Construction of multiplex PCR in variety identification of Porphyra haitanensis "Z-26" based on SCAR markers.

Q4 Environmental Science
W. Ting, Xu Yan, Xie Chaotian, J. Dehua, Chen Chang-sheng
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引用次数: 3

Abstract

Porphyra haitanensis is one of the most important economic marine crops of China.For any cultivar,the correct identification of species or forma of the cultivated strains is necessary to ensure a well-bred cultivation and good production quality.However,because the gametophytic blade of Porphyra is morphologically simple and marked variations can occur as environmental conditions change,it is very difficult to precisely identify the species or forma of cultivated strains based only on their morphological characteristics.With new advances in molecular biology,molecular markers and DNA fingerprinting techniques have become routine for the identification and classification of many crops,including seaweeds.The strain of "Z-26" of P.haitanensis was selected by the laboratory of germplasm improvement and the application of P.haitanensis in Jimei University which has the characters of high-temperature tolerance and high yield,and it has been widely cultivated in south China.In order to construct the technology of variety identification for "Z-26",firstly,300 primers of RAPD were used to scan the specific markers of 6 new strains of P.haitanensis and 9 specific RAPD markers of "Z-26" were selected.After cloning and sequencing,two specific RAPD markers of "Z-26" were transformed into the SCAR markers successfully,the length of the 2 SCAR markers was 540 and 242 bp,respectively.Secondly,after verification by 4 different experiments,we can affirm that the 2 SCAR markers were the specific and stable markers of "Z-26".At last,based on the 2 SCAR markers,after optimization of experimental conditions,the technology of multiplex PCR which was used to identify the variety of "Z-26" was constructed.The result supplied a simple,fast and reliable technigue for variety identification of "Z-26".
基于SCAR标记的海檀紫菜“Z-26”品种鉴定多重PCR构建
海紫菜是中国重要的海洋经济作物之一。对于任何一个栽培品种,正确鉴定栽培菌株的种类或形式是保证优良栽培和良好生产质量的必要条件。然而,由于紫菜的配子体叶片形态简单,且随环境条件的变化会发生明显的变化,因此仅根据其形态特征来精确鉴定栽培菌株的种类或形态是非常困难的。随着分子生物学的新进展,分子标记和DNA指纹技术已成为许多作物(包括海藻)鉴定和分类的常规技术。集美大学海棠种质改良与应用实验室选育的海棠“Z-26”品系具有耐高温、高产的特点,在华南地区广泛栽培。为了构建“Z-26”的品种鉴定技术,首先利用300条RAPD引物对6个海檀p.p haitanensis新品系的特异性标记进行扫描,筛选出9个“Z-26”的特异性RAPD标记。经过克隆和测序,成功地将“Z-26”的两个特异性RAPD标记转化为SCAR标记,2个SCAR标记的长度分别为540和242 bp。其次,经过4个不同实验的验证,我们可以肯定这2个SCAR标记是“Z-26”特异性和稳定性的标记。最后,在2个SCAR标记的基础上,通过优化实验条件,构建了多重PCR技术对“Z-26”品种进行鉴定。结果为“Z-26”的品种鉴定提供了一种简单、快速、可靠的方法。
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来源期刊
水产学报
水产学报 Environmental Science-Management, Monitoring, Policy and Law
CiteScore
1.40
自引率
0.00%
发文量
5213
期刊介绍: "Fisheries of" mainly reflects the results of scientific research and development of the direction of aquaculture for domestic and foreign academic exchanges Fisheries Service. Mainly basic research published in Fisheries, aquaculture and proliferation of fishing waters environmental protection, preservation of aquatic products processing and utilization, fishing equipment, and other aspects of mechanical papers, research briefings and reviewed.
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