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E. ANSARI, M. KHOSROWSHAHLI, A. ETMINAN, A.A. JAFARI
Primary screening of original diploid species and induced autotetraploids after induction of single node cutting of three populations of perennial diploid Medicago sativa ssp caerulea: (Karaj1, Karaj2 and Tehran) and five annual diploid alfalfa species: (M. lupulina, M. radiata, M. rijidula, M. truncatula, M. turbinata) with three concentrations (0.10, 0.50, 1 %) of colchicine counting the chloroplasts number in the stomata guard cells was done. In addition, the ploidy levels of species were determined using usual method of chromosomes counting in the metaphase cells of root tip meristems. Results of ANOVA showed significant effects of species, colchicine concentration and their interaction for survival and polyploidy induction rate (p < 0.01). The highest and lowest survival rate (80.2 and 8.9 %) and tetraploidy induction (74.3 and 20.8 %) were obtained in 0.10 and 1.0 % colchicine concentration, respectively. In comparison, between species, the highest survival and tetraploidy induction rate were observed in M. truncatula (80.2 and 100 %) followed by M. lupulina (73.6 and 73.6 %), respectively. In contrast, the lowest survival and tetraploidy induction rate were observed in M. turbinata with 34 and 27.4 %, respectively. Chloroplasts enumeration in stomata cell guards showed that in the survived species the number of chloroplasts of stomata guard cells were ranged between 8.07 to 9.15 in diploids and between 14.94 to 15.38 in tetraploids from induced diploids and five local tetraploid cultivated alfalfa cultivars, respectively. There was no significant difference between overall means of perennials and annual diploid alfalfa species for survival and tetraploidy induction. Ploidy levels have shown strong relationships with mean numbers of chloroplasts in stomata guard cells. It was concluded that enumeration of chloroplasts in stomata guard cells is an indirect and precise method for evaluating ploidy level in alfalfa.
Key words: alfalfa, ploidy, colchicine, chloroplasts, stomata guard cells
E-mail: mkhosrowchahli yahoo.com
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