CHARACTERIZATION OF A REACTIVE OXYGEN SPECIES-RESISTANT HIGH YIELDING MUTANT R3-1 OF PISUM SATIVUM CV. ‘ALASKA’ WITH HIGH TEMPERATURERESISTANT PHENOTYPE - Pisum Sativum - page 59

M ATERIALS AND M ETHODS

Preparation of Wild-Type and R3-1 and R3-2 Mutants

Wild-type of P. sativum cv. „Alaska‟ was prepared by self pollinated line
of wild type. The induction of ROS-resistant R3-1 and R3-2 mutants was as
described in Haque et al. (2008), which were obtained after 8-µM PQ
treatment of pea seedlings. The R3-2 was obtained as a branch of R3-1 and
therefore fundamentally the same line.

Morphological Characterization of Seedlings of Wild-Type
and the R3-1 Mutant

Thirty seeds of wild-type and the R3-1 mutants were sown in vermiculite
(1.5 L) moistened with 1L of 2x10 -3 diluted Hyponex. The plastic dishes
(33x28x7 cm) were incubated at 8 ℃ , 8-h light (160 µmol m -2 s -1 ), 16-h dark
regime for 30 days. (Figure 1a). The seedlings of earlier stages were
photographed at 14 days after sowing of seeds (Figure 1b).

Figure 1a. Early growth pattern of wild-type and the mutant R3-1. Seeds of wild-type
and the R3-1 mutant were sown in vermiculite moistened by 2x10 -3 diluted Hyponex
under 8-h light (160 µmol m -2 s -1 ) and 16-h dark regime. The cultures were maintained
for 30 days.