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作物学报 ›› 2026, Vol. 52 ›› Issue (2): 433-445.doi: 10.3724/SP.J.1006.2026.51060

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

小麦-华山新麦草二体异代换系16DH25-7的分子细胞遗传学及抗病性鉴定

王粤生(), 葛冬冬, 程兰斐, 陈春环, 王长有, 刘新伦, 李停栋, 邓平川, 吉万全(), 赵继新()   

  1. 西北农林科技大学农学院, 陕西杨凌 712100
  • 收稿日期:2025-06-16 接受日期:2025-10-30 出版日期:2026-02-12 网络出版日期:2025-11-12
  • 通讯作者: *吉万全 E-mail:jiwanquan2008@126.com;赵继新 E-mail:zhjx881@163.com
  • 作者简介:E-mail: wangyuesheng2022@126.com
  • 基金资助:
    杨凌种业创新中心“小麦远缘杂交与种质创新研发”项目(ylzy-xm-02)

Molecular cytogenetic and disease resistance characterization of the wheat- Psathyrostachys huashanica disomic substitution line 16DH25-7

Wang Yue-Sheng(), Ge Dong-Dong, Cheng Lan-Fei, Chen Chun-Huan, Wang Chang-You, Liu Xin-Lun, Li Ting-Dong, Deng Ping-Chuan, Ji Wan-Quan(), Zhao Ji-Xin()   

  1. College of Agriculture, Northwest A&F University, Yangling 712100, Shaanxi, China
  • Received:2025-06-16 Accepted:2025-10-30 Published:2026-02-12 Published online:2025-11-12
  • Contact: *Ji Wan-Quan E-mail: E-mail:jiwanquan2008@126.com;Zhao Ji-Xin E-mail:zhjx881@163.com
  • Supported by:
    Project “Research and Development on Wheat Distant Hybridization and Germplasm Innovation”(ylzy-xm-02) of Yangling Seed Industry Innovation Center

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摘要：

华山新麦草(Psathyrostachys huashanica Keng; 2n = 2x = 14, NsNs)作为小麦三级基因库的重要野生资源, 对白粉病和条锈病等生物胁迫表现出明显抗性, 在小麦遗传改良中具有重要应用价值。本研究在小麦-华山新麦草七倍体材料H8911与硬粒小麦D4286的杂交后代中筛选获得兼抗小麦白粉病和条锈病的衍生系16DH25-7。为解析该材料的遗传特性, 运用细胞遗传学分析、荧光原位杂交(FISH)、液相芯片技术及分子标记方法, 对其染色体组成进行了鉴定, 并对其农艺性状与抗病表型进行了系统评价。细胞遗传学分析表明, 16DH25-7的染色体数目为2n = 42。FISH检测显示, 该材料携带2条华山新麦草Ns染色体, 同时缺失2条小麦5D染色体。分子标记与SNP芯片联合分析证实, 缺失的5D染色体被华山新麦草5Ns染色体替代, 表明16DH25-7为小麦-华山新麦草5Ns (5D)二体异代换系。农艺性状调查表明, 16DH25-7表现出株高降低、分蘖数增多等优良特性; 抗病性鉴定显示其高抗小麦条锈病和白粉病。16DH25-7作为优异抗病种质资源, 为小麦抗病育种及遗传改良提供了新的候选种质。

关键词: 小麦, 华山新麦草, 原位杂交, 分子标记, 抗病性鉴定

Abstract:

Psathyrostachys huashanica Keng (2n = 2x = 14, NsNs), a valuable germplasm resource in the tertiary gene pool of wheat, exhibits remarkable resistance to biotic stresses such as powdery mildew and stripe rust, making it highly valuable for wheat genetic improvement. In this study, a derivative line, 16DH25-7, exhibiting resistance to both powdery mildew and stripe rust, was selected from the progeny of the wheat-P. huashanica heptaploid H8911 and durum wheat D4286. To elucidate the genetic background of this material, cytogenetic analysis, fluorescence in situ hybridization (FISH), liquid-phase chip technology, and molecular marker analysis were used to characterize its chromosomal composition. Cytogenetic analysis revealed that 16DH25-7 had a chromosome number of 2n = 42. FISH analysis showed that it carried two P. huashanica Ns chromosomes and simultaneously lacked both wheat 5D chromosomes. Combined analysis using molecular markers and SNP chip data confirmed that the missing 5D chromosomes were replaced by P. huashanica 5Ns chromosomes, indicating that 16DH25-7 is a wheat-P. huashanica 5Ns (5D) disomic substitution line. Agronomic evaluation demonstrated that 16DH25-7 possesses desirable traits such as reduced plant height and increased tiller number. Disease resistance assessments confirmed its high resistance to both stripe rust and powdery mildew. In conclusion, 16DH25-7 represents an elite disease-resistant germplasm that holds great potential for wheat resistance breeding and genetic improvement.

Key words: wheat, Psathyrostachys huashanica, in situ hybridization, molecular markers, disease resistance evaluation

图1

16DH25-7的细胞学观察与原位杂交鉴定 a: 根尖细胞在有丝分裂中期的染色体; b和d: 使用华山新麦草基因组DNA作为探针对16DH25-7进行GISH分析(红色箭头表示华山新麦草染色体); c: 使用Oligo-pSc119.2探针(绿色)和Oligo-pTa535探针(红色)对16DH25-7进行Mc-FISH分析; e: 7182和16DH25-7染色体2B、1D和5A荧光信号的比较; f: 使用Oligo-pSc119.2探针(绿色)和Oligo-pTa535探针(红色)对7182进行Mc-FISH分析。染色体用DAPI (蓝色)复染。标尺为10 μm。"

表1

16DH25-7中与华山新麦草染色体连锁的特异分子标记"

标记
Marker		 类型
Type		 引物序列 Sequence of primer (5′-3′) 染色体
Chromosome		 退火温度
Tm (℃)
正向引物Forward primer 反向引物Reverse primer
BF293016 ESTs CGTCCTCAAGTCCCTCCTCT GAACAGCTCAGCAGAATACGG 5AS, 5BS, 5DS 60
CD452608 ESTs TGATGTCTTGTCGTGGTCGT TTTTGGATGCAGCAAGACAG 5AS, 5BS, 5DS 58
BG607512 ESTs ATTGTCGAGGTGGTTCAAGG CAGTCTGCCACCTCCTCTTC 5BL, 5DL, 5AL 58
TNAC1624 PLUG TTCTCCTCGAGAAGTTTGCAG CCTCGTCACCAACTTCCTCTAC 4AL, 5BL, 5DL 58
TNAC1575 PLUG CAGCTCTAGCGAAAGACAAGG AGCGATATTACGCCTTCCACT 4AL, 5BL, 5DL 58
TNAC1567 PLUG ATGTTGGCTTTATACCAATGC AGGTGCGGCTTCACTATCTTT 5AL, 5BL, 5DL 58
RHS103 SCAR TCCGCATACCCGATAAT TCCGCATACCACCTGTT 5Ns 58
RHS141 SCAR CTCGGCACCATAAACTAT CTCGGCACTAGAGGAAAC 5Ns 58

图2

11个多态性标记对16DH25-7及其亲本的凝胶电泳分析 M: DL2000; 1: 7182; 2: D4286; 3: 华山新麦草; 4: 16DH25-7。a: BF293016; b: CD452608; c: BG607512; d: TNAC1624-TaqI; e: TNAC1624-HaeIII; f: TNAC1575-TaqI; g: TNAC1575-HaeIII; h: TNAC1567-TaqI; i: TNAC1567-HaeIII; j: RHS103; k: RHS141。箭头指示特异条带。"

图3

16DH25-7的SNP芯片分析 a: 普通小麦7182; b和c: 16DH25-7; 纵坐标为DMP。"

表2

华山新麦草5Ns染色体分子标记在试验材料中的多态性分析"

标记
Marker		 物种Species 引物序列
Sequence of primer (5′-3′)		 退火温度
Tm (℃)
7182 D4286 黑麦
Secale cereale		 长穗偃麦草
Thinopyrum ponticum		 华山新麦草
Psathyrostachys huashanica		 滨麦
Leymus mollis		 16DH
25-7
Ph#5Ns-1 + + + + + + + F: ACACTCTGACGCAGTTGATG 60
R: GAAACTGGCAGAAGAGGTCC
Ph#5Ns-2 - - - - + - + F: CCGCCGTGTCTACTTATTCG 62
R: CAGCAAACGCAACAGACAGG
Ph#5Ns-3 - - - - + - + F: GGTAATGCCCCTACTCCTGC 62
R: ACACACGCACTGATACGGGA
Ph#5Ns-4 + + + - + + + F: CTGCTCCCTCAAGTTCTCAA 60
R: CAGAATGGTGGCAGCAATAC
Ph#5Ns-5 - - - - + + + F: TCTTTCCTGAAGTTGGCTGG 60
R: CAAATCCCAGAATCCGAGC
Ph#5Ns-6 - - + + + + + F: AAGTGGTCGCACATTCCTCT 58
R: TTCGTGACCTTGATGTTGGA
Ph#5Ns-7 - - - - + + + F: GCCTGAAGACTTTCCTATCCT 58
R: TGAAGACAATGTTATCAAAGCA
Ph#5Ns-8 - - - - + - + F: CCCAAGGGTCTCCAATAAGG 60
R: GAGTGGTCTTTGGTTTCTTGC
Ph#5Ns-9 - - - - + - + F: CACCTAAGACGAGTAACCAA 58
R: AGCATTATCTGTAAGCGGAG
Ph#5Ns-10 + + + + + + + F: ACGAGCCAATCATCAGCATC 60
R: CCAGCAAGGAAACTTCTACA
Ph#5Ns-11 + + + + + + + F: GGTATGGCTTACGGGAGGTT 62
R: CTGAAGGCTGCCCAAGAGAA
Ph#5Ns-12 - - - + + + + F: TAATGGCGTGAGGTTTGAGG 60
R: CGGACACCATAAAGCACCAA
Ph#5Ns-13 - - - - + + + F: GCCATACTGCGTTCTCCACA 64
R: CTGCCCTCACGCTTCTCCTC
Ph#5Ns-14 + + + - + + + F: TTGATGTGCCAAAGGAACTC 58
R: CAACACTTACCTTGAATCCG
Ph#5Ns-15 - - - - + - + F: ACCTTTAGCCGAGTAGTGGA 62
R: CTCTCCAGCACAGCACTCAC
Ph#5Ns-16 - - - - + + + F: AGTTTGAATACCGAGAGGAG 56
R: ACCATTGATAAGATTGCTGC
Ph#5Ns-17 - - - - + - + F: TGAAGAAGAACAGGGGCAAG 60
R: TTCTCCCCTGAACCAAACTG
Ph#5Ns-18 - - - + + + + F: AAGTTTGCTTTTTTGTGCGT 56
R: CGGCTAAGGGTTATTATCAG
Ph#5Ns-19 + + + + + + + F: TGGAGCGAGATGGACTACGA 62
R: TAACATCCTGTATGGCACCG
Ph#5Ns-20 - - - - + + + F: CGCCTGAAGACAATACCACT 58
R: CAACTTTTTTAGAGTGGGCA
Ph#5Ns-21 - - - - + + + F: GTTTTGTTCACATTTTGGCG 60
R: GCAACAGAGAAGGCAGAGCG
Ph#5Ns-22 - - - - + + + F: TATCCGCTATGAGAATGCCT 58
R: GCTGAGAATCTACCCAAACC
Ph#5Ns-23 - - - - + + + F: TCCGTAGAAACAAGAGGCAA 58
R: ACTCCTTACCCCCTTCAAAA
Ph#5Ns-24 + + - + + + + F: GGAGTCCGAGAGAGGAACCG 64
R: TCGTTACCCTCAAAACCCCC
Ph#5Ns-25 + + + - + + + F: GCGTTGGTGGTTCCTTTCGT 62
R: CCTACGGGAAGAGAAACGGG
Ph#5Ns-26 - - - - + - + F: TCAGGAATCTCTATCGTGGG 60
R: CGGAAACCACAATGTCACTA
Ph#5Ns-27 - - + - + - + F: CCCCCTCCCCTCTCCTATCC 62
R: CTGCTGACCTTCACTCCCCG
Ph#5Ns-28 - - - - + + + F: ATAAGAGGTCTGGCTGTGGT 58
R: AAGTAATGTAAGAAGCGGCA
Ph#5Ns-29 - - - - + - + F: CTTATTAGCGGGTCCTTGGT 62
R: TCCTCGTCTGTGGGGTAGTC
Ph#5Ns-30 - + + - + - + F: GAGTCGTCGGCACAACATCC 62
R: ATCGTGCTCCTCATCTTCGC

图4

16个特异性标记在试验材料中的凝胶电泳分析 M: DL2000; 1: 7182; 2: D4286; 3: 黑麦; 4: 长穗偃麦草; 5: 华山新麦草; 6: 滨麦; 7: 16DH25-7。箭头指示特异条带。"

附表1

小麦供试材料2022-2023和2023-2024年度农艺性状调查表"

材料
Material		 2022-2023 2023-2024
株高
Plant height (cm)		 分蘖数Tiller number 穗长Spike length (cm) 小穗数Spikelets per spike 穗粒数Kernels per spike 千粒重Thousand-
kernel weight (g)		 株高Plant height (cm) 分蘖数
Tiller number		 穗长Spike length (cm) 小穗数Spikelets per spike 穗粒数Kernels per spike 千粒重Thousand-
kernel weight (g)
7182 76 12 9.6 20 52 37.65 75 8 10.5 20 58 37.21
78 12 11.7 23 55 38.12 79 9 11.4 21 58 37.68
80 10 11.5 19 46 37.91 76 9 12.1 22 50 38.89
81 9 12.3 22 47 38.24 80 12 11.7 21 48 39.08
75 8 11.1 22 50 39.16 82 12 11.3 23 57 38.47
79 10 12.5 23 61 38.51 77 12 11.0 22 52 38.33
78 9 11.7 20 55 38.37 79 10 11.9 23 54 38.62
79 9 11.4 22 56 37.55 83 11 11.6 22 53 37.95
84 12 10.9 22 55 36.92 78 11 10.8 21 55 38.02
80 11 10.7 20 58 37.76 81 10 12.4 22 56 38.15
D4286 90 9 7.6 14 44 45.65 91 13 7.5 15 43 46.32
91 11 8.5 16 42 46.17 87 11 8.4 17 41 45.41
85 14 8.7 16 39 46.26 87 16 7.6 14 40 46.55
88 16 8.1 15 40 45.77 85 9 8.3 16 44 45.20
87 11 7.9 18 45 46.12 90 11 8.2 18 38 46.01
D4286 90 11 7.7 17 43 45.11 89 15 7.8 15 42 45.93
85 14 8.0 16 42 44.98 83 13 7.9 17 45 46.47
86 10 8.0 15 40 45.63 86 10 8.0 16 39 45.76
91 12 7.5 16 37 46.87 88 12 8.1 14 41 45.09
81 14 7.4 14 39 46.28 88 14 7.9 16 39 46.24
华山新
麦草
P. huashanica		 62 — 8.2 16 42 3.72 63 — 8.4 17 43 3.68
63 — 8.0 18 41 3.44 61 — 7.8 19 40 3.21
62 — 8.1 18 42 3.12 62 — 8.3 16 42 3.85
61 — 8.5 17 39 2.96 63 — 8.1 15 41 3.05
63 — 7.9 16 40 3.56 61 — 8.2 18 39 3.53
62 — 8.1 16 45 3.91 62 — 7.9 17 44 3.37
61 — 8.2 15 41 3.51 63 — 8.5 16 41 3.74
60 — 8.2 18 42 3.62 61 — 8.1 18 43 3.18
63 — 8.5 17 42 3.39 62 — 7.9 17 38 3.59
63 — 7.7 17 38 3.72 62 — 8.4 17 42 3.45
16DH25-7 60 18 9.7 24 60 39.21 57 24 9.4 24 55 38.63
58 24 9.5 24 57 40.43 60 22 9.0 24 60 40.88
61 19 9.6 23 55 38.76 58 22 8.6 21 56 40.56
57 24 8.7 22 56 41.65 59 19 9.6 22 58 39.72
58 22 9.1 20 59 39.82 56 18 9.2 23 58 38.97
56 20 8.9 22 61 39.16 61 20 8.7 22 59 39.94
59 19 9.2 23 57 38.55 58 23 9.3 23 57 40.21
58 21 9.3 24 56 39.84 59 20 8.9 23 57 41.10
59 20 8.8 23 55 38.92 57 21 9.5 23 56 39.58
60 17 8.9 22 56 40.61 60 21 8.8 23 58 39.25

表3

16DH25-7及其亲本的主要农艺性状比较"

材料
Material		 株高
Plant height (cm)		 分蘖数
Tiller
number		 穗长
Spike length (cm)		 小穗数
Spikelets
per spike		 穗粒数
Kernels per spike		 千粒重
Thousand-kernel weight (g)		 芒性
Awn type
7182 79.0±2.49 Bb 10.3±1.42 Cc 11.41±0.71 Bb 21.7±0.95 Bb 53.8±4.02 Bb 38.13±0.58 Dd 长芒Long
D4286 87.4±2.76 Aa 12.3±2.15 Bb 7.96±0.35 Dd 15.8±1.32 Ee 41.2±2.35 Cc 45.94±0.53 Bb 长芒Long
华山新麦草
P. huashanica		 62.0±0.92 Dd 簇Clump 8.15±0.24 Dd 17.0±1.15 Dd 41.3±1.86 Cc 3.48±0.27 Ee 短芒Short
16DH25-7 58.6±1.50 Ee 20.7±2.08 Aa 9.14±0.34 Cc 22.8±0.92 Aa 57.3±1.78 Aa 39.79±0.88 Cc 长芒Long

图5

16DH25-7及其亲本的农艺性状和白粉病、条锈病抗性鉴定表型 a: 株高; b: 穗长; c: 小穗; d: 粒宽; e: 粒长; f: 白粉病抗性表型; g: 条锈病抗性表型。1: 7182; 2: D4286; 3: 华山新麦草; 4: 16DH25-7; 5: 陕优225; 6: 辉县红。"

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