The Culture Technique of Isolated Microspore in Tetraploid Chinese Cabbage

doi:10.16420/j.issn.0513-353x.2023-0046

Acta Horticulturae Sinica ›› 2023, Vol. 50 ›› Issue (12): 2641-2652.doi: 10.16420/j.issn.0513-353x.2023-0046

• Cultivation Physiology & Biochemistry • Previous Articles Next Articles

The Culture Technique of Isolated Microspore in Tetraploid Chinese Cabbage

LIU Xiaodong, WU Fang, MENG Chuan, MU Jingui, WANG Yuhai, WANG Mingqiu^**()

Institute of Cash Crops of Hebei Academy of Agricultural and Forestry Sciences，Hebei Vegetable Engineering Technology Centre，Shijiazhuang 050051，China

Received:2023-07-31 Revised:2023-10-16 Online:2023-12-25 Published:2023-12-29
Contact: WANG Mingqiu

Abstract

Abstract:

The gametes produced during the meiosis process of tetraploid Chinese cabbage are diverse. The purpose of carrying out free microspore culture of tetraploid Chinese cabbage is to obtain more new homozygous diploid resources with a large degree of variation. In this study，four easy embryogenic materials were selected from 14 early-maturing tetraploid Chinese cabbages. The technical system of isolated microspore culture of tetraploid Chinese cabbage was established in the aspects of flower bud phenotype，heat shock temperature，rooting hormone and ploidy identification. The results were as follows：For different genotypes of tetraploid Chinese cabbage，when petal/anther of flower buds was 0-0.5，the microspore was in the stage of mononuclear sideline，and the embryo rate was the highest at this time. When the heat shock temperature was 33.5 ℃ and dark culture for 24 h，they were beneficial to microspore embryogenesis. For rooting medium，the combination of indole butyric acid（IBA）and naphthylacetic acid（NAA）was better than single hormone. The optimum concentration was IBA 0.3 mg · L^-1and NAA 0.1 mg · L^-1. The ploidy was detected by flow cytometry，chromosome microscopy and cross pollination. The results showed that the probability of diploid was 94.87% and the probability of tetraploid was 5.13% in 39 regenerated plants.

Key words: Chinese cabbage, microspore culture, tetraploid, creation

LIU Xiaodong, WU Fang, MENG Chuan, MU Jingui, WANG Yuhai, WANG Mingqiu. The Culture Technique of Isolated Microspore in Tetraploid Chinese Cabbage[J]. Acta Horticulturae Sinica, 2023, 50(12): 2641-2652.

Figures/Tables 12

References 35

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编号 No.	生育期/d Days	叶球类型 Leafy head	编号 No.	生育期/d Days	叶球类型 Leafy head
D20	55 ~ 60	花心 Spreading type	D522	55 ~ 65	叠抱 Overlapping type
D21	55 ~ 60	合抱 Outward-curling type	D523	55 ~ 65	叠抱 Overlapping type
D22	55 ~ 60	花心 Spreading type	D524	55 ~ 65	叠抱 Overlapping type
D23	55 ~ 60	花心 Spreading type	D525	55 ~ 65	合抱Outward-curling type
D24	55 ~ 60	叠抱 Overlapping type	D526	55 ~ 65	花心 Spreading type
D520	55 ~ 65	花心 Spreading type	D527	55 ~ 65	合抱Outward-curling type
D521	55 ~ 65	花心 Spreading type	D528	55 ~ 65	合抱Outward-Curling Type

编号 No.	生育期/d Days	叶球类型 Leafy head	编号 No.	生育期/d Days	叶球类型 Leafy head
D20	55 ~ 60	花心 Spreading type	D522	55 ~ 65	叠抱 Overlapping type
D21	55 ~ 60	合抱 Outward-curling type	D523	55 ~ 65	叠抱 Overlapping type
D22	55 ~ 60	花心 Spreading type	D524	55 ~ 65	叠抱 Overlapping type
D23	55 ~ 60	花心 Spreading type	D525	55 ~ 65	合抱Outward-curling type
D24	55 ~ 60	叠抱 Overlapping type	D526	55 ~ 65	花心 Spreading type
D520	55 ~ 65	花心 Spreading type	D527	55 ~ 65	合抱Outward-curling type
D521	55 ~ 65	花心 Spreading type	D528	55 ~ 65	合抱Outward-Curling Type

NAA/ （mg · L^-1）	IBA/ （mg · L^-1）	根长/cm Root length	须根数量 Fibrous root	生根率/% Rooting rate	移栽成活率/% Transplantation survival rate
0	0	—	—	—	—
	0.1	3.51 ± 0.83 de	少 Few	50.0 a	36.7 b
0.1	0	3.15 ± 0.07 e	较少 Fewer	33.3 a	10.0 a
	0.1	3.86 ± 0.06 de	多 More	90.0 ab	90.0 c
	0.2	4.59 ± 1.41 d	多 More	100.0 b	90.0 c
	0.3	8.26 ± 1.05 c	多 More	100.0 b	100.0 c
	0.4	10.19 ± 0.39 b	少 Few	100.0 b	100.0 c
	0.5	11.51 ± 1.42 a	少 Few	93.3 b	93.3 c

NAA/ （mg · L^-1）	IBA/ （mg · L^-1）	根长/cm Root length	须根数量 Fibrous root	生根率/% Rooting rate	移栽成活率/% Transplantation survival rate
0	0	—	—	—	—
	0.1	3.51 ± 0.83 de	少 Few	50.0 a	36.7 b
0.1	0	3.15 ± 0.07 e	较少 Fewer	33.3 a	10.0 a
	0.1	3.86 ± 0.06 de	多 More	90.0 ab	90.0 c
	0.2	4.59 ± 1.41 d	多 More	100.0 b	90.0 c
	0.3	8.26 ± 1.05 c	多 More	100.0 b	100.0 c
	0.4	10.19 ± 0.39 b	少 Few	100.0 b	100.0 c
	0.5	11.51 ± 1.42 a	少 Few	93.3 b	93.3 c

编号 No.	鉴定植株数 Number of plant	二倍体数 Number of diploid	四倍体数 Number of autotetraploid	二倍体率/% Diploid rate	四倍体率/% Autotetraploid rate
D24	8	8	0	100.00	0
D522	18	16	2	88.89	11.11
D524	7	7	0	100.00	0
D528	6	6	0	100.00	0
合计Sum	39	37	2	94.87	5.13

The Culture Technique of Isolated Microspore in Tetraploid Chinese Cabbage

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小孢子再生植株 Microspore regenerated plants	授粉品种Varieties		小孢子再生植株 Microspore regenerated plants	授粉品种Varieties
小孢子再生植株 Microspore regenerated plants	冀白4号（2x） Jibai 4	多维462（4x） Duowei 462	小孢子再生植株 Microspore regenerated plants	冀白4号（2x） Jibai 4	多维462（4x） Duowei 462
XD24-1（2x）	3.19	0	XD522-13（2x）	4.14	0
XD24-2（2x）	2.00	0	XD522-14（2x）	4.73	0
XD24-3（2x）	1.63	0	XD522-15（2x）	1.05	0
XD24-4（2x）	1.55	0	XD522-16（2x）	2.87	0
XD24-5（2x）	1.97	0	XD522-17（2x）	1.56	0
XD24-6（2x）	2.66	0	XD522-18（2x）	1.51	0
XD24-7（2x）	2.48	0	XD524-1（2x）	1.04	0
XD24-8（2x）	3.06	0	XD524-2（2x）	2.29	0
XD522-1（4x）	0	1.15	XD524-3（2x）	0.99	0
XD522-2（4x）	0	0.83	XD524-4（2x）	3.06	0
XD522-3（2x）	5.01	0	XD524-5（2x）	2.68	0
XD522-4（2x）	7.69	0	XD524-6（2x）	2.77	0
XD522-5（2x）	3.23	0	XD524-7（2x）	3.64	0
XD522-6（2x）	5.11	0	XD528-1（2x）	0.87	0
XD522-7（2x）	2.24	0	XD528-2（2x）	2.96	0
XD522-8（2x）	1.17	0	XD528-3（2x）	3.49	0
XD522-9（2x）	4.38	0	XD528-4（2x）	1.98	0
XD522-10（2x）	2.57	0	XD528-5（2x）	1.86	0
XD522-11（2x）	2.36	0	XD528-6（2x）	3.59	0
XD522-12（2x）	5.52	0

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