A convenient and versatile hydroponic cultivation system for Arabidopsis thaliana

A versatile two-step cultivation procedure for Arabidopsis thaliana is described for the production of large quantities of leaf material suitable for biochemical and biophysical analysis. The first step comprises a miniature greenhouse made out of a plastic pipette box to grow the seedlings to the six-leaf stage. For continued growth, the seedlings are transferred to hydroponic cultivation using an opaque container covered by a styrofoam lid. Transfer of the small seedlings to hydroponic culture is facilitated by growth in separate pipette tips, which protects vulnerable roots from damage. The hydroponic cultivation system is easy to scale-up and produces large amounts of relatively large leaves and roots. This hydroponic system produces enough plant material to make Arabidopsis a feasible model for biochemical and biophysical experiments, which can be combined with the available genetic information to address various aspects of plant functional genomics.     

Molecular characterization of Haloxylon salicornicum Moq. in Saudi Arabia

The purpose of write this paper is to study the genetic variability between and within different Halosylon salicornicum populations in different regions of Saudi arabia Kingdom, using the determination of genetic fingerprint method by Inter Simple Sequence Repeat (ISSR). Because this plant highly vulnerable to depletion by humans in all places of existence, it is an economically valuable plant where raft is an important pastoral resource in central and northern Arabia. It also has multiple medical uses. It is a plant that can withstand abiotic stresses such as drought and high temperature, making it suitable for cultivation in marginal lands in arid areas. All of the above was a catalyst for plant characterization using a number of Haloxylon salicornicum samples, collected from different regions of Saudi Arabia to find out the genetic variation of species, genetic diversity in knowing the plant community is an essential step towards the design of programs for plant breeding as well as preserved from extinction. This was done using the Inter Simple Sequence Repeat (ISSR). The results showed there were significant differences and molecular differences between plant samples. The average polymorphism between the genetic inputs of the studied Haloxylon salicornicum samples was 53.7%, and this percentage of genetic variability is significant for progress in growth and plant regeneration in the face of unfair practices against it, in addition to adverse environmental conditions in most years. As evidenced by the percentage of matrix similarity. The ISSR results indicate that the genotype between five different regions genotypes ranged from 0.365 to 0.527, indicating that Haloxylon salicornicum is a local plant capable of surviving and adapting to the environmental conditions in Saudi Arabia through the positive change in the genetic makeup of this species.     

Suppression of disease in tomato infected by <Emphasis Type="Italic">Pythium ultimum</Emphasis> with a biosurfactant produced by <Emphasis Type="Italic">Pseudomonas koreensis</Emphasis>

The use of biosurfactants is a promising alternative in biological control of zoospore-producing oomycetes, which are a major plant pathogen world-wide in a wide variety of crops. Oomycetes are of particular concern in closed hydroponic cultivation systems. The present study investigated the efficacy of a biosurfactant produced by Pseudomonas koreensis and added as a crude extract against the oomycete Pythium ultimum in hydroponic tomato cultivation. A significant reduction in disease was observed. Biosurfactant addition did not affect the indigenous root microflora when evaluated as sole carbon source utilisation. Chemical analysis, using electrospray hybrid mass spectrometry (ESI-MSMS), of the biosurfactant indicated it to be lokisin, a cyclic lipopeptide. These results confirm that biosurfactants are important in developing sustainable biological control strategies for oomycetes.     

Hydroponics: A Versatile System to Study Nutrient Allocation and Plant Responses to Nutrient Availability and Exposure to Toxic Elements

Hydroponic systems have been utilized as one of the standard methods for plant biology research and are also used in commercial production for several crops, including lettuce and tomato. Within the plant research community, numerous hydroponic systems have been designed to study plant responses to biotic and abiotic stresses. Here we present a hydroponic protocol that can be easily implemented in laboratories interested in pursuing studies on plant mineral nutrition.This protocol describes the hydroponic system set up in detail and the preparation of plant material for successful experiments. Most of the materials described in this protocol can be found outside scientific supply companies, making the set up for hydroponic experiments less expensive and convenient.The use of a hydroponic growth system is most advantageous in situations where the nutrient media need to be well controlled and when intact roots need to be harvested for downstream applications. We also demonstrate how nutrient concentrations can be modified to induce plant responses to both essential nutrients and toxic non-essential elements.     

Antiproliferative Effect of Essential Oil Obtained from Oregano (Lippia palmeri S. Watson) Leaves Grown in Hydroponics and LED Light

Nowadays, light-emitting diodes (LED) provide an alternative source to sunlight with specific intensity and wavelength that promotes plant growth. The features offered by LED could also stimulate the production of secondary metabolites of pharmaceutical interest. This work analyzed the cultivation of oregano (Lippia palmeri S. Watson) in a floating root hydroponic system supplemented by full-spectrum LED artificial light. Growth indicators like height, diameter, number of shoots, and leaf length and width were measured. The essential oil (EO) composition from the leaves of wild and hydroponic conditions found thymol (41.8 %) as the main product for the former and carvacrol (47 %) in hydroponics. The antiproliferative activity of EOs on human colorectal cancer HCT-15 shows that 6.4 μg/ml for hydroponic and 7.4 μg/ml for the wild plant reduce more than 50 % the cell viability. Overall, this study indicates that hydroponic conditions and full spectrum LED modifies the composition of the EO of L. palmeri on compared with the wild plant, which effectively induces cell growth inhibition in human colorectal cancer.     

Characterization of cadmium uptake by the water lily Nymphaea aurora

This study characterizes cadmium (Cd) uptake by the waterlily Nymphaea aurora, (Nymphaeaceae) in two systems: a model hydroponic Cd solution and heavily polluted sludge from two sites in Israel. The uptake of Cd from hydroponic solution resulted in Cd storage in petioles and laminae of Nymphaea, as well as in the roots. The pH of the solution affected Cd solubility and availability, with pH 5.5 yielding maximum Cd content in the plant (140 mg Cd per g DW). Cd uptake was reduced by the addition of EDTA to the hydroponic growth medium, although EDTA enhanced heavy metal uptake by terrestrial plants. Nymphaea efficiently reduced the concentration of Cd in heavy metal polluted urban and industrial sludge and the amount of Cd uptake was enhanced by the addition of KCl to the sludge and by adjustment of the pH to 5.5. The inherent growth patterns of Nymphaea plants allowed Cd uptake by the shoot and root, and resulted in maximum contact between the various plant parts and the growth media. Thus, Nymphaea has potential as an optimal, highly effective phytoremediation tool for the removal of Cd from polluted waste sources.     

Influence of Clonal Variation on the Efficiency of Arundo donax Propagation Methods

The recent growth of interest in Arundo donax L. (Giant reed) as an energy crop is due to its great vegetative vigor and high biomass productivity. This perennial rhizomatous plant is able to grow in a wide range of pedo-climatic conditions and it has been employed by man for a great number of purposes. This has promoted its worldwide diffusion, despite its sexual sterility. Field establishment represents the most expensive phase of the whole cultivation cycle, because of the high cost of the propagules. In this work three agamic propagation methods, rhizome transplantation, hydroponic cultivation, and in vitro propagation, were compared with the aim of improving their efficiency, and reducing costs and time required. We focused our attention on in vitro and hydroponic cultivation, finding clones characterized by a high response to these propagation methods. In vitro propagation is the most widespread technique adopted in the large-scale production of propagules; according to our results hydroponic cultivation also appears to be a valid and cheap propagation method, making it possible to obtain in about 2 months, vigorous plants ready to be transplanted into the field. We found a strong effect of the clone used on the success rate of the specific propagation method, suggesting that clonal selection could be particularly useful in improving the efficiency of propagation techniques, contributing to cost reduction. Evaluation of field performance of plants propagated by tissue culture is an important matter, considering the possible residual effects of hormones on plants’ growth, and the importance of first year growth for field establishment. These experiments highlighted the lower biomass production from plants obtained by tissue culture. A novel cytokinin-like hormone, meta-topolin, was tested in vitro, in a preliminary study to evaluate its possible use for A. donax L. propagation.

     

Phytosiderophores revisited: 2′-deoxymugineic acid-mediated iron uptake triggers nitrogen assimilation in rice (Oryza sativa L.) seedlings

Poaceae plants release phytosiderophores into the rhizosphere in order to chelate iron (Fe), which often exists in insoluble forms especially under high pH conditions. The impact of phytosiderophore treatment at the physiological and molecular levels in vivo remains largely elusive, although the biosynthesis of phytosiderophores and the transport of phytosiderophore-metal complexes have been well studied. We recently showed that the application of 30 μM of the chemically synthesized phytosiderophore 2′-deoxymugineic acid (DMA) was sufficient for apparent full recovery of otherwise considerably reduced growth of hydroponic rice seedlings at high pH. Moreover, unexpected induction of high-affinity nitrate transporter gene expression as well as nitrate reductase activity indicates that the nitrate response is linked to Fe homeostasis. These data shed light on the biological relevance of DMA not simply as a Fe chelator, but also as a trigger that promotes plant growth by reinforcing nitrate assimilation.     

Role of soil microorganisms in improving P nutrition of plants

Phosphorus (P) is one of the major plant growth-limiting nutrients although it is abundant in soils in both inorganic and organic forms. Phosphate solubilizing micro-organisms (PSMs) are ubiquitous in soils and could play an important role in supplying P to plants in a more environmentally friendly and sustainable manner. Although solubilization of P compounds by microbes is very common under laboratory conditions, results in the field have been highly variable. This variability has hampered the large-scale use of PSMs in agriculture. Many reasons have been suggested for this variability, but none of them have been extensively investigated. In spite of the importance of PSMs in agriculture, the detailed biochemical and molecular mechanisms of P solubilization are not known. Recent work in our laboratory has shown that the conditions employed to isolate PSMs do not reflect soil conditions and that PSMs capable of effectively releasing P from soil are not so highly abundant as was suggested in earlier studies. These studies have also indicated that the mineral phosphate solubilizing (mps) ability of microbes could be linked to specific genes, and that these genes are present even in non P solubilizing bacteria. Understanding the genetic basis of P solubilization could help in transforming more rhizosphere-competent bacteria into PSMs. Further research should also focus on the microbial solubilization of iron (Fe) and aluminum (Al) phosphates, as well as mobilization of the organic phosphate reserves present in the soils.     

不同培育方式对橡胶树实生苗生理指标、养分及长势的影响

橡胶树(Heveabrasiliensis)种子催芽生长一般使用沙床培育,沙子是不可再生资源,为了选择一种适合橡胶树种子培育方式来替代对沙子的依赖,该研究通过水培、悬空培育和传统的沙培比较橡胶树实生苗第1蓬叶稳定时,苗木的生长势、生理指标及养分含量。结果表明,水培实生苗地上部株高、茎粗、叶面积的长势最佳,壮苗指数和生物量的含量最高,但其根太长,根相对较细。水培的叶、茎、根的可溶性糖、丙二醛、游离脯氨酸、超氧化物歧化酶的含量均较低;水培和悬空培育的叶片和茎的叶绿素、类胡萝卜素及根系活力的含量没有显著性差异,均高于沙培。水培的叶、茎、根中的氮和磷含量最低,沙培的最高;而水培实生苗根和茎中钾的含量较高,叶片中含量与悬空培育、沙培均没有显著性差异;悬空培育在叶、茎、根中钾的含量最低。水培促进了苗木的生长,降低干旱胁迫,提高养分利用率,但后续还需调控根系,建设良好根团。悬空培育的苗木长势较弱,还需进一步完善方法。     

Karyotype of continuous cell lines. I. The karyotype variability of M-HeLa cells cultured by static and roll-tube methods

A comparative investigation of three M-HeLa sublines, being of common origin but differing in cultivation technique (roller or static), was undertaken using G- and C-banding. It is shown that the M-HeLa sublines of a non-clonal origin differ in the level of their karyotype variability as compared to most human and animal lines studied earlier. A definite set of normal and marker chromosomes, as well as a typical karyotype, was revealed for either subline tested. The change of the static technique of cultivation into the roller one was accompanied with changes in the modal class, with the increase in polyploid cell number and proliferation index. It is postulated that the change of cultivation technique may result in selection of defined cell types due to the pre-existed karyotype variability.     

Analysing the role of soil properties, initial biomass and ozone on observed plant growth variability in a lysimeter study

This simulation study is based on a lysimeter experiment with juvenile beech trees (Fagus sylvatica L.) which were grown under ambient or doubled ambient atmospheric ozone concentrations. The aim of the study was to analyze the role of differences in soil properties, differences in initial biomass and ozone impacts on observed plant growth variability at the eight lysimeters of this experiment. For this purpose, we established a new simulation model based on the model system Expert-N by coupling soil water and nitrogen transport models with the plant growth model PLATHO, which was already tested and applied for juvenile beech. In order to parameterize the soil model, for all lysimeters soil hydraulic parameters as well as carbon and nitrogen stocks were measured. Simulation results reveal that the observed decreased growth rates under elevated ozone are due to ozone impacts on plant growth, whereas the high plant growth variability between lysimeters is to a major part the consequence of differences in soil hydraulic properties. Differences in initial biomass are of minor importance to explain plant growth variability in this experiment.     

Kinetics of littorine content in various developing stages of regenerates of Atropa belladonna

 Aseptically propagated regenerates were cultivated in a hydroponic apparatus, a phytotron or in the field, and their growth and littorine contents were investigated. No littorine was detected in aseptic regenerates cultured on solidified Murashige and Skoog medium, nor was it found in leaves under the three conditions tested. In roots, it was common features to all three conditions tested that littorine increased dramatically after transplantation from culture tubes and was a major alkaloid up to week 4; subsequently the littorine contents varied depending on the cultivation conditions. Roots cultivated in the field showed a marked thickening and rapid disappearance of littorine; those cultivated in the hydroponic apparatus were thin and maintained a high level of littorine for a long time. In a plant cultivated for 16 weeks in a pot, littorine content in the roots decreased with increasing root diameter. Received: 1 October 1999 / Revision received: 16 March 2000 / Accepted: 24 March 2000     

High‐yield production of a human monoclonal IgG by rhizosecretion in hydroponic tobacco cultures

Rhizosecretion of recombinant pharmaceuticals from in vitro hydroponic transgenic plant cultures is a simple, low cost, reproducible and controllable production method. Here, we demonstrate the application and adaptation of this manufacturing platform to a human antivitronectin IgG₁ monoclonal antibody (mAb) called M12. The rationale for specific growth medium additives was established by phenotypic analysis of root structure and by LC‐ESI‐MS/MS profiling of the total protein content profile of the hydroponic medium. Through a combination of optimization approaches, mAb yields in hydroponic medium reached 46 μg/mL in 1 week, the highest figure reported for a recombinant mAb in a plant secretion‐based system to date. The rhizosecretome was determined to contain 104 proteins, with the mAb heavy and light chains the most abundant. This enabled evaluation of a simple, scalable extraction and purification protocol and demonstration that only minimal processing was necessary prior to protein A affinity chromatography. MALDI‐TOF MS revealed that purified mAb contained predominantly complex‐type plant N‐glycans, in three major glycoforms. The binding of M12 purified from hydroponic medium to vitronectin was comparable to its Chinese hamster ovary (CHO)‐derived counterpart. This study demonstrates that in vitro hydroponic cultivation coupled with recombinant protein rhizosecretion can be a practical, low‐cost production platform for monoclonal antibodies.     

Conceptual design of LED-based hydroponic photobioreactor for high-density plant cultivation

A novel hydroponic photobioreactor is proposed for high-density cultivation of plants. This cultivation can be achieved by growing plants on a floatable platform, allowing the roots to directly contact a continuously aerated nutrient solution. Plant growth of Mentha x piperita (peppermint) can be shown to strongly correlate with the light intensity at incident light intensities between 0 and 650 &mgr;mol m(-)(2) s(-)(1). For a constant incident light intensity (I(0) = 420 &mgr;mol m(-)(2) s(-)(1)), the overall specific growth rates of these plants are found to be strongly dependent on the plant density. They range from 0.023 to 0.075 d(-)(1) for plants grown at a density range from 16 to 256 plants m(-)(2). A simple mathematical model is presented that allows one to predict these effects of light intensity and plant density on peppermint growth. Light delivery is derived from the modification of Beer-Lambert's law. From this, the relationship between the light extinction coefficient and plant density can be experimentally determined. The light transport can then be coupled with plant growth kinetics under light-limiting conditions. The predicted growth results agree reasonably well with most experimental results from a growth period of 17-20 days. On the basis of these simulation results, we suggest that a more efficient way of delivering light to this photobioreactor can be attained by supplying light from both the top and the bottom of the plant shoots. The proposed design takes advantage of the small size and low weight of light emitting diodes, which allow them to be mounted on platforms for delivering light closer to the plants.     

Fertiliser application positively affects plants performance but reduces seed viability in seashore mallow (Kosteletzkya pentacarpos): implication for biomass production and species conservation

Kosteletzkya pentacarpos seashore mallow, is recognised as an important plant species for several industrial applications, especially in North America and Asia. In Europe, seashore mallow is highly threatened and subject of conservation actions (e.g. reintroduction). In order to define appropriate multipurpose cultivation protocols, suitable for biomass production and for conservation purposes, we investigated the effect of varying levels of soil salt content and nutrient availability on seashore mallow. As expected, seashore mallow had the best performance in terms of growth rate, flowering and fruit production when fertiliser was applied, while salt had only limited effects. Interestingly, seeds produced by fertilised plants showed a lower germination performance and higher mortality than seeds developed from non‐fertilised plants. Our results highlight a trade‐off between parental plant growth and seed performance, the former being enhanced and the latter being reduced by fertiliser application. The causes of this trade‐off are related to a transgenerational maternal effect of fertiliser application that has important implications for seashore mallow cultivation. Biomass production benefits from fertiliser application but if the quality of seeds and the fitness of the next generation are major objectives (e.g. seed production and species conservation), fertiliser application is not recommended.     

黑杨水培中营养液抑菌处理研究

水培条件下进行快速繁殖是大量生产黑杨种苗的途径之一 ,营养液的抑菌处理是水培的一个重要方面。该研究进行了二硫氰甲烷、中生菌素、敌磺钠、链霉素、青霉素等五种杀细菌剂和多菌灵、福美双、三唑酮等三种杀真菌剂的抑菌试验 ,结果表明 ,杀细菌剂对微生物生长的抑制率要高于杀真菌剂 ,二硫氰甲烷和中生菌素能较好地抑制营养液中微生物的生长 ,浓度分别为 2 0mg/L和 1 5mg/L时 ,抑制率分别为 75 .60 %和78 94%。杀真菌剂和杀细菌剂复配对营养液中微生物生长的抑制表现出较好的加和作用 ,其中以中生菌素(1 5mg/L)与福美双 (5mg/L)复配的效果最好 ,抑制率为 87.48%。     

Growing rice in controlled environments

Rice (Oryza sativa) is less frequently used in basic research than Arabidopsis, although rice is a valuable model system for many monocot crops and possesses a high genetic variability in physiologically as well as agriculturally relevant features such as abiotic stress tolerance, nutrient efficiency and flower time control. A reason is the seemingly difficult cultivation of rice outside the rice production area. This review aims to assist newcomers to the field to develop cultivation protocols for their local controlled environment. The main challenges are high light demands, photoperiodicity and low micronutrient efficiency. The nutrient efficiency problem can be overcome by adding micronutrient fertiliser to potting substrates and keeping the soil waterlogged to increase micronutrient availability and mobility. Cultivation of rice on adjusted hydroponic solutions with high iron concentration provides the basis for successful heavy isotope labelling. Many rice cultivars need high light intensities in combination with short‐day conditions to complete their life cycle. However, some photoperiod‐insensitive cultivars will flower even under relatively low light intensities. In highly photoperiod‐sensitive cultivars, like Nipponbare, flowering can be induced by a limited period of short‐day treatment in the sensitive period, after which the cultivation can be continued in long‐day conditions. The life cycle of many cultivars is completed in 90 to 120 days, its length being thus comparable to Arabidopsis and shorter than in other cereals. In conclusion, with the right cultivation technique, rice is an amiable model species for researchers beyond the rice area too.     

赤霉素生物合成与信号传递对植物株高的调控

植物株高是影响作物产量和品质的重要农艺性状。赤霉素(Gibberellins,GAs)是调控植物株高的重要激素,GA相关株高基因的克隆与分子机制研究对于合理调控作物生长发育和农业生产具有极其重要的利用价值,在水稻、小麦等粮食作物育种中得到了广泛应用。为了促进GA在果树、花卉等园艺作物育种中的有效利用,文中在分子生物学水平上介绍GA生物合成和GA信号传递途径对植物株高的调控。     

Ara-Rhizotron: An Effective Culture System to Study Simultaneously Root and Shoot Development of Arabidopsis

Studying Arabidopsis thaliana (L.) Heynh. root development in situ at the whole plant level without affecting shoot development has always been a challenge. Such studies are usually carried out on individual plants, neglecting competition of a plant population, using hydroponic systems or Agar-filled Petri dishes. Those both systems, however, present some limitations, such as difficulty to study precisely root morphogenesis or time-limited culture period, respectively. In this paper, we present a method of Arabidopsis thaliana (L.) Heynh. cultivation in soil medium, named “Ara-rhizotron”. It allows the non-destructive study of shoot and root development simultaneously during the entire period of vegetative growth. In this system, roots are grown in 2D conditions, comparable to other soil cultures. Moreover, grouping several Ara-rhizotrons in a box enables the establishment of 3D shoot competition as for plants grown in a population. In comparison to a control culture grown in pots in the same environmental conditions, the Ara-rhizotron resulted in comparable shoot development in terms of dry mass, leaf area, number of leaves and nitrogen content. We used this new culture system to study the effect of irrigation modalities on plant development. We found that irrigation frequency only affected root partitioning in the soil and shoot nitrogen content, but not shoot or root growth. These effects appeared at the end of the vegetative growth period. This experiment highlights the opportunity offered by the Ara-rhizotron to point out tardy effects, affecting simultaneously shoot development and root architecture of plants grown in a population. We discuss its advantages in relation to root development and physiology, as well as its possible applications.