高温诱导牡丹产生未减数花粉

doi:10.16420/j.issn.0513-353x.2020-0815

摘要/Abstract

摘要：

以5年生‘凤丹’牡丹为试验材料,在掌握花粉母细胞减数分裂进程的基础上,使用“树木非离体枝芽加热处理装置”（ZL200610113448.X）施加高温处理,诱导花粉染色体加倍,建立了适宜牡丹花粉染色体加倍的技术体系。结果表明,花粉母细胞减数分裂进程和花蕾的外部形态及花药颜色变化有一定相关性,当花蕾处于“小风铃”至“大风铃”时期,即花蕾直径13 mm时,花粉母细胞减数分裂达到双线期—中期Ⅰ;此时期40 ℃高温处理4 h,可获得较高比率（21.98%）具有生活力的2n花粉。

关键词: 牡丹, 三倍体育种, 减数分裂进程, 2n花粉, 高温

Abstract:

In this research,five-year Paeonia suffruticosa‘Fengdan’was chosen as an experimental material. On the basis of mastering meiosis process of pollen mother cells,“a non-in-vitro branch bud heat treatment device of trees”（ZL200610113448.X）was used to implement pollen chromosome doubling of P. suffruticosa under high-temperature processing,a suitable technique for high temperature-induced pollen chromosome doubling in P. suffruticosa was established. The findings indicated that meiosis process of pollen mother cells had correlation with the external form of flower buds and anther color changes. When flower buds remained between“small wind bell”stage to“big wind bell”stage,namely the diameter of flower buds was 13 mm,meiosis process of pollen mother cells reached diplonema—metaphaseⅠ. The treatment combination of 40 ℃ for 4 h produced the greatest amount of diploid pollen（up to 21.98% of the total yield）.

Key words: Paeonia suffruticosa, triploid breeding, meiosis process, 2n pollen, high temperature

中图分类号:

S685.11

程世平, 姚鹏强, 耿喜宁, 刘春洋, 谢丽华. 高温诱导牡丹产生未减数花粉[J]. 园艺学报, 2022, 49(3): 581-589.

CHENG Shiping, YAO Pengqiang, GENG Xining, LIU Chunyang, XIE Lihua. High Temperature Treatment Generates Unreduced Pollen in Paeonia suffruticosa[J]. Acta Horticulturae Sinica, 2022, 49(3): 581-589.

图/表 5

图1 ‘凤丹’牡丹花蕾、花药发育时期及形态变化 a：小风铃期;b：减数分裂期;c：大风铃期;d：圆桃期。

Fig. 1 Development and morphological changes of flower bud and anther of Paeonia suffruticosa‘Fengdan’ a：Small wind bell stage;b：Meiosis stage;c：Big wind bell stage;d：Round peach stage.

图2 ‘凤丹’牡丹花粉母细胞减数分裂时期 a：花粉孢母细胞;b：细线期;c：粗线期;d：终变期;e：中期Ⅰ;f：后期Ⅰ; g：末期Ⅰ;h：前期Ⅱ;i：中期Ⅱ;j：后期Ⅱ;k：末期Ⅱ;l：四分体。

Fig. 2 Meiotic stages of microspore mother cells of Paeonia suffruticosa ‘Fengdan’ a：Microspore mother cells;b：Leptotene;c：Pachytene;d：Diakinesis;e：MetaphaseⅠ;f：AnaphaseⅠ;g：TelophaseⅠ;h：ProphaseⅡ;i：MetaphaseⅡ;j：Anaphase II;k：TelophaseⅡ;l：Tetrad.

表1 不同高温处理条件下的2n花粉诱导率

Table 1 2n pollen production in different treatments exposure to high temperature

温度/℃ Temperature	时间/h Duration	花粉总数 Total number of pollen	2n花粉总数 The number of 2n pollen	花粉直径/μm Diameter of pollen		2n花粉诱导率/% 2n pollen yield
温度/℃ Temperature	时间/h Duration	花粉总数 Total number of pollen	2n花粉总数 The number of 2n pollen	单倍型花粉 Haploid pollen	2n花粉 2n pollen	2n花粉诱导率/% 2n pollen yield
38	2	421	22	40.32 ± 0.24	56.50 ± 2.64	5.19 ± 0.58
	4	408	29	40.44 ± 0.64	57.16 ± 4.46	7.06 ± 0.77
	6	412	28	38.58 ± 0.36	58.32 ± 5.72	6.77 ± 0.64
40	2	406	45	38.72 ± 0.46	58.70 ± 2.64	11.17 ± 0.87
	4	414	91	40.60 ± 0.32	59.04 ± 3.18	21.98 ± 3.65
	6	403	67	37.56 ± 0.44	58.50 ± 2.54	16.58 ± 0.98
42	2	421	43	38.46 ± 1.80	59.16 ± 2.50	10.16 ± 0.95
	4	413	51	40.36 ± 1.50	59.52 ± 4.66	12.29 ± 0.67
	6	418	48	40.66 ± 1.23	59.76 ± 5.12	11.32 ± 1.74
44	2	423	32	36.22 ± 1.12	58.32 ± 2.70	7.50 ± 1.13
	4	416	41	39.26 ± 0.56	59.16 ± 4.42	9.86 ± 0.41
	6	409	36	40.24 ± 1.10	59.70 ± 2.64	8.97 ± 2.43
46	2	421	21	40.44 ± 0.70	58.24 ± 4.42	4.92 ± 0.68
	4	432	26	38.64 ± 0.50	59.76 ± 3.12	6.04 ± 0.36
	6	412	18	41.10 ± 1.36	59.56 ± 4.10	4.35 ± 0.40
对照Control	—	409	0	39.44 ± 0.82	—	0

图3 单倍型花粉和2n花粉的观察 a：花粉镜检;b：花粉萌发;c：激光共聚焦观察。黑色箭头表示2n花粉。

Fig. 3 Observation of the haploid pollen and 2n pollen a：Microscopic examination;b：Pollen germination;c：Laser scanning confocal observation. Arrows indicate 2n pollen.

表2 单倍型花粉与2n花粉萌发率及花粉管长度

Table 2 Germination rate and pollen tube length of haploid pollen and 2n pollen

类型Type	萌发率/% Germination rate	花粉管长度/μm Pollen tube length
单倍型花粉 Haploid pollen	70.94 ± 3.74	115.64 ± 36.94
2n花粉 2n pollen	82.27 ± 5.22	133.54 ± 30.02

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