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Jitlam Natenapit,Tetsuto Nagamidori,Seiichi Fukai 한국원예학회 2009 Horticulture, Environment, and Biotechnology Vol.50 No.3
Restoration of pollen germination ability using high temperatures was investigated in Oriental-Trumpet (OT) and Longiflorum-Asiatic (LA) triploid lily hybrids. The flower bud size relation to the pollen development stage was determined through microscopic observation of flower buds from plants grown in a growth chamber at 20℃. Pollen was collected from plants treated with 30℃ for 7 days and lowered to 20℃ until flowering. A sequence of events in meiosis was found in 20-32 ㎜ flower buds of ‘Vinadelmar’, 31-39 ㎜ of ‘Ventimiglia’, 32-38 ㎜ of ‘Manissa’, 32-40 ㎜ of ‘Conca d’Or’, and 14-22 ㎜ of ‘Ballroom’. Individual flowers showed considerable differences in developmental stages. Increased germination was shown as 8.4±1.2% in flower buds of 17-35 ㎜ in ‘Ventimiglia’; 2.7±1.0%, 21-35 ㎜ in ‘Manissa’; 4.6±1.3%, 21-35 ㎜ in ‘Conca d’Or’, respectively. Pollen was used in crossings with L. × formolongi, trumpet hybrids, Oriental hybrids, and diploid OT hybrids; a few plants were obtained but they died at the early stage of seedling development. These results demonstrate that high-temperature treatment is a promising method to restore pollen fertility. The flower bud is expected to have anthers in meiosis during high-temperature treatment; the appropriate flower bud size differed slightly depending on the genotype.
A new interspecififi c hybrid of Torenia obtained through ovule culture
Tanapoom Laojunta,Takako Narumi-Kawasaki,Takejiro Takamura,Seiichi Fukai 한국원예학회 2019 Horticulture, Environment, and Biotechnology Vol.60 No.3
Torenia is one of the most popular summer bedding plants in Japan. Commercially available hybrids are assumed to be interspecific hybrids derived from T. fournieri , T. concolor , and T. baillonii . Use of new Torenia species is required to producenovel fl ower types of interspecifi c hybrids. Three species from Thailand, T. bicolor , T. hirsutissima , and T. siamensis , wereselected and crossed with T. fournieri , T. concolor , and T. baillonii . The chromosome number of six Torenia species variedfrom 2n = 16 to 2n = 34. Mature seeds were obtained from only 3 of 30 reciprocal cross combinations: T. fournieri × T. bicolor ,T. fournieri × T. baillonii , and T. bicolor × T. baillonii . Ovule culture was applied to overcome the interspecifi c crossing barrierof Torenia , ovule culture was applied. Swollen ovules (250) were taken from pods at 10 days after crossing and cultured onMS medium. Progenies were obtained in 14 cross combinations, through ovule culture. This is the fi rst interspecifi c Toreniahybrid obtained using T. bicolor , T. hirsutissima , and T. siamensis. The hybridity of all the obtained plants was confi rmedusing PCR–RFLP analysis of rDNA. The obtained hybrids showed intermediate morphological characteristics and pigmentationof fl oral organs between parents. This study shows that ovule culture is a powerful tool for overcoming the crossingbarrier in interspecifi c crosses in Torenia.
Silva, Jaime A. Teixeira da,Fukai, Seiichi 한국식물학회 2003 Journal of Plant Biology Vol.46 No.2
Successful genetic transformation of plants requires non-chimeric selection of transformed tissues and their subsequent regeneration. With rare exceptions, most transformation protocols still rely heavily on antibiotics for selecting transgenic cells that contain an antibiotic-degrading selectable marker gene. Here, the morphogenic capacity of in-vitro explants of chrysanthemum and tobacco stems and leaves (control and transgenic) changed with the addition of aminoglycoside antibiotics (AAs). In a test of 6 Ah, phytotoxicity occurred at concentrations of 10 to 25 and 50 to 100 ㎍mL^(-1) in chrysanthemum and tobacco explants, respectively. light conditions as well as explant source and size also had significant effects, The use of transverse thin cell layers (tTCLs), in conjunction with high initial AA selection levels, supported the greatest regeneration of transgenic material (adventitious shoots or callus) and the lowest number of escapes. Flow-cytometricanalyses revealed no endoduplication in chrysanthemum, even at high AA levels. However, this phenomenon was observed in tobacco calli (8C or more), even at low AA concentrations (i.e., 5 to 10 ㎍mL^(-1).
Teixeira da Silva, Jaime A.,Fukai, Seiichi The Korean Society of Plant Biotechnology 2004 Plant molecular biology and biotechnology research Vol.6 No.1
Successful genetic transformation of plants requires non-chimeric selection of transformed tissues and their subsequent regeneration. With rare exceptions, most transformation protocols still rely heavily on antibiotics for selecting transgenic cells that contain an antibiotic-degrading selectable marker gene. Here, the morphogenic capacity of in-vitro explants of chrysanthemnum and tobacco stems and leaves (control and transgenic) changed with the addition of aminoglycoside antibiotics (AAs), In a test of 6 AAs, phytotoxicity occurred at concentrations of 10 to 25 and 50 to 100$\mu\textrm{g}$ $mL^{-1}$ in chrysanthemum and tobacco explants, respectively. Light conditions as well as explant source and size also had significant effects. The use of transverse thin cell layers (tTCLs), in conjunction with high initial AA selection levels, supported the greatest regeneration of transgenic material (adventitious shoots or callus) and the lowest number of escapes. Flow-cytometric analyses revealed no endodu-plication in chrysanthemum, even at high AA levels. However, this phenomenon was observed in tobacco calli(8C or more), even at low AA concentrations (i.e., 5 to 10 $\mu\textrm{g}$ mL$^{-1}$ ).
Crossing of Allotriploid LLO Hybrid and Asiatic Lilies (Lilium)
Jitlam Natenapit,Shin Taketa,Takako Narumi,Seiichi Fukai 한국원예학회 2010 Horticulture, Environment, and Biotechnology Vol.51 No.5
Triploid LLO hybrids were crossed with Asiatic hybrids. The triploid was derived by crossing somatic chromosome doubled LO hybrid (Longiflorum × Oriental) backcrossed with L. Genomic in situ hybridization showed that the LLO hybrid had 24 L chromosomes and 12 O chromosomes with no recombination. Crossing of the allotriploid LLO as the ovary parent with diploid Asiatic hybrid (A) produced some progenies, which were aneuploid, near-diploid, or -triploid containing L, O, and A chromosomes. Numbers of L and A chromosomes were stable (12 chromosomes) with one exception, which had 14A chromosomes. The number of O chromosomes varied from 3 to 10. One progeny (LLO × A) exhibited a chromosome recombination of L and O, indicating that recombination can occur in meiosis of an allotriploid (LLO) ovule.