An optimal SRAP-PCR system of Rhododendron hybridum and its genetic diversity analysis with SRAP marker

WU Yueyan; TAO Qiaojing; LI Bo; XU Danye

doi:10.11833/j.issn.2095-0756.2013.06.007

Volume 30 Issue 6

Nov. 2013

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WU Yueyan, TAO Qiaojing, LI Bo, XU Danye. An optimal SRAP-PCR system of Rhododendron hybridum and its genetic diversity analysis with SRAP marker[J]. Journal of Zhejiang A&F University, 2013, 30(6): 844-851. doi: 10.11833/j.issn.2095-0756.2013.06.007

Citation:

WU Yueyan, TAO Qiaojing, LI Bo, XU Danye. An optimal SRAP-PCR system of Rhododendron hybridum and its genetic diversity analysis with SRAP marker[J]. Journal of Zhejiang A&F University, 2013, 30(6): 844-851. doi: 10.11833/j.issn.2095-0756.2013.06.007

An optimal SRAP-PCR system of Rhododendron hybridum and its genetic diversity analysis with SRAP marker

doi: 10.11833/j.issn.2095-0756.2013.06.007

WU Yueyan^{1
,
,},
TAO Qiaojing^1,2,
LI Bo^1,2,
XU Danye¹

1.
College of Biology and Environment，Zhejiang Wanli University，Ningbo 315100，Zhejiang，China;
2.
College of Agriculture and Biotechnology，Zhejiang University，Hangzhou 310058，Zhejiang，China

More Information

Corresponding author: WU Yueyan
Received Date: 2012-11-14
Rev Recd Date: 2013-05-22
Publish Date: 2013-12-20

Abstract

In order to find the optimal sequence-related amplified polymorphism（SRAP）-PCR system of Rhododendron hybridum and analyse its genetic diversity，the functional leaves of Rhododendron hybridum were used to extract its genome DNA，using an improved cetyl trimethyl ammonium bromide（CTAB）method. Concentrations of Mg2+，Taq polymerase，dNTPs and primer, which maybe affect the SRAP-PCR reactions，were optimized by L16（44）orthogonal design experiments to establish the SRAP molecular marker system in Rhododendron hybridum. Then，an optimal, stable and repeatable SRAP-polymerase chain reaction（PCR）of 20.0 L containing 1.750 mmolL－1 Mg2+，0.175 mmolL－1 dNTPs，1.500 16.67 nkat Taq polymerase，and 0.200 molL－1 primer was established. Finally， genetic diversity and relationships of 24 Rhododendron hybridum cultivars were analyzed using an Unweighted Pair Group Method with Arithmetic Mean（UPGMA）cluster analysis with this optimized system. Results revealed 10 highly polymorphic and stable primer pairs selected from 88 pairs of SRAP primers with a total of 217 bands being detected from these 10 primer pairs. Of the detected bands，212 were polymorphic（a 97.35% average）having a genetic similarity coefficient ranging from 0.591 to 0.708. The cluster analysis divided the 24 cultivars into two groups at a 0.590 similarity level，and these were further delineated into two sub-groups at a 0.623 similarity level. These results demonstrated that this optimized SRAP-PCR system can be applied to cultivars identification and genetic diversity research on Rhododendron hybridum.［Ch，3 fig. 6 tab. 19 ref.］
- botany,
- Rhododendron hybridum,
- genetic diversity,
- SRAP-PCR,
- orthogonal design,
- system optimization,
- cluster analysis

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

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An optimal SRAP-PCR system of Rhododendron hybridum and its genetic diversity analysis with SRAP marker

1. College of Biology and Environment，Zhejiang Wanli University，Ningbo 315100，Zhejiang，China;

2. College of Agriculture and Biotechnology，Zhejiang University，Hangzhou 310058，Zhejiang，China

Corresponding author: WU Yueyan

Received Date: 2012-11-14

Rev Recd Date: 2013-05-22

Publish Date: 2013-12-20

Key words:

Abstract: In order to find the optimal sequence-related amplified polymorphism（SRAP）-PCR system of Rhododendron hybridum and analyse its genetic diversity，the functional leaves of Rhododendron hybridum were used to extract its genome DNA，using an improved cetyl trimethyl ammonium bromide（CTAB）method. Concentrations of Mg2+，Taq polymerase，dNTPs and primer, which maybe affect the SRAP-PCR reactions，were optimized by L16（44）orthogonal design experiments to establish the SRAP molecular marker system in Rhododendron hybridum. Then，an optimal, stable and repeatable SRAP-polymerase chain reaction（PCR）of 20.0 L containing 1.750 mmolL－1 Mg2+，0.175 mmolL－1 dNTPs，1.500 16.67 nkat Taq polymerase，and 0.200 molL－1 primer was established. Finally， genetic diversity and relationships of 24 Rhododendron hybridum cultivars were analyzed using an Unweighted Pair Group Method with Arithmetic Mean（UPGMA）cluster analysis with this optimized system. Results revealed 10 highly polymorphic and stable primer pairs selected from 88 pairs of SRAP primers with a total of 217 bands being detected from these 10 primer pairs. Of the detected bands，212 were polymorphic（a 97.35% average）having a genetic similarity coefficient ranging from 0.591 to 0.708. The cluster analysis divided the 24 cultivars into two groups at a 0.590 similarity level，and these were further delineated into two sub-groups at a 0.623 similarity level. These results demonstrated that this optimized SRAP-PCR system can be applied to cultivars identification and genetic diversity research on Rhododendron hybridum.［Ch，3 fig. 6 tab. 19 ref.］

An optimal SRAP-PCR system of Rhododendron hybridum and its genetic diversity analysis with SRAP marker

doi: 10.11833/j.issn.2095-0756.2013.06.007

Abstract

References

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通讯作者: 陈斌, bchen63@163.com

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