New roads towards chloroplast transformation in higher plants

The attempts to manipulate organelle DNA of higher plants have not yet been succesfull. Although some Agrobacterium mediated and direct gene transfer experi-ments suggested that chloroplasts of Nicotiana tabacum could be transformed, these experiments could not be reproduced. New transformation techniques have emerged: laser-injection, microinjection and high velocity microprojectiles. The biolistic ex-periments with Chlamydomonas made it clear that organelle DNA can support introduced DNA fragments by homologous recombination. In view of these devel-opments, we describe the parameters and new possibilities for chloroplast trans-formation of higher plants.     

Ecology and fishery management of reservoirs in India

India has 19 370 small reservoirs with a total water surface area of 3 153 366 ha. At least 100 of them have been subjected to scientific studies. Habitat variables responsible for a reservoir's productivity can be summed up into climatic, morphometric and hydro-edaphic factors. The peninsular reservoirs are characterized by a narrow range of fluctuations in water and air temperature across seasons, a phenomenon which prevents the formation of thermal stratification. Many reservoirs in the upper peninsula show thermal stratification during summer. Wind-induced turbulence facilitates the return of nutrients to the trophogenic zone. Most reservoirs on the mountain slopes of Western Ghats, Himalayas and the other highlands are deeper, with steeper basin walls, compared to irrigation impoundments. Mean depth does not show any direct correlation with productivity, either at primary or fish level. A high shoreline development index gives a better indication of productivity. Plankton, benthos and periphyton pulses of Indian reservoirs coincide with the months of least level fluctuations. Oligotrophic tendencies shown by some reservoirs are mainly due to poor nutrient status and other chemical deficiencies. In most cases, poor water quality is accountable to poor catchment soil. Low levels of phosphate and nitrate are not indicative of low productivity due to quick recycling of these nutrients. Specific conductivity reflects the production propensities of reservoirs satisfactorily. Almost all productive reservoirs have a klinograde oxygen curve and a vertical stratification of chemical variables such as pH, carbon dioxide, total alkalinity and specific conductivity. High seasonal rainfall and discharge of water during monsoon result in high flushing rates, which do not favour colonization by macrophytic communities. Similarly, inadequate availability of suitable substrata retards the growth of periphyton. Plankton constitutes the major link in the trophic structure. A rich plankton community with well-marked succession is the hallmark of Indian reservoirs with blue-green algae as the major component. The main factors that retard the growth of benthos are a rocky bottom, frequent water level fluctuation and rapid deposition of silt and other suspended particles. Large reservoirs, on average, harbour 60 species of fishes, of which at least 40 contribute to the commercial fisheries. Fast-growing Indian major carps are the prominent commercial fishes. Dam construction has adversely affected populations of many other species such as Tenualosa ilisha, Torspp. and Cirrhinus spp. Formulae for estimating fish yield potential and stocking density are described. While culture-based fisheries have been successfully practiced in many small reservoirs, the management norm followed in medium and large reservoirs is primarily on capture fishery. In large and medium reservoirs, stocking was successful only when stocked fishes bred. Indian experience on species enhancement and introductions is described. Environmental enhancement of small reservoirs has been attempted in some reservoirs of Tamil Nadu. Modeling, using standard population parameters, such as the density-dependent growth, size dependent mortalityand weight–length relationshipis discussed. Two exotic fishes viz., Oreochromis mossambicus and Cyprinus carpio have been introduced into Indian reservoir with discouraging results. Hypophthalmichthys molitrix, after an accidental introduction, has performed well in Gobindsagar, a reservoir with a distinct cold water regime. Reservoir fisheries in India are well poised for development, provided scientific management norms are adopted.     

Efficient plant regeneration through somatic embryogenesis from callus induced on mature rice embryos (Oryza sativa L.)

To obtain a reproducible efficient procedure for regeneration of rice plants through somatic embryogenesis from callus four published methods of callus induction and regeneration were compared. Callus was initiated from mature embryos of the Japonica cultivar Taipei 309 of rice (Oryza sativa L.). The number, mass and morphology of the callus formed on the scutellum were dependent on the medium used. A limited humidity and an optimal aeration of the culture vessels enhanced the frequency of embryogenesis and plant regeneration. A method described by Poonsapaya et al. (1989) was found to be the most efficient and was slightly modified. As a result 98% of the T309 embryos formed callus, of which 63% regenerated into plants. Each callus yielded an average of 6 plants. Plant morphology, fertility and seed set of the regenerants were found to be normal.Abbreviations 2,4-d 2,4-dichlorophenoxyacetic acid - IAA 3-indole-acetic acid - BA 6-benzyladeninepurine - S.E.M. standard error of mean     

The ubiquitous nature of the progesterone receptor binding factor-1 (RBF-1) in avian tissues

The avian oviduct receptor binding factor-1 (RBF-1) is a 10 kDa nuclear matrix protein that was originally identified through its ability to effect high affinity interaction of activated progesterone receptor (PR) with chromatin. In the present study, the RBF-1 is shown to not be restricted to reproductive tissues (e.g., oviduct) but present in all avian tissues examined by Western blot analysis with a monoclonal antibody prepared against purified RBF-1. The heart and pancreas had the highest and lowest RBF-1 levels, respectively; the concentration ranging by ～ 50-fold in these tissues. The 10 kDa size of the RBF-1 detected in all tissues suggests no significant tissue-specific differences in the protein. This was consistent with the finding that purified hepatic and oviductal RBF-1 have identical amino-terminal sequence. Using a recently isolated cDNA to RBF-1, the levels of RBF-1 mRNA were found to correlate well with the ubiquitous presence of the protein as well as tissue-specific differences in concentration. The presence of RBF-1 in non-progesterone responsive tissues suggests the possibility that RBF-1 may not be specifically involved in PR-DNA interactions but may play a more diverse role, possibly involving other steroid receptors such as the glucocorticoid receptor. © 1994 Wiley-Liss, Inc.     

Novel plasmid marker rescue transformation system for molecular cloning in Bacillus subtilis enabling direct selection of recombinants

Summary A versatile plasmid marker rescue transformation system was developed for homology-facilitated cloning in Bacillus subtilis. It is based on the highly efficient host-vector system 6GM15-pHPS9, which allows the direct selection of recombinants by means of -galactosidase -complementation. The system offers several advantages over previously described cloning systems: (1) the convenient direct selection of recombinants; (2) the ability to effectively transform B. subtilis competent cells with plasmid monomers, which allows the forced cloning of DNA fragments with high efficiency; (3) the availability of 6 unique target sites, which can be used for direct clone selection, SphI, NdeI, NheI, BamHI, SmaI and EcoRI; and (4) the rapid segregational loss of the helper plasmid from the transformed cells.     

Liposome-mediated gene delivery into plant cells

Liposomes may offer several advantages as vectors for gene delivery into plant cells: (1) enhanced delivery of encapsulated DNA by membrane fusion, (2) protection of nucleic acids from nuclease activity, (3) targeting to specific cells, (4) delivery into a variety of cell types besides protoplasts by entry through plasmodesmata, (5) delivery of intact small organelles. Realization of these advantages calls for the construction of efficient liposomes, for appropriate fusion conditions and for an understanding of the nature of liposome-cell interactions. Various characteristics and techniques of the liposome-cell system are described (mode of delivery, liposome types and composition, and means of promoting delivery of liposome contents). Data of liposome-mediated delivery of various macromolecules into plant cells, with special reference to protoplasts, calli and pollen are reviewed. This includes data obtained by the use of fluorescent probes, radioactive-labelled DNA, viral nucleic acids and expression of plasmid-DNA. Structure and characteristics of plant surfaces and plasmodesmata are discussed with respect to DNA entry. It is suggested that liposome-mediated gene delivery into plant cells, and not only protoplasts, will be advantageous in certain specific tissues and situations.     

Transformation and regeneration of the self-incompatible species Nicotiana alata Link & Otto

A transformation and regeneration system has been developed for Nicotiana alata, a plant which is being intensively studied as a model of gametophytic self-incompatibility. Plantlets can be regenerated efficiently from seedling hypocotyls. Kanamycin-resistant, transformed plants have been obtained by cocultivation of regenerating hypocotyls with Agrobacterium tumefaciens strain LBA4404 containing a binary vector. The transformation frequency was low with <1% of tissue explants regenerating transformed plants. The transformed plants contained from one to three copies of the introduced DNA. In most cases, the kanamycin resistance phenotype was transmitted to the offspring as a normal Mendelian factor. In one unusual case, none of the offspring inherited the kanamycin resistance of the transformed maternal parent. This plant may have been chimeric or the kanamycin resistance gene may have been inactivated.     

A pregnane ester diglycoside from periploca calophylla

A new pregnane ester diglycoside of ornogenin named as plocinine was isolated from the dried twigs of Periploca calophylla. On the basis of chemical and spectroscopic evidences its structure was established as 12,20-di-O-cinnamoyl sarcostin-3-O-α-L-oleandropyranosyl (1 → 4)-O-α-L-oleandropyranoside.