草业学报 ›› 2025, Vol. 34 ›› Issue (11): 114-124.DOI: 10.11686/cyxb2024515
• 研究论文 • 上一篇
张献芳(
), 聂刚, 黄思源, 余帅, 左粟田, 张新全(
)
收稿日期:2024-12-27
修回日期:2025-03-10
出版日期:2025-11-20
发布日期:2025-10-09
通讯作者:
张新全
作者简介:E-mail: zhangxq@sicau.edu.cn基金资助:
Xian-fang ZHANG(
), Gang NIE, Si-yuan HUANG, Shuai YU, Su-tian ZUO, Xin-quan ZHANG(
)
Received:2024-12-27
Revised:2025-03-10
Online:2025-11-20
Published:2025-10-09
Contact:
Xin-quan ZHANG
摘要:
采用传统杂交方法,以象草品种“GM1”为母本、新品系“F-B”为父本进行杂交,获得536个杂种后代,并从中选取150个有代表性的F1代株系群体进行变异分析。本研究结合表型性状和SSR分子标记,对这些后代进行杂种鉴定及表型变异分析,旨在为象草种质创新和品种改良提供技术支撑。结果表明:1)F1代群体较亲本表型变异丰富,其中叶长明显强于亲本,具有明显的杂种优势,性状存在显著差异,变异系数为5.90%~21.05%;2)筛选出的14对SSR引物共扩增得到219条清晰条带,其中多态性条带118条,多态性比率(PPB)54.18%;每个SSR位点的多态信息含量(PIC)为29.59%~44.63%,均值为35.40%;3)对亲本及杂交F1代进行聚类分析可知,基于表型数据的聚类分析将杂交后代分为3类;基于SSR分子标记的聚类分析将杂交后代分为4类,SSR分子标记进一步证实了母本在遗传上的主导地位;4)利用特异性引物可以准确鉴定出真杂种,鉴定率高达98%,大部分真杂种条带为双亲互补型,部分为父本型,表明F1代中存在父母本的遗传信息。通过对150个F1代株系群体的表型性状和筛选出的15对SSR核心引物分析,能有效鉴定杂种的真实性,为后续杂交育种选育新品系提供后备材料。
张献芳, 聂刚, 黄思源, 余帅, 左粟田, 张新全. 基于SSR标记的象草F1代杂种分子鉴定及表型变异分析[J]. 草业学报, 2025, 34(11): 114-124.
Xian-fang ZHANG, Gang NIE, Si-yuan HUANG, Shuai YU, Su-tian ZUO, Xin-quan ZHANG. Identification of hybrids and analysis of phenotypic variation in the F1 generation of Pennisetum purpureum based on SSR markers[J]. Acta Prataculturae Sinica, 2025, 34(11): 114-124.
| 亲本Parents | 材料来源Materials origin | 特征Characteristic |
|---|---|---|
| GM1 ♀ | 广西壮族自治区畜牧研究所Guangxi Zhuang Autonomous Region Institute of Animal Husbandry | 产量高,叶量丰富;披散型;被毛稀疏High yield, abundant leaf quantity; loose type; sparse coat |
| F-B ♂ | 四川农业大学Sichuan Agricultural University | 叶直立,分蘖性强;紧凑型;越冬性好,花粉活力强Leaf upright, strong tillering; compact; good overwintering, strong pollen activity |
表1 亲本材料相关信息
Table 1 Relevant information of parent materials
| 亲本Parents | 材料来源Materials origin | 特征Characteristic |
|---|---|---|
| GM1 ♀ | 广西壮族自治区畜牧研究所Guangxi Zhuang Autonomous Region Institute of Animal Husbandry | 产量高,叶量丰富;披散型;被毛稀疏High yield, abundant leaf quantity; loose type; sparse coat |
| F-B ♂ | 四川农业大学Sichuan Agricultural University | 叶直立,分蘖性强;紧凑型;越冬性好,花粉活力强Leaf upright, strong tillering; compact; good overwintering, strong pollen activity |
引物名称 Primer name | 上游引物 Forward primer (5'-3') | 下游引物 Reverse primer (5'-3') | 退火温度 Annealing temperature (Tm, ℃) |
|---|---|---|---|
| SSR4 | CATGCATATATGACTTTTCCCCAA | ATGATAAATAGGATAGCCATGCCG | 58 |
| SSR9 | CGTACCTTCTCGCAATAAGGAGAT | GAAGAAGGTTCACCTCTTTCGGTA | 58 |
| SSR23 | AGGTTCTGCTTCTGGAGGTGAC | TAGATTTGTTCATGATGATGCGTG | 58 |
| SSR40 | AAGCTCGATTTTGAGCATCTCC | GATCTGGACCACTGATCAACAGAC | 58 |
| SSR55 | AAGTCCACATCATCCCGGTC | GTGGCTTACCTGATCCGAGC | 58 |
| PaSSR3 | TCCATGGAGTACCCGAAGAG | ACATCAACCACTGCAACCAA | 50 |
| PaSSR9 | TGGATGGAGGACAGTGATGA | ACGACCAGGAAAGCCTTACA | 50 |
| PaSSR14 | GTCCACGAGAGAGGGAAGAG | GTAGCATATCCCGCCTGTGT | 50 |
| PaSSR23 | CAGCAAATGCAGCCTATCAA | CTGTTGGTCACTGGTCCCTT | 50 |
| PaSSR34 | TGCAGAGAAACCAATTGCAG | CCGGTTCATAAGCTGGTGTT | 50 |
| WSSR31 | TGTGTTTTTCATCCTCAAGTGTTGT | CAAATGTAAGATGATAAATGCAGCG | 58 |
| WSSR48 | CTTGTTGTCCAGGTCGAGACAC | ACGACTAGGTGCTTAGCTAGGGG | 58 |
| HSSR4 | GTAACTGGTTCTCCTGCTTCT | ACTAGGTTACTGCTGCTTGTG | 56 |
| C106112-g1 | CCAAGCGCTCCTTCTTCTGA | CACGGCACTGATCACTCTGT | 60 |
表2 供试引物信息
Table 2 Primer information of this test
引物名称 Primer name | 上游引物 Forward primer (5'-3') | 下游引物 Reverse primer (5'-3') | 退火温度 Annealing temperature (Tm, ℃) |
|---|---|---|---|
| SSR4 | CATGCATATATGACTTTTCCCCAA | ATGATAAATAGGATAGCCATGCCG | 58 |
| SSR9 | CGTACCTTCTCGCAATAAGGAGAT | GAAGAAGGTTCACCTCTTTCGGTA | 58 |
| SSR23 | AGGTTCTGCTTCTGGAGGTGAC | TAGATTTGTTCATGATGATGCGTG | 58 |
| SSR40 | AAGCTCGATTTTGAGCATCTCC | GATCTGGACCACTGATCAACAGAC | 58 |
| SSR55 | AAGTCCACATCATCCCGGTC | GTGGCTTACCTGATCCGAGC | 58 |
| PaSSR3 | TCCATGGAGTACCCGAAGAG | ACATCAACCACTGCAACCAA | 50 |
| PaSSR9 | TGGATGGAGGACAGTGATGA | ACGACCAGGAAAGCCTTACA | 50 |
| PaSSR14 | GTCCACGAGAGAGGGAAGAG | GTAGCATATCCCGCCTGTGT | 50 |
| PaSSR23 | CAGCAAATGCAGCCTATCAA | CTGTTGGTCACTGGTCCCTT | 50 |
| PaSSR34 | TGCAGAGAAACCAATTGCAG | CCGGTTCATAAGCTGGTGTT | 50 |
| WSSR31 | TGTGTTTTTCATCCTCAAGTGTTGT | CAAATGTAAGATGATAAATGCAGCG | 58 |
| WSSR48 | CTTGTTGTCCAGGTCGAGACAC | ACGACTAGGTGCTTAGCTAGGGG | 58 |
| HSSR4 | GTAACTGGTTCTCCTGCTTCT | ACTAGGTTACTGCTGCTTGTG | 56 |
| C106112-g1 | CCAAGCGCTCCTTCTTCTGA | CACGGCACTGATCACTCTGT | 60 |
性状 Traits | 平均值 Average | 范围 Range | 变异系数 Coefficient of variation (CV, %) | 多样性指数 Shannon-Wiener (H′) | F值 F value | ||
|---|---|---|---|---|---|---|---|
| 母本Female line (GM1) | 父本Male line (F-B) | F1代 F1 generation | |||||
| 株高Plant height (PH, cm) | 297.04±5.40 | 396.34±10.86 | 334.93±22.49 | 276.78~396.92 | 6.71 | 1.841 | 8.422** |
| 叶片数Number of leaves (NL, No.) | 16.00±1.58 | 24.00±2.86 | 19.88±1.58 | 16.40~24.00 | 7.95 | 1.615 | 6.146** |
| 茎节数Number of stem segments (NS, No.) | 10.00±0.71 | 15.00±1.14 | 13.30±1.66 | 9.00~25.00 | 12.39 | 1.604 | 9.722** |
| 叶长Leaf length (LL, cm) | 110.06±5.23 | 108.58±4.65 | 111.00±6.55 | 94.34~129.78 | 5.90 | 1.808 | 9.075** |
| 叶宽Leaf width (LW, mm) | 38.03±2.36 | 40.08±2.48 | 36.19±5.96 | 23.31~62.19 | 16.45 | 1.764 | 14.456** |
| 茎粗Stem diameter (SD, mm) | 11.30±0.89 | 17.05±0.26 | 13.70±2.89 | 9.20~26.65 | 21.05 | 1.661 | 14.758** |
| 节间长Internode length (IL, cm) | 14.42±1.02 | 16.22±1.67 | 15.13±1.45 | 10.41~18.62 | 9.60 | 1.826 | 4.969** |
| 叶绿素Chlorophyll (CHI, spad) | 49.36±3.33 | 58.84±0.95 | 45.64±5.06 | 24.15~58.56 | 11.05 | 1.824 | 4.709** |
表3 象草亲本及杂交F1代群体农艺性状的变异性分析
Table 3 Variability analysis of agronomic traits of parent and hybrid F1 generation population of P. purpureum
性状 Traits | 平均值 Average | 范围 Range | 变异系数 Coefficient of variation (CV, %) | 多样性指数 Shannon-Wiener (H′) | F值 F value | ||
|---|---|---|---|---|---|---|---|
| 母本Female line (GM1) | 父本Male line (F-B) | F1代 F1 generation | |||||
| 株高Plant height (PH, cm) | 297.04±5.40 | 396.34±10.86 | 334.93±22.49 | 276.78~396.92 | 6.71 | 1.841 | 8.422** |
| 叶片数Number of leaves (NL, No.) | 16.00±1.58 | 24.00±2.86 | 19.88±1.58 | 16.40~24.00 | 7.95 | 1.615 | 6.146** |
| 茎节数Number of stem segments (NS, No.) | 10.00±0.71 | 15.00±1.14 | 13.30±1.66 | 9.00~25.00 | 12.39 | 1.604 | 9.722** |
| 叶长Leaf length (LL, cm) | 110.06±5.23 | 108.58±4.65 | 111.00±6.55 | 94.34~129.78 | 5.90 | 1.808 | 9.075** |
| 叶宽Leaf width (LW, mm) | 38.03±2.36 | 40.08±2.48 | 36.19±5.96 | 23.31~62.19 | 16.45 | 1.764 | 14.456** |
| 茎粗Stem diameter (SD, mm) | 11.30±0.89 | 17.05±0.26 | 13.70±2.89 | 9.20~26.65 | 21.05 | 1.661 | 14.758** |
| 节间长Internode length (IL, cm) | 14.42±1.02 | 16.22±1.67 | 15.13±1.45 | 10.41~18.62 | 9.60 | 1.826 | 4.969** |
| 叶绿素Chlorophyll (CHI, spad) | 49.36±3.33 | 58.84±0.95 | 45.64±5.06 | 24.15~58.56 | 11.05 | 1.824 | 4.709** |
图1 基于表型性状的聚类分析图中GM1♀表示母本,F-B♂表示父本,其余数字表示采样编号,下同。In the figure, GM1♀ represents the female parent, F-B♂ represents the male parent, and the remaining numbers indicate the sample numbers. The same below.
Fig.1 Cluster analysis based on phenotypic traits
引物名称 Primer name | 扩增总条带数Total number of amplified bands | 多态性条带数Number of polymorphic bands | 多态条带比率Polymorphism band ratio (PPB, %) | 多态信息含量Polymorphism information content (PIC, %) |
|---|---|---|---|---|
| SSR4 | 13 | 6 | 46.15 | 36.07 |
| SSR9 | 10 | 8 | 80.00 | 44.63 |
| SSR23 | 9 | 5 | 55.56 | 30.47 |
| SSR40 | 21 | 15 | 71.43 | 41.48 |
| SSR55 | 7 | 1 | 14.29 | 29.93 |
| PaSSR-3 | 17 | 12 | 70.59 | 36.15 |
| PaSSR-9 | 17 | 6 | 35.29 | 38.89 |
| PaSSR-14 | 19 | 11 | 57.89 | 29.66 |
| PaSSR-23 | 9 | 3 | 33.33 | 29.59 |
| PaSSR-34 | 24 | 7 | 29.17 | 36.37 |
| C106112-g1 | 11 | 9 | 81.82 | 32.46 |
| WSSR31 | 14 | 9 | 64.29 | 36.92 |
| WSSR48 | 16 | 12 | 75.00 | 39.49 |
| HSSR4 | 32 | 14 | 43.75 | 33.52 |
| 总计Total | 219 | 118 | / | / |
| 平均Average | 15.64 | 8.43 | 54.18 | 35.40 |
表4 象草SSR条带的多样性
Table 4 Polymorphism of SSR bands in P. purpureum
引物名称 Primer name | 扩增总条带数Total number of amplified bands | 多态性条带数Number of polymorphic bands | 多态条带比率Polymorphism band ratio (PPB, %) | 多态信息含量Polymorphism information content (PIC, %) |
|---|---|---|---|---|
| SSR4 | 13 | 6 | 46.15 | 36.07 |
| SSR9 | 10 | 8 | 80.00 | 44.63 |
| SSR23 | 9 | 5 | 55.56 | 30.47 |
| SSR40 | 21 | 15 | 71.43 | 41.48 |
| SSR55 | 7 | 1 | 14.29 | 29.93 |
| PaSSR-3 | 17 | 12 | 70.59 | 36.15 |
| PaSSR-9 | 17 | 6 | 35.29 | 38.89 |
| PaSSR-14 | 19 | 11 | 57.89 | 29.66 |
| PaSSR-23 | 9 | 3 | 33.33 | 29.59 |
| PaSSR-34 | 24 | 7 | 29.17 | 36.37 |
| C106112-g1 | 11 | 9 | 81.82 | 32.46 |
| WSSR31 | 14 | 9 | 64.29 | 36.92 |
| WSSR48 | 16 | 12 | 75.00 | 39.49 |
| HSSR4 | 32 | 14 | 43.75 | 33.52 |
| 总计Total | 219 | 118 | / | / |
| 平均Average | 15.64 | 8.43 | 54.18 | 35.40 |
图2 引物的SSR图谱类型A: 双亲互补型Complementary pattern of both parents, C106112-g1; B: 父本型Male-specific pattern, HSSR4; C: 母本型Female-specific pattern, WSSR31; D: 其他型Other types, PaSSR-34. ♀: 母本Female parent; ♂: 父本Male parent; F1: 杂交子代Hybrid offspring.
Fig.2 SSR fingerprint pattern
图4 引物WSSR31对部分杂交后代的扩增M: 2000 bp Marker; 箭头所指条带为父本特征带The strip pointed by the arrow is the male parent feature strip; “+”所标注的条带为真杂种Bands marked with ‘+’ are true hybrids. 下同The same below.
Fig.4 Amplification of some hybrid progeny by primer WSSR31
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