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3 Journal of Applied Genetics

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Plant regeneration from immature inflorescence-derived callus of Italian ryegrass (Lolium multiflorum Lam.): chromosome number and fertility of regenerated plants

100%

Zwierzykowski Z. , Rybczynski J. J.

tom 39

nr 3

217-224

Twenty field-grown genotypes of diploid Italian ryegrass (Lolium multiflorum Lam., 2n = 2x = 14) were tested for their ability to induce callus and regenerate plants. Callus cultures were initiated from segments of immature inflorescences cultured on the MS medium supplemented with 4.0 mg L-1 2,4-D. The calluses were subcultured first on the maintaining medium (MS medium with 2.0 mg L-1 2,4-D) and later on the rooting medium (MS medium with 0.2 mg L-1 2,4-D). The frequency of callus induction varied depending on the source of explant and the initial genotype. A total of 473 green plantlets were regenerated, of which 420 were established in the soil. All these plants had the morphological characteristics of Italian ryegrass. Among 372 regenerants analysed cytologically, 302 (81.2%) had the expected diploid chromosome number (2n = 2x = 14), 65 (17.5%) were tetraploid (2n = 4x = 28); several aneuploids and mixoploids were also observed. All diploid and tetraploid regenerants were male and female fertile. However, a great variation of female fertility within and between both groups of regenerants was observed.

Chromosome pairing in triploid intergeneric hybrids of Festuca pratensis with Lolium multiflorum, revealed by GISH

88%

Kosmala A. , Zwierzykowska E. , Zwierzykowski Z.

tom 47

nr 3

215-220

Genomic in situ hybridisation (GISH) was used to reveal chromosome pairing in two partly fertile, triploid (2n = 3x = 21) hybrids obtained by crossing the diploid (2n = 2x = 14) Festuca pratensis Huds. (designated FpFp), used as a female parent, with the autotetraploid (2n = 4x = 28) Lolium multiflorum Lam. (designated LmLmLmLm), used as a male parent. The pattern of chromosome pairing calculated on the basis of the mean values of chromosome configurations identified in all 100 PMCs analysed, was: 0.71I Lm + 2.24I Fp + 2.18II Lm/Lm + 0.54II Lm/Fp + 4.18III Lm/Lm/Fp. A relatively high number of Lm/Lm bivalents and Fp univalents, and a low number of Lm/Fp bivalents and Lm univalents indicated that the pairing was preferential between L. multiflorum chromosomes. Other observations regarding chromosome pairing within the Lm/Lm/Fp trivalents also confirmed this preferential pairing in the analysed triploids, as the Fp chromosome was not randomly located in the chain- and frying-pan-shaped trivalents. The similarities and differences in chromosome pairing at metaphase I and the level of preferential pairing between Lolium chromosomes in the different triploid Lolium-Festuca hybrids are discussed.

Effects of interaction between pollen coat eluates and pistil at the molecular level in self-compatible and self-incompatible plants of Lolium multiflorum Lam.

75%

Kalinowski A. , Radlowski M. , Bocian A.

tom 47

nr 4

319-329

Two-dimensional electrophoresis (2-DE) of soluble proteins and enzymes was performed and specific activities of 5 enzymes (esterase, pectinesterase, acid phosphatase, protease and diaphorase) were determined in stigmas of Lolium multiflorum (Italian ryegrass) treated with self or foreign pollen coat eluates (pc). Also, a low-molecular-weight fraction of the treated self-compatible (SC) and self-incompatible (SI) stigmas was analyzed by high-pressure liquid chromatography (HPLC). The treatment of stigmas with foreign pollen induced the loss of 42% of the control sample proteins in SC plants but only of 5.5% in SI plants. In contrast, the treatment of stigmas with foreign pollen induced the loss of 15% proteins in SC plants and of 29% in SI plants. Specific activities of esterase, pectinesterase and diaphorase were higher in SC than in SI stigmas. The 2-DE enzyme patterns indicated qualitative relationships between the presence of some isoforms of acid phosphatase or protease and the treatment with self or foreign pc in SC and SI stigmas. No changes were observed in HPLC profiles of the low-molecular-weight fraction from SC and SI stigmas treated or not with pc. The presented results revealed different reactions of SC and SI stigmas to the treatment with self or foreign pc. Further investigations may explain if any of the observed reactions represent specific reorientations in the style, facilitating cross- or self-pollination.