[1]高兵,安文杰,孙俊.苹果愈伤组织SSR-PCR反应体系优化[J].北方园艺,2018,42(10):53-56.[doi:10.11937/bfyy.20173431]
　GAO Bing,AN Wenjie,SUN Jun.Optimization of SSR-PCR Reaction System for Apple Callus[J].Northern Horticulture,2018,42(10):53-56.[doi:10.11937/bfyy.20173431]
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苹果愈伤组织SSR-PCR反应体系优化

《北方园艺》[ISSN:1001-0009/CN:23-1247/S] 卷: 第42卷 期数: 2018年10 页码: 53-56 栏目: 出版日期: 2018-05-25
Title:
Optimization of SSR-PCR Reaction System for Apple Callus
作者:
高兵1; 2; 安文杰1; 孙俊2; 3
（1.山西林业职业技术学院 园艺系，山西 太原 030009；2.南京农业大学 园艺学院，江苏 南京 210095； 3.安徽农业大学 园艺学院，安徽 合肥 230036）
Author(s):
GAO Bing1; 2; AN Wenjie1; SUN Jun2; 3
(1.Department of Horticulture,Shanxi Forestry Vocational and Technical College,Taiyuan,Shanxi 030009；2.College of Horticulture,Nanjing Agricultural University,Nanjing，Jiangsu 210095;3.College of Horticulture,Anhui Agricultural University,Hefei，Anhui 230036)
关键词:
苹果; 愈伤组织; SSR-PCR; 体系优化
Keywords:
apple;  callus;  SSR-PCR;  system optimization
DOI:
10.11937/bfyy.20173431
文献标志码:
A
摘要:
以“嘎啦”苹果组培苗叶片诱导的愈伤组织作为DNA模板提取材料，采用正交设计，摸索了SSR反应体系中Mg2+、Taq DNA聚合酶、引物、模板DNA、dNTP的最适浓度，可为苹果愈伤组织的遗传变异研究奠定基础。结果表明：各因素不同水平变化对反应体系的影响为Taq DNA聚合酶>dNTP 用量>Mg2+用量>引物用量=模板DNA浓度。确立了苹果愈伤组织SSR-PCR最佳反应体系总体积25 μL，其中4 μL 25 mmol?L-1 Mg2+，0.25 μL 5 U?μL-1 Taq DNA聚合酶，3 μL 0.01 mmol?L-1 SSR引物，1 μL 30 ng?μL-1 模板DNA，4 μL 2.5 mmol?L-1dNTP，2.5 μL 10×PCR Buffer，10.25 μL超纯水。
Abstract:
In order to establish the basis of genetic variation research of apple callus，the ‘Gala’ induction of callus was DNA template extraction materials.the Mg2+，Taq DNA polymerase，primer dosage，template DNA concentration and dNTP which were suitable for concentration or dosage were explored.The results showed that the effect of different levels on reaction system was Taq DNA polymerase>dNTP dosage>Mg2+ dosage>primer dosage=template DNA concentration.The optimal reaction system of apple callus was established,the optimal reaction system of apple callus was that 25 μL for total volume，which were 4 μL 25 mmol?L-1 Mg2+，0.25 μL 5 U?μL-1Taq DNA polymerase，3 μL 0.01 mmol?L-1 SSR，1 μL 30 ng?μL-1 template DNA，4 μL 2.5 mmol?L-1dNTP，2.5 μL 10×PCR Buffer，10.25 μL ultra-pure water.

参考文献/References:

 

[1]王爱德,李天忠，许雪峰，等.苹果品种的SSR分析[J].园艺学报，2005，32（5）：875-877.

[2]金万梅,毛海亮,张强，等.苹果矮化砧木SH系的SSR标记鉴定[J].园艺学报，2010,37(2)：303-306.

[3]王刚刚,王飞,赵政阳,等.苹果果实酸/低酸性状的SSR标记[J].西北农林科技大学学报（自然科学版）,2010,38(6):133-137.

[4]董研,张军,任亚超，等.中国新疆野苹果天然群体遗传多样性SSR分析[J].植物遗传资源学,2013,14(5):1-7.

[5]ASHMORE S E.Status report on the development and application of in vitro techniques for conservation and use of plant genetic resources[J].International Plant Genetic Resources Institu Rome,1997，90:307-313.

[6]ENGELMANN F.In vitro conservation methods[M]//CALLOW J A,FORD-LLOYD,B V NEWBURY H J.Biotechnology and plant Genetic Resources.Oxon：CABI,1997:119-161.

[7]KALIA R K,RAI M K,KALIA S,et al.Microsatellite markers:An over-view of the recent progress in plants[J].Euphytica，2011,177(3):309-334.

[8]陈大明,张上隆,金勇丰.一种木本果树基因组DNA提取方法研究[J].浙江农业大学学报,1997,23（6）：621-624.

[9]续九如，黄智慧.林业试验设计[M].北京：中国林业出版社，1998:65-79.

[10]杨水云,李续娥，吴明宇,等.正交实验在PCR反应条件优化中的应用[J].生物数学学报,2005,20(2):202-206.

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备注/Memo

第一作者简介：高兵（1979-），女，山西太原人，硕士，讲师，现主要从事园艺教学等工作。E-mail：754025816@qq.com.基金项目：国家自然科学基金资助项目（30200190）；港澳台科技合作专项资助项目（2015DFT30090）。收稿日期：2018-03-12

更新日期/Last Update: 2018-06-01