Effects of the Microspore Development Stage and Cold Pre-treatment of Flower Buds on Embryo Induction in Apple (Malus × domestica Borkh.) Anther Culture

Chunfen Zhang; Tatsuro Tsukuni; Miyuku Ikeda; Mamoru Sato; Hatsuhiko Okada; Yoshitaka Ohashi; Hideyuki Matsuno; Toshiya Yamamoto; Masato Wada; Nobuyuki Yoshikawa; Shogo Matsumoto; Jijun Li; Naozumi Mimida; Manabu Watanabe; Akira Suzuki; Sadao Komori

doi:10.2503/jjshs1.82.114

ORIGINAL ARTICLES

Effects of the Microspore Development Stage and Cold Pre-treatment of Flower Buds on Embryo Induction in Apple (Malus × domestica Borkh.) Anther Culture

Chunfen Zhang, Tatsuro Tsukuni, Miyuku Ikeda, Mamoru Sato, Hatsuhiko Okada, Yoshitaka Ohashi, Hideyuki Matsuno, Toshiya Yamamoto, Masato Wada, Nobuyuki Yoshikawa, Shogo Matsumoto, Jijun Li, Naozumi Mimida, Manabu Watanabe, Akira Suzuki, Sadao Komori

Author information

Chunfen Zhang
The United Graduate School of Agricultural Science, Iwate University
Tatsuro Tsukuni
Faculty of Agriculture, Iwate University
Miyuku Ikeda
Faculty of Agriculture, Iwate University
Mamoru Sato
Fruit Tree Research Center, Fukushima Agricultural Technology Centre
Hatsuhiko Okada
Fruit Tree Research Center, Fukushima Agricultural Technology Centre
Yoshitaka Ohashi
Fruit Tree Research Center, Fukushima Agricultural Technology Centre
Hideyuki Matsuno
Agricultural College, Fukushima Agricultural Technology Centre
Toshiya Yamamoto
NARO Institute of Fruit Tree Science
Masato Wada
Apple Research Station, NARO Institute of Fruit Tree Science
Nobuyuki Yoshikawa
Faculty of Agriculture, Iwate University
Shogo Matsumoto
The Graduate School of Bioagricultural Science, Nagoya University
Jijun Li
Faculty of Agriculture, Iwate University
Naozumi Mimida
Faculty of Agriculture, Iwate University
Manabu Watanabe
The Field Science Center, Faculty of Agriculture, Iwate University
Akira Suzuki
Faculty of Agriculture, Iwate University
Sadao Komori
Faculty of Agriculture, Iwate University

Keywords: flower bud stage, microspore stage, SSR marker

JOURNAL OPEN ACCESS

2013 Volume 82 Issue 2 Pages 114-124

DOI https://doi.org/10.2503/jjshs1.82.114

Details

Abstract

To establish better embryo induction procedures in anther culture of apple, the relationships between the microspore stage and the flower bud stage, and between embryo induction and the developmental stage of microspores during cold pre-treatment of flower buds were investigated. We classified the flower bud stages into four: “Tight cluster”, “First pink”, “Pink”, and “Full pink”. Each stage was defined by a morphological characteristic. The microspore stages were classified into seven: “tetrads”, “early uninucleate”, “mid-uninucleate”, “late uninucleate”, “early binucleate”, “late binucleate”, and “mature grain”. The morphological characteristic in each stage was clarified. The “Tight cluster” flower bud stage mainly corresponded with the microspore stage of tetrad to mid-uninucleate. The “First pink” of the flower cluster mainly corresponded with the mid-uninucleate and late uninucleate microspore. The “Pink” and “Full pink” of flowering corresponded respectively with the late binucleate and the mature grain. Cold pre-treatment for more than 25 days of flower buds was found to be an efficient means of increasing embryo formation in anther culture. The microspore of late uninucleate and early binucleate stages grew into embryos efficiently after 25 days of cold pre-treatment. To achieve that condition, flower buds at late “Tight cluster” to early “First pink” stage were collected because the microspore stage progresses during 25 days of cold pre-treatment. Most microspores were early uninucleate to early binucleate stages at the “Tight cluster” to “First pink” flower bud stages. The shoots regenerated from embryos were analyzed using SSR markers. The investigation revealed that all shoots had a haploid origin.

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