Highly polymorphic DNA markers to specify strains of the ectomycorrhizal basidiomycete Tricholoma matsutake based on σmarY1, the long terminal repeat of gypsy-type retroelement marY1
Highly polymorphic DNA markers to specify strains of the ectomycorrhizal basidiomycete Tricholoma matsutake based on σmarY1, the long terminal repeat of gypsy-type retroelement marY1
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DOI：
10.1007/s00572-004-0319-0
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发表时间：
2005-05
期刊：
影响因子：
3.9
通讯作者：
H. Murata;K. Babasaki;A. Yamada
中科院分区：
文献类型：
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作者：
H. Murata;K. Babasaki;A. Yamada
外生菌根真菌松口蘑（Tricholoma matsutake）在松属（Pinussp.）森林本文报道了一种基于σ marY 1序列设计的外向引物PCR方法，该引物是基于吉普赛型逆转录元件marY 1的长末端重复序列σ marY 1设计的。松茸。根据σ marY 1 5′端保守的22 bp序列设计引物进行PCR扩增，共扩增出73条可靠的条带，其谱型与菌株有关。MatsutakeandT. magnivelare，后者被称为“美国松茸”。该PCR体系在其他菌种中均未检测到条带。bakamatsutakeandT. fulvocastaneum，与T.松茸以及寄主植物赤松。同样地，根据σ marY 1变异体的48-73 bp和281-308 bp的26-bp和28-bp序列设计的一组引物，仅在T菌株中产生90条可靠的条带。松茸。从理论上讲，用22 bp引物进行PCR可以产生273种或9.4×1021种多态性，而用26 bp和28 bp引物组合进行PCR可以产生290种或1.2× 1027种多态性。将两个不同分离株错误指定为同一菌株的概率<1/1021。而基于σ marY 1 5′端的引物所赋予的多态性则表现出一组T.与松茸相比，那些由基于内部序列的引物产生的结果更清楚地表明了种内多样性。这两个系统都揭示了T.松竹种内分化。外生菌根形成于P. densiflora和T.利用本研究建立的PCR体系对松茸进行了鉴定。该方法以σ marY 1为遗传标记，可用于分析T.松茸，监测单个菌根的行为，并指定在市场上交易的子实体的生态背景。
The ectomycorrhizal basidiomyceteTricholoma matsutakeproduces commercially valuable fruit bodies—matsutake—inPinussp. forest. Here we report that PCR with outward facing primers designed based on sequences comprising σmarY1, the long terminal repeat of thegypsy-type retroelementmarY1, specifies strains ofT. matsutake. PCR with a primer based on the 22-bp sequence conserved at the 5′-end of σmarY1conferred 73 reliable bands overall whose profiles depend upon strains ofT. matsutakeandT. magnivelare, the latter known as ‘American matsutake’. This PCR system gave no detectable band in any other species ofTricholomatested, includingT. bakamatsutakeandT. fulvocastaneum, symbionts closely related toT. matsutake, as well as a host plant,Pinus densiflora. Similarly, PCR with a set of primers based on 26-bp and 28-bp sequences at bp 48–73 and bp 281–308 of σmarY1, internal regions that are mutated in a variant ofmarY1, conferred 90 reliable bands only in strains ofT. matsutake. Theoretically, PCR with the 22-bp primer would allow generation of 273, or 9.4×1021, types of polymorphism, and PCR with a combination of 26- and 28-bp primers, 290, or 1.2×1027types. The probability of falsely specifying two different isolates as the same strain is <1/1021. While polymorphisms conferred by the primer based on the 5′ end of σmarY1rather exhibit genetic conservation of a group ofT. matsutake, those resulting from primers based on the internal sequences more clearly demonstrate intra-specific diversification. Both systems revealed thatT. matsutakeis divergent within the species. Ectomycorrhizas formed betweenP. densifloraandT. matsutakewere identified by the PCR systems developed in the present study. This method, using σmarY1as a genetic marker, is useful in analyzing the diversity ofT. matsutake, monitoring the behavior of individual mycorrhizas, and specifying the ecological background of fruit bodies traded in markets.