高温诱导‘凤丹’牡丹2n雌配子创制三倍体

doi:10.16420/j.issn.0513-353x.2022-0699

摘要/Abstract

摘要：

以5年生‘凤丹’牡丹（Paeonia ostii‘Fengdan’）为试验材料，使用“树木非离体枝芽加热处理装置”（ZL200610113448.X）施加高温处理诱导产生2n雌配子。大孢子母细胞减数分裂进程与花蕾的外部形态和小孢子发育进程存在时序相关性，当花蕾发育至“大风铃”期前后，小孢子发育至单核花粉期后3 ~ 4 d，40 ℃高温处理花蕾4 h，自然授粉后代群体中可获得高比率（2.45%）三倍体植株。

关键词: 牡丹, 高温诱导, 三倍体, 2n雌配子

Abstract:

High temperature treatment with“a non-in-vitro branch bud heat treatment device of trees”（ZL200610113448.X）was used to induce 2n female gametes of five-year-old Paeonia ostia‘Fengdan’in order to obtain polyploid germplasm. The results showed that the meiosis process of megaspore mother cell was time series correlated to the outward morphology of flower bud and microspore development state. Triploid plants with the highest ratio（2.45%）could be obtained in the natural pollinated progeny population，when the flower buds were treated at 40 ℃ for four hours in the“big wind-bell”stage and three to four days after the mononuclear pollen stage of the microspores.

Key words: Paeonia ostii, high temperature inducation, triploid, 2n female gamete

刘春洋, 彭朝凤, 程世平, 姚鹏强, 耿喜宁, 谢丽华. 高温诱导‘凤丹’牡丹2n雌配子创制三倍体[J]. 园艺学报, 2023, 50(7): 1455-1466.

LIU Chunyang, PENG Chaofeng, CHENG Shiping, YAO Pengqiang, GENG Xining, XIE Lihua. Creation of Triploid Germplasm in Paeonia ostii‘Fengdan’Through 2n Female Gametes Inducing with High Temperature Treatment[J]. Acta Horticulturae Sinica, 2023, 50(7): 1455-1466.

图/表 5

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发育时期 Development stage	花蕾形态 Morphology of flower bud	小孢子发育 Development progress of microspore	大孢子发育 Development progress of megaspore	胚珠形态 Morphology of ovule	染色体加倍处理 High temperature treatment
花粉母细胞减数分裂期 Meiosis stage of the pollen mother cells	介于大、小风铃期之间 Between big wind-bell and small wind -bell	进入减数分裂期 In meiosis period	—	胚珠原基分化出珠柄，珠心开始形成 Funiculus was formed by ovule primordium, nucellus formation initially	小孢子染色体加倍 Chromosomes doubling of microspore
3 ~ 5 d后（花粉母细胞四分体至单核花粉期） Uni-nucleate microspore stage from tetrad stage after 3 ~ 5 d	接近大风铃期 Approximately in big wind-bell stage	单核花粉形成，可见四分体 Uni-nucleate pollen formation with visible tetrad	减数分裂前期 Ⅰ Prophase I of meiosis	珠被隆起，珠心顶端与两侧珠被围成120 ~ 150°角，整体呈元宝形 Integument expanding，120-150° angle was formed by the tip cells of nucellus and two sides of integument, as a ingot	大孢子染色体加倍开始 The beginning of chromosomes doubling of Macrospore
单核花粉形成3 ~ 5 d后 After 3 ~ 5 d of Uni-nucleate pollen formation	大风铃期稍后 After big wind-bell stage	单核靠边期 Late uni-nucleate microspore	减数分裂末期Ⅰ至前期Ⅱ From anaphase I to prophaseⅡ	珠被半包被，珠心顶端与两侧珠被围成150 ~ 180°角 Nucellus was half covered by integument，150-180° angle was formed by the tip cells of nucellus and two sides of integument	大孢子染色体加倍接近结束 Nearby the end of chromosomes doubling of Macrospore

发育时期 Development stage	花蕾形态 Morphology of flower bud	小孢子发育 Development progress of microspore	大孢子发育 Development progress of megaspore	胚珠形态 Morphology of ovule	染色体加倍处理 High temperature treatment
花粉母细胞减数分裂期 Meiosis stage of the pollen mother cells	介于大、小风铃期之间 Between big wind-bell and small wind -bell	进入减数分裂期 In meiosis period	—	胚珠原基分化出珠柄，珠心开始形成 Funiculus was formed by ovule primordium, nucellus formation initially	小孢子染色体加倍 Chromosomes doubling of microspore
3 ~ 5 d后（花粉母细胞四分体至单核花粉期） Uni-nucleate microspore stage from tetrad stage after 3 ~ 5 d	接近大风铃期 Approximately in big wind-bell stage	单核花粉形成，可见四分体 Uni-nucleate pollen formation with visible tetrad	减数分裂前期 Ⅰ Prophase I of meiosis	珠被隆起，珠心顶端与两侧珠被围成120 ~ 150°角，整体呈元宝形 Integument expanding，120-150° angle was formed by the tip cells of nucellus and two sides of integument, as a ingot	大孢子染色体加倍开始 The beginning of chromosomes doubling of Macrospore
单核花粉形成3 ~ 5 d后 After 3 ~ 5 d of Uni-nucleate pollen formation	大风铃期稍后 After big wind-bell stage	单核靠边期 Late uni-nucleate microspore	减数分裂末期Ⅰ至前期Ⅱ From anaphase I to prophaseⅡ	珠被半包被，珠心顶端与两侧珠被围成150 ~ 180°角 Nucellus was half covered by integument，150-180° angle was formed by the tip cells of nucellus and two sides of integument	大孢子染色体加倍接近结束 Nearby the end of chromosomes doubling of Macrospore

单核花粉产生后天数/d Days after formation of mononuclear pollen	处理温度/℃ Treatment temperature	处理时间/h Treatment duration	处理花蕾数 Flower bud number	收获种子数 Seed number	单果结实数 Seed number per follicle	成苗数 Seedling number	三倍体株数 Triploid number	三倍体得率/% Rate of triploid
1	38	2	18	395	21.94	205		0
		4	17	340	20.00	160		0
		6	19	275	14.47	141		0
	40	2	18	351	19.50	186		0
		4	20	376	18.80	192		0
		6	17	175	10.29	91		0
	42	2	16	295	18.44	160		0
		4	20	256	12.80	126		0
		6	18	156	8.67	80		0
	44	2	17	176	10.35	83		0
		4	19	93	4.89	49		0
		6	15	40	2.67	18		0
2	38	2	22	463	21.05	246		0
		4	18	365	20.28	183		0
		6	19	247	13.00	126	1	0.79
	40	2	17	361	21.24	190		0
		4	19	366	19.26	172	1	0.58
		6	20	216	10.80	115		0
	42	2	22	420	19.09	225		0
		4	22	271	12.32	133		0
		6	20	171	8.55	90		0
	44	2	18	173	9.61	83		0
		4	16	82	5.13	40		0
		6	16	43	2.69	26		0
3	38	2	19	396	20.84	216		0
		4	18	386	21.44	199		0
		6	21	217	10.33	110	1	0.91
	40	2	20	408	20.40	191		0
		4	15	278	18.53	151	2	1.32
		6	18	189	10.50	107	1	0.93
	42	2	17	304	17.88	145	1	0.69
		4	16	206	12.88	112		0
		6	19	162	8.53	90		0
	44	2	19	170	8.95	88		0
		4	17	83	4.88	45		0
		6	20	—	—	—		0
4	38	2	19	400	21.05	211		0
		4	19	366	19.26	191		0
		6	20	254	12.70	136	1	0.74
	40	2	21	405	19.29	210	1	0.48
		4	15	286	19.07	144	2	1.39
		6	18	291	16.17	144	1	0.69
	42	2	16	308	19.25	161	1	0.62
		4	18	196	10.89	100		0
		6	17	176	10.35	101		0
	44	2	15	116	7.73	55		0
		4	16	73	4.56	39		0
		6	21	39	1.86	20		0
5	38	2	16	368	23.00	200		0
		4	18	343	19.06	165		0
		6	19	246	12.95	130		0
	40	2	21	425	20.24	205		0
		4	20	383	19.15	183	1	0.55
		6	19	206	10.84	111	1	0.90
	42	2	19	348	18.32	184		0
		4	19	261	13.74	140		0
		6	18	167	9.28	75		0
	44	2	16	149	9.31	60		0
		4	17	89	5.24	43		0
		6	19	29	1.53	18		0
对照Control			30	736	24.53	369		0
总计Total			1 128	15 565	—	7 969	15	-

单核花粉产生后天数/d Days after formation of mononuclear pollen	处理温度/℃ Treatment temperature	处理时间/h Treatment duration	处理花蕾数 Flower bud number	收获种子数 Seed number	单果结实数 Seed number per follicle	成苗数 Seedling number	三倍体株数 Triploid number	三倍体得率/% Rate of triploid
1	38	2	18	395	21.94	205		0
		4	17	340	20.00	160		0
		6	19	275	14.47	141		0
	40	2	18	351	19.50	186		0
		4	20	376	18.80	192		0
		6	17	175	10.29	91		0
	42	2	16	295	18.44	160		0
		4	20	256	12.80	126		0
		6	18	156	8.67	80		0
	44	2	17	176	10.35	83		0
		4	19	93	4.89	49		0
		6	15	40	2.67	18		0
2	38	2	22	463	21.05	246		0
		4	18	365	20.28	183		0
		6	19	247	13.00	126	1	0.79
	40	2	17	361	21.24	190		0
		4	19	366	19.26	172	1	0.58
		6	20	216	10.80	115		0
	42	2	22	420	19.09	225		0
		4	22	271	12.32	133		0
		6	20	171	8.55	90		0
	44	2	18	173	9.61	83		0
		4	16	82	5.13	40		0
		6	16	43	2.69	26		0
3	38	2	19	396	20.84	216		0
		4	18	386	21.44	199		0
		6	21	217	10.33	110	1	0.91
	40	2	20	408	20.40	191		0
		4	15	278	18.53	151	2	1.32
		6	18	189	10.50	107	1	0.93
	42	2	17	304	17.88	145	1	0.69
		4	16	206	12.88	112		0
		6	19	162	8.53	90		0
	44	2	19	170	8.95	88		0
		4	17	83	4.88	45		0
		6	20	—	—	—		0
4	38	2	19	400	21.05	211		0
		4	19	366	19.26	191		0
		6	20	254	12.70	136	1	0.74
	40	2	21	405	19.29	210	1	0.48
		4	15	286	19.07	144	2	1.39
		6	18	291	16.17	144	1	0.69
	42	2	16	308	19.25	161	1	0.62
		4	18	196	10.89	100		0
		6	17	176	10.35	101		0
	44	2	15	116	7.73	55		0
		4	16	73	4.56	39		0
		6	21	39	1.86	20		0
5	38	2	16	368	23.00	200		0
		4	18	343	19.06	165		0
		6	19	246	12.95	130		0
	40	2	21	425	20.24	205		0
		4	20	383	19.15	183	1	0.55
		6	19	206	10.84	111	1	0.90
	42	2	19	348	18.32	184		0
		4	19	261	13.74	140		0
		6	18	167	9.28	75		0
	44	2	16	149	9.31	60		0
		4	17	89	5.24	43		0
		6	19	29	1.53	18		0
对照Control			30	736	24.53	369		0
总计Total			1 128	15 565	—	7 969	15	-

单核花粉产生后天数/d Days after formation of mononuclear pollen	处理花蕾数 Flower bud number	收获种子数 Seed number	单果结实数 Seed number per follicle	成苗数 Seedling number	三倍体株数 Triploid number	三倍体得率/% Rate of triploid
1	30	583	19.43	286	1	0.35
2	30	531	17.70	250	3	1.20
3	30	539	17.97	245	6	2.45
4	30	564	18.80	247	4	1.62
5	30	554	18.47	273	0	0
对照Control	50	1130	22.60	603	0	0
总计Total	200	3 901	-	1 904	14