resistance based on aspects of the host-parasite relationship

Summary The relationship between the twospotted spider mite and cucumber has been studied on plants and on leaf disks of a number of varieties with different levels of resistance. Existing laboratory tests are critically discussed and it appears that they are only reliable if many factors are taken into account. A new, more efficient laboratory test for resistance, measuring acceptance and reproduction is described.     

Assessment of somaclonal variability in hop (Humulus lupulus L.) in vitro meristem cultures and clones by molecular methods

In vitro meristem tissue cultures are used for production of virus-free rootstocks of hop (Humulus lupulus L.). Because use of plant tissue cultures is associated with occurrence of somaclonal variability, we assessed somaclonal variability in hop meristem in vitro cultures before and after thermotherapy by different molecular methods (RFLP, RAPD, STS, ISSR and AFLP) and compared it with existing clonal variability of Osvald's clones 31, 72 and 114. No molecular differences were observed between mother plants and in vitro mericlones by RFLP and STS analyses. Amplified molecular differences were found in RAPD and ISSR products of one from five in vitromericlones cvs. Eroica (E5) and Southern Brewer (SB2), respectively. Similarities with mother plants were 0.965 and 0.913 (JSC), respectively. Specific amplified polymorphic products were found for every mericlone and mother plant in AFLP reactions and variability of DNA sequence ranged from 0.824 to 0.993 (JSC). This variability was very similar to determined intra-clonal variability within Osvald's clones 31, 72 and 114 by AFLP analysis. Inter-clonal variability of DNA sequence was exactly higher than intra-clonal variability of DNA sequence in these clones. The molecular differences between Osvald's clone 72 normal and meristem derived were not verifiable. Thermotherapy increased frequency of molecular changes, since amplified differences were found in 14 from 20 in vitro mericlones of cv. Eroica, in 6 from 11 in vitro mericlones of cv. Yeoman and in 15 from 23 in vitro mericlones of cv. Southern Brewer by RAPD and ISSR analyses.     

甲醛的快速检测方法之研究

针对危害食品安全与生活环境的甲醛测定方法进行了详尽研究 ,提出了在不同产品中检测甲醛含量的快速、高效、合理的几种方法。