Screening of NaCl-Tolerant Robinia pseudoacacia L. Callus through in Vitro Culture

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  • Chen R.
    Department of Forestry, Faculty of Agriculture, Kyushu University
  • Gyokusen Koichiro
    Department of Forestry, Faculty of Agriculture, Kyushu University
  • Saito Akira
    Department of Forestry, Faculty of Agriculture, Kyushu University

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Other Title
  • ニセアカシア(Robinia pseudoacacia L.)の耐塩性カルスの選抜
  • ニセアカシア Robinia pseudoacacia L.ノ タイエンセイ

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Abstract

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.

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