|Table of Contents|

Optimization of SRAP Reaction System and Genetic Relationship in Paeonia

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Issue:
2019年10
Page:
93-101
Research Field:
Publishing date:

Info

Title:
Optimization of SRAP Reaction System and Genetic Relationship in Paeonia
Author(s):
SA Na; WANG Pengyao; JU Fuzhu; HAO Qing
(College of Landscape Architecture and Forestry，Qingdao Agricultural University，Qingdao,Shandong 266109)
Keywords:
Paeonia; SRAP; reaction system; optimization; genetic relationship
PACS:
-
DOI:
10.11937/bfyy.20183403
Abstract:
In order to establish stable and optimal SRAP-PCR protocol for plants of Paeonia，the factors of reaction system and procedure for PCR were optimized using cultivars of ‘Oriental Gold’ and ‘Hui He’ as materials．With the system,genetic relationship of 65 cultivars in Paeonia was analyzed.The results showed that most suitable protocol was an initial denaturation at 94 ℃ for 3 min，eight cycles of three steps,30 s of denaturing at 94 ℃，30 s of annealing at 37 ℃ and 90 s of elongation at 72 ℃，the temperature for annealing was increased 1 ℃ after each cycle．Followed by 32 cycles，at 94 ℃ for 30 s，50 ℃ for 30 s，72 ℃ for 90 s．With a final elongation at 72 ℃ for 7 min．The optimal concentration of Mg2+ was 2.5 mmol?L-1，0.2 mmol?L-1 of dNTPs，1 U of Taq DNA polymerase，each prime 0.3 μmol?L-1 for final 20 μL reaction system．A total of 1 229 polymorphic bands were amplified used 16 primer pairs,the polymorphism was as high as 100%,the herbaceous peony and the hybrids were clustered in a class,and the tree peony was clustered in a class,which proved that the hybrids were more closely related to the herbaceous peony.The newly established SRAP-PCR system of Paeonia was fully reproducible and stable,and suitable for the study of SRAP in Paeonia plants.It provided a foundation for further studies on genetic diversity of germplasm resources,cultivars identification and molecular marker assisted selection breeding of Paeonia．

References:

[1]王士泉.芍药属两物种染色体结构变异杂合性研究[D].北京:中国科学院研究生院(植物研究所),2006.[2]郝青,刘政安,舒庆艳,等.中国首例芍药牡丹远源杂交种的发现及鉴定[J].园艺学报,2008,3(6):853-858.[3]史倩倩.中原牡丹传统品种遗传多样性研究[D].北京:中国林业科学研究院,2012.[4]唐琴,曾秀丽,廖明安,等.大花黄牡丹遗传多样性的SRAP分析[J].林业科学,2012,48(1):70-76.[5]BUDAK H,SHEARMAN R C,PARMAKSIZ I,et al.Comparative analysis of seeded and vegetative biotype buffalograsses based on phylogenetic relationship using ISSRs,SSRs,RAPDs,and SRAPs[J].Theoretical & Applied Genetics,2004,109(2):280-288.[6]FERRIOL M,PICO B,CORDOVA P F,et al.Molecular diversity of a germplasm collection of squash（Cucurbita moschata） determined by SRAP and AFLP markers[J].Plant Genetic Resources,2004,44:653-664．[7]LI G,GAO M,YANG B,et al.Gene for gene alignment between the Brassica and Arabidopsis genomes by direct transcriptome mapping[J].Theoretical and Applied Genetics,2003,107:168-180．[8]RIAZ A,POTTER D,STEPHEN M.Genotyping of peach and nectarine cultivars with SSR and SRAP moleculars markers[J].Journal of the American Society for Horticultural Science,2004,129:204-211．[9]郭大龙,侯小改,张静,等.牡丹SRAP反应体系的建立及正交设计优化[J].河南农业科学,2008,37(12):110-113.[10]苏美和,赵兰勇.牡丹杂交品系SRAP-PCR反应体系优化及引物筛选[J].中国农学通报,2012,28(19):189-193.[11]王燕青,季孔庶.利用正交设计优化牡丹SRAP-PCR反应体系[J].分子植物育种,2009,7(1):199-203.[12]孙宪芝,王晓菡,马燕,等.应用正交设计建立芍药的SRAP反应体系[J].华北农学报,2008,23(s2):198-200.[13]王晓菡.中国芍药品种遗传多样性SRAP分析和核心种质的初步构建[D].泰安:山东农业大学,2010.[14]LI G，QUIROS C F.Sequence-related amplified polymorphism（SARP） a new marker system based on a simple PCR reaction:Its application to mapping and gene tagging in Brassica[J].Theor Appl Genet，2001,103:455-461．[15]PAN J S,WANG G,LI X Z,et al.Construction of a genetic map with SRAP markers and localization of the gene responsible for the first-flower-node trait in cucumber(Cucumis sativus L.)[J].Progress in Natural Science,2005,15(5):407-413.[16]HAO Q,LIU Z A,SHU Q Y,et al.Studies on Paeonia cultivars and hybrids identication based on SRAP analysis[J].Hereditas,2008,145:38-47.[17]张栋.牡丹远缘杂交及部分杂交后代的AFLP分子标记鉴定[D].北京:北京林业大学,2008.

Memo

Memo:
-
Last Update: 2019-06-27