Determination of Optimum Pollen Developmental Stage for Inducing Callus in Anther Culture of Rice

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Other Title
  • イネのやく培養におけるカルス形成に最適の花粉発育時期の決定
  • イネの葯培養におけるカルス形成に最適の花粉発育時期の決定
  • イネ ノ ヤクバイヨウ ニ オケル カルス ケイセイ ニ サイテキ ノ カフン ハツイク ジキ ノ ケッテイ

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Description

Pollen developmental stage is critical for successful induction of callus in anther culture. In rice, the uninucleate pollen stage has been found to be suitable for callus induction. However, it remained to be determined which period of the uninucleate stage is suitable for callus induction. In the present study, we demonstrated that the use of anthers in a certain period of the uninucleate stage was the best method for callus induction. Two rice cultivars differing in their ability of plant regeneration from anther were used, Kitaake with a high ability and Kirara 397 with a low ability. One of six anthers in a spikelet was used to identify the pollen developmental stage and the remaining five were cultured. The uninucleate pollen stage was divided into three substages and more than 500 anthers were cultured at each of the three substages. Anthers were cultured in liquid medium to detect clearly differences in the callus formation rate among the pollen stages, because callus is induced with a higher frequency in liquid medium than in agar medium. Callus was induced with the highest frequency in the anthers containing mid-uninucleate pollen grains in both cultivars. It could not be confirmed whether the difference in the pollen stages affected the rates of plant regeneration from pollen callus and the rates of spontaneous doubled-haploids in regenerated green plants. Consequently, it was considered that the pollen developmental stage affected strongly the callus induction rate, and the mid-uninucleate pollen substage was found to be optimum for inducing callus. The miduninucleate substage can be identified under the microscope by the presence of a double wall, exine and intine, and by the location of the nucleus in the center of pollen cells filled with dense cytoplasm.

Journal

  • Breeding Research

    Breeding Research 2 (2), 73-79, 2000

    Japanese Society of Breeding

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