滇杨性别连锁的SSR标记

李启少; 雷瀚; 沈伟祥; 赵文植; 王飞; 徐剑; 瞿绍宏; 董章宏; 辛培尧

doi:10.19303/j.issn.1008-0384.2023.08.009

滇杨性别连锁的SSR标记

doi: 10.19303/j.issn.1008-0384.2023.08.009

李启少^{1, 2,},
雷瀚³,
沈伟祥^{1, 2},
赵文植^{1, 2},
王飞^{1, 2},
徐剑²,
瞿绍宏⁴,
董章宏^{1, 2},
辛培尧^{1, 2, ,}

1.
西南林业大学国家林业和草原局西南风景园林工程技术研究中心，云南　昆明，650224
2.
西南林业大学西南地区生物多样性保育国家林业和草原局重点实验室，云南　昆明，650224
3.
北京市农业农村部科技发展中心，北京　102600
4.
开远市白土墙国有林场，云南　开远　661600

基金项目: 云南省科技人才与平台计划项目（202205AF150022）

详细信息

作者简介:
李启少（1999 —），男，硕士研究生，主要从事林木遗传育种与良种繁育研究，E-mail：975025664@qq.com

通讯作者:
辛培尧（1975 —），男，博士，教授，博士生导师，主要从事植物遗传育种与分子生物学研究，E-mail：xpytgyx@163.com

中图分类号: S792
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出版历程
- 收稿日期: 2023-01-12
- 修回日期: 2023-06-26
- 网络出版日期: 2023-08-16
- 刊出日期: 2023-08-28

SSR Markers Associated with Sex-difference of Populus yunnanensis Dode

LI Qishao^{1, 2
,},
LEI Han³,
SHEN Weixiang^{1, 2},
ZHAO Wenzhi^{1, 2},
WANG Fei^{1, 2},
XU Jian²,
QU Shaohong⁴,
DONG Zhanghong^{1, 2},
XIN Peiyao^{1, 2
, ,}

1.
Southwest Research Center for Landscape Architecture Engineering of State Forestry and Grassland Administration, Southwest Forestry University, Kunming,Yunnan　650224, China
2.
Key Laboratory of National Forestry and Grassland Administration on Biodiversity Conservation in Southwest China, Southwest Forestry University, Kunming, Yunnan　650224, China
3.
Beijing Science and Technology Development Center of Ministry of Agriculture and Rural Affairs, Beijing　102600, China
4.
Kaiyuan Baituqiang National Forest Farm, Kaiyuan, Yunnan　661600, China

摘要

摘要: 目的在滇杨幼苗时期和开花前利用形态差异对雌雄株进行性别鉴定极为困难，因此亟需探明对其性别进行快速鉴定的分子标记方法。方法以滇杨雌雄各30株为材料，利用简单重复序列（SSR）筛选与滇杨性别相关的分子标记，并利用所获得的SSR标记对未知性别滇杨幼苗进行鉴定，分析滇杨苗期的性别分化情况。结果从已报道的相关文献中选出15对与植物性别连锁的SSR引物，经试验筛选出1对（BPCA90）在琼脂糖凝胶检测中显现清晰条带且反应稳定的SSR引物，并将这对引物经聚丙烯酰胺凝胶电泳进行复筛，经PCR扩增后在滇杨雄株中出现1条特异性条带，大小在680 bp左右。利用这对SSR引物对30株滇杨幼苗进行性别鉴定，结果表明雄株明显多于雌株，且比例为13∶2。结论试验筛选所得到的SSR引物可以用于滇杨早期进行性别鉴定。研究结果可作为滇杨早期性别鉴定的遗传标记，为今后滇杨在生产中的分性别利用提供技术支持。
- 滇杨 /
- SSR /
- 引物筛选 /
- 性别鉴定
Abstract: Objective Means utilizing molecular markers to differentiate rapidly and accurately male and female Populus yunnanensis plants at young stages was investigated. Method Using simple sequence repeats (SSR), molecular markers were screened for those might associate with the sex of P. yunnanensis at seedling stages and before flowering. Result Fifteen pairs of SSR primers were selected from the reported literatures. One of them, coded BPCA90, showed clear bands and stable reaction in agarose gel detection. After rescreening the primer by polyacrylamide gel electrophoresis, a specific band of approximately 680 bp appeared in the PCR amplification of male P. yunnanensis. Subsequent testing on 30 plants using these SSR primers showed significantly more males than females in a ratio of 13 to 2. Conclusion Short of a visible morphological sign for the determination, the SSR primers screened by this study appeared to suffice the general purpose of differentiating the sex of P. yunnanensis at early growth stage.
- Populus yunnanensis Dode /
- SSR /
- primer screening /
- gender identification

HTML全文

图 1 部分样品DNA质量检测

Figure 1. Determination on DNA quality of some specimens

下载: 全尺寸图片幻灯片

图 2 SSR引物BPCA90琼脂糖凝胶检测结果

Figure 2. Agarose gel test on SSR primer, BPCA90

下载: 全尺寸图片幻灯片

图 3 SSR引物BPTGG82琼脂糖凝胶检测结果

Figure 3. Agarose gel test on SSR primer, BPTGG82

下载: 全尺寸图片幻灯片

图 4 SSR引物BPCA90与样品扩增产物的变性聚丙烯酰胺凝胶检测图

Figure 4. Modified polyacrylamide gel detection on amplification products of BPCA90 and specimens

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图 5 SSR引物BPCA90检测未知性别滇杨

Figure 5. Detection on P. yunnanensis of unknown sex using SSR primer, BPCA90

下载: 全尺寸图片幻灯片

表 1 滇杨样品采集信息

Table 1. Specimen collection of P. yunnanensis

编号 Code	性别 Sex of the individual	采集地 Location
1-10	雄性（♂）	香格里拉
11-20	雄性（♂）	大理
21-25	雄性（♂）	禄劝
25-30	雄性（♂）	昆明
1-15	雌性（♀）	香格里拉
16-28	雌性（♀）	大理
29-30	雌性（♀）	禄劝

下载: 导出CSV

表 2 SSR扩增引物信息

Table 2. Primers for SSR amplification

引物名称 Locus name	引物序列 Primer sequence(5′-3′)	退火温度 Ta(annealing temperature)/℃	参考文献 Reference
Bmsat95	F:ATTGTAACCGATTTGAGAGA R:ATTCGCACAATAAGTTCACT	54.0	NAGARAJA^[19]
Bmsat153	F:TGCTGTCGTCTGCTTCCTAA R:CACGGTGCTGACTGTTGTTT	60.0	NAGARAJA^[19]
Bmsat155	F:AGGGATGATGGGTAAAGAGC R:GCAGTAGGCATTTGGAAGGAG	60.0	NAGARAJA^[19]
Bmsat156	F:CTCCTTATCCATCCGTTT R:CTCTCGGATCATAGATACG	56.0	NAGARAJA^[19]
Bmsat159	F:ATCTGGTGCTCAAAAACGGA R:CGGAACCAAACAAGAACGAT	58.0	NAGARAJA^[19]
Bmsat208	F:ACATGAAATGGGCAAACGACG R:GCTCATATTTGCTTGCCGGTT	55.8	NAGARAJA^[19]
SSR-8	F:TGCATATCGTCCAGGGTGTTTTC R:CCCAGCCAGGGTCTTGTTTC	64.0	SHAN^[20]
BPCA90	F:CCTAGCCTTCATTCTCATTCAGC R:GGTTGCTAGTCAGCTTCTTACC	58.4	PAKULL B^[21]
BPTTG60	F:CAAGAACTCAGACATGATCAGATC R:CTTTGCACGTTAATAAGGAGACTG	56.9	PAKULL B^[21]
BPTGG82	F:CTTGAAGAGCGAAAACTCAGCAG R:CTCTAAATCCAAGGTTGGTTACC	58.4	PAKULL B^[21]
BPTG50	F:CTAGCTGTCGAACGTAATTGGCAC R:CTTGCACGAGCCTCCATCACTC	61.8	PAKULL B^[21]
BPTC66	F:GGCCGTGATTCTCATCATGATG R:CTTCTTCATCAACAACTGCGTCC	58.4	PAKULL B^[21]
CADEx2/Intr2	F:CAAGGTTATCAGCTGTGG R:ATATTGTGTGCGTGTTCG	54.0	PAKULL B^[22]
HIAGA7	F:ACAAGCAGTAATGATGAGGA R:TCCAAGTCTCTCAATTAGGAA	56.0	JAKSE^[23]
35	F:GAGGCCGACAGGATCGTAC R:TACGACGTACTCCGGTGGTTTT	62.0	CHEN^[24]

下载: 导出CSV

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