1.

Breeding and Propagation Techniques and Plant Breeders’ Rights of Lilies in the Netherlands

Tso-Chi Yang ¹⁾, Junne-Jih Chen ¹⁾, Tzay-Fa Sheen ¹⁾, J. M. Van Tuyl ²⁾ and C. J.  Barendrecht ²⁾

¹⁾ Taiwan Seed Improvement and Propagation Station, COA, Taichung, Taiwan.

²⁾ Plant Research International, Wageningen, The Netherlands.

In the Netherlands, the most famous land of flowers, lily is the fourth important segment of cut flower industry, exceeded only by roses, chrysanthemums, and tulips. During the last 35 years, the acreage of lily bulb production in the Netherlands has increased from less than 100 ha in 1965 to 4503 ha in 1999. Total more than 1,200 million lily bulbs were produced, which were used worldwide for cut-flower production. The production acreage of cut-flower lily grown in the glasshouse has increased slightly annually to 249 ha in 1999, about 80% of them were exported. Plant research International (formerly named CPRO-DLO) maintains lily germplasm collections containing more than 1000 lily genotypes to support breeding and research. Traditional breeding is considered to be the only source of new cultivars and varieties for the ornamental markets. For lily breeding the cut style method was applied successfully to overcome pre-fertilization barriers. Cross-pollination barriers have been also overcome by polyploidization techniques restoring fertility of sterile interspecific diploid hybrids. Post-fertilization barriers are mainly caused by degeneration of the endosperm leading to the abortion of the embryo, and these barriers in lilies can be overcome by embryo-rescue methods. New varieties of lily can be assessed by flow cytometry, disease resistance testing and in situ hybridization to ascertain the major characters. For molecular breeding, transformation by Agrobacterium should make appropriately the combination of Agrobacterium strain and regeneration protocol for the generation of transgenic lilies. Confirmation of transgenic lily plants after bombardment was determined by PCR amplification and Southern blot analysis. For each newly developed cultivar, thirty lily bulbs in flowering sizes are requested by the Dutch Board for Plant Breeders’ Rights for DUS (Distinctness, Uniformity and Stability) testing. Once breeder’s right was granted, the new variety is allowed to be propagated and the following health certification is also conducted according to the scheme of EPPO standards. Finally the bulbs in flowering size were harvested, processed and stored for marketing all over the world for cut flower production. In 2000 there were 508 cultivars, which were granted Plant Breeders’ Rights certificates by the Dutch Board for PBR during 1988 to 2000, have been commercially used.

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2.

Effects of water and salt stress on seed germination of Bidens pilosa L. var. radiata Sch. from different sources

Chin Jin Hou^1)*and Shu Min Wang²⁾

¹⁾ Instructor, Department of Agronomy, National Chiayi University, Chiayi, Taiwan, Republic of China.

²⁾ Former Student, Department of Agronomy, National Chiayi University, Chiayi, Taiwan, Republic of China.   

* Corresponding author.

Experiments were conducted using seeds of Bidens pilosa L. var. radiata Sch. which were collected from different sources (seaside, plain, middle altitude and high altitude). Seeds were incubated at alternating temperature of 25/20℃ (12/12h, day/night) for investigating germination after seeds treated with different water potentials (0 Mpa, -0.2 MPa, -0.4 MPa, -0.8 MPa , -1.0 MPa, -1.2 MPa) and different levels of salt stress (0 %, 0.05 %, 0.1 %, 0.2 %, 0.4 %, 0.8 %, 1.2 %, 2.0 %) of NaCl.

Water potentials at -0.8 MPa was critical for seed germination of B. pilosa var. radiata, and seeds failed to germinate at -1.0 Mpa or higher. Under stress condition of -0.4 Mpa and -0.8 Mpa, seeds from high altitude showed higher level of germination than the other sources. High altitude seeds also showed higher germination percentage, speeds of germination and shorter mean days of germination.

Seeds were tolerant to salt stress up to 1.2 % of NaCl, and were unable to germinate exceed 2.0 %. High altitude seeds showed higher germination percentage, speeds of germination and shorter mean days of germination than middle altitude, plain and seaside seeds under level of NaCl exceed 0.2 %.

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3.

Tetrazolium test for the seeds of Apium graveolens L. var. secalinum

Jen-Chuan Chang¹⁾、Yu Sung²⁾、Mei-Shiuan Lin³⁾

¹⁾ Graduate Student,²⁾ and Associate professor of the Department of Horticulture, National Chung Hsing University, Taichung 402, Taiwain.

³⁾ Leader, Seed Testing Laboratory, Council of Agriculture.

Procedures and evaluation of the tetrazolium test on the seeds of Apium graveolens L. var. secalinum were investigated. Seeds were soaked in distilled water for 24 hours at 25 ℃. After being cut along ridge, seeds were soaked in 1 %TTC solution and incubated at 30 ℃ for 18, 24 and 30 hours. Six staining patterns were recognized. The viable categories of each pattern were determined by the root mean square method. Four patterns represented viable seeds and two patterns as non-viable seeds.

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4.

Development of New Spray Chrysanthemum (Dendranthema × grandiflorum (Ramat) Kitamura) by Callus Culture Irradiated γ-ray

Chien-Young Chu¹⁾, Ming-Chang Huang²⁾, Chun-Ya Chiang³⁾

¹⁾ The professor and corresponding author, ²⁾ professor and ³⁾ research assistant, Department of Horticulture, National Chung Hsing University, Taichung, Taiwan, R.O.C.

 The calli of 7 spray mums, such as ‘Red Beauty’ etc., induced from leaf explants were cultured on a regeneration medium, then were irradiated once or twice by γ-ray at the dose of 1~8Gy on the 7 ^th or 14^th date of subculture. After 5~9 weeks of subculture, the adventitious shoots of 2 cm in length were cut and propagated ex-vitro. When the cuttings flowered, the plants were invested and selected. There was no tendency about the dose effect of γ-ray on the regeneration of each cultivar. In the suitable range of irradiation dose, when the total dose was the same, callus irradiated by twice was better than irradiated by once for adventitious shoot regeneration, except of ‘Pink A-Lai’. In this experiment, there was no chimera mutant. Most of the mutants regenerated from ‘Red Beauty’, ‘Lucky Pink’ and ‘Yellow Papaya’ were in the second. In the former two cultivars, the mutation included flower color and inflorescence variation, while ‘Yellow Papaya’ only had the variation in size and number of ray florets. In ‘Pink Flame’, ‘Red Windmill’, ‘Pink A-Lai’ and ‘Ching Hsing Huang’, no mutant was found.

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5.

Somatic Embryogenesis of Poinsettia (Euphorbia pulcherrima Wild. ex Klotzsch)

Chien-Young Chu and Yu-Chen Chao ¹⁾

¹⁾ Professor (corresponding author) and graduate student, Department of Horticulture, National Chung Hsing University, Taichung, Taiwan, R.O.C.

The leaf, petiole and stem explants of four cultivars of poinsettia (Euphorbia pulcherrima) ‘Nobel Star’, ‘Picacho’, ‘Red Splendor’ and ‘Pepride’ were used. Calli were induced from leaf, petiole and stem explants. The most of calli induced from explants cultured on a full strength MS medium containing 0.4 or 0.8 mg/l 2,4-D and 0.2 or 0.4 mg/l BA. Among explants, stem had the strongest response for callus formation. However, ‘Pepride’ had the least response. Petiole-derived calli cultured on medium with higher BA concentration proliferated more calli and somatic embryos along with better shoot development, especially on ‘Picacho’. Stem-derived calli of ‘Nobel Star’ cultured on medium with 0.2 mg/l BA or 0.1 mg/l kinetin combined with 0.2 mg/l NAA forming more somatic embryos developing to plantlets.

Key words：poinsettia, leaf, petiole, callus, somatic embryo

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