两类木蹄层孔菌在酶蛋白水平上的遗传分化

木蹄层孔菌（Ｆｏｍｅｓ ｆｏｍｅｎｔａｒｉｕｓ）在中国东北和日本存在子实体形态不同、寄主范围明显有别的两个类型：小型和大型。用多位点分析方法研究了两者间的遗传分化状况。从两类４１个菌株的４种酶系统中检测出９个基因位点,其中８个位点上的绝大多数酶谱型都不为两者所共有,显示两个类型间的基因交流已极少发生。根据酶谱型频率得出两者间的遗传距离Ｄ＝０．９６５,属于典型的种间遗传分化。上述酶基因分化证据结合子     

木蹄层孔菌不同居群间生长特性、木质素降解酶与SRAP标记遗传多样性

以帽儿山、长白山、凉水、本溪木蹄层孔菌(Fomes fomentarius) 4个居群为研究对象,采用菌丝长度测量法比较4个地点木蹄层孔菌菌株在PDA固体培养基上的生长速度,采用菌丝体干重法比较4个地点木蹄层孔菌菌株在PDA液体培养基中生物量的变化,结果显示木蹄层孔菌在23 ℃下生长速度显著高于28 ℃,说明木蹄层孔菌的生长对温度较敏感,23 ℃更适合木蹄层孔菌的生长。在同一温度下培养,4个地点的木蹄层孔菌生长速度无显著差异。通过比色法检测4个地点的木蹄层孔菌木质素降解相关酶(木质素过氧化物酶,锰过氧化物酶,漆酶)活性差异,结果显示同一种酶酶活性在4个地点间没有显著差异;在不同培养基上培养时,3种酶在PDA培养基上的活性均显著高于完全培养基。同时,采用序列相关扩增多态性(Sequence-related amplified polymorphism, SRAP)技术初探了木蹄层孔菌4个居群的遗传多样性和遗传分化, 结果表明木蹄层孔菌4个居群中多态位点比率最高的是本溪,其次是帽儿山和凉水,而长白山最低;AMOVA分析结果显示,居群间的遗传分化为24.74%,居群内的遗传分化为75.26%,木蹄层孔菌的遗传分化主要发生在居群内部。根据Nei's遗传距离对木蹄层孔菌4个居群进行UPGMA聚类分析,结果显示帽儿山和本溪居群最先聚类,其次聚类的是长白山居群,最后是凉水居群。     

三种白腐菌生物学特性与木质纤维素酶基因遗传多样性

以采自东北林业大学帽儿山实验林场的3种白腐真菌——木蹄层孔菌(Fomes fomentarius)、鲍姆鲍姆木层孔菌(Phellinus baumiibaumii)和火木层孔菌(Phellinus igniarius)为材料,用菌落直径测量法比较3种白腐菌在马铃署葡萄糖固体培养基上的生长速度,采用菌丝体干重法比较其在马铃署葡萄糖液体培养基中的生物量变化。结果显示:马铃薯葡萄糖固体培养基上3种白腐菌均为快速生长类型,其生长速度木蹄层孔菌火木层孔菌鲍姆鲍姆木层孔菌;马铃署葡萄糖液体培养基中生物量增长速度木蹄层孔菌鲍姆鲍姆木层孔菌火木层孔菌。用比色法测量其木质纤维素酶活性,结果显示:木蹄层孔菌产锰过氧化物酶和漆酶量较高,鲍姆鲍姆木层孔菌和火木层孔菌产木质素过氧化物酶量较高;木蹄层孔菌、鲍姆鲍姆木层孔菌和火木层孔菌3种白腐菌的3种主要木质素酶(锰过氧化物酶、漆酶和木质素过氧化物酶)的表达量,种间差异显著(F=3.75*、5.20**、3.01*),白桦木屑诱导处理与对照间差异显著(F=3.84*、4.19*、5.28*);两种主要纤维素酶(葡聚糖内切酶、葡聚糖外切酶)的表达量,种间差异不显著,受碳源影响作用显著(F=3.99*、4.04*)。筛选29对引物组合,对3种白腐菌几种主要木质纤维素酶基因进行TRAP-PCR分子标记检测,比较3菌种间遗传差异,扩增总条带数为357条,多态性条带数为255条,多态性条带的比例为71.43%,其中木质素降解酶基因总多态位点比率为73.77%,纤维素降解酶基因总多态位点比率为68.97%。3种白腐菌的木质纤维素降解酶基因在种间均存在较高的遗传差异。因此,特定基因的TRAP分子标记可以用于木腐菌的遗传变异分析。     

4种形态类型诸葛菜的等位酶变异及遗传多样性分析

以采自江苏省南京市4个样点的4种不同形态类型(紫花毛果、紫花光果、白花光果和白花毛果)诸葛菜〔Orychophragmus violaceus(L.)O.E.Schulz〕63个单株为研究对象,采用不连续聚丙烯酰胺凝胶电泳技术(SDS-PAGE)对过氧化物酶同工酶(POD)、乙醇脱氢酶(ADH)、谷氨酸脱氢酶(GDH)、超氧化物歧化酶(SOD)和多酚氧化酶(PPO)酶谱进行了分析,并基于5个等位酶的酶谱分析结果对4种形态类型诸葛菜的遗传多样性进行了研究。结果表明:5个等位酶系统共包含12个位点32个等位基因,其中POD-1、POD-4、POD-5、POD-6、ADH-1、PPO-1、SOD-1和SOD-2为多态性位点;除具有共有谱带外,不同类型诸葛菜还具有各自特有的酶谱特征,其中,GDH-2的b带和c带分别是紫花类型和白花类型的特有谱带。紫花毛果、紫花光果和白花毛果类型的多态位点百分率(PPL)均为58.33%,而白花光果的PPL为66.67%;紫花毛果、紫花光果、白花光果和白花毛果每个位点的平均等位基因数分别为2.42、2.25、2.42和1.83,平均期望杂合度分别为0.35、0.30、0.38和0.29。4种形态类型诸葛菜的总基因多样度平均值为0.61,类型内基因多样度平均值为0.49,明显大于类型间基因多样度平均值(0.12);类型间的基因分化系数平均值为0.195,表明总基因多样性的80.5%来源于类型内。紫花光果和白花毛果类型间的遗传一致度最低(0.724 4)且遗传距离最大(0.322 5),而2个白花类型间的遗传一致度最高(0.954 1)且遗传距离最小(0.047 1)。研究结果显示:诸葛菜的种内变异程度很大、遗传分化程度较高,但在各类型内具有较高的遗传相似性。     

泡沙参同工酶基因位点的遗传分析

利用聚丙烯酰胺凝胶电泳技术 ,对来自天然群体 (居群 )的泡沙参 (Adenophora potaninii Korsh.)及其人工杂交子代进行了 8种同工酶的电泳检测和谱带遗传分析 ,以确定编码这些酶系统的基因位点和等位基因。选用 4种不同的凝胶缓冲系统 ,对下列不同酶系统进行了酶谱的遗传分析 :天冬氨酸转氨酶 (AAT)、酯酶 (EST)、甲酸脱氢酶 (FDH)、谷氨酸脱氢酶 (GDH)、异柠檬酸脱氢酶 (IDH)、乳酸脱氢酶(LDH)、苹果酸酶 (ME)和超氧化物歧化酶 (SOD)。结果表明 ,这 8种酶系统至少由 1 8个基因位点编码 ,其中 1 2个位点为遗传稳定的等位酶位点 ,是可靠的遗传标记。酶谱的分离式样表明 ,EST为单聚体结构 ,AAT、FDH、IDH、SOD为二聚体结构 ,GDH为六聚体结构。最后对同工酶的器官和发育特异性以及同工酶基因位点的遗传分析进行了讨论     

亚洲小车蝗不同地理种群遗传多样性的等位酶分析

采用等位酶电泳技术对亚洲小车蝗8个地理种群遗传多样性和遗传分化进行了研究,并对8种等位酶系统:苹果酸脱氢酶(MDH)、苹果酸酶(ME)、乙醇脱氢酶(ADH)、谷氨酸脱氢酶(GDH)、谷氨酸-草酰乙酸转氨酶(GOT)、异柠檬酸脱氢酶(IDH)、腺苷酸激酶(AK)和己糖激酶(HK)进行聚丙烯酰胺凝胶电泳分析。结果表明:在亚洲小车蝗8个地理种群中共检测到14个基因位点,其中7个位点为多态位点,检测到25个等位基因;种群总体水平多态位点比率P=42.86%,平均有效基因数A=1.786,平均期望杂合度He=0.072,种群平均遗传距离为0.069～0.235。聚类分析表明,遗传距离与地理距离存在一定相关性。亚洲小车蝗8个种群的遗传分化系数Fst=0.086,基因流Nm=3.142,表明亚洲小车蝗种群间有一定程度的遗传分化及基因交流。     

桦纤孔菌有性生殖相关特征及交配型位点分析

对桦纤孔菌菌株MDJCBS88的显微形态、菌丝及担孢子核相进行了观察。采用棉籽壳培养基对担孢子萌发形成的菌株进行栽培试验,筛选出不形成子实体或子实体发育不完整的菌株,将这些菌株在平板上进行了亲和试验,分析桦纤孔菌的有性生殖方式;并基于基因组序列进行交配型基因克隆验证,分析桦纤孔菌的交配型位点结构。显微观察发现,桦纤孔菌菌丝没有锁状联合结构,菌丝细胞无核到多核;子实层担孢子可含0-4个不等的细胞核,不同时期弹射的担孢子含有的细胞核数量不同。桦纤孔菌担孢子萌发率极低,能萌发的担孢子多为早期弹射的担孢子;培养基也影响担孢子的萌发率,与PDA培养基和CYM培养基相比,桦木屑培养基最适合桦纤孔菌担孢子萌发,萌发率为4.55%。从担孢子萌发的96个菌株中获得了2个不结实菌株和9个结实不产孢菌株,占11.5%,这些菌株间亲和试验出现不同的表现特征,包括形成产孢子实体,产生菌丝纽结,相互融合和相互拮抗等现象,认为桦纤孔菌的有性生殖以次级同宗结合为主,并受交配型基因控制。交配型位点克隆测序后分析发现,桦纤孔菌交配型A位点共14 034 bp,含有一个MIP基因和两组HD1和HD2基因;交配型B位点包含3个疑似信息素受体基因和1个信息素前体编码基因。     

龙牙木天然种群遗传多样性及遗传分化

采用水平切片淀粉凝胶电泳测定东北地区分布的4 个龙牙木种群的遗传结构。统计分析了10 个酶系统的11 个位点, 结果表明:龙牙木种群内存在着丰富的遗传变异, 多态位点百分率72.73%, 等位基因平均数为1.72, 期望杂合度为0.5617, 遗传一致度和遗传距离为0.9865和0.0133。这个结果与前文对其形态特征和薄层色谱特征分析所得结果基本一致, 充分说明区域龙牙木属于同一种物种的两个变种。     

龙牙楤木天然种群遗传多样性及遗传分化

采用水平切片淀粉凝胶电泳测定东北地区分布的4个龙牙楤木种群的遗传结构。统计分析了10个酶系统的11个位点,结果表明：龙牙楤木种群内存在着丰富的遗传变异,多态位点百分率72.73%,等位基因平均数为1.72,期望杂合度为0.5617,遗传一致度和遗传距离和0.9865和0.0133。这个结果与前文对其形态特片和薄层色谱特征分析所得结果基本一致,充分说明区域龙牙楤木属于同一种物种的两个变种。     

松毛虫属（Dendrolimus) 部分种类亲缘关系与遗传分化的等位酶分析

采用等位酶聚丙烯酰胺凝胶电泳技术对松毛虫属5个种和亚种的野生种群进行了亲缘关系和遗传变异的研究.8种等位酶系统(乳酸脱氢酶LDH、苹果酸脱氢酶MDH、苹果酸酶ME、乙醇脱氢酶ADH、甲酸脱氢酶FDH、谷氨酸脱氢酶GDH、过氧化物酶POD、过氧化氢酶CAT)共检测到12个基因位点,其中6个位点为多态位点,检测到15个等位基因.松毛虫属5个种和亚种的总体水平多态位点比率P＝50%,平均有效基因数A = 1.917,平均期望杂合度He =0.267,平均遗传距离为0.0730～0.5701.遗传参数表明松毛虫属昆虫种间存在较高程度的遗传变异,聚类图和遗传距离数据表明赤松毛虫与马尾松毛虫亲缘关系最近,落叶松毛虫与思茅松毛虫亲缘关系最远.     

Topography-specific emergence of fungal fruiting bodies in warm temperate evergreen broad-leaved forests on Yakushima Island,Japan

We conducted line route censuses of fungal fruiting bodies from August to September in 2005 and 2006 along ridges and valleys and compared the differences in the encounter rates of fungal fruiting bodies (= fruiting bodies seen per census kilometer) between types of topography and between fungal functional groups (i.e., ectomycorrhizal and saprobic fungi) in warm temperate evergreen broad-leaved forests on Yakushima Island, Japan. We found 251 fungal fruiting bodies (26 families, 50 genera, and 65 species) in total, including 51 bodies from Tricholomataceae, 41 from Russulaceae, 25 from Boletaceae, and 19 from Amanitaceae. The encounter rate of ectomycorrhizal fungi was greater at the ridge route (26.7 unit/km) than at the valley route (8.7 unit/km) and that of saprobic fungi was greater at the valley route (25.0 unit/km) than at the ridge route (12.5 unit/km). In addition, we conducted 7-year intermittent sampling and identified 40 families, 96 genera, and 142 species. The topography-specific emergence pattern of the intermittent sampling method was similar to that of the line census method. The fungal species composition in this study was possibly affected by a topographic gradient for both fungal functional groups through soil moisture, nutrient availability, and host tree distribution.     

Diversity and host associations of dipteran insects exploiting fungal fruiting bodies in Hokuriku,central Japan

To clarify the diversity and host associations of dipteran insects exploiting fungal fruiting bodies, we collected fruiting bodies at 18 localities in Hokuriku region, central Japan, from 2012 to 2015 and examined them for the emergence of insects. In total, 14,107 dipteran individuals belonging to 20 families emerged from fungi of 8 orders, 25 families, 49 genera and 129 species. Approximately 79% of dipteran individuals belonged to three families, Phoridae, Muscidae and Drosophilidae. The faunal similarity at the family level was relatively high between central (warm‐temperate) and northern (cool‐temperate) areas of Japan. However, the species composition of Drosophilidae was much different between central and northern Japan. The difference in the species composition was discussed in relation to the climatic conditions and fungal flora. None of the species from Drosophilidae, Phoridae, Muscidae, Mycetophilidae, Lonchaeidae and Chloropidae were specialists (they exploited more than one species of fungi), but they showed differences their fungi preference. Adults of some families, especially Drosophilidae, were frequently collected from fruiting bodies, but those of other families were seldom collected, probably reflecting differences in adult feeding ecology.     

Genetic differentiation in populations of the polymorphic fernCeratopteris thalictroides in Japan

Analysis of isozyme variation was carried out for 27 natural populations ofCeratopteris thalictroides in Japan. Of fifteen enzyme loci examined, eight loci were genetically polymorphic. At six loci,Lap, Pgi-2, Pgm-3, Pgm-4, Idh-2, and Skd-2, a marked genetic differentiation was observed between populations to the south of Okinawa Island and those to the north of the island. Okinawa Island contained a mixture of both southern and northern variants. Thus, two genetically distinct types (the south type and the north type) ofC. thalictroides occur allopatrically in Japan. Nei's genetic identity (I) between the two was 0.64, which was within the range of the I values between congeneric pteridophyte species. Regional fixation of a null allele was detected for one duplicated PGI locus in the north type ofC. thalictroides. This finding supports the recent hypothesis of genetic diploidization of polyploids through gene silencing.     

Genetic variation of oaks (<Emphasis Type="Italic">Quercus</Emphasis> spp.) in Switzerland. 3. Lack of impact of postglacial recolonization history on nuclear gene loci

Quercus petraea, Quercus pubescens and Quercus robur are closely related and interfertile white oaks native to Switzerland. The three species are known to share identical cpDNA haplotypes, which are indicative of the postglacial recolonization history of populations. Only two haplotypes are common in Switzerland. We compared variation of cpDNA and of isozymes in 28 oak populations from Switzerland in order to assess the impact of the postglacial population history on current genetic structures of nuclear controlled isozyme gene loci. Species delineation was based on Principal Component Analysis of leaf morphological traits. The species status of populations was reflected at isozyme gene loci, but differentiation between populations with different cpDNA haplotypes and hence different recolonization history was very low at enzyme gene loci for all species. Thus, glacial and postglacial population history was not reflected at nuclear gene loci on the temporal and spatial scale covered by the present study. Extensive gene flow through pollen among populations is likely to have blurred a previously existing genetic differentiation at biparentally inherited gene loci that possibly evolved in the different glacial refugia of the above mentioned cpDNA haplotypes.     

Genetic comparison between Pinus sylvestris and P. mugo using isozymes and chloroplast DNA

Pinus sylvestris and P. mugo populations from Poland and Czechoslovakia were compared using genetic variability at isozyme markers, chloroplast DNA variation, and mating system measurements. Two isozyme loci were found to differ between the species. P. mugo was as variable at isozyme loci as P. sylvestris. Diagnostic cpDNA fragments were found using the restriction enzyme Bcl-I. Populations that were morphologically classified as hybrids were found to be pure species, based both on isozyme and cpDNA results.     

二倍体型与三倍体型卫氏并殖吸虫同工酶谱的发育变异及其分类学意义

采用生趣板状聚丙烯酰胺胶是民泳法分析比较了二倍体型与三倍体型卫氏并殖吸虫童虫、成虫的醛缩酶、已糖激酶和葡萄糖－６－磷酸脱氢酶的同工酶谱,结果显示２ｎ型与３ｎ型卫氏并殖吸虫在不同发育期之间及两型吸虫相应各发育期之间ＡＬＤ、ＨＫ及Ｇ６ＰＤ同工酶谱均显著变异,变异表现为两种方式,一种是在同一个基因座位区的同工酶的酶带数目     

Interspecific hybridization and genetic variability of Phlebotomus sandflies

The first successful hybridization is reported between Phlebotomus papatasi and P. duboscqi, two important Old World sandfly vectors of leishmaniasis and other diseases. Laboratory strains of P. papatasi and P. duboscqi were separable by six diagnostic enzyme loci: Est-3, Idh-1, Mdh-2, Mpi, Tre-1 and Tre-3. Hybrids between the two species were verified by the recovery of heterozygous isozyme patterns for the diagnostic loci. No F2 or backcross progeny were obtained. P. papatasi was separated from P. bergeroti by three diagnostic enzyme loci: Est-3, Mpi and Pgd. The isozyme patterns of P. bergeroti contain elements of both P. duboscqi and P. papatasi, although seven diagnostic loci (Est-3, Idh-1, Me, Mpi, Pgd, Tre-1 and Tre-3) separated P. bergeroti from P. duboscqi. Genetic variability profiles of the three species were established for 20 enzyme loci. Three geographically distant strains of P. papatasi from Calcutta, Maharashtra and Israel had isozyme genetic distances of < 0.05. The recently established Calcutta strain showed an unexpectedly low genetic variability with only one (Idh-2) of 20 loci being polymorphic (average heterozygosity of 1.9%) in contrast to 5-8 polymorphic loci (10-12% heterozygosity) in the Maharashtra and Israel strains. Mass and single pair crosses between the three P. papatasi strains were fertile with normal progeny numbers. Thus we found no signs of speciation in P. papatasi.     

Genetic differences between two allopatric sibling species of the genus Polydora (Polychaeta: Spionidae) from the West Pacific

Polydora vulgaris Mohammad, 1972, a commensal borer of the oysters Pinctada margaritifera and Hyotissa hyotis from the South China Sea, was investigated by means of starch gel electrophoresis. Polydora vulgaris and the allopatric sibling Polydora glycymerica Radashevsky, 1993, a commensal borer of the bivalve Glycymeris yessoensis from the Sea of Japan, were compared with respect to their allozymic variation and number of isozyme loci. Interspecific differences in the number of gene loci coding for three enzymes: alanopine dehydrogenase, glucose-6-phosphate isomerase and -iditol dehydrogenase were revealed suggesting that we are dealing with two valid species. Two different modes of origin duplicate loci in polydorids are dicussed—polyploidization and regional gene duplication. The use of gene number as a character for discriminating between morphologically indistinguishable allopatric polydorid taxa is outlined.     

Isozyme differences between the developmental stages of the parthenogenetic gall wasp Diplolepis rosae

The value of electrophoretic analysis of enzymes as an aid in connecting a morphologically deviating form to one of several possible species is dependent on the proportion of shared enzyme loci between the forms. In order to determine this proportion for the different instars of the parthenogenetic gall wasp Diplolepis rosae, 16 different enzyme systems were analysed. Out of the 37 loci detected, 25 (68%) were active in all instars. The larvae, pupae, and imagines had two, one and five unique loci, respectively. Larvae and pupae shared four loci not expressed in the imagines. In the ten enzymes analysed for comparison of males and females a total of 22 loci were detected, 21 found in both sexes and one unique to the females. The difference in isozyme pattern was found to be much more pronounced between pupa and imago than between larva and pupa. The chance to find a locus in an instar when already detected in another was calculated to 87%. If this estimate is valid for other forms within species, differential activity should not present a problem when electrophoretic analysis is used in order to connect such forms.     

Isozyme variation and the reproductive biology of Hydrocharis morsus-ranae L. (Hydrocharitaceae)

Electro-phoretically detectable isozyme variation was studied for 17 enzyme systems in several N American populations of the introduced aquatic plant Hydrocharis morsus-ranae and in two European populations. Twenty-nine loci were inferred from progeny, adult and turion enzyme banding patterns with 28 of these loci homozygous in all individuals studied. Malate dehydrogenase-1 (MDH-1) was the only locus which could be interpreted as multi-allelic and heterozygous. Twenty-seven of 76 seedlings assayed showed an age specific expression for an alcohol dehydrogenase locus (ADH-2) never seen in adults or turions. Since all adults sampled were phenotypically identical at all loci assayed, it is possible that only one isozyme genotype of this species is present in N America. European turion data further indicated that the populations studied were identical to N American plants sampled at all loci except EST. Therefore, although H. morsus-ranae is dioecious, outcrossing appears to involve substantial inbreeding. Connections between extensive inbreeding and the failure of effective sexual reproduction are considered.