Articles
INDUCTION OF 2N GAMETES AND 4N EMBRYO IN LILIUM (LILIUM × FORMOLONGI HORT.) BY NITROUS OXIDE GAS TREATMENT
Article number
855_36
Pages
243 – 248
Language
English
Abstract
The effect of nitrous oxide gas (N2O) treatment on the induction of 2n male gametes and 4n embryo in Lilium × formolongi hort. was examined. Lilium × formolongi hort. is an artificial hybrid between Lilium longiflorum Thunb. and Lilium formosanum Wall.
In general the members in the genus Lilium are propagated vegetatively.
However, Lilium × formolongi hort. is exceptionally propagated by seeds and blooms 6-8 months after sowing.
Seeds of Lilium times; formolongi hort. cultivars Raizan was sown in a plastic tray with 200 holes on 5 February 2008 and seedlings bearing 2-3 leaves were transplanted in 10.5 cm (in diameter) pots on 5 May, grown under ambient conditions in a green house.
When the first flower bud of each plant were reached 15-30 mm in length, these buds were sampled consecutively and observed by the aceto-carmine squash method to determine the meiotic stage of pollen mother cells.
The buds ranging from 19-23 mm in length contained pollen grains at prophase I to tetrad.
Immediately after the staging of pollen grains, plants with the buds in the same length were treated with N2O gas for 48h at 6 atm at room temperature.
The diameter of mature pollen grains in metaphase, obtained from the plants with buds of 22 mm in length at the time of exposure to N2O gas, ranged from 76 to 169 μm, although those from untreated ones ranged from 62 to 95 μm.
For the induction of 4n embryo, plants were treated with N2O gas for 72h at 6 atm 13 days after the pollination.
Plants with pollinated ovaries grew visually normal after the treatment with N2O gas and these capsules were harvested 75-85 days after pollination.
We obtained many plantlets from these capsules.
Of 49 randomly selected seedlings, 22 were revealed to be tetraploid by flow cytometry.
We could demonstrate that N2O gas treatment was useful for the manipulation of both male gamete as well as zygotic embryo for breeding Lilium × formolongi on polyploid level.
In general the members in the genus Lilium are propagated vegetatively.
However, Lilium × formolongi hort. is exceptionally propagated by seeds and blooms 6-8 months after sowing.
Seeds of Lilium times; formolongi hort. cultivars Raizan was sown in a plastic tray with 200 holes on 5 February 2008 and seedlings bearing 2-3 leaves were transplanted in 10.5 cm (in diameter) pots on 5 May, grown under ambient conditions in a green house.
When the first flower bud of each plant were reached 15-30 mm in length, these buds were sampled consecutively and observed by the aceto-carmine squash method to determine the meiotic stage of pollen mother cells.
The buds ranging from 19-23 mm in length contained pollen grains at prophase I to tetrad.
Immediately after the staging of pollen grains, plants with the buds in the same length were treated with N2O gas for 48h at 6 atm at room temperature.
The diameter of mature pollen grains in metaphase, obtained from the plants with buds of 22 mm in length at the time of exposure to N2O gas, ranged from 76 to 169 μm, although those from untreated ones ranged from 62 to 95 μm.
For the induction of 4n embryo, plants were treated with N2O gas for 72h at 6 atm 13 days after the pollination.
Plants with pollinated ovaries grew visually normal after the treatment with N2O gas and these capsules were harvested 75-85 days after pollination.
We obtained many plantlets from these capsules.
Of 49 randomly selected seedlings, 22 were revealed to be tetraploid by flow cytometry.
We could demonstrate that N2O gas treatment was useful for the manipulation of both male gamete as well as zygotic embryo for breeding Lilium × formolongi on polyploid level.
Publication
Authors
T. Sato, K. Miyoshi, K. Okazaki
Keywords
embryo culture, giant pollen, lily, polyploidy
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