兼抗白粉病和条锈病的小麦-中间偃麦草7JS附加系CH71的分子细胞学鉴定

doi:10.3864/j.issn.0578-1752.2025.16.013

中国农业科学 ›› 2025, Vol. 58 ›› Issue (16): 3304-3316.doi: 10.3864/j.issn.0578-1752.2025.16.013

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

兼抗白粉病和条锈病的小麦-中间偃麦草7J^S附加系CH71的分子细胞学鉴定

张树伟¹(), 史文宇¹, 李冰洁¹, 常松鹤¹, 乔麟轶¹, 李欣¹, 常利芳¹, 陈芳¹, 郭慧娟¹, 畅志坚¹, 张晓军¹^,²^,^*()

¹ 山西农业大学农学院（作物科学研究所），太原 030031
² 农业农村部有机旱作农业重点实验室（部省共建），太原 030031

收稿日期:2025-02-25 接受日期:2025-04-22 出版日期:2025-08-11 发布日期:2025-08-11
通信作者:
张晓军，E-mail：zxjemail@163.com
联系方式: 张树伟，E-mail：zshuwei@sxau.edu.cn。
基金资助:
山西省小麦育种联合攻关项目(NYGG26); 山西农业大学优秀博士启动项目(2024BQ49); 山西省自然科学基金(202103021224140); 山西农业大学农学院育种工程专项(YZ2021-0)

Molecular Cytogenetic Characterization of Wheat-Thinopyrum intermedium 7J^S Addition Line CH71 with Resistance to Both Powdery Mildew and Stripe Rust

ZHANG ShuWei¹(), SHI WenYu¹, LI BingJie¹, CHANG SongHe¹, QIAO LinYi¹, LI Xin¹, CHANG LiFang¹, CHEN Fang¹, GUO HuiJuan¹, CHANG ZhiJian¹, ZHANG XiaoJun¹^,²^,^*()

¹ College of Agriculture (Institute of Crop Sciences), Shanxi Agricultural University, Taiyuan 030031
² Key Laboratory of Sustainable Dryland Agriculture (Co-Construction by Ministry and Province), Ministry of Agriculture and Rural Affairs, Taiyuan 030031

Received:2025-02-25 Accepted:2025-04-22 Published:2025-08-11 Online:2025-08-11

摘要/Abstract

摘要：

【目的】中间偃麦草是小麦的三级基因库，利用远缘杂交将中间偃麦草含优异基因的染色体导入小麦背景创制新型种质资源，探究其对小麦抗病性及农艺性状的影响，为小麦育种改良提供理论依据。【方法】以普通小麦烟农999与小麦-中间偃麦草部分双二倍体TAI8047杂交BC₁F₆代选育的小麦新品系CH71为试验材料，利用Oligo-pSc119.2、Oligo-pTa535、Oligo-B11、Oligo-pDb12H等探针通过非变性荧光原位杂交（non-denaturing fluorescence in situ hybridization，ND-FISH）进行核型分析，明确CH71及其亲本的染色体组成；通过Synt探针的Oligo-FISH painting技术，鉴定CH71外源染色体同源群归属，并结合中间偃麦草特异STS标记进行PCR扩增验证；利用国内流行的白粉菌小种E09和条锈菌混合小种CYR32、CYR33、CYR34对CH71进行抗病性鉴定，并测定其株高、穗长、小穗数、千粒重、粒长、粒宽等农艺性状。【结果】小麦亲本烟农999含有21对小麦染色体；部分双二倍体亲本TAI8047含有58条染色体，包括21对小麦染色体和8对外源染色体；CH71含有44条染色体，包括21对小麦染色体和1对外源染色体J^S-1；利用Synt7探针对CH71进行Oligo-FISH painting鉴定，表明CH71附加的J^S-1染色体可能为中间偃麦草7J^S染色体；进一步利用中间偃麦草第7同源群的183对STS引物对CH71进行PCR扩增，筛选到12个可以同时在中间偃麦草、TAI8047和CH71中扩增出特异条带的STS标记，表明J^S-1为中间偃麦草7J^S染色体；人工接种我国广泛流行的白粉菌小种E09和条锈菌混合小种CYR32、CYR33、CYR34，抗性鉴定表明，CH71中抗白粉病、免疫条锈病，遗传分析表明，其对白粉病和条锈病的抗性可能均来自中间偃麦草7J^S染色体；农艺性状调查表明，CH71的株高比小麦亲本烟农999增加24.0 cm、穗长增加3.44 cm、小穗数增加1.6个、粒长增加0.68 mm，但千粒重降低6.78%、粒宽减少2.70%。【结论】新品系CH71为小麦-中间偃麦草7J^S异附加系，兼抗小麦白粉病和条锈病，可作为有价值的种质资源用于小麦抗病育种及外源抗病基因克隆；筛选到12个STS标记可为小麦背景下中间偃麦草7J^S染色体的快速鉴定提供便捷。

关键词: 小麦, 中间偃麦草, 异附加系, 荧光原位杂交, STS标记

Abstract:

【Objective】Thinopyrum intermedium, a tertiary gene pool of wheat, harbors valuable genetic resources for wheat improvement. This study aimed to develop novel wheat germplasms by transferring elite chromosomes from Thinopyrum intermedium into wheat via distant hybridization, investigating their impacts on disease resistance and agronomic traits to establish a theoretical foundation for wheat breeding. 【Method】A wheat-Thinopyrum intermedium disomic addition line, CH71 (2n=44), was developed from the BC₁F₆ progeny of a cross between common wheat cultivar Yannong 999 and the partial amphidiploid TAI8047 (2n=58). Non-denaturing fluorescence in situ hybridization (ND-FISH) with oligonucleotide probes (Oligo-pSc119.2, Oligo-pTa535, Oligo-B11, Oligo-pDb12H) was employed for karyotype analysis. Synteny-based (Synt) Oligo-FISH painting and Th. intermedium-specific STS markers were utilized to identify the homologous group and validate the origin of the alien chromosome. Disease resistance to powdery mildew (race E09) and stripe rust (mixed races CYR32, CYR33, CYR34) was evaluated under artificial inoculation. Agronomic traits, including plant height, spike length, spikelet number, thousand-grain weight, grain length, and grain width, were systematically measured.【Result】The wheat parent Yannong 999 exhibited a standard karyotype of 21 wheat chromosome pairs. TAI8047 contained 58 chromosomes (21 wheat pairs + 8 alien pairs), while CH71 harbored 44 chromosomes (21 wheat pairs + 1 alien pair, J^S-1). Synt-FISH and PCR amplification using 183 STS primers specific to Th. intermedium chromosome 7J^S confirmed J^S-1 as a 7J^S-derived chromosome. Twelve STS markers consistently amplified diagnostic bands in Th. intermedium, TAI8047, and CH71. CH71 displayed moderate resistance to powdery mildew and immunity to stripe rust, with genetic analysis indicating both resistances were conferred by the 7J^S chromosome. Compared to Yannong 999, CH71 exhibited significant increases in plant height (+24.0 cm), spike length (+3.44 cm), spikelet number (+1.6), and grain length (+0.68 mm), but reductions in thousand-grain weight (-6.78%) and grain width (-2.70%). 【Conclusion】The novel disomic addition line CH71 (wheat-Th. intermedium 7J^S) demonstrates dual resistance to powdery mildew and stripe rust, serving as a valuable germplasm resource for disease-resistant breeding and the cloning of alien resistance genes. The 12 STS markers identified herein provide an efficient molecular tool for rapid tracking of the 7J^S chromosome in wheat backgrounds.

Key words: wheat, Thinopyrum intermedium, alien addition line, fluorescence in situ hybridization, STS markers

张树伟, 史文宇, 李冰洁, 常松鹤, 乔麟轶, 李欣, 常利芳, 陈芳, 郭慧娟, 畅志坚, 张晓军. 兼抗白粉病和条锈病的小麦-中间偃麦草7J^S附加系CH71的分子细胞学鉴定[J]. 中国农业科学, 2025, 58(16): 3304-3316.

ZHANG ShuWei, SHI WenYu, LI BingJie, CHANG SongHe, QIAO LinYi, LI Xin, CHANG LiFang, CHEN Fang, GUO HuiJuan, CHANG ZhiJian, ZHANG XiaoJun. Molecular Cytogenetic Characterization of Wheat-Thinopyrum intermedium 7J^S Addition Line CH71 with Resistance to Both Powdery Mildew and Stripe Rust[J]. Scientia Agricultura Sinica, 2025, 58(16): 3304-3316.

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探针名称 Probes	修饰类型 Modification types	探针序列 Probe sequences (5′-3′)	参考文献 References
Oligo-B11	5′6-FAM	TCCGCTCACCTTGATGACAACATCAGGTGGAATTCCGTTCGAGGG	[22]
Oligo-pDb12H	5′TAMRA	TCAGAATTTTTAGGATAGCAGAAGTATTCGAAATACCCAGATTGC TACAG	[23]
Oligo-pSc119.2	5′6-FAM	CCGTTTTGTGGACTATTACTCACCGCTTTGGGGTCCCATAGCTAT	[24]
Oligo-pTa535	5′TAMRA	AAAAACTTGACGCACGTCACGTACAAATTGGACAAACTCTTTCGGAGTATC AGGGTTTC	[24]

序号 No.	引物名称 Primer name	染色体 Chromosome	正向引物 Forward sequence (5′-3′)	反向引物 Reverse sequence (5′-3′)	退火温度 Annealing temperature (℃)
1	C19-08	7J^S	CTTAGTCTGCAGCCTCTGTCC	AGAGCATCGTCAATAGCCCAG	60
2	C19-22	7J^S	AAACATAGCCTAAGACGGGC	GGTCATTGGGTGCAGAATAG	60
3	C19-39	7J^S	GAATCTGCGGTGTGCTATTT	TCGGTCGACAACATAGTGTG	60
4	C19-54	7J^S	ACGCAAGTTTCAGATAAACAAGACT	TCTCTTGATCGGTTGGACACC	58
5	C19-61	7J^S	CGTGTATCGAACGTGAAATG	CAGTTGGAAATCCAGGAATG	60
6	C19-68	7J^S	CTCTTGTCGTCGAGGGTCT	GTAGAGGTACTTAGCGGCGA	58
7	C19-83	7J^S	TTTGCTCACTCTCTGGTTCC	GAAAGTTGGAGAACGCTCAA	60
8	C19-103	7J^S	GGATGAAGCATCTCCACAAC	ATCAGGAAGCTCAGCATCAG	58
9	C19-105	7J^S	TTCTCATGTGTGTGACCCGC	TTGGTAGAACGGCCACAAGA	60
10	C19-109	7J^S	AGTAGTGCTGAACTTGCCACT	GCAAACGTCTTCGGCGTATC	60
11	C19-136	7J^S	GCTCACCATGAGAAGGAAAA	GGATGAATGTTAATGGCGAG	60
12	C19-138	7J^S	CCTAGAAGTTCACGGCCTC	GGCACTTCTGTAGCTTGCAT	60

材料名称 Material name	染色体数目 Number of chromosome	外源染色体 Alien chromosome	条锈菌小种 CYR32/CYR33/CYR34 Stripe rust species CYR32/CYR33/CYR34	白粉菌小种E09 Powdery mildew species E09
材料名称 Material name	染色体数目 Number of chromosome	外源染色体 Alien chromosome		苗期 Seedling resistance	成株期 Adult-plant resistance
太原768 Taiyuan 768	42	—	4	4	4
系76（64）Line 76(64)	42	—	4	4	4
TAI8047	58	J+J^S+St	0	0;	0
CH71	44	7J^S	0;	2	2
烟农999 Yannong 999	42	—	4	4	4
SY95-71	42	—	—	4	4
TC29	42	—	4	—	—

材料名称 Material name	外源染色体 Alien chromosome	株高 Plant height (cm)	穗长 Spike length (cm)	小穗数 Spikelet number	千粒重 Thousand grain weight (g)	粒长 Grain length (mm)	粒宽 Grain width (mm)
太原768 Taiyuan 768	—	76.60±3.51a	8.88±0.43a	20.80±1.10a	49.43±1.67b	6.84±0.17a	3.75±0.11c
系76（64）Line76 (64)	—	77.20±3.90a	9.06±0.36a	20.40±0.89a	49.03±1.21b	6.86±0.14a	3.72±0.08bc
TAI8047	J+J^S+St	94.80±4.44b	17.20±0.57c	21.60±1.67a	29.89±3.09a	8.10±0.17c	2.95±0.09a
烟农999 Yannong 999	—	77.40±3.96a	8.30±0.35a	20.40±0.89a	52.39±5.09b	6.82±0.24a	3.70±0.12bc
CH71	7J^S	101.40±7.78b	11.74±1.28b	22.00±1.33a	48.84±2.43b	7.50±0.18b	3.60±0.09b

兼抗白粉病和条锈病的小麦-中间偃麦草7J^S附加系CH71的分子细胞学鉴定

Molecular Cytogenetic Characterization of Wheat-Thinopyrum intermedium 7J^S Addition Line CH71 with Resistance to Both Powdery Mildew and Stripe Rust

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兼抗白粉病和条锈病的小麦-中间偃麦草7JS附加系CH71的分子细胞学鉴定

Molecular Cytogenetic Characterization of Wheat-Thinopyrum intermedium 7JS Addition Line CH71 with Resistance to Both Powdery Mildew and Stripe Rust

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兼抗白粉病和条锈病的小麦-中间偃麦草7J^S附加系CH71的分子细胞学鉴定

Molecular Cytogenetic Characterization of Wheat-Thinopyrum intermedium 7J^S Addition Line CH71 with Resistance to Both Powdery Mildew and Stripe Rust