Alternation of generations and experimental design: a guided-inquiry lab exploring the nature of the her1 developmental mutant of Ceratopteris richardii (C-Fern).

Inquiry-based labs have been shown to greatly increase student participation and learning within the biological sciences. One challenge is to develop effective lab exercises within the constraints of large introductory labs. We have designed a lab for first-year biology majors to address two primary goals: to provide effective learning of the unique aspects of the plant life cycle and to gain a practical knowledge of experimental design. An additional goal was to engage students regardless of their biology background. In our experience, plant biology, and the plant life cycle in particular, present a pedagogical challenge because of negative student attitudes and lack of experience with this topic. This lab uses the fern Ceratopteris richardii (C-Fern), a model system for teaching and research that is particularly useful for illustrating alternation of generations. This lab does not simply present the stages of the life cycle; it also uses knowledge of alternation of generations as a starting point for characterizing the her1 mutation that affects gametophyte sexual development. Students develop hypotheses, arrive at an appropriate experimental design, and carry out a guided inquiry on the mechanism underlying the her1 mutation. Quantitative assessment of student learning and attitudes demonstrate that this lab achieves the desired goals.

Cytokinins induce photomorphogenic development in dark-grown gametophytes of Ceratopteris richardii.

The cytokinins benzylaminopurine, kinetin and isopentenyladenine induce photomorphogenesis in dark-grown gametophytes of the fern Ceratopteris richardii. At sub-nanomolar concentrations each altered the rate and pattern of cell division, elongation and differentiation, mimicking aspects of the light-mediated transition from filamentous to prothallial growth. Untreated dark-grown gametophytes grow as narrow, elongate, asexual filaments with an apical meristem. Cytokinin treatments as low as 10(-12) M reduced the length-to-width ratio through decreased cell elongation, increased periclinal cell division and induced the formation of rhizoid initials in the cells immediately below the apical meristem. Higher concentrations (10(-9)-10(-8) M) induced conversion of the meristem from apical to notch morphology. Cytokinins induced both red- and blue-light-mediated photomorphogenic events, suggesting stimulation of both phytochrome and cryptochrome signaling; however, cytokinin treatment only partially substituted for light in that it did not induce hermaphroditic sexual development or spore germination in the dark. Additionally, cytokinins did not increase chlorophyll synthesis in dark-grown gametophytes, which unlike angiosperms are able to produce mature chloroplasts in the dark. Cytokinin treatment had only slight effects on light-grown gametophytes. These results suggest evolutionary conservation between angiosperms and pteridophytes in the role of cytokinins in regulating photomorphogenesis.

A comparison of oligogalacturonide- and auxin-induced extracellular alkalinization and growth responses in roots of intact cucumber seedlings.

Oligogalacturonic acid (OGA) affects plant growth and development in an antagonistic manner to that of the auxin indole-3-acetic acid (IAA), the mechanism by which remains to be determined. This study describes the relationship between IAA and OGA activity in intact cucumber (Cucumis sativus) seedlings. Both OGA and IAA induced rapid and transient extracellular alkalinization; however, the characteristics of the OGA and IAA responses differed in their kinetics, magnitude, calcium dependence, and region of the root in which they induced their maximal response. IAA (1 microM) induced a saturating alkalinization response of approximately 0.2 pH unit and a rapid reduction (approximately 80%) in root growth that only partially recovered over 20 h. OGAs, specifically those with a degree of polymerization of 10 to 13, induced a maximal alkalinization response of 0.48 pH unit, but OGA treatment did not alter root growth. Saturating concentrations of OGA did not block IAA-induced alkalinization or the initial IAA-induced inhibition of root growth but allowed IAA-treated roots to recover their initial growth rate within 270 min. IAA-induced alkalinization occurs primarily in the growing apical region of the root, whereas OGA induced its maximal response in the basal region of the root. This study demonstrates that OGA and IAA act by distinct mechanisms and that OGA does not simply act by inhibition of IAA action. These results also suggest that IAA-induced extracellular alkalinization is not sufficient to account for the mechanism by which IAA inhibits root growth.