ニセアカシア(Robinia pseudoacacia L.)の耐塩性カルスの選抜

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ニセアカシアの胚軸からのカルスの誘導，誘導カルスからの不定芽の分化及び不定根の分化に適した培地の組成，ホルモン濃度及び培養環境を検討し，カルス培養系を確立した．すなわち，カルス誘導にはMS（Murashige et al., 1962）培地が最も過しており，2，4－D（2，4－ジクロロフェノキシ酢酸）1μMとBAP（6－ベンジルアミノプリン）10μMを加えた培地で誘導されたカルスの不定芽分化率が高かった．カルスからの不定芽分化には，NAA（α-ナフタレン酢酸）0．1μMとBAP6μMから10μMを組み合わせた培地が適しており，不定根の分化にはIAA（3－インドール酢酸）単独で6μMから9μM加えた培地が適していた．この培養系に基づいて，中国の3産地から採取された種子を類型（無縞，有縞）別にカルスを誘導した．更に誘導されたカルスを，NaClを0．15M，0．20M及び0．25M添加した耐塩性選抜用培地で培養した．その結果，産地間，類型間で耐塩性に差があり，懐仁産無縞類型のカルスで耐塩性が強かった．カルスで得られた耐塩性は，それぞれの実生苗の耐塩性と類似しており，カルスからの耐塩性選抜は有効な手法であると考えられた．The method for the regeneration of plantlets from callus culture obtained from the hypocotyl of Robinia pseudoacacia L. was established. It was found that MS (Murashige et al. 1962) medium was most suitable for callus culture. The callus induced on MS medium supplemented with 1μM 2,4-dichlorophenoxyacetic acid (2,4-D) in combination with 10 μM benzylaminopurine (BAP) was considered highly efficient for shoot regeneration. Shoots differentiating from the callus were observed only on the MS medium containing 6 - 10 μM BAP and 0.1 μM naphthaleneacetic acid (NAA). The shoots produced roots on 1/2 strength MS medium containing 6-10 μM 3-indoleacetic acid (IAA) alone. Based on these findings, calli from dark and spotted seed types from 3 provenances were induced on the best suitable callus inducement medium (MS medium containing 1μM 2,4-D and 10μM BAP). The calli were then transplanted to the NaCl-tolerance screening media, the best suitable callus inducement medium supplemented with different concentrations of NaCl (0.15M, 0.20M and 0.25M), to investigate the difference in the NaCl-tolerance among provenances, between seed types of each provenance and among individuals of each seed type, and to determine the suitable NaCl concentration and exposure time for NaC1-tolerance screening. There were significant differences among provenances and between seed types but no significant difference among individuals. The dark seed type of Huairen provenance had the strongest NaCl-tolerance in this testing. These results were very similar to the findings on the NaCl-tolerance test on seedlings. Therefore, this method was considered useful for NaCl-tolerant callus screening. It seemed very difficult to regenerate plantlets from the NaCl-tolerant calli. The calli could not differentiate into shoots when they were transferred directly to the adventitious shoot inducement medium (MS medium containing 6-10 μM BAP and 0.1 μM NAA) and had to be transferred to the best suitable callus inducement medium for 2 weeks for refreshing after screening. However, the shoot growth was unhealthy and abnormal morphology was observed. This problem must be addressed in next experiment.
九州大学の論文

ニセアカシア(Robinia pseudoacacia L.)の耐塩性カルスの選抜

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