1971 487

Chromosomes of Some West African Orchids

A. H. ar-Rushdi

Department of Botany, University of Ibadan,

Ibadan, Nigeria

Received February 10, 1970

The "Flora of West Tropical Africa" lists over 400 species of orchids.
About 300 of these are estimated to occur in Nigeria and West Cameroun.
In contrast to the wide range of data on American and Asian orchids,
cytological information is available for only a few West African species. An
opportunity for increasing such information was provided by Mr. W. W.
Sanford's living collection of over 2,000 orchids, mainly from Nigeria and
some from West Cameroun. His assistance in the identification of over 200
collected species and in making available the right material for cytological
studies is duly acknowledged.

Materials and methods

The main source of material is Mr. Sanford's collection at the University

of Ife. These are designated in Table 1 by the prefix WS. Additional
material. collected by the staff of the Botany Department, University of Ibadan,
appears in the table as herbarium number with the prefix UIH. As far as
possible, more than one specimen was examined.
Root-tip cytology in orchids, especially of subterranean roots, is beset
with difficulties. It was therefore decided to use the sporogenous cells, since
flowers were readily available. Post-meiotic metaphase in the tetrad of micro
-spores was found to be a suitable and easily prepared stage for haploid counts.
One pollinium was directly stained, the other being kept in case the spores
were found premature for metaphase studies. Such pollinia were cultured in
distilled water at room temperature for periods of 3-12 hours, depending on
their stage of division. Culturing was successful only for pollinia with micro
- spores in late prophase or prometaphase stages.
The stain used was a preparation of 1% (w/v) synthetic orcein, dissolved
by gentle heating in 40% propionic acid, cooled, filtered, and diluted with
the acid as required. Spores were teased out of the pollinium into a drop
of stain and gently covered. The cover was temporarily sealed to the slide,
which was examined, photographed and often made permanent by the freezing
method. The same staining procedure was followed in preparing smears of
pollen-mother-cells but the pollinia were fixed for 15 minutes in glacial acetic
alcohol (2:1) prior to staining.
1 Present Address: Biology Department, Pahlavi University, Shiraz. Iran.

33*
488 A. H. ar-Rushdi Cytologia 36

Table 1. Chromosome numbers

1971 Chromosomes of Some West African Orchids 489

Table 1. (continued)
490 A. H. ar-Rushdi Cytologia 36
1971 Chromosomes of Some West African Orchids 491

Results and discussion

Sixty-three plants, derived from about 32 different species belonging to

eight genera, were studied. Table 1 shows the record of their sporophytic
chromosome numbers. Plate 1 illustrates some representative photomicro
graphs.
In the genus Polystachya, Jones (1966) reported the chromosome numbers
of about 21 species. At least 14 species are reported here, confirming the
constancy of the basic number (n=20) for the genus. A polyploid number,
hitherto not found in West African species of Polystachya, is reported for
P. odorata (WS-4450), (Figs. 7 and 8). Its chromosomal association in meiosis
was not observed but lack of variability in 58 spores examined from the plant
indicates its chromosomal stability. The plant was collected from the summit
of Mt. Orosun. Morphologically, it could not be separated from the other
diploid members of the species.
In the genus Eulophia (where six species were investigated), the basic
chromosome number varies, and basic numbers of n=20, 22, 23, 31, and
possibly 24, have been observed.
Jones (1967) reports the chromosome numbers in species belonging to
ten genera of the subtribe Aerangidinae. He concludes that the predominant
basic number is x=25, that most of the species are diploid but polyploid
species do occur. He also records two exceptions, Ae. rhodostica (2n=42)
and Cyrorchis sp. (2n=46), the latter collected from Southern Nigeria.
Ancistrohyncus clandestinus (2n=48) is another West African exception.
In all species studied, tetrad analysis of the chromosomes was readily
possible as the spores remained attached to one another and their stages of
division were synchronised. Furthermore, in the majority of the species, a
high degree of synchronisation was also observed between tetrads within a
pollinium. Tetrad analysis was useful in detecting trisomics, triploidy and
occasional distributional errors of meiosis in diploids. The trisomic condition
in Eulophia euglosa (UIH 10288), suspected from tetrad analysis (Figs. 3
and 4) and from the occurrence of micronuclei in some tetrads was verified
by meiotic observations. In Polystachya rhodoptera (WS-203-66), the trisomic
plant also showed complementary counts of 20 and 21 in the tetrad, but its
meiotic associations were not examined. Plant UIH 10285 was recognised
as a triploid by the highly variable counts in the spores and by the occur
rence of micronuclei of variable sizes and chromosome content. It was further
verified as an autotriploid by examining the first meiotic metaphase where 22

Figs. 1 and 3-11. Metaphase chromosomes, ca. •~2,000 in the microspores of: 1, Eulophia

gracilis (n=22). 3, Eulophia euglossa (n=20). 4, Eulophia euglossa (n+1=21). 5, Eulophia

cristata (n=23). 6, Eulophia guineensis (n=23). 7, diploid Polystachya odorata (n=20).

8, tetraploid Polystachya odorata (n=40). 9, Ancistrorhynchus clandestinus (n=24). 10,

Euclophidium saundersianum (n=29). 11, Eulophia quartiniana (n=48). Fig. 2. Chromo

some configurations at metaphase I of meiosis in triploid Eulophia gracilis (22III). •~1,600.

 492 A. H. ar-Rushdi Cytologia 36

trivalents invariably occurred (Fig. 2). The fertility of this plant was much
impaired, yet three capsules of seeds were set in two flowering seasons.
Observed errors of meiosis in diploids consisted of occasional non
- disjunction of individual chromosomes, recognised as exceptional counts of
n-1 and n+1 in the tetrad, and of non-reduction of the chromosome number.
Pooling the data from all the diploid species investigated, non-disjunctional
counts were recorded for 0.22% of the spores. Data from individual plants
were considered insufficient to associate such error with specific plants or
species.
Restitutional dyads were observed in individual plants belonging to different
species and genera. Their incidence was rare, but their occurrence in each
of the three plants of Polystachya rhodoptera investigated, including the
trisomic plant, is perhaps noteworthy. Restitution was also observed to have
occurred in one of the products of the first meiotic division, giving rise to a
"Triad" with one diploid and two haploid spores .
Barber (1942) drew attention to the physiological implications of the
spores of orchids remaining attached in tetrads. He suggested that the
possibility of nuclear interaction between adherent spores made the group of
four cells, rather than the individual cell, act as a unit of development. He
concluded that the synchronisation of cell division was a climax of the unitary
control of cell processes within the tetrad. He also pointed out the significance
of the condition in permitting the division of chromosomally deficient spores,
even of micronuclei with only a few chromosomes, in harmony with the
balanced members of the tetrad. These explanations appear to be corroborated
by the present observations. Furthermore, Barber's data as well as the
observation made by Miduno (1940) on a haploid plant of Bletilla striata,
indicated no pollen abortion of chromosomally unbalanced spores. It would
therefore appear that the natural occurrence of chromosomal variants in spores
and the lack of stringent selection against such spores, coupled with prevalent
vegetative reproduction, provide a combination of features permitting variation
in chromosome number in orchids. This probably explains the diversity of
basic chromosome numbers reported for numerous genera of orchids, and may
be a means for exploiting the profusion of habitats available in the tropics.

References

Barber, H. N. 1942. The pollen-grain division in the Orchidaceae. J. Genet. 43: 97-103.
Jones, K. 1966. The chromosomes of orchids I. Polystachya Hook. Kew Bull. 20: 357
- 359.-
1967. The chromosomes of orchids II. Vandeae Lindl. Kew Bull. 21: 151-156.
Miduno, T. 1940. Chromosomenstudien an Orchidazeen III. Uber das Vorkommen von
haploiden Pflanzen bei Bletilla striata Reichb. f. var. gebina Reichb. f. Cytologia
11: 156.