• Journal of Korean Society of Forest Science
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• v.23 no.1
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• pp.9-16
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• 1974

The anthers of late uninucleate microspore or early binucleate microspore stage of Paeonia suffruticosa Andr. (economic tree) were cultured on the modified Murashige and Skoog's medium suppliment with Keinetin, 2,4-D, and NAA for inducing haploid plants. The results are as follows; 1. Callus were induced from both anther locule and anther wall, where that from anther locule was identified as haploid. 2. Among 2,000 anthers cultured, fourteen haploid callus were developed. These haploid callus were clearly identified to be originated from the microspore in anther locule. 3. Diploid callus were induced only 0.5 percent from the callus of endothelium of anther wall, septium of two neighboring anther locule, parenchyma tissues of connectives and or anther locules. 4. In the anther locule of Paeonia suffruticosa Andr. cultured in medium, swollen microspores, polynucleate microspores, multicellurar pollen grains, or callus mass was frequently observed. And the haploids were seemed to be caused by the callus originated from the reduced microspore.

• Korean Journal of Plant Tissue Culture
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• v.25 no.4
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• pp.251-256
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• 1998

The effect of several factors on efficiency of anther culture of broccoli was studied. The medium supplemented with 14% sucrose, 125mg/L of silver nitrate and 0.1mg/L of each of NAA and 2,4-D was best in embryogenesis in all varieties tested. The effect of auxin was affected by silver nitrate, that is, it lowered embryogenesis when the concentration of NAA and 2,4-D was low, but it enhanced when their concentrations were high. Benzyl aminopurine increased embryogenesis in some varieties, but decreased in others. Although microspore embryogenesis was observed in anthers of which heigest was 1 to 0.6 - 1.0 of petal heigest in the bud, which corresponds to the early uninucleate to early binucleate stage of the microspores, it was much better when the ratio of heigest was 1 : 0.8. More embryos ware inducted from culture in March than in February in general. Because there was a big yearly variation, it was assumed that microspore embryogenesis was influenced greatly by the culture condition and the physiological condition of donor plants.

• Journal of Korean Society of Forest Science
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• v.43 no.1
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• pp.6-13
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• 1979

In order to substract the time and cost of propagation for inducing the haploid plants per each species. 500 anthers of late uninucleate microspore on early binucleate microspore stage of Robinia pseudoacacia (Fuel tree) Punius granatum (Ornamental tree). Aleurites fordii (Faty tree) and Styrax japonica (Silvicultural tree) were cultured on the modified Murashige and Skoog's medium supplemented with Kinetin, 2.4-D and NAA as growth regulators. And I observed the samples of cultured anthers under the microscope which were made by Microtoming method and Paraffin method. The results were summarized as follows: 1) Among 500 cultured anthers per each species, anther numbers inducing the diploid callus were as follow: Styrax japonica 20 (4% for the species total); Aleurites fordii 10 (2% for the species, total) and Punica granatum 45 (9% for the species total) were showed. 2) 2n Callus were induced from anther wall. but haploid callus were induced from anther locule. 3) Haploid callus were induced only in 25 anthers (5% for the species total) of Robinia pseudoacacia. 4) These haploid callus were not originated from body cell of anther wall tissue, but from reduced microspores, 5) Since already reported many herbaceous haploid plants were induced from the callus which were originated from reduced microspores, I conclude that the anther of woody plant which induced the haploid callus also will be cultured haploid plant.

• Journal of Plant Biotechnology
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• v.49 no.1
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• pp.74-81
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• 2022

Brassica napus, an oil crop that produces rapeseed oil, is an allotetraploid (AACC, 2n = 38) produced by natural hybridization between B. rapa and B. oleracea. In this study, microspore was cultured using the F₁ developed from a cross between 'EMS26' line with high oleic acid content and 'J8634-B-30' lines. The flower bud size showing the nuclear development at the late uninucleate and binucleate stage with high embryogenesis rate was 2.6 ~ 3.5 mm. Microspores were cultured using only this size and after then most microspore embryo developed into secondary embryos and then regeneration plants obtained from the developed multilobe. The analysis of the ploidy of the plants revealed that 66.7% and 27.8% of the total lines were tetraploids and octoploids, respectively. The sizes of stomatal cells in tetraploids, octoploids, and diploids were 25.5, 35.6, and 19.9 ㎛, respectively, indicating that ploidy level was positively correlated with cell size. Furthermore, 62 tetraploid doubled haploid (DH) lines were selected. The average oleic acid (C18:1) and linolenic acid (C18:3) concentrations of DH were 72.3% and 6.2%, respectively. Oleic acid and linolenic acid concentrations exceeded the two parental values in 5 and 14 DH lines, respectively, suggesting that these two fatty acids had transgressive segregation. Therefore, the DH population can be utilized for the biosynthesis of unsaturated fatty acids in rapeseed and related genes. It can also be used as a breeding material for varieties with high oleic acid concentrations.