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. 2022 Sep;72(4):275-284.
doi: 10.1270/jsbbs.21100. Epub 2022 Aug 26.

Induction of 2n pollen with colchicine during microsporogenesis in Phalaenopsis

Ting Wu  1   2 , Xin Zhao  1 , Shuhua Yang  1 , Jiahui Yang  1 , Jun Zhu  1 , Yaping Kou  1 , Xiaonan Yu  2 , Hong Ge  1 , Ruidong Jia  1
Affiliations

Induction of 2n pollen with colchicine during microsporogenesis in Phalaenopsis

Ting Wu et al. Breed Sci. 2022 Sep.

Abstract

The induction of 2n pollen is an important technique for breeding polyploid plants. Here, we observed meiosis in the pollen mother cells (PMCs) of six Phalaenopsis cultivars and attempted to induce 2n pollen. The meiotic stage was related to flower bud length. During meiosis, Phalaenopsis cultivars with flower widths of approximately 20-40 mm and 50-60 mm had bud lengths of approximately 3-8 mm and 5-13 mm, respectively. The duration of meiosis ranged from 4.2 to 14 d. This was the first study to characterize meiosis of the PMCs of Phalaenopsis. The natural generation frequency of 2n pollen varied from 0.68% to 1.78%. Meiotic stage and colchicine concentration significantly affected the induction of 2n pollen. The most effective treatment for obtaining 2n pollen was 0.05% colchicine in the leptotene to zygotene stage for 3 d, which achieved a 2n pollen frequency of 10.04%.

Keywords: 2n pollen; colchicine; flower bud growth; meiosis stage.

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Figures

Fig. 1.
Fig. 1.
Morphology of Phalaenopsis cultivars. (A) Phalaenopsis equestris, (B) Phalaenopsis equestris var. coerulea, (C) Phalaenopsis Anna-Larati Soekardi, (D) Phalaenopsis Tzu Chiang Sapphire, (E) Phalaenopsis Queen Beer ‘Red Sky’, and (F) Phalaenopsis Purple Crystal. Scale bar = 10 mm.
Fig. 2.
Fig. 2.
Length of the flower buds of different Phalaenopsis cultivars at different meiotic stages of PMCs.
Fig. 3.
Fig. 3.
External morphology of the flower buds of Phalaenopsis cultivars at different developmental stages. Scale bar = 10 mm. (A) Phalaenopsis equestris, (B) Phalaenopsis equestris var. coerulea, (C) Phalaenopsis Anna-Larati Soekardi, (D) Phalaenopsis Tzu Chiang Sapphire, (E) Phalaenopsis Queen Beer ‘Red Sky’, and (F) Phalaenopsis Purple Crystal. (a) (Leptotene, Zygotene), (b) (Pachytene, Diakinesis), (c) (Metaphase I, Telophase I), (d) (Prophase II, Telophase II), (e) Tetrad, (f) Mitosis, and (g) Mature pollen.
Fig. 4.
Fig. 4.
Meiotic stages of Phalaenopsis Anna-Larati Soekardi PMCs. (A) Pollen mother cell, (B) Leptotene, (C) Zygotene, (D) Pachytene, (E) Diplotene, (F) Diakinesis, (G) Metaphase I, (H) Anaphase I, (I) Telophase I, (J) Prophase II, (K) Metaphase II, (L) Anaphase II, (M) Telophase II, (N) Tetrad, (O) Prophase of mitosis, (P) Metaphase of mitosis, (Q) Anaphase of mitosis, (R) & (S) Telophase of mitosis, and (T) Mature pollen.
Fig. 5.
Fig. 5.
Abnormal behavior and asynchrony during the meiotic process in the PMCs of Phalaenopsis cultivars. (A) Phalaenopsis equestris, (B) Phalaenopsis equestris var. coerulea, (C) Phalaenopsis Anna-Larati Soekardi, (D) Phalaenopsis Tzu Chiang Sapphire, (E) Phalaenopsis Queen Beer ‘Red Sky’, and (F) Phalaenopsis Purple Crystal.
Fig. 6.
Fig. 6.
Flower bud length of different Phalaenopsis cultivars over time.
Fig. 7.
Fig. 7.
Formation of unreduced male gametes in Phalaenopsis cultivars. (A) Phalaenopsis equestris, (B) Phalaenopsis equestris var. coerulea, (C) Phalaenopsis Anna-Larati Soekardi, (D) Phalaenopsis Tzu Chiang Sapphire, (E) Phalaenopsis Queen Beer ‘Red Sky’, and (F) Phalaenopsis Purple Crystal.
Fig. 8.
Fig. 8.
Occurrence of unreduced male gametes in Phalaenopsis equestris before and after colchicine treatment. (A) Control Phalaenopsis equestris; (B) Phalaenopsis equestris treated with colchicine.
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