Somatic embryogenesis in sugarcane—An addendum to the invited review ‘sugarcane biotechnology: The challenges and opportunities,’ in vitro cell. Dev. Biol. Plant 41(4):345–363; 2005

Summary Since the 1960s, numerous studies on sugarcane plant regeneration have been reported. Essentially, successful culture and regeneration of plants from protoplasts, cells, callus, and various tissue and organs, have been achieved in this crop. Although plant regeneration from callus cultures had been reported since the 1960s, definitive proof of somatic embryo development was not available until 1983. Since then, considerable progress has been made in understanding and refining somatic embryogenesis and plant regeneration in sugarcane, for which development of an efficient embryogenic system was critical for the application of transgenic technology. Recent research in Australia and South Africa has led to the development of direct somatic embryogenic systems, which may improve transgenesis in sugarcane.     

Chilli peppers — A review on tissue culture and transgenesis

Biotechnology techniques involving plant tissue culture and recombinant DNA technologies are powerful tools that can complement conventional breeding and expedite Capsicum improvement. The rate of progress in Capsicum is relatively slower than other members of Solanaceae because of its high genotypic dependence and recalcitrant nature. Capsicum is a recalcitrant plant in terms of in vitro cell, tissue and organ differentiation, plant regeneration and genetic transformation which makes it difficult to apply recombinant DNA technologies aimed at genetic improvement against pests, diseases and abiotic stress. Despite this, application of tissue culture and genetic transformation have led to significant development in chilli pepper plants, and studies are underway to achieve the targets of pre-harvest improvement and post-harvest characterization for value addition to this crop. This review presents a consolidated account of in vitro propagation and focuses upon contemporary information on biotechnological advances made in Capsicum.     

Plant cryopreservation: a continuing requirement for food and ecosystem security

This issue of In Vitro Cellular and Developmental Biology—Plant is dedicated to current developments in liquid-nitrogen cryopreservation methods and their use in plant biology and germplasm preservation. The development of cryopreservation for storage of plant cells, tissues, and organs began in the 1960s and continues to this day. Long-term storage of in vitro cultures of secondary metabolite cell cultures, embryogenic cultures, clonal germplasm, endangered species, and transgenic products remains an important requirement for many scientists, organizations, and companies. The continued development of cryopreservation techniques and their application to new plants is the subject of this issue.     

Biotechnological applications in in vitro plant regeneration studies of broccoli (<Emphasis Type="Italic">Brassica oleracea</Emphasis> L. var. <Emphasis Type="Italic">italica</Emphasis>), an important vegetable crop

Biotechnology holds promise for genetic improvement of important vegetable crops. Broccoli (Brassica oleracea L. var. italica) is an important vegetable crop of the family Brassicaceae. However, various biotic and abiotic stresses cause enormous crop yield losses during commercial cultivation of broccoli. Establishment of a reliable, reproducible and efficient in vitro plant regeneration system with cell and tissue culture is a vital prerequisite for biotechnological application of crop improvement programme. An in vitro plant regeneration technique refers to culturing, cell division, cell multiplication, de-differentiation and differentiation of cells, protoplasts, tissues and organs on defined liquid/solid medium under aseptic and controlled environment. Recent progress in the field of plant tissue culture has made this area one of the most dynamic and promising in experimental biology. There are many published reports on in vitro plant regeneration studies in broccoli including direct organogenesis, indirect organogenesis and somatic embryogenesis. This review summarizes those plant regeneration studies in broccoli that could be helpful in drawing the attention of the researchers and scientists to work on it to produce healthy, biotic and abiotic stress resistant plant material and to carry out genetic transformation studies for the production of transgenic plants.     

Plant cell and biotechnology studies in <Emphasis Type="Italic">Linum usitatissimum</Emphasis> – a review

The species Linum usitatissimum (flax/linseed) has been the focus of a great deal of both basic and applied research effort in plant cell and biotechnology studies in recent years. In this review we consider applications of the techniques of plant biotechnology in this species under several distinct headings. Plant cell and tissue regeneration strategies and applications are discussed, and the applications of the techniques of somatic embryogenesis, protoplast isolation, culture and fusion and cell suspension cultures in this species are described. A major area of study is the use of anther and microspore culture where clear advantages to breeding programmes could be applied. In addition, embryo and ovary culture studies have resulted in significant findings. The more recent technologies of gene transfer and expression by genetic transformation are reviewed, and a section on strategies for improvements in technological quality is also included. Finally we propose conclusions and future prospects for this ancient, but still highly relevant crop.     

Application of plant tissue cultures in phytoremediation research: Incentives and limitations

The aim of this review is to critically assess the benefits and limitations associated with the use of in vitro plant cell and organ cultures as research tools in phytoremediation studies. Plant tissue cultures such as callus, cell suspensions, and hairy roots are applied frequently in phytoremediation research as model plant systems. In vitro cultures offer a range of experimental advantages in studies aimed at examining the intrinsic metabolic capabilities of plant cells and their capacity for toxicity tolerance. The ability to identify the contributions of plant cells to pollutant uptake and detoxification without interference from microorganisms is of particular significance in the search for fundamental knowledge about plants. However, if the ultimate goal of plant tissue culture experiments is the development of practical phytoremediation technology, the limitations inherent in the use of in vitro cultures as a representative of whole plants in the field must be recognized. The bioavailability of contaminants and the processes of pollutant uptake and metabolite distribution are likely to be substantially different in the two systems; this can lead to qualitative as well as quantitative differences in metabolic profiles and tolerance characteristics. Yet, many studies have demonstrated that plant tissue cultures are an extremely valuable tool in phytoremediation research. The results derived from tissue cultures can be used to predict the responses of plants to environmental contaminants, and to improve the design and thus reduce the cost of subsequent conventional whole plant experiments. Biotechnol. Bioeng. 2009;103: 60–76.     

The convergence of haemodynamics,genomics, and endothelial structure in studies of the focal origin of atherosclerosis

The completion of the Human Genome Project and ongoing sequencing of mouse, rat and other genomes has led to an explosion of genetics-related technologies that are finding their way into all areas of biological research; the field of biorheology is no exception. Here we outline how two disparate modern molecular techniques, microarray analyses of gene expression and real-time spatial imaging of living cell structures, are being utilized in studies of endothelial mechanotransduction associated with controlled shear stress in vitro and haemodynamics in vivo. We emphasize the value of such techniques as components of an integrated understanding of vascular rheology. In mechanotransduction, a systems approach is recommended that encompasses fluid dynamics, cell biomechanics, live cell imaging, and the biochemical, cell biology and molecular biology methods that now encompass genomics. Microarrays are a useful and powerful tool for such integration by identifying simultaneous changes in the expression of many genes associated with interconnecting mechanoresponsive cellular pathways.     

Cotton ovule culture: A tool for basic biology, biotechnology and cotton improvement

Summary Nearly 30 years ago the conditions for culturing immature cotton ovules were established to serve as a working research tool for investigating the physiology and biochemistry of fiber development. Not only has this tissue culture method been employed to characterize the biochemistry of plant cell expansion and secondary cell wall synthesis, but ovule cultures have contributed to numerous other aspects of plant cell physiology and development as well. In addition to basic studies on fiber development, cotton ovule cultures have been used to examine plant-fungal interactions, to model low temperature stress responses, to elucidate the pathways responsible for pigment formation in naturally pigmented fiber and to probe how cytoskeletal elements regulate cell wall organization. Success in rescuing Gossypium interspecific hybrids was dependent on ovule culture media formulations that could support early embryo development in ovulo. As tissues produced in culture are analyzed by increasingly more sophisticated techniques, there appear to be some differences between ovule growth in planta and ovule growth in vitro. Discerning how ovule culture fiber development is different from fiber development in field-grown plants can contribute valuable information for crop improvement. Cotton ovule cultures are an especially attractive model system for studying the effects of gravity on cell elongation, cellulose biosynthesis and embryo development and are excellent targets for examining transient expression of introduced gene constructs. With only minor modification, the procedure originally described by C. A. Beasley and I. P. Ting for growing cotton ovules in vitro will continue to be useful research tool for the foreseeable future.     

BRCA1 and BRCA2 associated breast cancer and the roles of current modelling systems in drug discovery

For a drug candidate to be fully developed takes years and investment of hundreds of millions of dollars. There is no doubt that drug development is difficult and risky, but vital to protecting against devastating disease. This difficulty is clearly evident in BRCA1 and BRCA2 related breast cancer, with current treatment options largely confined to invasive surgical procedures, as well as chemotherapy and radiotherapy regimes which damage healthy tissue and can leave remnant disease. Consequently, patient survival and relapse rates are far from ideal, and new candidate treatments are needed. The preclinical stages of drug discovery are crucial to get right for translation to hospital beds. Disease models must take advantage of current technologies and be accurate for rapid and translatable treatments. Careful selection of cell lines must be coupled with high throughput techniques, with promising results trialled further in highly accurate humanised patient derived xenograft models. Traditional adherent drug screening should transition to 3D culture systems amenable to high throughput techniques if the gap between in vitro and in vivo studies is to be partially bridged. The possibility of organoid, induced pluripotent stem cell, and conditionally reprogrammed in vitro models is tantalising, however protocols are yet to be fully established. This review of BRCA1 and BRCA2 cancer biology and current modelling systems will hopefully guide the design of future drug discovery endeavours and highlight areas requiring improvement.     

短端粒小鼠与正常小鼠间充质干细胞的分离培养比较

目的建立从小鼠微粒骨中获取间充质干细胞（MSC）方法,并观察比较野生型小鼠（WT鼠）和端粒酶基因敲除小鼠（Terc-/-鼠）MSC生物学特性的差异。方法采用体外细胞培养技术,从小鼠自体微骨片中分离纯化WT鼠和Terc-/-鼠的间充质干细胞,通过培养和传代,研究其增殖及生长特征。结果原代培养及传代培养显示,WT鼠自体骨间充质干细胞比Terc-/-鼠具有活跃的增殖倍增能力。结论从鼠的微骨片获取MSC的方法重复性好,细胞纯度和细胞数量优于以往从骨髓获取MSC的方法,Terc-/-鼠MSC生长增殖能力明显弱于WT鼠,其机制的研究有待深入。     

Use of in vitro cultures of Nicotiana benthamiana for the purification of grapevine virus A

Seedlings of Nicotiana benthamiana were inoculated with grapevine virus A (GVA). Three weeks later, upon systemic symptom expression, cultures were established in vitro using single nodes from these plants. GVA was purified from these cultures and from leaves of GVA-infected N. benthamiana plants maintained in the greenhouse. More virus was obtained from the proliferating in vitro node cultures than from the leaves. The use of in vitro cultures for virus purification represents a valuable practical application of plant tissue culture techniques.     

植物细胞悬浮培养影响因子研究

植物细胞悬浮培养已广泛应用于育种和生产次生代谢产物,在生物工程中发挥着重要作用。本文综述了愈伤组织继代改良的措施,影响胚性悬浮细胞系建立的几个因素,存在问题及前景展望。     

The myth of plant transformation

Technology development is innovative to many aspects of basic and applied plant transgenic science. Plant genetic engineering has opened new avenues to modify crops, and provided new solutions to solve specific needs. Development of procedures in cell biology to regenerate plants from single cells or organized tissue, and the discovery of novel techniques to transfer genes to plant cells provided the prerequisite for the practical use of genetic engineering in crop modification and improvement. Plant transformation technology has become an adaptable platform for cultivar improvement as well as for studying gene function in plants. This success represents the climax of years of efforts in tissue culture improvement, in transformation techniques and in genetic engineering. Plant transformation vectors and methodologies have been improved to increase the efficiency of transformation and to achieve stable expression of transgenes in plants. This review provides a comprehensive discussion of important issues related to plant transformation as well as advances made in transformation techniques during three decades.     

Frontiers in mammalian cells culture

Summary For the past 60 years, fundamental discoveries in eukaryotic biology using mammalian cell cultures have been significant but modest relative to the enormous potential. Combined with advances in technologies of cell and molecular biology, mammalian cell culture technology is becoming a major, if not essential tool, for fundamental discovery in eukaryotic biology. Reconstruction of the milieu for cells has progressed from simple salt solutions supporting brief survival of tissues outside the body to synthesis of the complete set of structurally defined nutrients, hormones and elements of the extracellular matrix needed to reconstruct complex tissues from cells. The isolation of specific cell types in completely defined environments reveals the true complexity of the mammalian cell and its environment as a dynamic interactive physiological unit. Cell cultures provide the tool for detection and dissection of the mechanism of action of cellular regulators and the genes that determine individual aspects of cell behavior. The technology underpins advances in virology, somatic cell genetics, endocrinology, carcinogenesis, toxicology, pharmacology, hematopoiesis and immunology, and is becoming a major tool in develomental biology, complex tissue physiology and production of unique mammalian cell-derived biologicals in industry. This article is the first of a series of invited reviews aimed at identifying fundamental contributions and current challenges associated with research activities in subdiscriplines of cell and developmental biology in vitro. This treatise is dedicated to Dr. Brian Kimes, Program Director at the National Cancer Institute, whose vision, encouragement and support have contributed significantly to modern developments in mammalian cell culture.     

Variation from plant tissue cultures: biotechnological application to improving salinity tolerance

Breeding for salt tolerance in crop plants is envisaged as one way to combat a worldwide problem of increasing soil salinity in agricultural land. Tissue culture techniques may prove valuable as a means of achieving this goal. In this review, reports of the selection and characterization of plant cell cultures tolerant to excess salt are assessed, in the context of variability from tissue culture and the significance of cellular physiological adaptation to salinity. The examples of plant regeneration from salt grown cell cultures are also outlined, with emphasis on correlation to the effect of salt on cell cultures, genetic variability for salt tolerance in vitro, and the value of regenerates in the development of salt tolerant plants.     

细胞打印技术及应用

细胞打印技术是一种在体外构造具有生物活性的三维多细胞体系的先进技术。近年来,有关细胞打印技术的研究引起广泛的关注,其原因在于该领域具有明显的学科交叉与渗透融合的特点,它处于生命科学与快速成型技术、生物制造技术、生物科学和材料科学的交汇点。更加值得关注的是它为组织工程学突破二维研究的局限性,在三维尺度上精确控制与人体组织或器官相似的三维构造体方面的研究提供了一种新的思路。基于这一技术不仅在三维组织工程,还对细胞生物学、高通量药物筛选及细胞传感器等方面的前沿问题均有广阔的研究应用前景,介绍了近年来开发用于细胞打印的技术及其潜在的应用。     

Plant cell cultivation as a biotechnological method

An overview is given about some aspects of history, principles and application of plant cell and tissue cultivation. The main topics of application discussed in the paper are aspects of plant propagation, plant breeding, techniques for in vitro cultivation, in vitro production of secondary plant products, biotransformations, application of immobilized cells as well as economic aspects of plant cell and tissue culture.     

香蕉转基因研究进展

香蕉转基因研究逐渐成为国内外植物分子生物学研究的热点。人们从 2 0世纪 60年代开始就进行了香蕉的组织培养 ,70年代茎尖培养成功 ,近 1 0年来相继在微繁殖、单克隆抗体、分子生物学和遗传转化等方面开展了工作。香蕉转化受体系统已从茎尖发展到胚状体和愈伤组织受体系统 ,遗传转化方法也在不断改进 ,从单一的方法发展为根癌农杆菌感染法与基因枪法的结合。从香蕉组织培养、遗传转化受体系统、基因转化方法、筛选方法的研究等方面进行综述 ,并提出了一些看法。     

Characterization of photosynthetically active duckweed (<Emphasis Type="Italic">Wolffia australiana</Emphasis>) in vitro culture by Respiration Activity Monitoring System (RAMOS)

The feasibility of oxygen transfer rate (OTR) measurement to non-destructively monitor plant propagation and vitality of photosynthetically active plant in vitro culture of duckweed (Wolffia australiana, Lemnaceae) was tested using Respiration Activity Monitoring System (RAMOS). As a result, OTR proofed to be a sensitive indicator for plant vitality. The culture characterization under day/night light conditions, however, revealed a complex interaction between oxygen production and consumption, rendering OTR measurement an unsuitable tool to track plant propagation. However, RAMOS was found to be a useful tool in preliminary studies for process development of photosynthetically active plant in vitro cultures.