Article(id=1237016046385820550, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1237016039171608726, articleNumber=null, orderNo=null, doi=10.3969/j.issn.1000-2561.2025.09.015, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1741795200000, receivedDateStr=2025-03-13, revisedDate=null, revisedDateStr=null, acceptedDate=1746633600000, acceptedDateStr=2025-05-08, onlineDate=1772857208105, onlineDateStr=2026-03-07, pubDate=1758729600000, pubDateStr=2025-09-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1772857208105, onlineIssueDateStr=2026-03-07, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1772857208105, creator=13701087609, updateTime=1772857208105, updator=13701087609, issue=Issue{id=1237016039171608726, tenantId=1146029695717560320, journalId=1235980609244409860, year='2025', volume='46', issue='9', pageStart='2031', pageEnd='2286', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=null, createTime=1772857206385, creator=13701087609, updateTime=1773049161445, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1237821157118890427, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1237016039171608726, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1237821157118890428, tenantId=1146029695717560320, journalId=1235980609244409860, issueId=1237016039171608726, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=2173, endPage=2180, ext={EN=ArticleExt(id=1237016048029987748, articleId=1237016046385820550, tenantId=1146029695717560320, journalId=1235980609244409860, language=EN, title=Analysis of Genetic Diversity of Pitaya Canker Pathogen Neoscytalidium dimidiatum in Guangdong and Guangxi, columnId=1236292524264968282, journalTitle=Chinese Journal of Tropical Crops, columnName=Plant Protection & Bio-safety, runingTitle=null, highlight=null, articleAbstract=

Guangdong and Guangxi are the major production regions of pitaya and the common area for pitaya canker. Understanding the genetic diversity of Neoscytalidium dimidiatum is of great significance for the disease prevention and control. This research focused on four geographical population of N. dimidiatum mainly collected from Guangxi and Guangdong. To obtain the genetic fingerprints of N. dimidiatum, the isolates were amplified using ISSR genetic molecular markers and further analyzed the genetic diversity. The tested N. dimidiatum isolates were amplified with eight ISSR primers, resulting in 203 bands and the percentage of polymorphic sites was 31.7%, with the genetic similarity coefficient for any two germplasm resources ranged from 0.7619-1.0000. When the genetic similarity coefficient was 0.90 on the UPGMA cluster, all isolates could be divided into three groups, but the genetic groups were not closely related to the geographical distribution. The results of principal component analysis indicated considerable overlap between the four geographical groups and there were no relatively independent geographical poputation. This study proved that there existed lower level of genetic polymorphism in the isolates and there was frequent exchange of inoculum source between different places. The study would provide the scientific basis for researching the genetic architecture and pathogen surveillance of pitaya canker pathogen N. dimidiatum.

, correspAuthors=Liping QIN, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, authorCompany=null, fund=null, authors=null, authorsList=Shanyu LIN, Jinling HUANG, Xiuhong LU, Liping QIN), CN=ArticleExt(id=1237016050831781994, articleId=1237016046385820550, tenantId=1146029695717560320, journalId=1235980609244409860, language=CN, title=广东和广西火龙果溃疡病菌新暗色柱节孢遗传多样性分析, columnId=1236292524520820846, journalTitle=热带作物学报, columnName=植物保护与生物安全, runingTitle=null, highlight=null, articleAbstract=

广东和广西是我国火龙果主要产区,也是火龙果溃疡病的常发区,明确这些地区火龙果溃疡病菌新暗色柱节孢(Neoscytalidium dimidiatum)的遗传多样性对溃疡病的防治具有重要意义。本研究以主要来源于广西、广东的4个火龙果溃疡病菌地理群体为试验对象,利用ISSR分子标记扩增以获得其指纹图谱,并进一步分析溃疡病菌的遗传分化情况。供试的火龙果溃疡病菌经过8条ISSR引物扩增,得到203条条带,多态性位点为31.7%,菌株间遗传相似系数在0.7619~1.0000之间。在UPGMA聚类树上遗传相似系数为0.90时可分为3个类群,但遗传类群与地理分布上并无紧密联系。主成分分析结果显示,4个地理群体重叠性较高,不存在相对独立的地理种群。本研究证明我国火龙果溃疡病菌遗传多样性水平较低,各地之间存在频繁的菌源交流,该结果可为新暗色柱节孢的种群遗传结构和火龙果溃疡病病原菌监测的研究提供科学依据。

, correspAuthors=覃丽萍, authorNote=null, correspAuthorsNote=
* 覃丽萍(QIN Liping),E-mail:
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林珊宇(1983—),女,博士,助理研究员,研究方向:植物病理学。

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林珊宇(1983—),女,博士,助理研究员,研究方向:植物病理学。

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林珊宇(1983—),女,博士,助理研究员,研究方向:植物病理学。

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Geographic population delineation of sampling sites

, figureFileSmall=null, figureFileBig=null, tableContent=
群体
Group
采集地点(样本)
Sampling location(Sample)
菌株数
Number of strains
G1广州(H38,S1)、阳江(YJ10-1,YJ11-1,YJ12-1,YJ13-1,YJ15-1,YJH1-1,YJH2-2,YJH3-2)、东莞(DG6,DG7,DG8,DG10,PY3)、惠州(HZ1-1,HZ7-1,HZ8-1,HZ10-1)、江门(JM1-1,JM2-1,JM3-1,JM4-2,JM4-4,JM5,JM6-2,JM13-1)、揭阳(JY2-1,JY3-1,JY10-1,JY13-1,JY15-1)、茂名(GZ1-1,GZ2-1)、梅州(MZH1-1,MZH2-1)、深圳(SZ2-1,SX3-2)和漳州(LH2-1)39
G2南宁(LA1,LJ3,LX1,LY2,NT1,SQ2,YLY1,KM1)、贵港(GG1-1,GG6-1)、桂林(GL2)、梧州(CX3,WZ3)和玉林(BL3,BL5,BL8,XY4,XY9,RX1,RY2)20
G3湛江(ZJ1)、北海(BH5,BH9,HF2-5,HF4)、钦州(MP2-2,MP2-7,MP2-9,MP10-2,MP2-3)和防城港(FQ4)11
G4崇左(NM2,NM3,NM8)、百色(PG2,BS4)、玉溪(YN5-2)和黔南(QN1)7
), ArticleFig(id=1237023461487473268, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1237016046385820550, language=CN, label=表1, caption=

采样点地理群体划分

, figureFileSmall=null, figureFileBig=null, tableContent=
群体
Group
采集地点(样本)
Sampling location(Sample)
菌株数
Number of strains
G1广州(H38,S1)、阳江(YJ10-1,YJ11-1,YJ12-1,YJ13-1,YJ15-1,YJH1-1,YJH2-2,YJH3-2)、东莞(DG6,DG7,DG8,DG10,PY3)、惠州(HZ1-1,HZ7-1,HZ8-1,HZ10-1)、江门(JM1-1,JM2-1,JM3-1,JM4-2,JM4-4,JM5,JM6-2,JM13-1)、揭阳(JY2-1,JY3-1,JY10-1,JY13-1,JY15-1)、茂名(GZ1-1,GZ2-1)、梅州(MZH1-1,MZH2-1)、深圳(SZ2-1,SX3-2)和漳州(LH2-1)39
G2南宁(LA1,LJ3,LX1,LY2,NT1,SQ2,YLY1,KM1)、贵港(GG1-1,GG6-1)、桂林(GL2)、梧州(CX3,WZ3)和玉林(BL3,BL5,BL8,XY4,XY9,RX1,RY2)20
G3湛江(ZJ1)、北海(BH5,BH9,HF2-5,HF4)、钦州(MP2-2,MP2-7,MP2-9,MP10-2,MP2-3)和防城港(FQ4)11
G4崇左(NM2,NM3,NM8)、百色(PG2,BS4)、玉溪(YN5-2)和黔南(QN1)7
), ArticleFig(id=1237023461588136573, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1237016046385820550, language=EN, label=Tab. 2, caption=

Screening results of ISSR analysis primer annealing temperature

, figureFileSmall=null, figureFileBig=null, tableContent=
引物
Primer
退火温度
Annealing temperature/℃
引物序列(5′–3′)
Primer sequence(5′–3′)
(GTG)559GTGGTGGTGGTGGTG
U0260DHBCGACGACGACGACGA
UBC 88046GATAGATAGACAGACA
VCA1351AGAGAGAGAGAGAGAGS
VCA2160GTCGTCGTCGTCGTCGTC
VCA2261GTGGTGGTGGTGGTGGTG
VCA2554CACCACCACCACGC
YHY(GT)7G48YHYGTGTGTGTGTGTGTG
), ArticleFig(id=1237023461697188483, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1237016046385820550, language=CN, label=表2, caption=

ISSR分析引物退火温度的筛选结果

, figureFileSmall=null, figureFileBig=null, tableContent=
引物
Primer
退火温度
Annealing temperature/℃
引物序列(5′–3′)
Primer sequence(5′–3′)
(GTG)559GTGGTGGTGGTGGTG
U0260DHBCGACGACGACGACGA
UBC 88046GATAGATAGACAGACA
VCA1351AGAGAGAGAGAGAGAGS
VCA2160GTCGTCGTCGTCGTCGTC
VCA2261GTGGTGGTGGTGGTGGTG
VCA2554CACCACCACCACGC
YHY(GT)7G48YHYGTGTGTGTGTGTGTG
), ArticleFig(id=1237023461789463180, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1237016046385820550, language=EN, label=Tab. 3, caption=

Genetic information of 77 N. dimidiatum isolates amplified by 8 ISSR primers

, figureFileSmall=null, figureFileBig=null, tableContent=
引物
Primer
等位基因观测数Na有效等位基因数NeNei's基因多样性指数HShannon信息指数I位点数loci多态性条带数NP多态性位点百分率P/%
U021.1431.0240.0210.0397114.3
M131.5001.2590.1450.2204250.0
(GTG)51.6671.1030.0880.1696466.7
VCA131.7501.0690.0600.1244375.0
VCA211.0001.0000.0000.000500
AVC221.3331.0530.0420.0766233.3
VCA251.1431.0750.0490.0757114.3
YHY(GT)7G1.0001.0000.0000.000400
平均值1.3171.0730.0510.0885.3751.62531.7
), ArticleFig(id=1237023461919486613, tenantId=1146029695717560320, journalId=1235980609244409860, articleId=1237016046385820550, language=CN, label=表3, caption=

8个ISSR引物扩增77株新暗色柱节孢获得的遗传信息

, figureFileSmall=null, figureFileBig=null, tableContent=
引物
Primer
等位基因观测数Na有效等位基因数NeNei's基因多样性指数HShannon信息指数I位点数loci多态性条带数NP多态性位点百分率P/%
U021.1431.0240.0210.0397114.3
M131.5001.2590.1450.2204250.0
(GTG)51.6671.1030.0880.1696466.7
VCA131.7501.0690.0600.1244375.0
VCA211.0001.0000.0000.000500
AVC221.3331.0530.0420.0766233.3
VCA251.1431.0750.0490.0757114.3
YHY(GT)7G1.0001.0000.0000.000400
平均值1.3171.0730.0510.0885.3751.62531.7
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广东和广西火龙果溃疡病菌新暗色柱节孢遗传多样性分析
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林珊宇 , 黄金玲 , 陆秀红 , 覃丽萍 *
热带作物学报 | 植物保护与生物安全 2025,46(9): 2173-2180
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热带作物学报 | 植物保护与生物安全 2025, 46(9): 2173-2180
广东和广西火龙果溃疡病菌新暗色柱节孢遗传多样性分析
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林珊宇, 黄金玲, 陆秀红, 覃丽萍*
作者信息
  • 广西作物病虫害生物学重点实验室/广西壮族自治区农业科学院植物保护研究所/农业农村部华南果蔬绿色防控重点实验室,广西南宁 530007
  • 林珊宇(1983—),女,博士,助理研究员,研究方向:植物病理学。

通讯作者:

* 覃丽萍(QIN Liping),E-mail:
Analysis of Genetic Diversity of Pitaya Canker Pathogen Neoscytalidium dimidiatum in Guangdong and Guangxi
Shanyu LIN, Jinling HUANG, Xiuhong LU, Liping QIN*
Affiliations
  • Guangxi Key Laboratory of Biology for Crop Diseases and Insect Pests / Plant Protection Research Institute, Guangxi Academy of Agricultural Sciences / Key Laboratory of Green Prevention and Control on Fruits and Vegetables in South China, Ministry of Agriculture and Rural Affairs, Nanning, Guangxi 530007, China
出版时间: 2025-09-25 doi: 10.3969/j.issn.1000-2561.2025.09.015
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广东和广西是我国火龙果主要产区,也是火龙果溃疡病的常发区,明确这些地区火龙果溃疡病菌新暗色柱节孢(Neoscytalidium dimidiatum)的遗传多样性对溃疡病的防治具有重要意义。本研究以主要来源于广西、广东的4个火龙果溃疡病菌地理群体为试验对象,利用ISSR分子标记扩增以获得其指纹图谱,并进一步分析溃疡病菌的遗传分化情况。供试的火龙果溃疡病菌经过8条ISSR引物扩增,得到203条条带,多态性位点为31.7%,菌株间遗传相似系数在0.7619~1.0000之间。在UPGMA聚类树上遗传相似系数为0.90时可分为3个类群,但遗传类群与地理分布上并无紧密联系。主成分分析结果显示,4个地理群体重叠性较高,不存在相对独立的地理种群。本研究证明我国火龙果溃疡病菌遗传多样性水平较低,各地之间存在频繁的菌源交流,该结果可为新暗色柱节孢的种群遗传结构和火龙果溃疡病病原菌监测的研究提供科学依据。

火龙果溃疡病  /  新暗色柱节孢  /  遗传多样性  /  ISSR

Guangdong and Guangxi are the major production regions of pitaya and the common area for pitaya canker. Understanding the genetic diversity of Neoscytalidium dimidiatum is of great significance for the disease prevention and control. This research focused on four geographical population of N. dimidiatum mainly collected from Guangxi and Guangdong. To obtain the genetic fingerprints of N. dimidiatum, the isolates were amplified using ISSR genetic molecular markers and further analyzed the genetic diversity. The tested N. dimidiatum isolates were amplified with eight ISSR primers, resulting in 203 bands and the percentage of polymorphic sites was 31.7%, with the genetic similarity coefficient for any two germplasm resources ranged from 0.7619-1.0000. When the genetic similarity coefficient was 0.90 on the UPGMA cluster, all isolates could be divided into three groups, but the genetic groups were not closely related to the geographical distribution. The results of principal component analysis indicated considerable overlap between the four geographical groups and there were no relatively independent geographical poputation. This study proved that there existed lower level of genetic polymorphism in the isolates and there was frequent exchange of inoculum source between different places. The study would provide the scientific basis for researching the genetic architecture and pathogen surveillance of pitaya canker pathogen N. dimidiatum.

pitaya canker  /  Neoscytalidium dimidiatum  /  genetic diversity  /  ISSR
林珊宇, 黄金玲, 陆秀红, 覃丽萍. 广东和广西火龙果溃疡病菌新暗色柱节孢遗传多样性分析. 热带作物学报, 2025 , 46 (9) : 2173 -2180 . DOI: 10.3969/j.issn.1000-2561.2025.09.015
Shanyu LIN, Jinling HUANG, Xiuhong LU, Liping QIN. Analysis of Genetic Diversity of Pitaya Canker Pathogen Neoscytalidium dimidiatum in Guangdong and Guangxi[J]. Chinese Journal of Tropical Crops, 2025 , 46 (9) : 2173 -2180 . DOI: 10.3969/j.issn.1000-2561.2025.09.015
火龙果溃疡病在全球范围内对火龙果的品质和产量造成严重威胁[1]。在高温高湿的季节,火龙果溃疡病易于流行,病原菌分生孢子穿透表层建立寄生关系后,最初形成褪绿斑,并伴随黄色晕圈,随着病程发展,病斑突出枝条表面形成硬痂,末期病变死亡组织脱落后,枝条上留下缺刻,在高湿条件下,损伤的细胞壁软化和溶解,形成空洞和塌陷[2]。火龙果溃疡病主要为害我国台湾、广西、广东、海南、贵州和云南的火龙果园,在美国、伊朗、马来西亚、以色列、泰国、越南等国家也有不同程度的发生[3-6]。据报道佛罗里达州一些火龙果园感病率超过70%[5],2012年广东省清远市火龙果溃疡病的发病率高达60%[7],广西防城港市高发病区火龙果溃疡病的发病率在50%左右,造成产量下降10%~50%,商品价值受到严重损害[8]。2012年台湾首次报道葡萄座腔菌科(Botryosphaeriaceae)成员新暗色柱节孢(N. dimidiatum)引起火龙果茎溃疡[9],通过不断地形态学观察、多基因联合构建系统发育树分析,现在已经明确葡萄座腔菌科的一些真菌引起火龙果溃疡病的病原菌,其中新暗色柱节孢是最重要的一种[110]
真菌遗传信息的可变性使病原菌能够忍受环境压力,并通过多种生活方式在种群中形成不同的生存模式,作物病害暴发往往伴随着病原菌的遗传变异[11],因此研究新暗色柱节孢群体遗传构成信息和遗传变化,对作物的抗病育种、病害流行监测和防控具有重要意义。与物种鉴定类似,遗传多样性也可以通过形态学、生物化学和分子方法来确定。分子方法可以在不受环境因素干扰的情况下快速获得结果[12]。目前多个ISSR、SSR和SRAP标记已被开发并用于葡萄座腔菌中葡萄溃疡病菌(Botryosphaeria dothidea)等的遗传多样性分析,并证实了该种群具有较高的遗传变异[13-14]。中国火龙果栽培区地域宽广、环境多样、物种丰富,不同地理环境、多样化栽培模式可能给我国火龙果溃疡病菌群体的遗传多样性带来影响。因此,本研究拟通过对主要来自广东、广西的火龙果溃疡病病原菌新暗色柱节孢进行ISSR指纹图谱分析,以明确其优势种的遗传结构,为深入研究火龙果溃疡病及防治等提供理论基础。
于2018—2022年,从南宁、北海、钦州、贵港、梧州、玉林,广州、茂名、湛江、阳江、江门、惠州、梅州、揭阳、深圳等地收集火龙果溃疡病样品,进行常规病菌组织分离,也采集了距离较远的福建漳州、云南玉溪、贵州黔南的3个病样作为参考。纯化的菌株经过形态学和分子生物学鉴定后,获得77株新暗色柱节孢,并根据采样地点分布及地形地势,分为4个地理群体(表1):广东和福建群体(G1),该区域海拔低,地势平坦开阔,夏秋易受台风干扰,溃疡病发生较严重;广西中、北部群体(G2),该区域多为丘陵,气候湿润;北部湾沿海群体(G3),该区域气候特征与G1相似,部分地形为丘陵;广西西部和云南、贵州群体(G4),该区域以丘陵和喀斯特地貌为主,冬天平均气温低于沿海,溃疡病发病较轻。
使用天根植物基因组DNA提取试剂盒(DP305)提取DNA,步骤按说明书进行。提取的总DNA用琼脂糖凝胶电泳分析DNA降解程度以及是否有污染;使用nanodrop测定OD260、OD280,并计算OD260/OD280比值,检测DNA的纯度。并将DNA溶液稀释至50 ng/μL用于后续试验。
从ISSR引物库中随机选取44条引物并由生工生物工程(上海)股份有限公司合成,采用地理差距较大的4株新暗色柱节孢(LQ6、NT1、QN1和ZJ1)DNA作为扩增模版来筛选多态性好的引物,用无菌ddH2O作阴性对照。选择扩增条带清晰、多态性好和具有重复性的引物用于后续新暗色柱节孢基因组ISSR-PCR扩增。
以优选的ISSR引物理论退火温度Tm值为中间值,按1 ℃设置温度梯度,共选取5个温度对候选的ISSR引物退火温度进行筛选,试验重复2次。
采用上述优选的ISSR-PCR扩增的退火温度,对新暗色柱节孢分离株DNA进行扩增。扩增体系为:94 ℃预变性5 min;94 ℃变性30 s,最佳退火温度45 s,72 ℃延伸1.5 min,共35个循环;最后72 ℃延伸10 min,4 ℃保存。
在对ISSR-PCR的谱带的统计分析中,首先确定条带的大小,然后根据相应位置条带的有无,分别记为1和0。8个引物扩增出的每一个条带都作为一个遗传位点进行统计,构建菌株和条带的1和0二元矩阵,从而将图像信息转化为数据信息,并将其保存成Excel文件格式。然后,利用POPGENE version 1.32软件计算参数,反映基因多少和状况的遗传参数为多态性条带数(number of polymorphic loci,NP),多态性位点百分率(proportion of polymorphic loci,P),等位基因观测数(observed number of alleles,Na),有效等位基因数(effective number of alleles,Ne);反映基因多样性信息的遗传参数为Nei's基因多样性指数(gene diversity,H),Shannon信息指数(Shannons information index,I)。最后计算遗传距离,并用NTsys-2.0软件计算遗传相似系数,用非加权算术平均聚类(Unweighted Pair-Group Mean Average,UPGMA)方法进行聚类分析,并通过Tree plot模块生成聚类图,使用GenAlEx软件进行主成分分析。
选用来自广西南宁(NT1,LQ6)、广东湛江(ZJ1)和贵州黔南(QN1)的新暗色柱节孢,用44条ISSR引物进行PCR扩增。扩增结果表明,不同的引物对新暗色柱节孢扩增出的条带数目存在差异,一些引物扩增产物的条带数目较少,如引物VCA02、VCA27等;一些引物则无扩增条带,如(GATA)4,这些引物均不适合用于对新暗色柱节孢群体遗传多样性的分析。最终从供试的44条ISSR引物中筛选出(GTG)5、M13、U02、VCA13、VCA21、VCA22、VCA25、YHY(GT)7G 8条引物,其ISSR-PCR扩增产物的多态性良好,条带清晰稳定,可用于对新暗色柱节孢的遗传结构研究。
引物退火温度筛选结果表明,退火温度一定程度上能够影响ISSR-PCR扩增效果,退火温度过低,有时会降低ISSR扩增的多态性,退火温度过高则不利于形成清晰的条带。部分引物的最佳退火温度与Tm值存在偏离,重复2次后,筛选获得条带清晰稳定的温度作为随后各引物扩增的最佳退火温度,选用引物退火温度的筛选结果见表2
77株新暗色柱节孢株经过ISSR-PCR扩增,获得不同引物的扩增图谱,8个引物共扩增出43个位点,其中多态性位点数为13个,多态性位点百分率为31.7%,所获得的遗传多样性信息见表3。结果表明新暗色柱节孢遗传多样性在各群体之间具有一定差异,但遗传分化程度较低。
可视化热图(图1)可以展现不同指标、不同样本之间的相似性,颜色越偏红,说明相似性越大,越偏蓝说明相似性越低。经比较,大部分菌株之间的遗传相似系数在0.95以上(图1),说明新暗色柱节孢群体遗产变异较低。
从UPGAM聚类树(图2)可以看出,在遗传相似系数为0.90水平上,77个菌株被聚类成3个类群,第Ⅰ类群包含广西北海、南宁和梧州的3个菌株,第Ⅱ个类群仅有来自广西玉林的1个菌株,其余73个菌株被聚在第Ⅲ个类群。聚类结果表明,77个火龙果溃疡病菌之间具有较高的遗传相似性,但是种内仍存在一定的遗传变异;同一地理来源的菌株归入不同的类群或者亚类群中,说明菌株的遗传多样性与其地理来源无明显的相关性。但小范围来说,菌株的聚集具有一定的特点,例如来自不同区域的27个菌株聚集在遗传相似系数约为0.976的A分支上,广东和福建沿海的G1群体菌株在B和C分支中占主导地位。来自云南玉溪、贵州黔南和福建漳州的3个菌株也未和广东和广西菌株分离。
主成分分析可知,G2、G3和G4群体都在G1群体范围内。虽然G2、G3和G4大部分区域互相重叠,但G2和G3重叠的部分更多,说明来自广西中、北部群体和北部湾沿海的菌株亲缘关系更为密切。由图3可知,所有菌株大致可分为4个类群,每个类群都包含了来自不同地理分组的菌株,表明不同地理来源之间菌系交流频繁,遗传关系相近。
本研究选取来自广东、广西和周边的77株新暗色柱节孢按地理位置分成4个群组,进行种群的遗传多样性分析。葡萄座腔菌科真菌作为水果和林木上的重要病原菌,为揭示其遗传关系,人们对它们的遗传多样性进行了一些探索,但目前尚无火龙果溃疡病菌新暗色柱节孢遗传多样性研究的报道。基于ISSR的遗传多样性研究结果显示新暗色柱节孢菌株间的遗传相似系数范围为0.7619~1.0000,Shannon's信息指数(I)为0.088;基因多样性指数(H)为0.051,表明各菌株之间遗传距离比较近,相似性较高。通过UPGMA方法构建的火龙果溃疡病菌遗传关系的聚类分析结果显示,来自同一地理区域的菌株分布在不同的聚类群中,遗传系数高于0.91的分支Ⅲ包含不同地域的菌株,来自云南玉溪、贵州黔南和福建漳州的3个菌株也未和广东和广西菌株有显著差异,说明火龙果溃疡病菌的遗传亲缘关系与地理来源之间并无明显的相关性。主成分分析说明来自广西中、北部群体和北部湾沿海的菌株亲缘关系更为密切,推测某些适应性强的主流菌株已经分布在规模化的火龙果种植区。总体上,来自内陆的菌株表现出更高的遗传分化,例如广西南宁的NT1、YLY1,广西玉林的RY2、广西梧州的CX3,贵州黔南的QN1,这几个菌株位于遗传系数低于0.94的分支上。较高的温度利于火龙果溃疡病的发生,可能纬度较高的菌株为了适应冬季较低的温度,遗传信息发生了改变,而病原菌的遗传变异给病害防控提出了更高要求。
研究病原菌的遗传多样性有利于了解群体的起源和演变,如果病原菌是最近的一次引入,那么病原体群体将在遗传上是同质的,表现出与源群体相似或相同的遗传变异[15]。相反,如果病原体群体是由不同来源重组(地方性源群体与引进群体重组)产生的,那么它将表现出较高的遗传多样性[16]。例如CHETHANA等[17]利用全基因组序列数据开发的新型微卫星标记探索了葡萄座腔菌(B. dothidea)的遗传多样性和群体结构,结果表明,从中国葡萄品种中分离出的B. dothidea,不同地域菌株具有相同的等位基因,并表现出高度的遗传多样性,表明中国葡萄藤上的B. dothidea群体起源复杂,包括本地寄主转移、外来引进以及本地和引进分离株之间的重组。火龙果起源于热带中南美洲地区,后传入越南、泰国等东南亚国家和中国台湾[18-19]。从1998年开始,我国陆续从越南引种火龙果,并随着火龙果规模化种植,苗木调运十分频繁[20-21]。2012年我国台湾首次发现火龙果茎溃疡由新暗色柱节孢引起,随后在很短的一段时间内,中国、伊朗、以色列、马来西亚、美国和泰国等火龙果种植区均发现了新暗色柱节孢引起火龙果溃疡病,从火龙果溃疡病菌较高的遗传相似性,可推测侵染我国火龙果的新暗色柱节孢种群可能是随着苗木输入和转运传播到各地。这一结果还预示着新暗色柱节孢可以通过气流或昆虫在2个地区之间传播,因为这2种方式的传播距离比雨水飞溅的孢子传播距离更长。迁徙的病菌携带着菌源地的基因型,造成引入地病害的流行,该结果与王璠[22]对湖北桃流胶病病原菌B. dothidea遗传多样性与地理来源无关的结论相似。
遗传多样性分析能够获得物种之间的遗传变异、物种进化以及它们对不同环境条件和生活方式的特殊适应的线索[23]。众所周知,生殖模式与遗传变异有关,经历有性生殖的种群更有可能表现出更大的遗传多样性[24]。葡萄座腔菌(B. dothidea)和小新壳梭孢(Neofusicoccum parvum)是非常重要的植物病原菌,种群表现出较高的遗传多样性,特别在本地寄主中表现出更高的遗传分化,显著的连锁平衡表明菌株之间发生了基因重组或有性生殖[25]。这能够增加病原菌子代适应自然选择的能力,在促进病害显症、地理和寄主扩张方面发挥重要作用。新暗色柱节孢虽然为携带MAT1-1的异宗交配型真菌[26],但尚未发现它的有性生殖,繁殖主要依赖于田间条件下的菌丝断裂形成节孢子,遗传物质交换不够充分,遗传分化较为保守,因此新暗色柱节孢寄主专化性较高,对木本植物的致病力有限,这符合GARCIA等[27]对新暗色柱节孢致病性的描述和田间病害发生的实际情况。
本研究使用ISSR标记探讨了我国火龙果主要产区溃疡病菌新暗色柱节孢的遗传多样性。研究结果显示,虽然新暗色柱节孢种群显示出一定的遗传分化,但多样性指数相对较低,自然地域种群的遗传亲缘关系与其地理来源之间并无明显的相关性,这为研究我国南方火龙果溃疡病的传播规律以及致病力差异提供理论依据。未来还需开展新暗色柱节孢致病机制研究,为病害的预防和治理提供科学指导。
  • 广西农业科学院基本科研业务专项(桂农科2021ZX25)
  • 广西农业科学院基本科研业务专项(桂农科2021YT062)
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doi: 10.3969/j.issn.1000-2561.2025.09.015
  • 接收时间:2025-03-13
  • 首发时间:2026-03-07
  • 出版时间:2025-09-25
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  • 收稿日期:2025-03-13
  • 录用日期:2025-05-08
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广西农业科学院基本科研业务专项(桂农科2021ZX25)
广西农业科学院基本科研业务专项(桂农科2021YT062)
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    广西作物病虫害生物学重点实验室/广西壮族自治区农业科学院植物保护研究所/农业农村部华南果蔬绿色防控重点实验室,广西南宁 530007

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2种不同金属材料的力学参数

Family
属数
Number of
genus
种数
Number of
species
占总种数比例
Percentage of
total species (%)

Genus
种数
Number of
species
占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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