Identification of SSR marker linked to gynoecious loci in cucumber (Cucumis sativus L.)
Gynoecy plays an important role in cucumber (Cucumis sativus L.) heterosis breeding and identification of the markers linked to this character will facilitate selection of gynoecious cucumber line in breeding program. Traditional selection for cucumber cultivars with gynoecious line has required eva...
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Zhejiang University Press
2013-05-01
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| Series: | 浙江大学学报. 农业与生命科学版 |
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| Online Access: | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2012.11.141 |
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| author | ZHOU Shengjun ZHANG Peng ZHU Yuqiang CHEN Xinjuan CHEN Liping |
| author_facet | ZHOU Shengjun ZHANG Peng ZHU Yuqiang CHEN Xinjuan CHEN Liping |
| author_sort | ZHOU Shengjun |
| collection | DOAJ |
| description | Gynoecy plays an important role in cucumber (Cucumis sativus L.) heterosis breeding and identification of the markers linked to this character will facilitate selection of gynoecious cucumber line in breeding program. Traditional selection for cucumber cultivars with gynoecious line has required evaluation in complicated environments over several years, which is long period, time and labor consuming. Molecular markers offer a faster and more accurate way for breeding, as selection can be based on genotype rather than phenotype. The use of molecular markers for indirect selection of important agronomic characters, or marker-assisted selection (MAS) can improve the efficiency of traditional breeding. Many studies developed a lot of SSR markers, which had greatly facilitated MAS in cucumber breeding. Now some studies showed that some markers were connected with gynoecious gene but the distances were not compact, so few were availably applied to breeding.The aim of this study was genetic analysis of gynoecy and identification of molecular marker associated with gynoecious gene using gynoecious line, monoecious line, and SSR marker.The genetic analysis of cucumber gynoecious was evaluated with a gynoecious line 240-1-2-2-3-1, monoecious line 3-5-1-3-2-1-1-1-1-2 and their F<sub>1</sub>, F<sub>2</sub>, BC<sub>1</sub> P<sub>1</sub>, BC<sub>1</sub> P<sub>2</sub> populations in the present study. Total DNA of parents and F<sub>2</sub> were isolated from freeze-dried leaf tissue by the CTAB method. SSR markers were analyzed with gynoecious line 240-1-2-2-3-1, monoecious line 3-5-1-3-2-1-1-1-1-2, F<sub>2</sub>, and 699 pairs of SSR primers. SSR analysis was performed with the primers. PCR was performed in 20 µL reaction containing 2 μL genomic DNA (20 ng), 10 pmol/μL primers 0.4 μL, respectively, 2.5 mmol/L Mg<sup>2+</sup>2 μL, 2 mmol/L dNTP 1 μL, 5 U/μL Taq DNA polymerase enzyme 0.1 µL, 10×buffer 2 µL and double distilled water. The amplification profiles were 5 min at 94 ℃, followed by 35 cycles of 30 s at 94 ℃; 1 min at 55 ℃, 1 min at 72 ℃; then 10 min at 72 ℃. After amplification, the PCR products were mixed with loading buffer (2.5 mg/mL bromophenol blue, 2.5 mg/mL diphenylamine blue, 10 mmol/L EDTA, 95% (V/V) formamide), denatured for 5 min at 94℃ and put on ice for 5 min. The denatured PCR products were separated on 6%(W/V) denaturing polyacrylamide gel at 100 W power and visualized by silver straining. Polymorphic fragments of primers were cloned and sequenced. Linkage analysis used the software of Mapmaker V3.0.During analysing the separated rate of F<sub>1</sub> and F<sub>2</sub>, the results showed that the gynoecy in 240 -1-2-2 - 3 -1 was controlled by oligogene with some background genes modified. Inheritance of gynoecy was accord with the additive-dominant-epistatic model. From 699 pairs of SSR primer, two pair of stable SSR markers (CSWCT25 and SSR18956), 331 bp and 145 bp in bands size were obtained respectively during PCR products of two SSR markers cloned and sequenced, and linkage analysis indicated that its genetic distance to the gynoecious loci was 7.7 cM and 6.8 cM, respectively. Two SSR markers are tightly linked to gynoecious loci on the chromosome 6.In sum, knowledge of location of gynoecious gene in cucumber and its related traits in crosses will be helpful to the design of more effective selection schemes to develop gynoecious cucumber genotypes. Progress in breeding gynoecious lines is still slow because of the complex inheritance of this character. However, gynoecious cultivars and the markers identified in this study can contribute to improving gynoecious line. Two SSR markers could be used effectively for molecular marker-assisted selection in breeding programs to develop cucumber gynoecious line breeding. |
| format | Article |
| id | doaj-art-d52309f5c9a241e991186a4094ce71e4 |
| institution | Kabale University |
| issn | 1008-9209 2097-5155 |
| language | English |
| publishDate | 2013-05-01 |
| publisher | Zhejiang University Press |
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| series | 浙江大学学报. 农业与生命科学版 |
| spelling | doaj-art-d52309f5c9a241e991186a4094ce71e42025-08-20T03:58:18ZengZhejiang University Press浙江大学学报. 农业与生命科学版1008-92092097-51552013-05-013929129810.3785/j.issn.1008-9209.2012.11.14110089209Identification of SSR marker linked to gynoecious loci in cucumber (Cucumis sativus L.)ZHOU ShengjunZHANG PengZHU YuqiangCHEN XinjuanCHEN LipingGynoecy plays an important role in cucumber (Cucumis sativus L.) heterosis breeding and identification of the markers linked to this character will facilitate selection of gynoecious cucumber line in breeding program. Traditional selection for cucumber cultivars with gynoecious line has required evaluation in complicated environments over several years, which is long period, time and labor consuming. Molecular markers offer a faster and more accurate way for breeding, as selection can be based on genotype rather than phenotype. The use of molecular markers for indirect selection of important agronomic characters, or marker-assisted selection (MAS) can improve the efficiency of traditional breeding. Many studies developed a lot of SSR markers, which had greatly facilitated MAS in cucumber breeding. Now some studies showed that some markers were connected with gynoecious gene but the distances were not compact, so few were availably applied to breeding.The aim of this study was genetic analysis of gynoecy and identification of molecular marker associated with gynoecious gene using gynoecious line, monoecious line, and SSR marker.The genetic analysis of cucumber gynoecious was evaluated with a gynoecious line 240-1-2-2-3-1, monoecious line 3-5-1-3-2-1-1-1-1-2 and their F<sub>1</sub>, F<sub>2</sub>, BC<sub>1</sub> P<sub>1</sub>, BC<sub>1</sub> P<sub>2</sub> populations in the present study. Total DNA of parents and F<sub>2</sub> were isolated from freeze-dried leaf tissue by the CTAB method. SSR markers were analyzed with gynoecious line 240-1-2-2-3-1, monoecious line 3-5-1-3-2-1-1-1-1-2, F<sub>2</sub>, and 699 pairs of SSR primers. SSR analysis was performed with the primers. PCR was performed in 20 µL reaction containing 2 μL genomic DNA (20 ng), 10 pmol/μL primers 0.4 μL, respectively, 2.5 mmol/L Mg<sup>2+</sup>2 μL, 2 mmol/L dNTP 1 μL, 5 U/μL Taq DNA polymerase enzyme 0.1 µL, 10×buffer 2 µL and double distilled water. The amplification profiles were 5 min at 94 ℃, followed by 35 cycles of 30 s at 94 ℃; 1 min at 55 ℃, 1 min at 72 ℃; then 10 min at 72 ℃. After amplification, the PCR products were mixed with loading buffer (2.5 mg/mL bromophenol blue, 2.5 mg/mL diphenylamine blue, 10 mmol/L EDTA, 95% (V/V) formamide), denatured for 5 min at 94℃ and put on ice for 5 min. The denatured PCR products were separated on 6%(W/V) denaturing polyacrylamide gel at 100 W power and visualized by silver straining. Polymorphic fragments of primers were cloned and sequenced. Linkage analysis used the software of Mapmaker V3.0.During analysing the separated rate of F<sub>1</sub> and F<sub>2</sub>, the results showed that the gynoecy in 240 -1-2-2 - 3 -1 was controlled by oligogene with some background genes modified. Inheritance of gynoecy was accord with the additive-dominant-epistatic model. From 699 pairs of SSR primer, two pair of stable SSR markers (CSWCT25 and SSR18956), 331 bp and 145 bp in bands size were obtained respectively during PCR products of two SSR markers cloned and sequenced, and linkage analysis indicated that its genetic distance to the gynoecious loci was 7.7 cM and 6.8 cM, respectively. Two SSR markers are tightly linked to gynoecious loci on the chromosome 6.In sum, knowledge of location of gynoecious gene in cucumber and its related traits in crosses will be helpful to the design of more effective selection schemes to develop gynoecious cucumber genotypes. Progress in breeding gynoecious lines is still slow because of the complex inheritance of this character. However, gynoecious cultivars and the markers identified in this study can contribute to improving gynoecious line. Two SSR markers could be used effectively for molecular marker-assisted selection in breeding programs to develop cucumber gynoecious line breeding.https://www.academax.com/doi/10.3785/j.issn.1008-9209.2012.11.141cucumber (<italic>Cucumis sativus</italic> L.)gynoeciousmolecular marker |
| spellingShingle | ZHOU Shengjun ZHANG Peng ZHU Yuqiang CHEN Xinjuan CHEN Liping Identification of SSR marker linked to gynoecious loci in cucumber (Cucumis sativus L.) 浙江大学学报. 农业与生命科学版 cucumber (<italic>Cucumis sativus</italic> L.) gynoecious molecular marker |
| title | Identification of SSR marker linked to gynoecious loci in cucumber (Cucumis sativus L.) |
| title_full | Identification of SSR marker linked to gynoecious loci in cucumber (Cucumis sativus L.) |
| title_fullStr | Identification of SSR marker linked to gynoecious loci in cucumber (Cucumis sativus L.) |
| title_full_unstemmed | Identification of SSR marker linked to gynoecious loci in cucumber (Cucumis sativus L.) |
| title_short | Identification of SSR marker linked to gynoecious loci in cucumber (Cucumis sativus L.) |
| title_sort | identification of ssr marker linked to gynoecious loci in cucumber cucumis sativus l |
| topic | cucumber (<italic>Cucumis sativus</italic> L.) gynoecious molecular marker |
| url | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2012.11.141 |
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