物种名 Species | 编号 No. | 材料 Material | 来源国家 Country of origin |
苜蓿菟丝子 Cuscuta approximata | 1 | “至仁同济”苜蓿菟丝子 Zhiren Tongji dodder | 中国 China |
2 | 榆中苜蓿菟丝子 Yuzhong dodder | 中国 China | |
3 | “惠仁堂”苜蓿菟丝子 “Huirentang” dodder | 中国 China | |
紫花苜蓿 Medicago sativa | 1 | 中苜2号 Zhongmu No.2 | 中国 China |
2 | 陇东3号 Longdong No.3 | 中国 China | |
3 | 陇东苜蓿 Longdong | 中国 China | |
4 | 瑞西斯 Resistador | 丹麦 Denmark | |
5 | 巨人4号 Ameristand No.4 | 美国 America | |
6 | 阿尔冈金 Algonguin | 加拿大 Canada | |
7 | 巨人3号 | 美国 America | |
8 | 金皇后 Golden-queen | 美国 America |

Citation: WU Y G, ZHOU Q, JIA C L, LIU Z P. A universal molecular method for rapid identification of alfalfa and dodder seeds. Pratacultural Science, 2022, 39(7): 1-8 doi:

快速鉴定紫花苜蓿混入苜蓿菟丝子的分子方法及其通用性
English
A universal molecular method for rapid identification of alfalfa and dodder seeds
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凯时66
图 1 紫花苜蓿和苜蓿菟丝子叶绿体DNA部分rbcL基因区域的序列比对
Figure 1. Sequence alignments of a portion of the rbcL regions of alfalfa and dodder chloroplast DNA
水平黑色箭头表示用于PCR扩增的引物位置。垂直的黑色箭头表示在引物位置处由单核苷酸多态性 (single-nucleotide polymorphisms, SNPs)和小于50 bp的片段插入或缺失(insertions and deletions, InDels)引起的序列差异。
The horizontal black arrows indicate the locations of the primers used for PCR amplification. The vertical black arrows indicate sequence differences resulting from SNPs (single-nucleotide polymorphisms) and InDels (insertions and deletions) at primer locations.
图 2 紫花苜蓿和苜蓿菟丝子种子
Figure 2.
Seeds of Medicago sativa and Cuscuta approximata 红色箭头表示在形态上和菟丝子种子相似的紫花苜蓿种子。
The red arrows indicate alfalfa seeds that are morphologically similar to dodder seeds.
图 3 寄生在紫花苜蓿上的菟丝子
Figure 3. Cuscuta approximata twining around Medicago sativa
A: 菟丝子和紫花苜蓿建立的寄生系统,红色箭头表示菟丝子吸器;B: 成熟后的菟丝子种子和紫花苜蓿种子混杂在一起。
A: The parasitic system established between dodder and alfalfa. The red arrow indicates the haustorium of dodder, Scale bar = 5 mm; B: The intermingled mature seeds of dodder and alfalfa.
图 4 苜蓿菟丝子和紫花苜蓿种质间的电泳结果
Figure 4. The electrophoresis results obtained for dodder and alfalfa accessions
苜蓿菟丝子的1 − 3编号表示3种不同来源的苜蓿菟丝子种子(编号1 − 3,同表1),紫花苜蓿的1 − 8编号表示紫花苜蓿的8个不同种质(编号1 − 8,同表1)。M: 2 000 DNA marker; H2O:负对照。
Lanes 1 − 3: dodder (Nos. 1 − 3, Table 1), dodder seed from three different habitats; Lanes 1 − 8: alfalfa (Nos. 1 − 8, table 1), eight alfalfa accessions. M: 2 000 DNA marker; H2O: negative control.
图 5 苜蓿菟丝子种子和紫花苜蓿种子不同比例DNA混合物的电泳结果
Figure 5. The electrophoresis results obtained for mixtures of dodder and alfalfa seed DNA
A:紫花苜蓿种子中混有不同比例苜蓿菟丝子种子的电泳结果;B:苜蓿菟丝子种子和紫花苜蓿种子DNA比例为1 ꞉ 5 000的二次PCR扩增电泳结果;C:苜蓿菟丝子种子和紫花苜蓿种子DNA比例为1 ꞉ 10 000的二次PCR扩增电泳结果。M: 2 000 DNA marker; H2O:负对照。
A: The electrophoresis results obtained for alfalfa seeds mixed with dodder seeds in different proportions; B: The DNA ratio of dodder and alfalfa seed was 1 ꞉ 5 000 for the second PCR amplification; C: The DNA ratio of dodder and alfalfa seed was 1 ꞉ 10 000 for the second PCR amplification. M: 2 000 DNA marker; H2O: negative control.
图 6 苜蓿菟丝子和紫花苜蓿不同比例种子粒数混合的电泳结果
Figure 6. The electrophoresis results obtained for different numbers of dodder seeds mixed with alfalfa seeds
紫花苜蓿种子中混有不同粒数苜蓿菟丝子种子的电泳结果。M: 2 000 DNA marker; H2O:负对照。
The electrophoresis results obtained for alfalfa seeds mixed with different number of dodder seeds. M: 2 000 DNA marker; H2O: negative control.
图 7 鉴定种子形态相似牧草的总结流程图
Figure 7. A summary flowchart for the discrimination of forage seeds with similar morphologies
右侧为该流程的时间轴,可同时进行PCR和琼脂糖胶的制备,整个实验流程可在大约5 h完成。
The time required is shown on the right, with polymerase chain reaction (PCR) and agarose gel preparation being performed simultaneously, and the total protocol can be completed in approximately 5 h.
表 1 紫花苜蓿和苜蓿菟丝子种质资源信息
Table 1. Germplasm accessions of Medicago sativa and Cuscuta approximata used in this study
下载: 导出CSV
表 2 8份紫花苜蓿种质和3份不同产地苜蓿菟丝子的4项种子形态指标
Table 2. Statistical analyses of four morphological indices between the seeds of eight Medicago sativa accessions and Cuscuta approximata from different habitats
指标
Parameter项目
Item苜蓿菟丝子
Cuscuta
approximata紫花苜蓿
Medicago
sativa种子长度
Seed length/mm变幅 Range 1.3~1.8 1.7~2.1 均值 Mean 1.4 ± 0.14 1.2 ± 0.13 种子宽度
Seed width/mm变幅 Range 1.0~1.4 0.9~1.3 均值 Mean 1.2 ± 0.13 1.9 ± 0.09 种子周长
Seed perimeter/mm变幅 Range 3.6~4.8 4.4~5.4 均值 Mean 4.1 ± 0.34 5.0 ± 0.26 种子面积
Seed area/mm2变幅 Range 1.0~1.6 1.3~2.0 均值 Mean 1.3 ± 0.20 1.7 ± 0.17 n = 30. 下载: 导出CSV
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