Meiotic behaviour and pollen fertility of F1, F2 and BC1 progenies of Iris dichotoma and I. domestica

Pollen characteristics are very important for Iris interspecific hybridisation. In this study, the pollen viability and male meiosis were studied in yellow-flowered Iris dichotoma (Y2), I. domestica (S3) and their hybrids F1, F2 and BC1 (BC1-Y and BC1-S). The BC1-Y hybrids showed higher pollen viability than that of F1, F2 and BC1-S hybrids, which were between I. dichotoma (26.1%) and I. domestica (35.1%). Two sterile hybrids, F2-1 and BC1-S-1, exhibited more meiotic abnormalities (57.3% and 58.7%) than other individuals. During the first meiotic division, a diffuse diplotene stage was observed for the first time in the genus Iris. The meiotic abnormalities included non-congressed chromosomes, chromosome bridges, lagging chromosomes, unequal division, abnormally oriented spindle fibres, nonsynchronous division and polyad, and resulted in reduced pollen fertility. The relatively high frequency of 2n pollen grains was found in hybrids of BC1-Y-2, BC1-Y-1, BC1-S-2, BC1-S-3 and BC1-S-4. Our research provides a new resource for meiotic behaviour and pollen fertility of the genus Iris.

Ultraviolet microbeam irradiation of chromosomal spindle fibres in Haemanthus katherinae endosperm. I. Behaviour of the irradiated region

We used an ultraviolet microbeam to irradiate chromosomal spindle fibres in metaphase Haemanthus endosperm cells. An area of reduced birefringence (ARB) was formed at the position of the focussed ultraviolet light with all wavelengths we used (260, 270, 280, and 290 nm). The chromosomal spindle fibre regions (kinetochore microtubules) poleward from the ARBs were unstable: they shortened (from the ARB to the pole) either too fast for us to measure or at rates of about 40 microns per minute. The chromosomal spindle fibre regions (kinetochore microtubules) kinetochore-ward from the ARBs were stable: they did not change length for about 80 seconds, and then they increased in length at rates of about 0.7 microns per minute. The lengthening chromosomal spindle fibres sometimes grew in a direction different from that of the original chromosomal spindle fibre. The chromosome associated with the irradiated spindle fibre sometimes moved off the equator a few micrometers, towards the non-irradiated half-spindle. We discuss our results in relation to other results in the literature and conclude that kinetochores and poles influence the behaviour of kinetochore microtubules.

Rhodamine-labelled phalloidin stains components in the chromosomal spindle fibres of crane-fly spermatocytes and Haemanthus endosperm cells

In crane-fly spermatocytes and Haemanthus endosperm, all metaphase and anaphase chromosomal spindle fibres were stained with rhodamine-labelled phalloidin. In crane-fly spermatocytes, each kinetochore was stained with rhodamine-labelled phalloidin at diakinesis of prophase and after colcemid caused metaphase spindles to depolymerize. Since phalloidin stains actin filaments, the distributions of rhodamine-labelled phalloidin-stained material in crane-fly spermatocytes and Haemanthus endosperm suggest that actin filaments might interact with microtubules to produce forces that move chromosomes during cell division, either directly or via an intermediate motor molecule.Key words: phalloidin, chromosomal spindle fibres, kinetochores.

The behaviour of microtubules in chromosomal spindle fibres irradiated singly or doubly with ultraviolet light

Areas of reduced birefringence (ARBs) produced by ultraviolet microbeam irradiation are areas of depolymerized microtubules. ARBs probably move poleward either by microtubule subunit addition at the kinetochore and loss at the pole, or by microtubule subunit addition at one edge of the ARB and loss from the other edge. In this paper we have used two approaches to try to distinguish between these two models. First, we determined whether the edges of the ARB move at the same rate; if ARB motion is due solely to addition at the kinetochore and loss at the pole, with the ARB edges unable to exchange subunits, then the two edges of each ARB should move at the same rate. On the other hand, if the exchange is at the ARB edges, then, from data from microtubules in vitro, the poleward edge should move much faster than the kinetochoreward edge. We found that the two edges of the ARB move at the same rate about half the time, but half the time they do not. Second, we studied the behaviour of two ARBs on a single fibre. If ARB motion is due solely to subunit addition at the kinetochore and loss at the pole, then the two ARBs must move poleward together. We found that after two ARBs are formed on a single fibre the region between the ARBs is unstable and rapidly depolymerizes. These results do not fit either model and suggest that influences of kinetochores and poles or other factors need to be considered that are not duplicated in experiments on microtubules in vitro.