Fast and cost-effective preparation of plant cells for scanning electron microscopy (SEM) analysis.

The analysis of plant cell structure provides valuable information about its morphological, physiological, and biochemical characteristics. Nowadays, scanning electron microscope (SEM) is widely used to provide high-resolution images at the surface of biological samples. However, biological specimens require preparation, including dehydration and coating with conductive materials for imaging by SEM. There are several techniques for providing images with maximum maintenance of cell structure and minimum cellular damage, but each requires the use of expensive and hazardous materials, which can be damaging to the cell in many cases. Therefore, the provision of new and effective preparation methods based on maintaining cell structure for imaging can be very practical. In the present study, a fast and cost-effective protocol was first performed for chemical fixation and preparation of the plant cells for imaging by SEM. Taxus baccata and Zhumeria majdae cells were chemically fixed using glutaraldehyde and then successfully dried with different percentages of ethanol including 70, 80, 90, and 100%. In addition, SEM was performed for imaging the cell surface in different micro-scales. This protocol can be used by plant cell biologists and biotechnologists who are interested in studying structural and biochemical responses of treated or stressed plant cells by SEM.

Equivalence of three allocation currencies as estimates of reproductive allocation and somatic cost of reproduction in Pinguicula vulgaris.

Which is the most appropriate currency (biomass, energy, water, or some mineral nutrient) for expressing resource allocation in plants has been repeatedly discussed. Researchers need to assess to which extent interindividual, interpopulational, or interspecific comparisons of resource allocation could be affected by the allocation currency chosen. The "currency issue" is relevant to at least three related aspects of resource allocation to reproduction: (a) reproductive allocation (RA), (b) size-dependence of reproductive allocation, and (c) somatic cost of reproduction (SCR). Empirical tests have mostly dealt with the first aspect only. We examined the equivalence of estimates for the three aspects above across three different allocation currencies (dry mass, N, P) in 11 populations of PINGUICULA VULGARIS. For RA we studied the equivalence of allocation currencies at three scales: among individuals of the same population, between populations of the same species, and among species. Equivalence of currencies in the ranking of RA for individuals within populations was high ( RS >/= 0.43) and did not strongly decrease when comparing populations or species. Excepting for size-dependence of RA, ranking of RA, or SCR between populations was equivalent for biomass and N, but not for P. Our study gives two positive guidelines for empirical plant reproductive ecologists facing the "currency issue": (1) become increasingly concerned about the "currency issue" as you increase the scale of your comparison from individuals to populations to species, and (2) avoid estimating allocation in redundant currencies (biomass and N in our case) and choose preferentially "complementary" currencies that provide a broader view of allocation patterns (biomass and P in our case).