Meiotic mutants of Medicago sativa show altered levels of α- and β-tubulin

We have analysed the level of accumulation of α- and β-tubulin polypeptides in flowers collected from different meiotic mutants of alfalfa (Medicago sativa L.). The H33 mutant previously identified as a producer of male and female gametes with the somatic chromosome number (2n gametes) as a result of defective spindle orientation or, more rarely, abnormal cytokinesis, showed a higher level of α- and β-tubulin compared to control diploid plants and approximately the same level as control tetraploid plants. A higher level of tubulin was likewise observed in diploid plants displaying abnormalities in spindle orientation and cytokinesis, which had gone through 3-4 cycles of phenotypic recurrent selection to increase 2n gamete production. A similar analysis was performed on another class of Medicago meiotic mutants characterized by production of 4n pollen (jumbo pollen, due to the absence of cytokinesis at the end of meiosis) and 2n eggs. Again, the level of α- and β-tubulin was found to be higher in the mutants than in diploid controls. We conclude that meiotic defects, such as abnormal spindle orientation or cytokinesis leading to the formation of 2n gametes, determine an increased level of tubulin, the main constituent of plant microtubules (MTs). Key words: meiotic mutants, 2n gametes, cytoskeleton, α- and β-tubulin.

Genetics, cytology, and crossing behavior of an alfalfa (Medicago sativa) mutant resulting in failure of the postmeiotic cytokinesis

During pollen investigations on diploid alfalfa (Medicago sativa L.) several plants were identified which produced "jumbo" pollen. The cause of the jumbo pollen is failure of the postmeiotic cytokinesis. These plants produce a single, four-nucleate microspore from one microspore mother cell (MMC) rather than the normal four, single-nucleate microspores from one MMC. Subsequent gametophyte development is characterized by fusion of the four nuclei into a single nucleus in most cases (range of 80 to 100%), followed by a developmental sequence comparable to normal alfalfa. Mature 4n male gametophytes are thus formed from 2n sporophytes. Genetic control of the postmeiotic cytokinesis failure is by a single recessive gene, designated jp. Although a low frequency of jumbo pollen does germinate (range of 3.1 to 37.8%), crossing studies demonstrate jumbo pollen is incapable of effecting fertilization. The use of the jp mutant in breeding studies, and interspecific hybridization research, is discussed.