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Materials and Methods

Three accessions of Roegneria tsukushiensis (Honda) 13.R. Lu et J.L. Yang (2n=42, SSHHYY) used in this study were collected from Yaan and Yibin, Sichuan Province respectively. Two accessions of Psathyrostachys huashanica Keng (2n=14, NhNh) were collected from Mt. Huashan, Shaanxi Province, China. All these accessions were cultivated at Triticeae Research Institute, Sichuan Agricultural University in 1990.

Hybridization was made by pollinating hand-emasculated spikes of R. tsukushiensis with pollen of P. huashanica. Three F1 seeds were germinated in petri-dish, then transplanted into pots.


For cytological study, young spikes of the three F1 hybrids derived from three different accessions of R. tsukushiensis crossed with one accession of P. huashanica and their parents were fixed in Carnoy's fluid for 24 hours. They were transferred to 70% ethanol and stored at 4oC. Microsporogensis was studied on slides prepared by standard acetocarmine squashing.


Results and Discussion

The intergeneric F1 hybrid of R. tsukushiensis with P. huashanica is theoretically expected to have 28 chromosomes. Chromosome numbers counted at metaphase 1 of the pollen mother cells (PMCs) met the expectation (
Fig. 1-1). The meiotic data indicated that R. tsukushiensis shared no common genome with P. huashanica (Sun et al. 1993a).

Abnormal microspore formation was observed in microsporogenesis of this intergeneric hybrid. Chromatin materials were transferred through conjugation opening among neighbouring cells from the time before meiosis until the young pollen grains stage (Fig. 1-2~7). The amount of chromatin materials transferred varied among cells. The process of chromatin migration among cells causes coenocytes, multiplication and diminution of chromosomes greatly, and variation in cell size. Fig. 1-2 shows PMC a and PMC b in contact with each other and forming a fused big opening. The numbers of anaphase chromosomes in these two cells are unequal. Anaphase chromosomes also show non- synchronous separation. In cell a, there are 9 anaphase chromosomes and 7 chromatids; in cell b there are 2 anaphase chromosomes and 3 chromatids. In Fig. 1-3, two PMCs at chromonema stage contacted each other, chromonemata migration takes place through the conjugation opening. In Fig. 1-4, cell a has two separate synchronous nuclei at chromonema stage; cell b with small cell size has only one chromosome and two chromatids. The chromosome diminution might be caused by uneven distribution of chromosomes before cytokinesis of two conjugated PMCs as shown in Fig. 1-2. After cytokinesis, this kind of small size PMC has occurred as a result a diminished number of chromosomes. In cell c, two chromonema stage nuclei are mixed together. A nucleus has twice as many chromosomes as the usual one. Polyploidy or aneuploidy will occur. A bud-like structure has a micronucleus (Fig. 1-4). The young pollen grains are shown in Figures 1-5~7. We can see that the resting stage nuclei are just migrating through the conjugation opening between two young pollen grains.

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