Research on the Double Fertilization of Adonis amurensis

doi:10.11983/CBB16179

Abstract

Abstract: The pollen tube growth and duration of double fertilization in Adonis amurensis were studied by paraffin section and fluorescence microscopy observation. A. amurensis has a wet column cap. At 1-2 h after pollination, the pollen grain is recognized by the stigma; at 2-4 h after pollination, the pollen grain begins germination; at 4-6 h after pollination, the pollen tube enters into the stigma. A. amurensis is porogamous. About 10 h after pollination, the sperm is released; about 30 h after pollination the egg nucleus and sperm nucleus begin fusing; at 7 d after pollination, a sperm fuses with the egg cell to form a zygote; dormant stage of the zygote is about 8 days long and at 15 d after pollination, the first mitotic division of zygote takes place. The two polar nuclei are not fused before fertilization. At 14-16 h after pollination, the sperm nucleus fuses with a polar nucleus; at 20-22 h after pollination, the sperm nucleus fuses with another polar nucleus and forms a primary endosperm nucleus. The type of double fertilization is premitotic syngamy. This study provides an understanding for the morphological change and corresponding timing of male and female cells in A. amurensis fertilization, confirms the duration of zygote dormancy, and enriches the knowledge of A. amurensis embryological information, which has great significance for A. amurensis breeding and genetic modification.

Ying Sun, Lei Wang, Xue Yang, Axiang Wang, Miao He. Research on the Double Fertilization of Adonis amurensis[J]. Chinese Bulletin of Botany, 2017, 52(4): 480-486.

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URL: https://www.chinbullbotany.com/EN/10.11983/CBB16179

https://www.chinbullbotany.com/EN/Y2017/V52/I4/480

Figures/Tables 4

Figure 1 Pollen germinates on the stigma of Adonis amurensis^(A), (B) Pollen on stigmas (1-2 h after pollination); (C), (D) Pollen germination and pollen tube growth (2-4 h after pollination); (E), (F) Pollen tube enters into the stigma and grows toward the ovary (4-6 h after pollination). Bar=20 μm

Figure 2 Egg fertilization of Adonis amurensis^(A) The pollen falls onto the stigma; (B), (C) Pollen germinates, and the head of pollen tube expands; (D) Degenerated synergid contains 2 sperm cells; (E) Sperm moves to the egg cell; (F) The sperm nucleus membrane attaches to the egg cell membrane; (G), (H) Fusion of sperm and egg cell membrane; (I), (J) Sperm nucleus gradually moves to the egg nucleus; (K), (L) Sperm nucleus attaches to the egg nucleus membrane like convex lens; (M) Chromatin in the sperm nucleus is dispersed into the egg nucleus, sperm nucleus fuses with egg nucleus; (N)-(P) A small male nucleolus appears and enlarges in the egg nucleus; (Q), (R) Zygote; (S), (T) There are two small male nucleolus in zygote. ec: Egg cell; fne: Free nuclear endosperm; mn: Male nucleolus; pt: Pollen tube; sp: Sperm nucleus; zyg: Zygote; vn: Vegetative nucleus. Bar=10 μm

Figure 3 Polar nuclei fertilization of Adonis amurensis^(A) Two sperm cells move to the polar nucleus and the egg nucleus, respectively; (B), (C) Sperm nucleus attaches on polar nucleus; (D)-(F) Male nucleolus appears and enlarges in polar nucleus; (G) The fusion of the male nucleus and the polar nucleus; (H), (I) The nuclear membrane of 2 polar nuclei disappear and the karyoplasm fuses; (J)-(L) Two nucleoli gradually come near and fuse; (M) When the two polar nuclei merge, the egg cell is still unfertilized; (N) Primary endosperm nucleus; (O) Antipodal cells degenerate when two polar nuclei fusing; (P) When the primary endosperm nucleus forms, the egg cell is still unfertilized. ec: Egg cell; mn: Male nucleolus; pn: Polar nucleus; pen: Primary endosperm nucleus; ant: Antipodal cell; sp: Sperm nucleus. Bar=10 μm

Table 1 The duration of double fertilization in Adonis amurensis

After pollination	Egg fertilization	After pollination	Polar nuclei fertilization
1-2 h	Pollen recognition	1-2 h	Pollen recognition
2-4 h	Pollen germination	2-4 h	Pollen germination
4-6 h	Pollen tube enters into stigma	4-6 h	Pollen tube enter into stigma
10 h	Sperm enters into synergid	10 h	Sperm enters into synergid
10-20 h	Sperm moves to egg cell	10-12 h	Sperm moves to polar nucleus
20 h	The sperm nucleus attaches to the egg cell membrane	12-14 h	The sperm nuclei attache to the polar nucleus membrane
30 h	Egg nucleus fuses with sperm nucleus	14-16 h	Egg nucleus fuses with sperm nucleus
72 h	Male nucleolus appears in the egg nucleus	16-18 h	Male nucleolus appears in the polar nucleus
7 d	Zygote formed	18-20 h	Fertilized polar nucleus formed
15 d	The division of zygote	20-22 h	The endosperm nucleus formed

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