ZHAO Cuimin,ZHAO Cen,ZHANG Lanying,et al.Genetic Diversity and Population Structure Analysis of Ganoderma lucidum Based on ITS and ISSR Markers[J].Northern Horticulture,2024,(3):103-111.[doi:10.11937/bfyy.20232329]
基于ITS和ISSR标记的灵芝遗传多样性和群体结构分析
- Title:
- Genetic Diversity and Population Structure Analysis of Ganoderma lucidum Based on ITS and ISSR Markers
- 文章编号:
- 1001-0009(2024)03-0103-09
- Keywords:
- Ganoderma lucidum; internal transcribed spacer (ITS); inter-simple sequence repeats(ISSR); genetic diversity
- 分类号:
- S 567.3+1
- 文献标志码:
- A
- 摘要:
- 以来自不同地理区域的48个灵芝种质资源为试材，采用ITS和ISSR分子标记的方法，研究了灵芝遗传多样性，以期利用遗传信息数据较理想地显示出不同灵芝菌株的遗传多样性。结果表明：通过进行ITS序列扩增测序，将48个灵芝菌株分为赤芝（35个）、无柄灵芝（11个）、四川灵芝（1个）、古巴栓孔菌（1个）。5条ISSR引物共扩增得到53条条带，多态性条带比率为96.23%，Nei′s基因多样性为0.27，Shannon′s信息指数为0.43。ISSR标记遗传相似系数约为0.70，将48个灵芝菌株分为3组，其中第1组为11个无柄灵芝，第2组为菌株29“盆景1”，其他菌株为第3组，说明2种标记结合分析能够更加准确分析不同菌株间的亲缘关系。菌株16和17同为赤芝，且ISSR分子标记遗传相似系数达0.98，推测可能为同一品种，为生产中出现的同物异名现象。该研究选取的用于PCR扩增的ISSR引物，多态性较好、稳定性较高，能有效鉴别出无柄灵芝与其他灵芝，并揭示种质的遗传多样性和群体遗传结构，可为灵芝种质资源保护及优质种质材料选育提供参考依据，以期更好地推动灵芝产业健康发展。
- Abstract:
- 48 Ganoderma lucidum germplasm resources from different geographical regions were used as samples，and the genetic diversity of Ganoderma lucidum was studied by using ITS and ISSR molecular markers，in order to better display the genetic diversity of different Ganoderma lucidum strains by using genetic information data.The results showed that 48 strains of Ganoderma were divided into Ganoderma lucidum (35)，Ganoderma resinaceum (11)，Ganoderma sichuanense (1) and Trametes cubensis(1) by ITS sequence amplification and sequencing.A total of 53 bands were amplified by 5 ISSR primers，the percentage of polymorphic bands was 96.23%，the Nei′s gene diversity was 0.27，the Shannon′s information index was 0.43.The genetic similarity coefficient of ISSR marker was about 0.70，and 48 strains of Ganoderma were divided into three groups.The first group was 11 strains of Ganoderma lucidum，the second group was 29 ‘Penjing 1’，and the remaining strains were the third group，indicated that the combination of two markers could explain the genetic relationship between different strains in more detail.Strains 16 and 17 were both Ganoderma lucidum，and the genetic similarity coefficient of ISSR molecular markers reached 0.98，suggested that they might be the same variety，which was a synonym phenomenon in production.The ISSR primers selected for PCR amplification in this study had good polymorphism and high stability，which could effectively distinguish sessile Ganoderma lucidum from other Ganoderma lucidum，and reveal the genetic diversity and population genetic structure of the germplasm，which could provide reference basis for the protection of Ganoderma lucidum germplasm resources and the selection of high-quality germplasm materials，so as to better promote the healthy development of Ganoderma lucidum industry.
参考文献/References:
［1］RAMADAN H A I，NABIH A.Biological identifications through DNA barcodes［M］.Biodiversity conservation and utilization in a diverse world.IntechOpen，2012.［2］胡佳,戴丹,彭新红,等.灵芝属9个野生菌株的分子鉴定与系统发育关系分析［J］.食用菌，2023，45(1)：29-34.［3］谭秀梅,阿地力·沙塔尔,朴春根,等.无柄灵芝遗传多样性的SRAP、ITS、TEF1-α和LSU分析［J］.微生物学通报,2016,43(12):2667-2677.［4］耿晓进,蒋秋梦,张腾洋,等.秦皇岛地区三种野生食用菌分子鉴定与系统发育分析［J］.北方园艺，2021(8)：111-116.［5］孟虎,孙国琴,睢韡,等.ISSR技术在食用菌研究上的应用［J］.北方园艺,2016(5):207-210.［6］陆欢,尚晓冬,宋春艳,等.基于ISSR标记和农艺性状的金顶侧耳种质资源遗传多样性分析［J］.食用菌学报，2022，29(6)：14-24.［7］吕锐玲,陈实,王杰,等.基于ISSR标记的我国羊肚菌栽培品种遗传多样性分析［J］.北方园艺，2023(8)：107-114.［8］郭金英,史灵燕,张慧,等.冀中南毛木耳栽培菌株的鉴定和优良菌株筛选［J］.北方园艺，2019(14)：129-134.［9］李慧,夏伟伟,胡子钰,等.利用ISSR-SCAR标记鉴定金针菇双-单杂交子［J］.北方园艺,2022(16):121-127.［10］KUMAR S,STECHER G,TAMURA K.MEGA7：Molecular evolutionary genetics analysis version 7.0 for bigger datasets［J］.Molecular Biology and Evolution，2016，33(7)：1870-1874.［11］ROHLF F J.NTSYS-pc：Microcomputer programs for numerical taxonomy and multivariate analysis［J］.The American Statistician，1987，41(4)：330.［12］CHEN C,CHEN H,ZHANG Y,et al.TBtools:an integrative toolkit developed for interactive analyses of big biological data［Z］.Molecular Plant,2020,13(8):1194-1202.［13］邢佳慧.灵芝属的物种多样性、分类与系统发育研究［D］.北京:北京林业大学，2019.［14］戚元成,张小强,高玉千,等.中国栽培糙皮侧耳品种体细胞不亲和性实验与RAPD分析结果的比较［J］.菌物学报，2010，29(3)：379-388.［15］袁滨,柯丽娜,黄艺宁,等.应用拮抗和ISSR方法综合鉴定灰树花菌株［J］.福建农业学报，2019，34(4)：393-399.［16］汪乔,王祥锋,郭玉峰,等.肺形侧耳菌株的遗传差异分析与农艺性状评价［J］.中国农学通报，2022，38(10)：46-52.［17］唐传红,唐庆九,张劲松,等.基于rDNA ITS序列和ERIC-PCR研究虫草真菌无性分离物的遗传多样性［J］.南京农业大学学报，2014，37(2)：146-152.［18］XING X K,MA X T,HART M M,et al.Genetic diversity and evolution of Chinese traditional medicinal fungus Polyporus umbellatus (Polyporales，Basidiomycota)［J］.PLoS One，2013，8(3)：e58807.［19］张宇,黄国弟,莫永龙,等.CDDP结合SRAP标记分析杧果品种的遗传多样性［J］.中国南方果树，2022，51(2)：57-63.
相似文献/References:
[1]王瑞霞,甄红伟,牟建楼,等.灵芝红枣复合饮料的研制[J].北方园艺,2013,37(04):151.
[2]朱会霞.灵芝真菌发酵产灵芝酸培养基优化研究[J].北方园艺,2013,37(22):154.
ZHU Hui-xia.Study on the Optimization of Culture Medium of Ganoderma Fermentation Before Extraction of Ganoderic Acid[J].Northern Horticulture,2013,37(3):154.
[3]董平,赵培宝,任爱芝.蝴蝶兰灰霉病菌的分离鉴定[J].北方园艺,2014,38(04):109.
DONG Ping,ZHAO Pei-bao,REN Ai-zhi.Isolation and Identification of Botrytis cinerea?in Phalaenopsis[J].Northern Horticulture,2014,38(3):109.
[4]朱会霞.不同培养方式对灵芝中麦角固醇产量的影响研究[J].北方园艺,2014,38(04):127.
ZHU Hui-xia.Effect of Different Fermentation Methods on the Yield of Ergosterol for Ganodorma lucidum [J].Northern Horticulture,2014,38(3):127.
[5]孙金旭.溶氧控制条件对深层灵芝发酵生产灵芝酸产量的影响[J].北方园艺,2014,38(05):130.
SUN Jin-xu.Effect of Dissolved Oxygen Controlling Conditions on Production of Ganoderic Acids by Ganoderma[J].Northern Horticulture,2014,38(3):130.
[6]朱会霞.灵芝发酵覆盆子生产总黄酮的培养基优化研究[J].北方园艺,2013,37(15):169.
ZHU Hui-xia.Study on Optimization of Medium for Total Flavnoids Production from Rubus idaeus L. by GanodermaFermentation[J].Northern Horticulture,2013,37(3):169.
[7]欧立军,孙海军.基于ITS序列的蔷薇纲二十六个物种遗传多样性分析[J].北方园艺,2013,37(11):94.
OU Li-jun,SUN Hai-jun.Genetic Diversity Analysis of Twenty-six Species in Rose Class Based on ITS Sequences[J].Northern Horticulture,2013,37(3):94.
[8]杨海军,周小苹.辣椒粉固态培养灵芝菌丝体的培养基优化研究[J].北方园艺,2013,37(07):161.
YANG Hai-jun,ZHOU Xiao-ping.Study on Optimization of Solid Medium Containing Chili Powder for Ganoderma lucidum Culture[J].Northern Horticulture,2013,37(3):161.
[9]刘 洋,孙来玉,陆云华,等.九株蛹虫草菌株的生物学性状研究[J].北方园艺,2013,37(20):140.
LIU Yang,SUN Lai-yu,LU Yun-hua,et al.Study on the Biological Characteristics of Nine Cordyceps militaris Strains[J].Northern Horticulture,2013,37(3):140.
[10]王昱,王义,王康宇,等.灵芝原生质体的制备与再生研究[J].北方园艺,2013,37(16):184.
WANGYu,WANGYi,WANGKang-yu,et al.StudyonPreparationandRegenerationoftheProtoplastsfrom Ganodermalucidum[J].Northern Horticulture,2013,37(3):184.
备注/Memo
第一作者简介：赵翠敏（1992-），女，硕士，助理农艺师，现主要从事食用菌遗传育种等研究工作。E-mail：zcm1014@163.com.责任作者：李敏敏（1991-），女，博士，高级农艺师，现主要从事食用菌遗传育种等研究工作。E-mail：liminmin527@163.com.基金项目：山东省聊城市重点研发计划（农业良种工程）资助项目（2021LZ01）。收稿日期：2023-07-11