S. Dutta Gupta

Plant tissue culture engineering

Part 1: Machine Vision Evaluation of photosynthetic capacity in micropropagated plants by image analysis/Y. Ibaraki Monitoring gene expression in plant tissues/J.J. Finer, S.L. Beck, M.T. Buenrostro-Nava, Y. Chi, P.P. Ling Applications and potentials of artificial neural networks in plant tissue culture/V.S.S. Prasad, S. Dutta Gupta Evaluation of plant suspension cultures by texture analysis/Y. Ibaraki Part 2: Bioreactor Technology Bioengineering aspects of bioreactor application in plant propagation/S. Takayama, M. Akita Agitated, thin-films of liquid media for efficient micropropagation/ J. Adelberg Design, development and applications of mist bioreactors for micropropagation and hairy root culture/M.J. Towler, Y. Kim, B.E. Wyslouzil, M.J. Correll, P.J. Weathers Bioreactor engineering for recombinant protein production using plant cell suspension culture/W.W. Su Types and designs of bioreactors for hairy root culture/Y-E Choi, Y-S Kim, K-Y Paek Oxygen transport in plant tissue culture systems/W.R. Curtis, A.L. Tuerk Temporary immersion bioreactor/F. Afteen Design and use of the wave bioreactor for plant cell culture/R. Eibl, D. Eibl Part 3: Mechanized Micropropagation Integrating automation technologies with commercial micropropagation/C.J. Sluis Machine vision and robotics for the separation and regeneration of plant tissue cultures/P.H. Heinemann, P.N. Walker Part 4: Engineering Cultural Environment Closed systems for high quality transplants using minimum resources/T. Kozai Aeration in plant tissue culture/S.M.A. Zobayed Tissue culture gel firmness: measurement and effects on growth/S.I. Cameron Effects of dissolved oxygen concentration on somatic embryogenesis/K. Kurata, T. Shimazu A commercialized photoautotrophic micropropagation system/T. Kozai, Y. Xiao Intelligent inverse analysis for temperature distribution in a plant culture vessel/H. Murase, T. Okayama, Suroso Part 5: Physical Aspects of Plant Tissue Engineering Electrical control of plant morphogenesis/C.G. Carmen The uses of ultrasound in plant tissue culture/V. Gaba, K. Kathiravan, S. Amutha, S. Singer, X. Xiaodi, G. Ananthakrishnan Acoustic characteristics of plant leaves using ultrasonic transmission waves/M. Fukuhara, S. Dutta Gupta, L. Okushima Physical and engineering perspectives of in vitro plant cryopreservation/E.E. Benson, J. Johnston, J. Muthusamy, K. Harding

Antioxidant Enzyme Activities during in vitro Morphogenesis of Gladiolus and the Effect of Application of Antioxidants on Plant Regeneration

Activity of antioxidant enzymes was evaluated during somatic embryogenesis and shoot organogenesis from cultured leaf segments of Gladiolus hybridus Hort. The effect of exogenous antioxidants on somatic embryogenesis and shoot organogenesis has also been monitored. Activity of superoxide dismutase (SOD) gradually increased during somatic embryogenesis. while activities of catalase (CAT) and peroxidase (POX) decreased. In contrast, increase in CAT and POX activity and a concomitant decrease in SOD activity were noted during shoot organogenesis. Exogenous application of antioxidants such as glutathione (GSH), α-tocopherol and ascorbate (AA) inhibited somatic embryogenesis but stimulated shoot organogenesis. The frequency of somatic embryogenesis increased with the addition of H2O2. However, H2O2 inhibited shoot organogenesis.

Enhancement of peroxidase, polyphenol oxidase and superoxide dismutase activities by triadimefon in NaCl stressed Raphanus sativus L.

The activities of peroxidase, polyphenol oxidase and superoxide dismutase was significantly lower in roots and leaves of NaCl stressed radish (Raphanus sativus L.) plants. Addition of triadimefon to the NaCl stressed plants increased peroxidase, polyphenol oxidase and superoxide dismutase activities, and thereby ameliorated the negative effect of NaCl stress.

Fundamentals and applications of light-emitting diodes (LEDs) in in vitro plant growth and morphogenesis

Advances in plant tissue culture methods with regard to lighting requirements are currently focused on the improved features of light-emitting diodes (LEDs). Over the years, the steady development of LED technology, with the emergence of new types of semi-conductor materials, has made it possible to apply it in an increasing number of new areas. As an alternative to conventional lighting systems, LED has been demonstrated to be an artificial flexible lighting source for plant tissue culture. Numerous studies have been conducted in order to investigate the effects of LED on plants, which have led to many satisfactory results. Various morphological, anatomical, and physiological attributes, such as shoot elongation, axillary shoot formation, somatic embryo induction, rhizogenesis, leaf anatomy, and photosynthetic abilities of plants grown in vitro have found to be regulated by spectral properties of LEDs. The present review gives an overview of the fundamentals of LEDs and describes their effects on in vitro plant growth and morphogenesis and their future potentials.

High-frequency plant regeneration through cyclic secondary somatic embryogenesis in black pepper (Piper nigrum L.)

A high-frequency plantlet regeneration protocol was developed for black pepper (Piper nigrum L.) through cyclic secondary somatic embryogenesis. Secondary embryos formed from the radicular end of the primary somatic embryos which were originally derived from micropylar tissues of germinating seeds on growth regulator-free SH medium in the absence of light. The process of secondary embryogenesis continued in a cyclic manner from the root pole of newly formed embryos resulting in clumps of somatic embryos. Strength of the medium and sucrose concentration influenced the process of secondary embryogenesis and fresh weight of somatic embryo clumps. Full-strength SH medium supplemented with 1.5% sucrose produced significantly higher fresh weight and numbers of secondary somatic embryos while 3.0 and 4.5% sucrose in the medium favored further development of proliferated embryos into plantlets. Ontogeny of secondary embryos was established by histological analysis. Secondary embryogenic potential was influenced by the developmental stage of the explanted somatic embryo and stages up to “torpedo” were more suitable. A single-flask system was standardized for proliferation, maturation, germination and conversion of secondary somatic embryos in suspension cultures. The system of cyclic secondary somatic embryogenesis in black pepper described here represents a permanent source of embryogenic material that can be used for genetic manipulations of this crop species.

Estimation of the chlorophyll content of micropropagated potato plants using RGB based image analysis

A method has been developed for rapid and non invasive determination of chlorophyll content of leaves of micropropagated potato plants using RGB based image analysis. Among the trichomatic colors, R and G negatively correlated with the chlorophyll content, while a positive correlation was observed with B chromate. Compared to mean brightness value, the use of mean brightness ratio considerably improved the relationship of the tricolors with chlorophyll content. The brightness values and ratios of the primary colors are modeled as linear correlation functions for chlorophyll content. A significant correlation was observed between the model predicted chlorophyll content with the chlorophyll content measured by chlorophyll content meter. Spectral properties such as luminosity and saturation were also found to be negatively correlated with the chlorophyll content. The relationship was improved by combining the mean brightness ratio at B band region with luminosity. The potential of the imaging system in micropropagation has been discussed.

In vitro differentiation of multiple shoot clumps from intact seedlings of switchgrass

SummaryAn efficient and reproduciblein vitro culture system has been developed for regeneration of multiple shoot clumps from intact seedlings of both lowland and upland cultivars of switchgrass (Panicum virgatum L.). The multiple shoots were induced on Murashige and Skoog medium supplemented with various combinations of 2,4-dichlorophenoxyacetic acid (2,4-D) and 1-phenyl-3-(1,2,3-thiadiazol-5YL)-urea (thidiazuron or TDZ). Maximum response was obtained with 4.5 μM 2,4-D and 18.2 μM TDZ. These shoots proliferated and rooted efficiently on MS medium without growth regulators. The developmental pattern of the multiple shoots indicated their origin from the enlarged shoot apex via proliferation of axillary buds and subsequent reprogramming of shoot meristems followed by secondary differentiation of adventitious shoots The simplicity of the protocol and direct production of multiple shoots make this a potential system that is highly attractive and amenable for microprojectile-mediated gene transfer.

DIRECT SHOOT ORGANOGENESIS AND PLANT REGENERATION IN SAFFLOWER

SummaryAdventitious shoot buds were induced directly on the adaxial surface of the cotyledons of eight safflower cultivars after 14 d of culture initiation on Murashige and Skoogs (MS) medium supplemented with various levels of 6-benzylaminopurine (BA). Maximum shoot organogenesis of 54.4% with 10.2 shoots per responding cotyledon was obtained with 8.87 μM BA in the cv. S-144. Regenerated shoots were classified into three groups on the basis of their morphological features and were found to be correlated with the levels of BA. The highest number of normal shoots was obtained from 2.2 μM BA treatment. The regenerated shoots of group I (normal shoots) were rooted on half-strength MS medium supplemented with 5.3 μM α-naphthaleneacetic acid, 3% sucrose and 0.8% bacto-agar. Rooted plantlets were successfully transferred to soil and appeared morphologically normal. Histological studies revealed that shoot buds originated adventitiously from subepidermal cells.

Direct somatic embryogenesis and plantlet regeneration from cotyledonary leaves of safflower

Somatic embryos were induced directly on adaxial surface of cotyledonary leaves within 8–10 days of culture on Murashige and Skoog medium containing 5.37 to 10.74 μM 1 — napthaleneacetic acid and 2.22 μM benzyl adenine. Germinated embryos with shoot axes developed into complete plants after transfer onto half stength Murashige and Skoog medium containing 1.07 μM 1 — napthaleneacetic acid. Histological studies suggested direct origin of somatic embryos with broad-base attachment.

Somatic embryogenesis of safflower: influence of auxin and ontogeny of somatic embryos

Influence of auxin type and concentration on somatic embryogenesis from cotyledonary explants of safflower was investigated. Embryogenic frequency as well as number of somatic embryos was dependent on auxin type and concentration. NAA at 10.74 μM (2 mg l−1) was optimum for high frequency of somatic embryos, whereas IAA provided the maximum number of somatic embryos per responding culture. Somatic embryo development was asynchronous and strongly affected by auxin type and concentration. Maximum numbers of well developed somatic embryos at the cotyledonary stage were obtained with 5.37 μM (1 mg l−1 ) NAA + 2.22 μM (0.5 mgl −1) BA. Histological studies confirmed the unicellular origin of somatic embryos from the adaxial epidermis of the cotyledon. Broad base attachment of somatic embryos to the epidermis indicated the absence of a suspensor.