Many enzymes in plants have isozymes because the same catalytic reaction is often present in several subcellular compartments, most frequently the plastids and the cytosol. The number and subcellular locations of the isozymes appear to be highly conserved in plant evolution. However, gene duplication in diploid species and the addition of genomes in polyploid species have increased the number of isozymes.

https://science.sciencemag.org/content/sci/216/4544/373.full.pdf

Conservation and duplication of isozymes in plants.

Green fluorescent protein (GFP) has emerged as a powerful new tool in a variety of organisms. An engineered sGFP(S65T) sequence containing optimized codons of highly expressed eukaryotic proteins has provided up to 100-fold brighter fluorescence signals than the original jellyfish GFP sequence in plant and mammalian cells. It would be useful to establish a non-invasive, quantitative detection system which is optimized for S65T-type GFP, one of the brightest chromophore mutants among the various GFPs. We demonstrate here that highly fluorescent transgenic Arabidopsis can be generated, and the fluorescence intensity of whole plants can be measured under non-disruptive, sterile conditions using a quantitative fluorescent imaging system with blue laser excitation. Homozygous plants can be distinguished from heterozygous plants and fully fertile progenies can be obtained from the analyzed plants. In the case of cultured tobacco cells, GFP-positive cells can be quantitatively distinguished from non-transformed cells under non-selective conditions. This system will be useful in applications such as mutant screening, analysis of whole-body phenomena, including gene silencing and quantitative assessments of colonies from microorganisms to cultured eukaryotic cells. To facilitate the elucidation of protein targeting and organelle biogenesis in planta, we also generated transgenic Arabidopsis that stably express the plastid- or mitochondria-targeted sGFP(S65T). Etioplasts in dark-grown cotyledons and mitochondria in dry seed embryos could be visualized for the first time in transgenic Arabidopsis plants under normal growing conditions.

Non-invasive quantitative detection and applications of non-toxic, S65T-type green fluorescent protein in living plants

Eukaryotic cells arose through endosymbiotic uptake of free-living bacteria followed by massive gene transfer from the genome of the endosymbiont to the host nuclear genome. Because this gene transfer took place over a time scale of hundreds of millions of years, direct observation and analysis of primary transfer events has remained difficult. Hence, very little is known about the evolutionary frequency of gene transfer events, the size of transferred genome fragments, the molecular mechanisms of the transfer process, or the environmental conditions favoring its occurrence. We describe here a genetic system based on transgenic chloroplasts carrying a nuclear selectable marker gene that allows the efficient selection of plants with a nuclear genome that carries pieces transferred from the chloroplast genome. We can select such gene transfer events from a surprisingly small population of plant cells, indicating that the escape of genetic material from the chloroplast to the nuclear genome occurs much more frequently than generally believed and thus may contribute significantly to intraspecific and intraorganismic genetic variation.

https://www.pnas.org/content/pnas/100/15/8828.full.pdf

High-frequency gene transfer from the chloroplast genome to the nucleus.

Chitosan is a natural biopolymer modified from chitins which act as a potential biostimulant and elicitor in agriculture. It is non-toxic, biodegradable and biocompatible which favors potentially broad application. It enhances the physiological response and mitigates the adverse effect of abiotic stresses through stress transduction pathway via secondary messenger(s). Chitosan treatment stimulates photosynthetic rate, stomatal closure through ABA synthesis; enhances antioxidant enzymes via nitric oxide and hydrogen peroxide signaling pathways, and induces production of organic acids, sugars, amino acids and other metabolites which are required for the osmotic adjustment, stress signaling, and energy metabolism under stresses. It is also known to form complexes with heavy metals and used as tool for phytoremediation and bioremediation of soil. Besides, this is used as antitranspirant compound through foliar application in many plants thus reducing water use and ensures protection from other negative effects. Based on such beneficial properties, chitosan is utilized in sustainable agricultural practices owing to changing climates. Our review gathers the recent information on chitosan centered upon the abiotic stress responses which could be useful in future crop improvement programs.

/pdf/application-of-chitosan-on-plant-responses-with-special-4tlam8ubkf.pdf

Application of chitosan on plant responses with special reference to abiotic stress

A nuclear recessive mutant of Arabidopsis thaliana, arc5, has been isolated in which there is no significant increase in chloroplast number during leaf mesophyll cell expansion and in which there are only 13 chloroplasts per mesophyll cell compared with 121 in wild-type cells. Mature arc5 chloroplasts in fully expanded mesophyll cells are 6-fold larger than in wild-type cells. A large proportion of arc5 chloroplasts also show some degree of central constriction, suggesting that the mutation has prevented the completion of the chloroplast division process. To examine the interaction of arc loci, a double mutant was constructed between arc1, a mutant possessing many small chloroplasts, and arc5. A second double mutant was also constructed between arc3, a previously discovered mutant also possessing few large chloroplasts per cell, and arc1. Analysis of these double mutants shows that chloroplast number per mesophyll cell is greater when arc5 and arc3 mutations are expressed in the arc1 background than when expressed alone. The cell-specific nature of arc mutants was also analyzed. The phenotypic traits characteristic of arc3 and arc5 are a reduction in chloroplast number and an increase in chloroplast size in mesophyll cells: these changes are also observed in reduced form in the epidermal and guard cell chloroplasts of arc3 and arc5 plants. Analysis of parenchyma sheath cell chloroplasts suggests that in leaves of arc1 plants the normal developmental distinction between mesophyll and parenchyma sheath chloroplasts is perturbed. The relevance of these findings to the analysis of the control of chloroplast division in mesophyll cells is discussed.

http://www.plantphysiol.org/content/plantphysiol/104/1/201.full.pdf

John V. Possingham

Papers

Plastid replication and development in the life cycle of higher plants.