Cytological Observation and Genetic Analysis of the Nuclear Male Sterile Line Y37S of Broccoli

doi:10.16420/j.issn.0513-353x.2025-0148

Acta Horticulturae Sinica ›› 2026, Vol. 53 ›› Issue (1): 99-109.doi: 10.16420/j.issn.0513-353x.2025-0148

• Genetic & Breeding·Germplasm Resources·Molecular Biology • Previous Articles Next Articles

Cytological Observation and Genetic Analysis of the Nuclear Male Sterile Line Y37S of Broccoli

YAO Xueqin¹, ZHOU Wei¹^,², LIU Chunqing¹, HUANG Lei¹, JIANG Jing², GONG Jing¹, LI Guangqing¹^,^**(), XIE Zhujie¹^,^**

¹ Shanghai Key Laboratory of Facility Horticulture Technology， Institute of Horticulture，Shanghai Academy of Agricultural Sciences， Shanghai 311023， China
² Horticulture College， Shenyang Agricultural University， Shenyang 110161， China

Received:2025-04-11 Revised:2025-10-31 Online:2026-01-25 Published:2026-01-26
Contact: LI Guangqing, XIE Zhujie

Abstract

Abstract:

The nuclear male sterile material Y37S of broccoli is a naturally mutated sterile strain discovered in the inbred line Y37. To clarify its genetic characteristics and the causes of anther failure，studies were conducted from aspects such as morphology，cytology，genetics and molecular identification. The results showed that the stamens of the Y37S sterile line plants were degenerated，significantly lower than the stigma，and there was no pollen. The results of the scanning electron microscope showed that the anthers of Y37S were shriveled and twisted，presenting a triangular shape，with no pollen grains attached. The microspore viability of flower buds of different sizes was detected by fluorescein diacetate（FDA）. When the flower buds of sterile plants developed to 4.5-5.0 mm，the microspore viability was lost and there was no green fluorescence signal. Observation of the cytological characteristics of anthers revealed that in sterile plants，microspore collapse occurred during the mononuclear microspore stage，and the degradation of felt-like layer cells was delayed and incomplete，which affected the normal functions of anthers and pollen，ultimately leading to infertility. Six fertile strains within the same line were crossed with Y37S respectively. Among them，the ratios of self-pollinated fertile strains to sterile strains of Y37-1 and Y37-3 were 23︰7 and 17︰5，respectively，and the ratios of fertile strains to sterile strains of the hybrid offspring of Y37-1 and Y37-3 with Y37S were 15︰14 and 28︰30，respectively. It is preliminarily speculated that Y37S is controlled by a pair of recessive nuclear genes. To verify its genetic pattern，Y37S was crossed with the normal fertile inbred line B50. All F₁ generations were fertile. The ratio of fertile to sterile lines in the F₂ population was 107︰29，and in the test cross population，it was 67︰69. This conforms to Mendel’s law of inheritance and proves that Y37S is a nuclear male sterile line controlled by a pair of recessive genes. The sterile and fertile plants in the test population were amplified using the molecular markers of the single recessive nuclear sterility gene linkage of Chinese cabbage and cabbage that have been identified. No differential bands were amplified，suggesting that the gene controlling the fertility of the sterile line Y37S might be a new recessive nuclear sterility gene.

Key words: broccoli, nuclear male sterility, enetic analysis, molecular marker

YAO Xueqin, ZHOU Wei, LIU Chunqing, HUANG Lei, JIANG Jing, GONG Jing, LI Guangqing, XIE Zhujie. Cytological Observation and Genetic Analysis of the Nuclear Male Sterile Line Y37S of Broccoli[J]. Acta Horticulturae Sinica, 2026, 53(1): 99-109.

Figures/Tables 7

References 40

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不育基因 Sterile genes	引物名称 Primer name	正向引物（5′-3′） Forward primer sequence	反向引物（5′-3′） Reverse primer sequence
Brnnd1	HABS378	CCGAAGACTAGAACGTCCAA	ACAATGAACACACCGGTCTA
Brnnd1	HABS386	GAATTCTAGCAGGTAGCTGACG	GCGGAAACCTTTGTGAGTTT
BoTPD1	YIndel0116	TGCAGAGTTGAGTCACGATG	TGTTGGCTGTTCTGCGATAC
	YIndel012	CGCCTTGCTTCTATTCCAAG	TGTTGGCTGTTCTGCGATAC
	Ms3-dec	GAATGTATCAGGCGTGGAGG	TCACCGATTTGGTATGAGTTT

不育基因 Sterile genes	引物名称 Primer name	正向引物（5′-3′） Forward primer sequence	反向引物（5′-3′） Reverse primer sequence
Brnnd1	HABS378	CCGAAGACTAGAACGTCCAA	ACAATGAACACACCGGTCTA
Brnnd1	HABS386	GAATTCTAGCAGGTAGCTGACG	GCGGAAACCTTTGTGAGTTT
BoTPD1	YIndel0116	TGCAGAGTTGAGTCACGATG	TGTTGGCTGTTCTGCGATAC
	YIndel012	CGCCTTGCTTCTATTCCAAG	TGTTGGCTGTTCTGCGATAC
	Ms3-dec	GAATGTATCAGGCGTGGAGG	TCACCGATTTGGTATGAGTTT

群体 Group	可育株数 Number of fertile plants		分离比 Separation ratio	卡方值 Value	χ²_0.05,1
Y37S × Y37-1	15	14	1.00︰0.93	0.034	3.84
Y37S × Y37-2	18	0	全部可育All fertile	0	3.84
Y37S × Y37-3	28	30	1.00︰1.07	0.069	3.84
Y37S × Y37-4	52	0	全部可育All fertile	0	3.84
Y37S × Y37-5	47	0	全部可育All fertile	0	3.84
Y37S × Y37-6	28	0	全部可育All fertile	0	3.84
Y37-1 ⊗	23	7	3.28︰1.00	0.044	3.84
Y37-2 ⊗	31	9	3.44︰1.00	0.133	3.84
Y37-3 ⊗	17	5	3.40︰1.00	0.060	3.84
Y37-4 ⊗	37	0	全部可育All fertile	0	3.84
Y37-5 ⊗	22	0	全部可育All fertile	0	3.84
Y37-6 ⊗	26	0	全部可育All fertile	0	3.84
F₁（Y37S × B50）	34	34	全部可育All fertile	0	3.84
F₂（Y37S × B50）	107	29	3.68︰1.00	0.980	3.84
B₁（Y37S × F₁）	67	69	1.00︰1.02	0.029	3.84

群体 Group	可育株数 Number of fertile plants		分离比 Separation ratio	卡方值 Value	χ²_0.05,1
Y37S × Y37-1	15	14	1.00︰0.93	0.034	3.84
Y37S × Y37-2	18	0	全部可育All fertile	0	3.84
Y37S × Y37-3	28	30	1.00︰1.07	0.069	3.84
Y37S × Y37-4	52	0	全部可育All fertile	0	3.84
Y37S × Y37-5	47	0	全部可育All fertile	0	3.84
Y37S × Y37-6	28	0	全部可育All fertile	0	3.84
Y37-1 ⊗	23	7	3.28︰1.00	0.044	3.84
Y37-2 ⊗	31	9	3.44︰1.00	0.133	3.84
Y37-3 ⊗	17	5	3.40︰1.00	0.060	3.84
Y37-4 ⊗	37	0	全部可育All fertile	0	3.84
Y37-5 ⊗	22	0	全部可育All fertile	0	3.84
Y37-6 ⊗	26	0	全部可育All fertile	0	3.84
F₁（Y37S × B50）	34	34	全部可育All fertile	0	3.84
F₂（Y37S × B50）	107	29	3.68︰1.00	0.980	3.84
B₁（Y37S × F₁）	67	69	1.00︰1.02	0.029	3.84

Cytological Observation and Genetic Analysis of the Nuclear Male Sterile Line Y37S of Broccoli

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