Determination of Chromosome Number of Kuwaiti Flora I. The

C 1999 The Japan Mendel Society Cytologia 64: 181-196, 1999

Determination of Chromosome Number of Kuwaiti Flora I.

G. A. Malallah and G. Brown

Department of Biological Sciences, Faculty of Science, Kuwait University, Kuwait.

Accepted February 12, 1999

Summary This paper deals with the chromosome numbers (somatic or sporophytic and gametic) of some wild species of the Kuwaiti flora. All species were collected from the wild except Plantago lanceolata, which was cultivated. Some values are reported for the first time. The species and their respective chromosome numbers are as follows: Anthemis deserti 2n=18, Atractylis carduus 2n=20, Calendula arvensis 2n=36, 44, Koelpinia linearis 2n=54, 36, Launaea capitata 2n=18, L. mucronata 2n=16, Picris babylonica 2n=10, Reichardia tingitana 2n=16, Senecio glaucus 2n=20, Sonchus oleraceus 2n=32, Astragalus hauarensis 2n=48, A. schimperi 2n=16, Lotus halophilus 2n=14, 28, Medicago laciniata 2n=16, Plantago amplexicaulis 2n=10, P. boissieri 2n=10, P. ciliata 2n = 10, P. coronopus 2n= 10, P. lanceolata 2n= 12, P. ovata 2n =8. Karyotype analysis of P. babylonica, R. tingitana, P. boissieri, and P. ciliata is shown.

The chromosome set is not only one of the most characteristic attributes of a species, but also carries the bulk of essential information for controlling its organization and metabolism. Therefore, variations in number, size and form of the chromosomes between genera and related species, and sometimes between populations of the same species, are of importance for classification and certain evolutionary aspects (Sato 1962). Such variations represent different karyotypes (Jackson 1971). Numerical analysis of chromosomes is one of the processes in assessing the genomic status of a species. Chromosome numbers of species can provide useful information for various fields of research including karyotaxonomy, genetics, cytogenetics, plant breeding, ecology, biogeography and molecular biology. A number of workers have been involved in reporting the chromosome numbers of wild and cultivated plant species (Darlington and Wylie 1955, Bolkhovskikh et al. 1969, Love and Love 1974, 1975, Moore 1973a, b, 1977, Meikle 1977, Beuzenberg and Nair 1983, Goldblatt 1981a, b, 1985, Webb and Beuzenberg 1987, Majovsky and Murin 1987, Rao and Chandel 1991). On the basis of such information it has often been possible to interpret the genetic interrelationships existing within species (Berdahle and Barker 1991, Brochmann 1992, Stepankova 1993). Halwagy and Macksad (1972) give a historical account of the exploration of the Kuwaiti flora, but no information on chromosomal aspects has been reported. Indeed, there is a complete lack of knowledge regarding the karyology of wild plant species in Kuwait (Malallah et al. 1996). The main aim of this paper is to provide information on the chromosome number of various species of the Kuwaiti flora. In this initial study, 20 species from three different families were selected, and their mitotic and meiotic chromosome numbers determined as well as the mitotic index, karyotype analysis of 4 species is also considered.

Materials and methods

20 species from three families, Asteraceae, Fabaceae, and Plantaginaceae were selected for this study. Identification of the species was checked by Mrs. M. Al-Doseri, Biological Sciences- University of Kuwait. Nomenclature and synonyms are in accordance with Boulos and Al-Dosari (1994), Daoud (1985) and Al-Rawi (1987). Seeds of each species were collected from at least three different plants growing at various lo- 182 G. A. Malallah and G. Brown Cytologia 64

calities in Kuwait during November to April 1993 and 1998. They were germinated in a controlled

growth chamber for 12 h dark at 16•Ž and 12 h light at 22•Ž on moist filter paper enclosed in Petri dishes. Actively growing root tips were excised and fixed in Carnoy's fixative (1 : 3 acetic acid alcohol) until ready for use. For slide preparation, excised roots were washed with distilled water for 4-

5 min, hydrolyzed in 1N HCl for 10-12 min at 60•Ž and finally rinsed with distilled water for 4- 5 min. The roots were stained in 1% aceto-carmine for microscopic analysis. Some three-day old

seedlings were treated with 0.05% colchicine for 4 h or 20 h cold water treatment at 50•Ž in order to have c-metaphases.

Young flower buds were collected and kept in Carnoy's fixative until further use. After discard- ing unwanted tissues, anthers were squashed in a drop of aceto-carmine stain and covered with a

clean cover glass. Microscopic analysis was carried out with an Olympus BH-2 photomicroscope. Micrographs were taken when necessary from temporary preparations. Slides were made permanent by removing the cover slip with dry ice techniques followed by dehydration of the slides in increas- ing alcohol concentrations. Several well-spread metaphases were counted to confirm the validity of the chromosome number and for karyotype analysis. The mitotic index was calculated by recording the stage of every nucleus present in a field of view at 400 •~ magnification. When expressed as a percentage, the relative amount of time nuclei spent at each mitotic stage can be determined. Data on general distribution and various ecological aspects of individual species were taken from the literature, although we have added our own observations when more appropriate.

Karyotype analysis For diploid karyotype analysis, a minimum of 5 root tip cells from each case was analyzed.

Cells in which the chromosomes were well separated were photographed at high magnification. Photo-ideograms were prepared from the photographs by cutting out individual chromosomes and arranging them in a descending order of their length. On the basis of gross morphology and centromere position, the chromosomes were grouped to form homologous pairs. Averages of the paired chromosomes were taken for total length, arm ratio and total volume. The total length was ascertained by adding the length of all chromosomes in the karyotype. The ratio of the long arm (L) to the short arm (S) was used to identify centromere position for each chromosome. The nomenclature system of Levan et al. (1964) was used to determine the chromosome types as median (m), sub-median (sm) or sub-terminal (st). The chromosome volume was determined from the metaphase plate, assuming the chromosome as cylindrical structure by the formula ƒÎr2h, where r and h represent the radius and the length of the chromosome respec- tively. The total volume is determined by adding the volumes of all chromosomes.

Results and discussion

The mitotic and meiotic chromosome numbers and the mitotic index of each species are listed in Table 1 while the corresponding data from the literature are included in the text. Further details on morphological and ecological aspects of each species in Kuwait, are available from Al-Rawi

(1987) and Boulos (1987). The rate of mitotic division in the indicated species has not been investigated before. In some systems, the mitotic index or the proportion of cells in visible stages of division directly reflects the relative duration of mitotic stages (Dyer 1979). The procedure performed in the present work demonstrates an asynchronous fashion, and cannot therefore provide an ab- solute value for the duration of the individual stages, or the duration of the whole cycle. Further work is required to clarify. However, the mitotic index mean gives an indication of that mitosis oc- cupied a typical value for the total time (Brown and Bertke 1974). The life cycle of annual wild species in Kuwait is very short and the germination starts just several days after two or three show- ers of rain in the beginning of the season and develops into full plants in very short time period in- 1999 Determination of Chromosome Number of Kuwaiti Flora I. 183

Table 1. Chromosome number and mitotic index of some wild plant species in Kuwait

dicating that the main factor affecting the germination is the rain. The question is whether the life span of the wild species could be controlled or not and the duration of the cell cycle are the subjects of future work.

ASTERACEAE Anthemis deserti Boiss. Syn.: A. melamopodina subsp. deserti var. transiens Eig loc.cit.; A. diffusa Salzm. ex DC. Geographical distribution: Saharo-Arabian. Kuwaiti data: Found in many localities particularly common at Al-Sabiyah. Flowering time: Febru- ary-April. Literature data: No information. Comment: According to the literature, this species has not been investigated cytogenetically. This study therefore provides the first count of the chromosome number and the relative duration of mitosis. An interesting observation was that most cells were 2n=18, while a few others were 2n=24. A possible explanation is that some capitula had seeds of another species attached to them. We therefore consider 2n=18 to be the correct value.

Atractylis carduus (Forssk.) Syn.: A. flava Coss. and Dur.; Centaurea carduus Forssk. Geographical distribution: Saharo-Arabian. Kuwaiti data: Found in many sandy localities. Flowering time: March-April. Literature data: 2n=20 (Waisel 1962); A.flava 2n=20 (Brullo et al. 1990). Comment: The somatic number of 2n=20 is the third published karyological study for this species and the first undertaken on Kuwait material. This mitotic chromosome number was the same from the three different localities in Kuwait and which is consistent with the findings from the other countries indicating the high stability of the diploid number. Accordingly, the gametic number 184 G. A. Malallah and G. Brown Cytologia 64 n=10.

Calendula arvensis L. Syn.: C. sancta L.; C. persica C. A. Mey.; C. aegyptiaca Desf. Geographical distribution: Mediterranean, Irano-Turanian, and Saharo-Arabian. Kuwaiti data: It is an annual herb, which can be found in many localities in sandy places. Flowering time: February-April. Literature data: 2n=22 (Aparicio et al. 1989); C. persica C. A. Mey. 2n-22 (Ghaffari 1987a, b); 2n=36 (Negodi 1935, 1937); 2n=44 (Meusel and Ohle 1966, Janaki and Sobti 1962, Forissier 1973, Queiros 1973, Van Loon 1974, Heyn et al. 1974, Borgen 1975, Scrugli 1974, Gallego and Talvera 1983 (2 References in IPCN 1992-93); 2n=42 (Borgen 1974, Love and Kjellquist 1974); 2n=14 (Humphries et al. 1978); 2n=18 (Humphries et al. 1978); n=22 (Strother 1972); n=23 (Rashid 1974). Comment: This species seems to have variable counts of chromosome number and the variation in the counts reflects the variation at the ploidy level of this species. The Kuwaiti value supports those with 2n=36 and 2n=44, and the two counts are consistent with counts from different countries and show the high level of ploidy. This raises the question of the existence of the diploid plant in the Kuwaiti flora and the origin of the available species. No data were recorded for the gametic chromosome number before and the present account n=22 is the first.

Koelpinia linearis Pall. Syn.: Lapsana koelpinia L.; K. mucronata C. Winkl.; K. linearis Pall. var. mucronata (C. Winkl.) Geographical distribution: Saharo-Arabian. Kuwaiti data: This herb can be found on many localities in sandy, calcareous and saline soils. Flow- ering time: March-April. Literature data: 2n=14, 42, 56 (Nazarova 1981); K. mucronata C. Winkl. 2n=12 (Nazarova 1981); 2n=54 (Stebbins et al. 1953); 2n=40 (Podlech and Bader 1974). Comment: The counts of chromosomes of sporophytic are the hexaploid 54 and the tetraploid 36 and gametic number is 18 and all are first to be reported from Kuwaiti flora. The sporophytic number of 36 is the first to be reported while the sporophytic number of 54 is the second to be reported in the world. The basic number is x=9. More karyological studies need to be conducted to clarify the origin of this species.

Launaea capitata (Spreng.) Syn: Sonchus capitatus Spreng.; Lomatolepis glomerata Cass.; Zollikoferia glomerata (Cass.) Boiss.; Launaea glomerata (Cass.) Hook. F.; Microrhynchus glomeratus (Cass.) Jaub. & Spach. Geographical distribution: Saharo-Arabian, Irano-Turanian, Sudanian. Kuwaiti data: Found in many desert localities. Flowering time: February-April. Literature data: n = 9 (Amin 1973). Comment: Somatic count appears to be the first for this taxon. It has a diploid level of ploidy, with the gametic count of 9. The gametic count is in agreement with Amin (1973).

Launaea mucronata (Forssk.) Muschl. Syn.: Leontodon mucronatum Forssk.; Zollikoferia chondrilloides DC.; Z. mucronata Forssk. Boiss.; Sonchus candolleanus Jaub. & Spach. Geographical distribution: Saharo-Arabian, Irano-Turanian. Kuwaiti data: Found in many localities both in desert and in urban areas. Flowering time: March- April. Literature data: 2n=16 (Reese 1957); n= 8 (Ghaffari 1987b, Amin 1973). 1999 Determination of Chromosome Number of Kuwaiti Flora I. 185

Picris babylonica

Plantago boissieri

Plantago ciliata

Reichardia tingitana

Fig. 1. Karyotyping of Picris babylonica, Plantago boissieri, Plantago ciliata and Reichardia tingitana speices.

Comment: The counts for this species are the first for Kuwaiti flora and the second for the world flora according to the available plant chromosome indexes and both of them show the diploid level of this species.

Picris babylonica Hand.-Mazz Syn.: P. babylonica Hand.-Mzt. var. dimorphocarpa Eig. Geographical distribution: Saharo-Arabian, Irano-Turanian, Sudanian. Kuwaiti data: Widely distributed in many localities on sandy gravelly soil. It is one of the most common and conspicuous species, forming dense stands in favorable years. Flowering time: Febru- ary-May. Literature data: n=5 (El-Bayoumi 1973). Comment: The sporophytic chromosome number was first recorded by the first author in 1993 and is confirmed in 1998 for the same species. This count which shows the diploid level is the first from Kuwaiti flora and no previous count has been recorded for this species in literature. This work con- 186 G. A. Malallah and G. Brown Cytologia 64

Table 2. Karyotype analysis of five different cells of Picris babylonica 2n= 10

m=Median, sm=Submedian, P.O.C.=Position of centromere. firms the gametic number, which are 5. From Fig. 1 and Table 2 it is shown for the first time that P. babylonica has 5 pairs of average sized chromosomes in which the third pair has the centromere in median position while the rest pairs have the sub-median position. From the five analyzed karyotypes, it is shown that the first pair with satellite. Although the variation in the total length among the five pairs is not large, yet the mean volume of the chromosomes is proportion with the length to some extent.

Reichardia tingitana (L.) Roth Syn: Scorzonera tingitana L.; S. orientalis L.; Picridium tingitanum (L.) Desf.; P. orientale (L.) DC.; R. orientalis (L.) Hochreutiner. Geographical distribution: Saharo-Arabian, Irano-Turanian, Mediterranean. Kuwaiti data: Found in many localities. Flowering time: March-May. Literature data: 2n=16 (Brullo and Pavone 1978, Dalgaard 1986b, Siljak-Yakovlev 1982, Ober- preieler and Vogt 1993); n=8 (Gallego 1980, Aparicio 1989). Comment: The counts of somatic (2n=16) and gametic (n= 8) numbers are the first in Kuwaiti flora and all of them are consistent with the counts of other countries. However, no karyotype has been shown in the literature for this species. From the Fig. 1 and Table 4, it is shown that this species has 1999 Determination of Chromosome Number of Kuwaiti Flora I. 187

Table 3. Karyotype analysis of five different cells of Plantago boissieri 2n= 10

M, m= Median, P.O.C. = Position of centromere.

8 pairs of small sized chromosomes, and all pairs have the centromere in the median position except the third pair which is in the sub-median position. The variation in the total length among the pairs is very little.

Senecio glaucus L. subsp. coronopifolius (Maire) Alexander Syn: S. coronopifolius Desf.; S. desfontainei Druce. Geographical distribution: Saharo-Arabian, Irano-Turanian. Kuwaiti data: It is distributed in many localities, particularly in urban areas. Flowering time: Janu- ary-March. Literature data: 2n=20 (Alexander 1979, Kadereit 1984, Vogt and Oberpreieler 1993a); (Vogt and Oberpreieler 1993b); 2n=40 (Dalgaard 1987); n=20 (Mehra and Remanandan 1969, 1975); n=10 (Aryavand 1975). Comment: The gametic and sporophytic numbers were reported from Kuwait in 1993. The results of the present study are consistent with these data, as well as with Alexander (1979), Kadereit (1984) and Mehra and Remandan (1969, 1975). In 1998 the two counts are confirmed. The two values are the second to be reported according to the available plant chromosome indexes. However, from the gametic number it is accepted that the available species has the diploid level of ploidy. 188 G. A. Malallah and G. Brown Cytologia 64 1999 Determination of Chromosome Number of Kuwaiti Flora I. 189

Sonchus oleraceus L. Geographical distribution: Euro-Siberian, Mediterranean, Irano-Turanian. Kuwaiti data: Restricted to wasteland and garden beds in urban areas. Flowering time: February- April. Literature data: 2n=16 (5 References in IPCN 1955-67); 2n=32 (16 References in IPCN 1955-69, 8 References in IPCN 1967-71, 5 References in IPCN 1973-74, 2 References in IPCN 1975-78, 2 References in IPCN 1990-91); 2n= 18 (1 Reference in IPCN 1967-71, 4 References in IPCN 1992-93); n=9 (1 Reference in IPCN 1967-71, 1 Reference in IPCN 1975-78); n=16 (3 Refer- ences in IPCN 1967-71, 1 Reference in IPCN 1972, 2 References in IPCN 1973-74, 2 References in IPCN 1975-78, 2 References in IPCN 1990-91, 1 Reference in IPCN 1992-93); n=18 (1 Refer- ence in IPCN 1972); 2n=32 (Van Loon 1980, Kliphuis and Wieffering 1979, Peng and Hsu 1978, Pajaron Sotomajor 1982, Arohonka 1982, Gupta and Gill 1983, Nazarova 1984, 1989, Dalgaard 1986b, Hiremath and Chennaveeaiah 1985, Guerra and Dos 1986, Nishikawa 1984, Mathew and Mathew 1988); 2n = 36 (Bir and Sidhu 1980); 2n = 64 (Hiremath and Chennaveeraiah 1985); n = 16 (Jansen and Stuessy 1980, Gallego Cidoncha 1985, Gupta and Gill 1983, 1984, Mathew and Math- ew 1988, Gupta and Gill 1989); n=18 (Bir and Sidhu 1979, Sidhu 1979, Gupta and Garg 1987, Gupta et al. 1989). Comment: Due to the wide distribution of this species, there are variations in the chromosomal numbers, in both counts for somatic and gametic one. Kuwaiti material so far examined has 32 somatic and 18 gametic chromosome numbers, and therefore corresponds to the majority of the literature data cited above.

FABACEAE Astragalus hauarensis Boiss. Syn.: A. gyzensis Del. Geographical distribution: Saharo-Arabian. Kuwaiti data: Widely spread and frequent in many desert areas. Flowering time: March-April. Literature data: 2n=48 (Ledingham and Rever 1963). Comment: Only one determination of the chromosome number has been published for this species. Kuwait plants have the same number of chromosomes. However, it was difficult to obtain a reliable gametic count due to the congression of the chromosomes.

Astragalus schimperi Boiss. Geographical distribution: Saharo-Arabian. Kuwaiti data: Widely distributed and locally abundant on sandy soil. Flowering time: March-April. Literature data: 2n =16 (Maasoumi 1986, 1987). Comment: The somatic count is consistent with previous counts from other countries and shows the diploid level of the species. The gametic number n= 8 equals to the basic number.

Lotus halophilus Boiss. Syn.: L. villosus Forssk.; L. pusillus Viv.; L. pusillus Viv. var. major Boiss.; L. aucheri Boiss. Geographical distribution: Saharo-Arabian, Irano-Turanian, Mediterranean. Kuwaiti data: Widely distributed and locally abundant on sandy soils. Flowering time: February-April. Literature data: 2n =14 (Snogerup 1985, Bartolo et al. 1979). Comment: This is the first count from Kuwaiti flora and the value of 14 is consistent with other countries. The value 2n=28 was reported in some Kuwaiti plants in 1993, indicating that this is a tetraploid with the basic number of 7. 190 G. A. Malallah and G. Brown Cytologia 64

Medicago laciniata (L.) Mill. Syn.: M. rigidula L. var. laciniata L. Geographical distribution: Saharo-Arabian, Irano-Turanian. Kuwaiti data: Widely distributed and locally abundant on sandy soils. Flowering time: February- April. Literature data: 2n=16 (Abdelguerfi and Guittonneau 1979); M rigidula (L.) All. 2n=14 (Van- Loon 1980); and 2n=16 (Abdelguerifi and Guittonneau1979, Astanova 1981); n=8 (Agarwal and Gupta 1983). Comment: The mitotic number of chromosomes found in this study is consistent with the other reported values.

PLANTAGINACEAE Plantago amplexicaulis Cav ssp. bauphula (Edgew.) Syn.: P. bauphula Edgew. Geographical distribution: Saharo-Arabian. Kuwaiti data: With a very local distribution. Flowering time: February-April. Literature data: 2n=10 (3 References in IPCN 1955-67; Sharma et al. 1986); n=5 (Gonzalez and Silvestre 1980, Silvestre 1991). Comment: The sporophytic and gametic numbers were first reported for Kuwait in 1993, and confirmed in this study. These counts are consistent with those from other countries.

Plantago boissieri Hausskn. and Bornm. Syn.: P. albicans (non L.) Boiss.; P. cylindrica. Geographical distribution: Sahara-Arabian, Irano-Turanian. Kuwaiti data: Widely locally distributed on sandy areas. Flowering time: February-April. Literature data: P. albicans L. 2n=30 (Brullo et al. 1985); 2n=30+3B (Badr and El-Kholy 1987); P. albicans L. ssp. albicans 2n=10, 30, 31 (Puech 1987); n=5, 10 (Puech 1987); P. cylindrica 2n= 10 (Badr et al. 1987); P. cylindrica Forssk. 2n=10 (Badr and El-Kholy 1987); n=5 (El-Bayou- mi 1973). Comment: Despite the widespread occurrence of this species in Kuwait, this is the first time the somatic chromosome number has been determined. Kuwait plants so far investigated belong to the diploid number 2n=10. From Fig. 1 and the Table 3, it is shown that this species has five pairs of small sized chromosomes, all of them have the centromere in the centric position. The variation in the total length among the five pairs is very little.

Plantago ciliata Desf. Geographical distribution: Saharo-Arabian, Irano-Turanian. Kuwaiti data: Common on compact soils. Flowering time: February-April. Literature data: 2n=10 (Snogerup 1985); n=5 (El-Bayoumi 1973). Comment: The value obtained in this study is in agreement with the values collected from other countries. Despite the wide spread occurrence of this species in Kuwait, this is the first time the somatic chromosome number has been determined. Kuwait plants so far investigated belong to the diploid cytotype, which is 2n=10. Fig. 1 and Table 5 show that this species with five pairs of small sized chromosomes, all of them have the centromere in the centric position. The variation in the total length among the five pairs is very little.

Plantago coronopus L. Syn.: P.coronopus L. subsp. eu-coronopus Pilger.; P. coronopus L. subsp. coronopus. Geographical distribution: Saharo-Arabian, Mediterranean, Irano-Turanian. 1999 Determination of Chromosome Number of Kuwaiti Flora I. 191

Table 5. Karyotype analysis of five different cells of Plantago ciliata 2n=10

M, m= Median, P.O.C. = Position of centromere.

Kuwaiti data: Fairly commonly distributed on sandy soils. Flowering time: March-April. Literature data: 2n=10 (17 References in IPCN 1955-67, 3 References in IPCN 1967-71, 2 Refer- ences in IPCN 1973-74, 4 References in IPCN 1975-78); 2n=20 (1 Reference in IPCN 1955-69, 1 Reference in IPCN 1967-71, 1 Reference in IPCN 1975-78); 2n=11 (4 References in IPCN 1955- 67); 2n=10+0-1B (1 Reference in IPCN 1973-74); P coronopus subsp. commutata 2n=10 (1 Reference in IPCN 1972, 2 References in IPCN 1990-91; Gonzalez and Silvestre 1980, Van Den Brand et al. 1979, Brown and Jackson 1982); 2n=10+1B, 10+2B (Brullo et al. 1985); n=5 (1 Reference in IPCN 1990-91); n=10 (Gonzalez and Silvestre 1980); P coronopus L. subsp. commutata (Guss.) Pilger 2n=20 (Bartolo et al. 1980); P coronopus L. subsp. coronopus 2n=10 (Tor- nadore 1981); 2n=10, 13 (Bartolo et al. 1980); P coronopus n=5, 51I+1B (Kumar and Raghuvan- shi 1984). Comment: The counts from Kuwait are consistent with counts from other countries with 2n=10.

Plantago lanceolata L. Geographical distribution: Euro-Siberian, Mediterranean, Irano-Turranian. Kuwaiti data: Found in gardens in Kuwait city. Flowering time: March-May. Literature data: 2n=12 (Aryavand 1980, Strid and Franzen 1981, Gonzalez and Silvestre 1980, Van 192 G. A. Malallah and G. Brown Cytologia 64

Den Brand et al. 1979, Lessani and Chariat-Panahi 1979, Brown and Jackson 1982, Magulaev 1982, Castroviejo 1982, Dmitrieva and Parfernov 1985, Kartashova et al. 1974, Sharma et al. 1984, Strid and Andersson 1985, Dalgaard 1986a, D'Ovidio 1984, Badr and El-Kholy 1987, Sharma et al. 1986, Matsuo and Noguchi 1989, 46 Reference cf. Murin 1997); 2n=24 (Bartolo et al . 1980, 1 Reference cf. Murin 1997); 2n=12, 24 (Brullo et al. 1985); 2n=12 , 13 (Sharma et al. 1985); 2n=12+1 (Kliphuis and Wieffering 1979, 2 Reference cf. Murin 1997); n=6 (Gonzalez and Sil- vestre 1980, Arohonka 1982, Hill 1984, Sharma et al. 1984, 1 Reference in IPCN 1992-93); P. lanceolata 2n=12 (Badr et al. 1987; 4 References in IPCN 1990-91, 3 References in IPCN 1992- 93); n=6 (Sharma and Koul 1982). Comment: The data obtained from Kuwaiti material is consistent with the values reported from other countries. This is not surprising as P. lanceolata is a cosmopolitan weed, which has profited from the few localities in Kuwait city.

Plantago ovata Forssk. Syn.: P.decumbens Forssk.; P. trichophylla Nab. Geographical distribution: Saharo-Arabian, Irano-Turanian. Kuwaiti data: Widely distributed and locally abundant on sandy soils. Flowering time: February- April. Literature data: 2n=6 (6 References in IPCN 1955-67, 3 References in IPCN 1967-71, 1 Refer- ence in IPCN 1973-74, 1 Reference in IPCN 1992-93); n=4 (2 References in IPCN 1967-71); 2n=16 (Badr 1980); 2n=8 (Rahn 1979, Dube 1979, Singh 1986, Sharma et al. 1985a, b, 1986, 2 References in IPCN 1992-93); 2n=8, 16 (Badr et al. 1987, Badr and El-Kholy 1987); n=4 (Dube 1981). Comment: The diploid value is in agreement with counts from other countries of the world. It appears as if a tetraploid cytotype also exists (e.g. Badr 1980).

Acknowledgements

We would like to thank Professor Augustin Murin for detailed and helpful suggestions on the earlier version of this paper. We would like to express our gratitude to Mrs. Modi Al-Doseri for her help in the identification of the plant species in the field. Thanks also go to Mr . Muhammad Ma- sood for his professional technical assistance. This research was supported financially by Kuwait University, grant SO 072.

References

Abdelguerfi, A. and Guittoneau, G. G. 1979. IOPB Chromosome number reports LXIV. Taxon 28: 392-393. Agarwal, K. and Gupta P. K. 1983. Cytological studies in the genus Medicago Linn. Cytologia 48: 781-793. Alexander, J. C. M. 1979. The mediterranean species of Senecio sections Senecio and Delphinifolius. Notes Roy. Bot. Gard. Edinburgh 37: 387-428. Al-Rawi, A. 1987. Flora of Kuwait Vol. 2. Kuwait Univ. Oxford, i-xiii: 225-455 . Amin, J. 1973. Cytological studies on some Egyptian plant. United Arab Repub. J. Bot. 16: 501-506. Aparicio, A. 1989. Numeros chromosomaticos de plantas occidentales, 487-507 . Anales del Botanico de Madrid 45: 483-494. Arohonka, T. 1982. Chromosome counts of vascular plants of the island Seili in Nauvo, SW Finland. Turun Yliopiston Bi- ologian-Laitoksen Julkaisuja 3: 1-12. (In Finnish). Aryavand, A. 1975. Contribution a l'etude cytotaxonomique quelques Angiosperms de l'Iran. Botaniska Notiser, Lund 128: 299-311. ― 1980 . Chromosome number reports LXIX. Taxon 29: 704. Astanova, S. B. 1981. Chromosome numbers of Leguminosae of flora in Tajikistan. Doklady Akadamii Nauk Tadzikskoj SSR, Stalinabad 24: 61-63. (In Russian). 1999 Determination of Chromosome Number of Kuwaiti Flora I. 193

Badr, A. 1980. Chromosomal counts of six Egyptian plants. United Arab Repub , J. Bot. 23: 127-129. ― and El-Kholy , M. A. 1987. Chromosomal studies in the Egyptian flora II. Karyotype studies in the genus Plantago L. Cytologia 52: 725-731. ― Labani , R. and Elkington, T. T. 1987. Nuclear DNA variation in relation to cytological features of some species in the genus Plantago L. Cytologia 52: 733-737. Bartolo, G., Brullo, S. and Pavone, P. 1979. Numeri cromosomici per la flora Italiana: 617-631. Informatore Botanico Ital- iano, Firenze 11: 149-159.

― , ― Pavon, P. and Terrasi, M. C. 1980. Contributo alla cariologia del genere Plantago in Sicilia e Calabria meridionale . Giorn. Bot. Ital. 114: 100. Berdahl, J. D. and Barker, R. E. 1991. Characterization of autotetraploid Russian wild rye produced with nitrous oxide. Crop Sci. 31(5): 1153-1155. Beuzenberg, E. J. and Nair, J. B. 1983. Contributions to a chromosome atlas of the New Zealand flora-25 miscellaneous species. New Zealand J. Bot. 21(1): 13-20. Bir, S. S. and Sidhu, M. 1979. Cytological observations in weed flora of orchards of Paticla district, Punjab. Recent Res. in Plant Sci. (New Delhi) 7: 261-271. ― and ― 1980 . Cyto-palynological studies on weed flora of cultivable lands of Patiala district (Punjab). J. Palynology 16: 85-105. Bolkhovskikh, Z., Grif, V., Matveyeva, O. and Zakharyeva, O. 1969 . Khromosomnye Chisla Tsvetkovykh Rasteniy.- Leningard, 926 pp. Borgen, L. 1974. Chromosome numbers of Macaronesian flowering plants II . Norwegian J. Bot. 21: 195-210.

― 1975 . Chromosome numbers of vascular plants from Macaronesia. Norway J. Bot. 22: 71-76. Boulos, L. 1987. A Contribution to the flora of Kuwait. Candollea 42: 263-75 . ― and Al-Dosari , M. 1994. Checklist of the flora of Kuwait. J. Univ. Kuwait (Sci.) 21, 1994. Brochmann, C. 1992. Pollen and seed morphology of Nordic draba (Brassicaceae): Phylogenetic and ecological implica- tions. Nord. J. Bot. 21(1): 13-20. Brown, M. J. and Jackson, W. D. 1982. IOPB Chromosome number reports LXXV. Taxon 31: 367-368. Brown, W. V. and Bertke, E. M. 1974. Text Book of Cytology. Pub. Mosby, 420 pp. Brullo, S., Guglielmo, A., Pavone, P., and Terrasi, M. C. 1990. Chromosome counts of flowering plants from N. Cyrenaica. Candolea 45: 65-74. ― and Pavone , P. 1978. Numeri cromosomici per la Flora Italiana: 464-483. Inform . Bot. Ital. 10: 248-265. ― , ― and Terrasi, M. C. 1985. Considerazioni cariologiche sul genere Plantago in Sicilia. Candollea 40: 217-230. Castroviejo, S. 1982. Numeros crosomaticos de plantas occidentales: 157-163. Anales del Botanico de Madrid, Madrid. 38: 507-514. Cave, M. S. Index to Plant Chromosome Numbers 1958-1960. 1-4 and supplement, Univ. North Carolina Press, Chapel Hill. ― Index to Plant Chromosome Numbers 1961-1965.2(5-9). Univ. North Carolina Press, Chapel Hill. Dalgaard, V 1986a. Chromosome numbers in flowering plants from Madeira. Willdenowia 16: 221-240. ― 1986b . Chromosome studies in flowering plants from Macaronesia. Anales del Botanico de Madrid, Madrid 43: 83-111 . ― 1987 . Chromosome number reports XCVI. Taxon 36: 660. Daoud, H. S. 1985. Flora of Kuwait. Vol. 1: Dicotyledoneae. (Publisher. KPI. England). Darlington, C. D. and Wylie, A. P. 1955. Chromosome Atlas of flowering Plants. George Alen & Unwin, London. Dmitrieva, S. A. and Parfernov, V I. 1985. Kariologicheskaja kharakteristika nekotorykh vidov poleznykh rastenij flory Be- lorussii. Izvestija Akademmi Nauk Belorusskoj SSR, Minsk. Ser. Biol. Nauk (Baku) 6: 3-8. D'Ovidio, R. 1984. Numeri cromosomici per la flora Italiana: 1011-1021. Informatore Botanico Italiano, Firenze. 16: 251-260. Dube, K. G. 1979. Karyotype Plantago ovata Forsk. Proc. Indian Sci. Congr. Assoc. (III, C) 66: 80-81. ― 1981 . Meiotic studies in Plantago ovata Forsk. Proc. Indian Sci. Congr. Assoc. 68 (Scet. VI): 81. Dyer, A. E 1979. Investigating Chromosomes. Edward Arnold, London 117-118. El-Bayoumi, A. 1973. A study of meiosis on some wild plant species from Kuwait. Cytologia 38: 357-361 . Forissier, R. 1973. IOPB Chromosome number reports XLII. Taxon 22: 647-654. Gallego, M. J. 1980. Estudiocariologicode las espectes espanolas del genero Reichardia Roth (Compositae). Lagascalia 9: 149-158.

― and Talvera . 1983. Nota sobra las especies espanolas del genero Calendula. Lagascalia 11: 100-104. Gallego Cidoncha, M. J. 1985. Numeros 404-408, in Numeros cromosomicos para la flora espanola. 363-434. Lagascalia 13: 311-313. Ghaffari, S. M. 1987a. Chromosome number reports XCV. Taxon 36: 497-498. ― 1987b . Chromosome number reports XCVI. Taxon 36: 659. Goldblatt, P. 1981a. Index to Plant Chromosome Numbers 1975-1978. Monogr. Syst. Bot. Missouri Bot. Gard. 5. 194 G. A. Malallah and G. Brown Cytologia 64

― 1981b . Index to Plant Chromosome Numbers 1975-1978. Missouri Bot. Gard. 553 pp. ― 1984 . Index to Plant Chromosome Numbers 1979-1981. Monogr. Syst. Bot. Missouri Bot. Gard. 8.

― 1985 . Index to Plant Chromosome Numbers 1982-1983. Missouri Bot. Gard. 224 pp. ― and Johnson , D. E. 1994. Index to Plant Chromosome Numbers 1990-1991. Monogr. Syst. Bot. Missouri Bot. Gard. 51. ― and ― 1996 . Index to Plant Chromosome Numbers 1992-1993. Monogr. Syst. Bot. Missouri Bot. Gard. 58. Gonzalez, F. and Silvestre, S. 1980. Numeros chromosomicos para la flora Espanola. 121-182. Lagascalia 9: 249-284. Guerra, M. and Dos, S. 1986. Citogenetica de Angiospermas coletadas en Pernambuco, I. Revista Brasil. Genet. 9: 21-40. Gupta, R. C. and Garg, R. K. 1987. SOCGI plant chromosome number reports-IV [i.e., V]. J. Cytol. and Genet. 22: 162-163. ― and Gill , B. S. 1983. Cytology of family Compositae of the Punjab plains. Proc. Indian National Sci. Acad. B49: 359-370. ― and ― 1984 . Cytological investigations on central Indian Compositae. Cytologia 49: 427-435. ― and ― 1989 . Cytopalynology of north and central Indian Compositae. J. Cytol. and Genet. 24: 96-105. ― , ― and Grag, R. K. 1989. Chromosomal conspectus of western Himalayan Compositae. Aspects of Plant Scie. 11: 427- 437. Halwagy, R. and Macksad, A. 1972. A contribution towards a flora of the state of Kuwait and the Neutral zone. Bot. J. Lin- nean soc. London 65: 61-79. Heyn, C., Dagan, O. and Nachman, B. 1974. The annual Calendula species: taxonomy and relationships. Israel J. Bot. 23: 169-201. Hill, L. M. 1984. Chromosome number reports LXXXIV. Taxon 33: 536-539. Hiremath, B. S. and Chennaveeraiah, M. S. 1985. Cytological studies in Sonchus oleraceous Linn. Proc. Indian national sci. Acad. New Delhi. Plant sci. 95: 373-377. Humphries, C. J., Murray, B. G., Bocquet, G. and Vasudevan, K. N. 1978. Chromosome numbers of phanerogams from Mo- rocco and Algeria. Bot. Not. 131: 391-404. Jackson, R. C. 1971. The karyotype in systematics. Ann. Rev. Ecol. Syst. 2: 327-368. Janaki Ammal, E. K. and Sobti, S. N. 1962. Chromosome relationship in Calendula species. Proc. Indian National Sci. Acad. Sect. B 55: 128-130. Jansen, R. K. and Stuessy, T. E 1980. Chromosome counts of compositae from Latin America. Amer. J. Bot. 76: 585-594. Kadereit, J. W. 1984. Chromosome number reports LXXXII. Taxon 33: 126-134. Kartashova, N. N., Malakhova, L. A. and Koslova, A. A. 1974. Study of the chromosome of the representatives of the Ob re- gion flora. I. Number of the chromosomes of the Tomsk district. Nauchnyye Doklady Vyshey Shkoly, Biologich- eskikh Nauk 4: 114-119. (In Russian). Kliphuis, E. and Wieffering, J. H. 1979. IOPB Chromosome number reports LXIV. Taxon 28: 398-400. Kumar, G. and Raghuvanshi, S. S. 1984. The influence of B chromosomes on chiasma frequency in Plantago coronopus. J. Cytol. and Genet. 19: 106-112. Ledingham, G. E and Rever, B. M. 1963. Chromosome numbers of some southwest Asian species of Astragalus and Oxytropis (Leguminosae). Canad. J. Genet. and Cytol. 51: 18-32. Lessani, H. and Chariat-Panahi, S. 1979. IOPB Chromosome number reports LXV. Taxon 28: 635-636. Levan, A., Fredga, K. and Sandberg, A. A. 1964. Nomenclature for centromeric position on chromosomes. Institute of Ge- netics, Lund, Sweden, and Roswell park memorial institute, Buffalo, N.Y., U.S.A. Love, A. and Kjelquist. 1974. Cytotaxonomy of Spanish plants. IV Dicotyledons: Caeselpiniaceae-Ateraceae. Lagasclia 4: 153-211. ― and Love , D. 1974. Cytotaxonomical Atlas of the Slovenian Flora. Lehre, 1241 pp. ― and ― 1975 . IOPB Chromosome number reportsXLVIII. Taxon 24: 367-372. Maassoumi, A. A. 1986. Astragalus L., vol. 1, Annuals. Techn. Publ. Res. Inst. Forests Rangelands 47, Tehran.

― 1987 . Notes on the genus Astragalus in Iran I, cytotaxonomic studies on some species. Iranian J. Bot. 3: 117-128.

Magulaev, A. Y. 1982. The number of chromosome of the species of Asteraceae, Caryophyllaceae and Plantaginaceae of the North Caucasus. Scientific Reports of Higher School for Biological Sciences 11(227): 74-79. (In Russian).

Majovsky, J. and Murin, A. 1987. Karyotaxonomicky Prehl'ad Flory Slovenska.•\Veda, Bratislava, 436 pp. Malallah, G. A., Afzal, M., Attia, T. A. and Abraham, D. 1996. Tapetal cell nuclear characteristics of some Kuwaiti plants.

Cytologia 61: 259-267. Mathew, A. and Mathew, P. M. 1988. Cytological studies on the south Indian Compositae. Glimses Pl. Res. 8: 1-177.

Matsuo, K. and Noguchi, J. 1989. Karyotype analysis of several Plantago species in Japan, with reference to the taxonomic status of Plantago japonica. J. Phytogeography and Taxonomy 37: 27-35. Mehra, P. N. and Remanandan, P. 1969. IOPB Chromosome number reports XXII. Taxon 18: 433-442.

― and ― 1975 . Cytological investigations on Indian Compoitae. IV Tribes Senecioneae, Eupatrieae, Vernonieae and In- uleae. The Nucleus 18: 6-19. Meikle, R. D. 1977. Flora of Cyperus. Bentham-Moxon Trust, Roy. Bot. Gard. Kew. 1999 Determination of Chromosome Number of Kuwaiti Flora I. 195

Meusel, H. and Ohle, H. 1966. Zur Taxonomie und Cytologia der Gattung Calendula. Osterreich. Bot. Zeitschr. 113: 191- 210. Moore, R. J. 1970. Index to Plant Chromosome Numbers for 1968. Regnum Vegitabile 68.

― 1971 . Index to Plant Chromosome Numbers for 1969. Regnum Vegitabile 77.

― 1973a . Index to Plant Chromosome Numbers 1967-1971. Regnum Vegitabile 90. ― 1973b . Index to Plant Chromosome Numbers 1967-1971. Utrecht, Netherlands, 539 pp.

― 1974 . Index to Plant Chromosome Numbers 1972. Regnum Vegitabile 91.

― 1977 . Index to Plant Chromosome Numbers for 1973-1974. Utrecht, Netherlands, 257 pp. Murin, A. 1997. Karyotaxonomy of some medicinal and aromatic plants. Thaiszia T. Bot., Kosice 7: 75-88. Nazarova, E. A. 1981. Cytosystematic investigation of the genus Koelpinia (Asteraceae). Botanicheskiy Zhurnal. SSSR. 66(12): 1755-1758. (In Russian). ― 1984 . Chromosome numbers in the Caucasian representatives of the families Asteraceae, Brassicaceae, Fabaceae, Limo- niaceae. Boticheskiy Zhurnal SSSR. 69(7): 972-975. (In Russian).

― 1989 . Karological and palynological study of representatives of the genus Sonchus from the Caucasus. Botanicheskiy Zhurnal SSSR. 74: 886-969. Negodi, G. 1935. Reporti cariolgici su Fanerogame, Atti della Societa Naturali e Matematiche. Modena 67: 7-9.

― 1937 . Caratteri nucleari a morfologici in alcune Calendulae (Compositae-Tubuliflorae), Annali di Botanica (Roma) 21: 510-518. Nisikawa, T. 1984. Chromosome counts of flowering plants of Hokkaido (7). J. Hokkaido Univ. Education Section 2B 35: 31-42. Oberprierler, C. and Vogt, R. 1993. Chromosome numbers of north African phanerogams. II. Willdenowia 23: 211-238. Ornduff, R. 1967. Index to Plant Chromosome Numbers for 1965. Regnum Vegitabile 50.

― 1968 . Index to Plant Chromosome Numbers for 1966. Regnum Vegitabile 55. ― 1969 . Index to Plant Chromosome Numbers for 1967. Regnum Vegitabile 59. Pajaron Sotomayor, S. 1982. Numeroscrosmaticos de plantas occidentales, 169-175. Anales Jard. Bot. Madrid 38: 519-521. Peng, C. I. and Hsu, C. C. 1978. Chromosome number in Taiwan Compositae. Bot. Bull. Acad. Sinica, Taipei 19: 53-66. Podlech, D. and Badr, O.1974. Chromosomen Studien an Aphganischen Pflanzen II. Mitteilungen der Botanische. Puech, S. 1987. Fertilite, aptitudes germinatives et caryotype dans deux populations de Plantago albicans L. (Plantagi- naceae) en Tunisie. Bulletin de la Societe de France. Paris. Letters of Botany 134: 145-154. Queiros, M. 1973. Contrbuicao para o conhecimento citotaxonomico das spermatophyta de portugal. II Compositae, Suple- ment I. Boletim da Sociedade Broteriana, Coimbra 47 (Ser. 2): 299-314. Rahn, K. 1979. Plantago ser. Ovatae. A taxonomic revision. Botanisk Tidsskrift, Kobenhaven 74: 13-20. Rao, S. R. and Chandel, K. P. S. 1991. Karyomorphological studies in the cultivated and wild Vigna species in Indian Gene Centre. Cytologia 56: 47-57. Rashid, A. 1974. Chromosome numbers of some Mediterranean plants. Bangladesh J. Bot. 3: 75-82. Reese, G. 1957. Uber die Polyploidienspektren in der nordsanarischen Vustenpflanzen. Flora 144: 598-634. Sato, D. 1962. Law of karyotype evolution, with special reference to prokaryotype. Sci. papers coll. Gen. Educ.Univ. Tokyo 12: 173-210. Scrugli, A. 1974. Numeri cromosomici per la flora Italiana: 167-171, Informatore Botanico Italiano, Firenze 6: 37-43. Sharma, P. K. and Koul, A. K. 1982. Genetic diversity among Plantago, I. Desynapsis. Nucleus 25: 10-13. ― , ― and Langer, A. 1984. Genetic diversity among Plantago II. Karyotype of Plantago lanceolata L. with special empha- sis on nucleolar chromosomes. Cytologia 49: 351-357. ― , ― and ― 1985a. Genetic diversity among plantagos IV. Nucleolar organization in autotetraploid Plantago ovata Forsk. And allotetraploid P. major L. Beitrage Zur Biologie der Pflanzen, Berlin. 60: 243-253. ― , Langer, A. and Koul, A. K. 1985b. Nature and possible origin of trisomy in Plantago lanceolata L. Nucleus 28: 42-45. ― , ― and ― 1986. Studies on nucleolus and nucleolar chromosomes in angiosperms XI. Plantago L. Annals of Botany, London 57: 145-154. Sidhu, M. K. 1979. Distributional and cytological studies of the weed flora of cultivable fields of Patiala district (Punjab). Ph.D. Thesis, Patiala. 1-230.1979. Siljak-Yakovlev,S. 1982. IOPB Chromosome number reports LXXVII. 31: 768. Silvestre, S. 1991. Numeros cromosomaticos de plantas occidentales 639-648. Anales jard. Bot. Madrid 49: 95. Singh, R. K. 1986. Cytomixis in Plantago ovata. Curr. Sci. 55: 658-659. Snogerup, B. 1985. Chromosome Number Reports LXXXIX. Taxon 34: 727-730. Stebbins, G. L., Jenkins, J. A. and Walters, M. S. 1953. Chromosomes and phylogeny in the Compositeae, tribe Cichoriae. Univ. Calif Publ. in Bot. Los Angales 26: 401-430. Stepankova, I. 1993. Ploidy variation in the group of Myosotis palustris and M. laxa in the Czech Republic and Slovakia. Folia-Geobot. Phytotaxon 28(2): 113-128. 196 G. A. Malallah and G. Brown Cytologia 64

Strid, A. and Andersson, I. A. 1985. Chromosome numbers of Greek mountain plants. An annotated list of 115 species. Botanische Jahrbucher fur Systematik, Pflanzengeschichte und Pflanzengeographie, Leipzig, Stuttgart. 107: 203-228.

― and Franzen , R. 1981. Chromosome number reports LXXIII. Taxon 30: 829-842. Strother, J. L. 1972. Chromosome studies in Western North American Compositae. Amer. J. Bot. 59: 242-247. Tornadore, N. 1981. Numeri cromosomici per la flora Italiana: 806-813. Informatore Botanico Italiano, Firenze 13: 151-157. Van Den Brand, C., Van Meel, F. C. M. and Wieffering, J. H. 1979. IOPB Chromosome number reports LXIV. Taxon 28: 395-397. Van Loon, J. 1974. A cytological investigation of flowering plants from Canary islands. Acta Botanica Neederlandica, Am- sterdam 23: 113-124. Van Loon, J. C. 1980. Chromosome number reports LXIX. Taxon 29: 718-720. Vogt, R. and Oberprieler, C. 1993a. Chromosome numbers of north African phanerogams. I. Fl. Medit. 3: 187-210. ― and ― 1993b . Mediterranean chromosome number reports 3 (171-176). Fl. Medit. 3: 338-340. Waisel, Y. 1962. Ecotypic differentiation in the flora of Israel II. Section D. Botany 11 3: 174-176. Webb, C. J. and Beuzenberg, E. J. 1987. Contributions to a chromosome atlas of the New Zealand flora-corrections and ad- ditions to number 21 Umbeliferae (Hydrocotyle). New Zealand J. Bot. 25(3): 371-372.