基于ISSR分子标记分析陆川猪种群遗传差异

doi:10.16431/j.cnki.1671-7236.2019.12.019

摘要/Abstract

摘要： 试验旨在探究陆川猪种群之间的遗传差异，对陆川猪种群进行遗传多样性分析，为种质改良与创新提供参考。以29份陆川猪种及部分广西地方猪种样品为试验材料，采用了3条带型清晰、重复性好的ISSR引物，通过对PCR扩增及电泳得到的DNA片段条带进行聚类分析并构建聚类树状图，并对陆川猪种群进行遗传差异分析。结果显示，ISSR分子标记上共发现182条条带，其中84条多态性条带，3条ISSR引物扩增出来的总DNA片段条带与多态性DNA片段条带差异较大且分布不均匀，表明所选择的3个ISSR分子标记在陆川猪种群中具有丰富的多态性；陆川猪种群之间遗传相似系数变异范围为0.43~1.00，表明种群间的遗传差异变化较大，遗传背景丰富；UPGMA聚类分析表明，陆川猪与长陆二元杂猪、巴马香猪、环江香猪均可聚为一类，大陆二元杂猪、三元杂猪和龙宝猪聚为一类。本研究证实了所选的3条ISSR引物可作为有效的遗传标记用于陆川猪种群间的遗传差异分析，丰富了当前的陆川猪ISSR分子标记资源，同时也表明陆川猪种群具有丰富的遗传多样性。

关键词: 陆川猪; ISSR; 分子标记技术; 遗传相似系数; 聚类分析

Abstract: This study was aimed to investigate the genetic differences and genetic diversity in Luchuan pig populations,provide reference for germplasm improvement and innovation.A total of 29 samples of Luchuan pigs and some samples of Guangxi local pigs were used as experimental materials,three bands of clear and reproducible ISSR primers were used,and DNA fragments obtained by PCR amplification and electrophoresis were used to carry out cluster analysis and clustering trees were constructed to analyze the genetic differences in Luchuan pig populations.The results showed that a total of 182 bands were found on the ISSR molecular markers,84 of which were polymorphic.The total DNA fragment bands amplified by three ISSR primers were different from the polymorphic DNA fragments,which indicated that the selected three ISSR molecular markers were abundant in Luchuan pigs populations.The genetic similarity coefficient in Luchuan pig populations rangeed from 0.43 to 1.00,which indicated that the genetic differences in populations were larger,and the genetic background was rich.UPGMA cluster analysis results showed that Luchuan pigs and Chang Lu binary pigs,Bama Xiang pigs and Huanjiang Xiang pigs could be grouped together,and Da Lu binary pigs,ternary pigs and Longbao pigs were grouped together.This study confirmed that the selected three ISSR primers could be used as effective genetic markers for analyzing genetic difference in Luchuan pig populations,enriched the current ISSR molecular marker resources of Luchuan pigs,and also indicated the rich genetic diversity in Luchuan pig populations.

Key words: Luchuan pigs; ISSR; molecular marker technique; genetic similarity coefficient; cluster analysis

中图分类号:

S813.3

李中宇, 封毅, 黄丽, 王淼, 滕建文. 基于ISSR分子标记分析陆川猪种群遗传差异[J]. 中国畜牧兽医, 2019, 46(12): 3617-3626.

LI Zhongyu, FENG Yi, HUANG Li, WANG Miao, TENG Jianwen. Genetic Differences Analysis in Luchuan Pigs Populations Using ISSR Molecular Marker[J]. China Animal Husbandry and Veterinary Medicine, 2019, 46(12): 3617-3626.

参考文献

[1] 农业部农产品质量安全监管局.中华人民共和国农业部公告第1459号文件[EB/OL].http://www.moa.gov.cn/gk/tzgg_1/gg/201010/t20101018_1682633.htm,2010.9.13.SUPERVISION BUREAU OF AGRICULTURAL PRODUCT QUALITY AND SAFETY,MINISTRY OF AGRICULTURE.Document No.2468 of the Ministry of Agriculture of the People's Republic of China[EB/OL].http://www.moa.gov.cn/gk/tzgg_1/gg/201010/t20101018_1682633.htm,2010.9.13.(in Chinese)
[2] 何若钢,李琼华,段福君,等.陆川猪杂交优势选育的种质特征研究[J].畜牧与兽医,2008,40(10):58-60. HE R G,LI Q H,DUAN F J,et al.Germplasm characteristics of Luchuan pig hybrid breeding[J].Animal Husbandry & Veterinary Medicine,2008,40(10):58-60.(in Chinese)
[3] 何若钢,刘桂武,林德源,等.杜洛克与陆川猪杂交后代的肉质研究[J].黑龙江畜牧兽医,2011,12:37-39. HE R G,LIU G W,LIN D Y,et al.Study on the meat quality of the cross between Duroc and Luchuan pigs[J].Heilongjiang Animal Husbandry and Veterinary Medicine,2011,12:37-39.(in Chinese)
[4] RAKOCZY-TROJANOWSKA M,BOLIBOK H.Characteristics and a comparison of three classes of microsatellite-based markers and their application in plants[J].Cellular & Molecular Biology Letters,2004,9(2):221-238.
[5] REDDY M P,SARLA N,SIDDIQ E A.Inter simple sequence repeat (ISSR) polymorphism and its application in plant breeding[J].Euphytica,2002,128(1):9-17.
[6] 孙雪梅,赵才美,李友勇,等.云南茶树地方品种与野生茶遗传多样性的ISSR分析[J].西南农业学报,2018,31(5):30-35. SUN X M,ZHAO C M,LI Y Y,et al.Genetic diversity of tea landraces and wild tea germplasms in Yunnan revealed by ISSR analysis[J].Southwest China Journal of Agricultural Sciences,2018,31(5):30-35.(in Chinese)
[7] NUDIN N,ALI A M,NGAH N,et al.SSR marker-assisted genetic diversity analysis of Dioscorea hispida and selection of the best variety for sustainable production[J].Comptes Rendus Biologies,2017,340(8):359.
[8] ASKARI N,ABADI M M,BAGHIZADEH A,et al.SSR markers for assessing DNA polymorphism and genetic characterization of cattle,goat and sheep populations[J].Iranian Journal of Biotechnology,2011,9(3):222-229.
[9] STOLPOVSKIǏ IUA,KOL N V,EVSIUKOV A N,et al.Analysis of the genetic structure of Tuvinian short-fat-tailed sheep populations with the use of the ISSR-PCR method[J].Genetika,2010,46(12):1660-1669.
[10] 金玉兰,李婉丽,李松,等.ISSR分子标记鉴定巴马香猪[J].四川动物,2016,35(4):569-573. JIN Y L,LI W L,LI S,et al.Species authentication of Bama Miniature pig through inter-simple sequence repeat[J].Sichuan Journal of Zoology,2016,35(4):569-573.(in Chinese)
[11] 史子学,赵晓明,唐赛涌,等.应用ISSR-PCR技术鉴别梅山猪肉及其肉制品的研究[J].猪业科学,2016,33(4):132-133. SHI Z X,ZHAO X M,TANG S Y,et al.Identification of Meishan pork and its meat products by ISSR-PCR[J].Pig Science,2016,33(4):132-133.(in Chinese)
[12] 孙俊丽,张冰,马青艳,等.陆川猪mtDNA D-loop序列遗传多样性分析[J].中国畜牧兽医,2010,37(6):122-125. SUN J L,ZHANG B,MA Q Y,et al.Analysis of mtDNA partial D-loop sequences in Luchuan pig[J].China Animal Husbandry & Veterinary Medicine,2010,37(6):122-125.(in Chinese)
[13] 彭红元,廖龙,黄业,等.陆川猪种群线粒体DNA遗传多样性研究[J].广东农业科学,2012,39(2):91-94. PENG H Y,LIAO L,HUANG Y,et al.Genetic diversity of Luchuan pig inferred from the mitochondrial DNA control region[J].Guangdong Agricultural Sciences,2012,39(2):91-94.(in Chinese)
[14] 孙俊丽,张冰,潘天彪,等.基于微卫星位点研究7个广西地方猪种的遗传关系[J].西南农业学报,2016,29(6):1475-1480. SUN J L,ZHANG B,PAN T B,et al.Genetic diversity of seven Guangxi local pig breeds assessed with microsatellite markers[J].Southwest Agricultural Journal,2016,29(6):1475-1480.(in Chinese)
[15] 何瑜林,黄丽,封毅,等.ISSR分子标记鉴定陆川猪及其肉制品[J].肉类研究,2014,28(10):10-14. HE Y L,HUANG L,FENG Y,et al.Species authentication of Luchuan pig meat and meat products using ISSR molecular markers[J].Meat Research,2014,28(10):10-14.(in Chinese)
[16] 赵瑞杰,严海燕,谭艳平,等.CTAB法提取动物DNA[J].中国西部科技,2011,10(21):7-8. ZHAO R J,YAN H Y,TAN Y P,et al.Animal DNA extraction with CTAB method[J].Western China Science and Technology,2011,10(21):7-8.(in Chinese)
[17] 何建文,韩建林,罗玉柱.利用不同方法从深加工牦牛肉产品中提取基因组DNA效果的比较[J].生物技术通报,2010,10:162-167. HE J W,HAN J L,LUO Y Z.Comparison of quantity and quality of genomic DNAs extracted from highly processed Yak meat products using different methods[J].Biotechnology Bulletin,2010,10:162-167.(in Chinese)
[18] 张维铭.现代分子生物学实验手册[M].北京:科学出版社,2003. ZHANG W M.Modern Molecular Biology Experiment Manual[M].Beijing:Science Press,2003.(in Chinese)
[19] 高丹丹,陈燕,王迎华,等.动物肉制品基因组DNA的提取和纯化[J].食品科技,2007,8:42-44. GAO D D,CHEN Y,WANG Y H,et al.Extraction and purification of genomic DNA in the animal origin food[J].Food Science and Technology,2007,8:42-44.(in Chinese)
[20] 李武,倪薇,林忠旭,等.海岛棉遗传多样性的SRAP标记分析[J].作物学报,2008,34(5):893-898. LI W,NI W,LIN Z X,et al.Genetic diversity analysis of sea-island cotton cultivars using SRAP carkers[J].Acta Agronomica Sinica,2008,34(5):893-898.(in Chinese)
[21] 陈祥,丁玫,许厚强,等.贵州5个地方猪品种微卫星标记的遗传多样性[J].中国畜牧杂志,2015,51(7):1-5. CHEN X,DING M,XU H Q,et al.Genetic diversity of microsatellite markers in five domestic pig breeds in Guizhou province[J].Chinese Journal of Animal Science,2015,51(7):1-5.(in Chinese)
[22] 陈智武,沈前程,朱春红,等.鸡基因组DNA不同提取方法的比较[J].中国家禽,2017,39(17):64-66. CHEN Z W,SHEN Q C,ZHU C H,et al.Comparison of different extraction methods for chicken genomic DNA[J].China Poultry,2017,39(17):64-66.(in Chinese)
[23] 张雨欣,高秋,逄洪波,等.小白鼠6种组织部位总DNA不同提取方法的比较[J].江苏农业科学,2015,43(10):47-50. ZHANG Y X,GAO Q,PANG H B,et al.Comparison of different extraction methods for total DNA of 6 tissue parts in mice[J].Jiangsu Agricultural Sciences,2015,43(10):47-50.(in Chinese)
[24] LIAN C L,ZHOU Z H,HOGETSU T Z.A simple method for developing microsatellite markers using amplified fragments of inter-simple sequence repeat(ISSR)[J].Journal of Plant Research,2001,114(3):381-385.
[25] GONZALEZ A,WONG A,DELGADOSALINAS A,et al.Assessment of inter simple sequence repeat markers to differentiate sympatric wild and domesticated populations of common bean[J].Crop Science,2005,45(2):606-615.
[26] COSTA J C,FRACETTO G G M,FRACETTO F J C,et al.Genetic diversity of Desmanthus sp accessions using ISSR markers and morphological traits[J].Genetics & Molecular Research,2017,16(2):gmr16029667.
[27] MOUSAVIFARD S S,SAEIDI H,RAHIMINEJAD M R,et al.Molecular analysis of diversity of diploidTriticumspecies in Iran using ISSR markers[J].Genetic Resources and Crop Evolution,2015,62(3):387-394.
[28] PURNAMANINGRUM A,HANDAYANI N S N,TRIJOK O,et al.Assessment of genetic variation in outbreed and inbreed giant tiger shrimp (Penaeus monodon Fabricius,1798) using ISSR marker[A].Advances of Science and Technology for Society:Proceedings of the 1st International Conference on Science and Technology 2015(ICST-2015)[C].2016.
[29] 石洪玥,刘阳,王晓梅,等.七里海中华绒螯蟹遗传多样性的RAPD和ISSR分析[J].水产科学,2016,35(3):255-260. SHI H Y,LIU Y,WANG X M,et al.Analysis of genetic diversity of Chinese mitten handed crab eriocheir sinensis from Qilihai by RAPD and ISSR markers[J].Fisheries Science,2016,35(3):255-260.(in Chinese)
[30] 黄敏瑞,孙俊丽,潘天彪,等.6个微卫星DNA标记在陆川猪群体中的遗传多样性分析[J].广西畜牧兽医,2008,24(2):67-69. HUANG M R,SUN J L,PAN T B,et al.Analysis of genetic diversity of six microsatellite DNA markers in Luchuan pig population[J].Guangxi Animal Husbandry and Veterinary Medicine,2008,24(2):67-69.(in Chinese)