Cytoplasmic composition in Pinus densata and population establishment of the diploid hybrid pine

Sequence and restriction site analyses of the paternally inherited chloroplast rbcL gene and maternally inherited mitochondrial nad1 fragments from the same set of populations and individuals were used to investigate cytoplasmic composition and population establishment of Pinus densata, a diploid pine that originated through hybridization between P. tabuliformis and P. yunnanensis. Two variable sites and three chlorotypes (TT, TC and GC) were detected on the rbcL gene of the three pines. P. densata harboured the three chlorotypes, two of which (TT, GC) were characteristic of the parental species, respectively. The third chlorotype (TC) was distributed extensively in seven of the 10 P. densata populations analysed, and might represent a mutation type or have been derived from an extinct parent. The distribution of chlorotypes, together with that of mitotypes, indicated that significant founder effect and backcross happened during the population establishment of the hybrid pine. P. tabuliformis and P. yunnanensis had acted as both mother and father donors, i.e. bi-directional gene flow existed between the two parental species in the past. Population differentiation of P. densata is high, as detected from the cytoplasmic genomes: GST = 0.533 for cpDNA and GST = 0.905 for mtDNA. The differences in cytoplasmic composition among the hybrid populations suggest that the local populations have undergone different evolutionary histories.     

Influence of ammonium and nitrate supply on growth,nitrate reductase activity and N-use efficiency in a natural hybrid pine and its parents

Aims Selection of tree species with a high capacity to assimilate N and efficiently utilize N resources would facilitate the success of initial tree seedling establishment in infertile soils. The preference for N forms was tested using three pine species (Pinus densata, Pinus tabuliformis and Pinus yunnanensis). Pinus densata is a natural diploid hybrid between P. tabuliformis and P. yunnanensis .Methods Seedlings of three pine species were supplied with nitrate-N, ammonium-N (at two different pH regimes) or combined ammonium and nitrate as a nitrogen source in perlite culture in a controlled environment.Important findings Seedlings of P. densata had higher total biomass and net photosynthesis when supplied with nitrate-N and ammonium nitrate than with ammonium-N. In parental species, total biomass and net photosynthesis for P. yunnanensis seedlings was higher in ammonium-N than in nitrate-N, whereas the other parental species P. tabuliformis had the highest total biomass among species for all treatments except ammonium with CaCO 3. Most morphological traits in P. densata seedlings were intermediate between its two parental species. However, N-use efficiency and photosynthetic N-use efficiency of P. densata significantly exceeded both parents when supplied with nitrate-N and ammonium nitrate. The results suggested that the diploid hybrid tree species P. densata has a preference for nitrate and is not well adapted to ammonium-N as a sole nitrogen source regardless of the growth medium pH. Based on changes in environmental conditions, such as predicted future temperature increases in high altitude areas associated with climate change, P. densata is likely to be increasingly competitive and have wide adaptation in high altitude regions.     

同倍杂交种高山松与亲本种云南松的地理隔离研究

为了探讨横断山区同倍杂交种高山松与其亲本种云南松的地理隔离机制,采用实际的路线踏查方法,调查了高山松和云南松群体的地理分布情况和开花物候特征,并基于7个气象因子进行了聚类分析。调查研究表明高山松和云南松群体分布在不同的地貌类型中,以云南中甸为临界区,沿着金沙江河谷向北分布高山松,向南分布云南松。云南松的开花物候日期明显早于高山松,聚类分析表明两个物种分别聚为一支。在地质历史过程中,高大山系的形成以及海拔的升高将高山松和云南松从地理上隔离开来,两物种的适生区的气象因子的差异造成花期不遇,从而阻碍了两个物种问的基因交流,进而隔离开来。     

Chromosomal localization of 5S and 18S–5.8S–25S ribosomal DNA sites in five Asian pines using fluorescence in situ hybridization

Fluorescence in situ hybridization (FISH) was employed on mitotic metaphase chromosome preparations of five Asian Pinus species: Pinus tabuliformis, Pinus yunnanensis, Pinus densata, Pinus massoniana and Pinus merkusii, using simultaneously DNA probes of the 18S rRNA gene and the 5S rRNA gene including the non-transcribed spacer sequences. The number and location of 18S rDNA sites varied markedly (5-10 pairs of strong signals) among the five pines. A maximum of 20 major 18S rDNA sites was observed in the diploid genome (2n = 24) of P. massoniana. The 5S rDNA FISH pattern was less variable, with one major site and one minor site commonly observed in each species. The differentiation of rDNA sites on chromosomes among the five pines correlates well with their phylogenic positions in Pinus as reconstructed from other molecular data. P. densata, a species of hybrid origin, resembles its parents ( P. tabuliformis and P. yunnanensis), including some components characteristic of each parent in its pattern. However, the species is unique, showing new features resulting possibly from recombination and genome reorganization.     

Genetic composition and diploid hybrid speciation of a high mountain pine, Pinus densata, native to the Tibetan plateau

Pinus densata has been suggested to have originated from hybridization events involving P. tabulaeformis and P. yunnanensis. In this study, allozyme differentiation at 12 loci was studied in 14 populations of P. tabulaeformis, P. densata, and P. yunnanensis from China. The observed genetic composition of P. densata supported the hybrid hypothesis and showed varying degrees of contribution from P. yunnanensis and P. tabulaeformis among its populations. These data, together with previous chloroplast DNA results, indicated different evolutionary histories among P. densata populations. To examine the possibility of ongoing hybridization among the three species, we analyzed patterns of linkage disequilibria between allozyme loci in ovule, pollen, and zygote pools. None of these tests suggested that there is significant ongoing gene exchange, implying that populations of P. densata have a stabilized hybrid nature. The normal fertility and high fecundity of P. densata indicate that this hybrid is maintained through sexual reproduction. P. densata represents an example of diploid hybrid speciation in an extreme ecological habitat that is both spatially and ecologically separated from that of its parents.     

高山松及其亲本种群在油松生境下的苗期性状

高山松(Pinus densata)是油松(P. tabuliformis)和云南松(P. yunnanensis)的天然二倍体杂交种, 是为数不多的与亲本种没有严格生殖隔离的同倍体杂交种。为了检测3个种在油松生境下的苗期适应性和生长发育特点, 为高山松成种机制提供数据, 选取了代表高山松及其亲本种遗传多样性的25个种群, 在油松生境下对比分析了苗期11个适应性指标和3个生长发育指标。结果表明, 大部分适应性指标和生长发育指标在种间和种内群体间差异显著, 主要变异存在于种间及种内群体间; 出苗率、2011年11月和2012年10月的封顶率、二年生苗在2012年10月的保存率等指标的种间方差分量较大, 种内变异较小, 是体现种间苗期适应性和生长发育状况的重要指标。油松在大部分适应性指标和3个生长发育指标上均表现最好, 总体适合度高于云南松和高山松; 云南松在封顶率和保存率上都居于最低值, 大部分一年生苗木到11月底仍未出现封顶现象且黄苗比例最高, 二年生苗的存活率为0, 表明云南松在油松生境下适合度最低; 高山松除了在紫苗比例上表现出超亲优势外, 大部分性状居于亲本种之间。此外, 位于青藏高原东北部的高山松祖先种群在适应性上表现较好, 具有在油松生境下发展的潜力; 而位于青藏高原西部的高山松种群及东南部的康定种群对油松生境的适应性较差。研究揭示出生态选择在高山松的同倍性杂交物种形成中起到了关键作用。     

云南松及其近缘种的遗传变异与亲缘关系

The authors investigated the genetic diversity of 29 natural populations representing Pinus yunnanensis Franch. and its two close relatives, P. densata Mast. and P. kesiya Royle ex Gordn. var. langbianensis (A Chey.) Gaussen. Horizontal starch gel electrophoresis was performed for macrogametophytes collected from populations in Yunnan, Sichuan and Guangxi. Allozyme data for 33 loci of 14 enzymes demonstrated high levels of genetic variation at both population and species levels in comparison with other conifers, with the mean values for populations being P=0.694, A=2.0 and He=0.145 for P. yunnanensis; P=0.714, A=2.0 and He=0.174 for P. densata; and P=0.758, A=2.1 and He=0.184 for P. kesiya var. langbianensis. Based on Wright's F-statistics, the fixation index of P. yunnanensis, P. densata and P. kesiya var.langbianensis were 0.101, 0.054 and 0.143, respectively, indicating that the populations were largely under random mating. Based on Nei's genetic distance, the genetic differentiation was not obvious among the three species (i.e. the genetic distance was less than 0.075). Because of the wider distribution of P. yunnanensis with greater variety of habitats, it was shown that the genetic differentiation among the P. yunnanensis populations was larger than that of the populations of the other two species. According to morphological, geographic and allozymic evidences, the authors suggested that the three species be better treated as varieties under a single species. In addition, the extensive gene flow among the three pine species resulted in great genetic diversity and evolutionary potential. Also, high level of genetic variation of P. yunnanensis provides important basis for its genetic improvement and breeding in future.     

高山松及其亲本种油松和云南松DHAR基因的功能分化

高山松(Pinus densata)是油松(P. tabulaeformis)与云南松(P. yunnanensis)自然杂交产生的同倍性杂种, 分布于青藏高原东南缘, 占据了油松和云南松两个亲本种都不能正常生长的高海拔地带。为了揭示高山松、油松和云南松脱氢抗坏血酸还原酶(DHAR)基因的组成和功能分化, 分别从高山松、油松和云南松中克隆到2类DHAR基因(DHAR1与DHAR2)。组织表达模式分析表明, 这6个基因在根、韧皮部、叶和芽中均有表达; 通过系统发育分析发现, 高山松在物种形成过程中保留了油松的DHAR1拷贝以及云南松的DHAR2拷贝; 酶学性质分析则表明, 高山松与油松DHAR1蛋白对底物具有相似的催化活性、催化效率、最适pH和热力学稳定性, 但其催化活性比云南松DHAR1蛋白高约300倍。高山松DHAR2蛋白对底物的催化活性和热力学稳定性均高于油松DHAR2蛋白。高山松DHAR基因在生化功能上显示出优于或类似亲本DHAR, 这种优势功能的选择与杂种独特的生态适应性可能有重要的相关性。     

The postglacial history of Scots pine (Pinus sylvestris L.) in western Europe: evidence from mitochondrial DNA variation

The geographical structure of mitochondrial (mt)DNA variants (mitotypes) was investigated in 38 western European populations of Scots pine Pinus sylvestris using restriction fragment length polymorphism (RFLP) analysis of total DNA and a homologous cox1 probe. Three major mitotypes (designated a, b and d ) were detected. Within Spain all three major mitotypes were found, gene diversity was high, H_T = 0.586, and this diversity was distributed predominantly among rather than within populations (F_ST(M) = 0.813 for the seven Spanish populations). Mitotype d was present only in the most southerly population from the Sierra Nevada . Elsewhere in Europe, populations showed little or no mtDNA diversity within regions, but there were marked differences between regions. Italian populations were fixed for mitotype b ; populations from northern France, Germany, Poland, Russia and southern Sweden were fixed for mitotype a ; while populations in northern Fennoscandia were fixed for mitotype b . The isolated Scottish populations were predominantly of mitotype a , but mitotype b was present in three of the 20 populations scored. In Scotland, UK gene diversity (H_T = 0.120) and genetic differentiation among populations (F_ST(M) = 0.37) was much lower than in Spain. When interpreted in the light of complementary data from pollen analysis and nuclear genetic markers, the results suggest that present-day populations of P. sylvestris in western Europe have been derived from at least three different sources after glaciation.     

Colonization history of Scots pine in Eastern Europe and North Asia based on mitochondrial DNA variation

During Quaternary glaciations, the ranges of Northern Eurasia forest species periodically experienced contraction followed by subsequent re-colonizations in the interglacial intervals. However, unlike the broadleaf trees of temperate forests, taiga species seem not to have retreated fully to southern regions in unfavorable periods and possibly survived at mid-latitudes in multiple refugia. Here, we report a study of genetic variation of three mitochondrial DNA markers in 90 populations of Scots pine (Pinus sylvestris) located from Eastern Europe to Eastern Siberia. The geographic distribution of seven mitotypes demonstrated the split between western and eastern populations approximately along the 38th meridian. Genetic diversity in the western part was significantly higher than in the eastern one. Five mitotypes were western- and one eastern-specific. One mitotype was common in both regions, but in the eastern part it occurred only in the South Urals and adjacent areas. The geographic structure in the mitotype distribution supports a hypothesis of post-glacial re-colonization of the studied territory from the European and Ural refugia.     

Polymorphism of mitochondrial DNA and infection with symbiotic cytoplasmic bacterium Wolbachia pipientis in mosquitoes of the Culex pipiens complex from Russia

A total of 208 mosquitoes of the Culex pipiens complex from 15 basement and terrestrial populations collected in different regions of the European part of Russia and Siberia were examined by genetic methods. Among these, two major mitotypes, M and P, were identified. These mitotypes differed by six substitutions in the 246-bp mitochondrial DNA cytochrome oxidase I gene fragment examined. Urban basement mosquito ecotype C. pipiens (form molestus) were characterized by the presence of mitotype M and infection with the endosymbiotic bacteria of the genus Wolbachia. Mosquitoes of the C. pipiens complex inhabiting opened biotopes harbored mitotype P, or its variety, mitotype P1, and were not infected with Wolbachia. Thus, in natural conditions marked linkage disequilibrium between cytoplasmic elements, mitochondrial DNA and Wolbachia, can be observed. Similarity of mitotypes in form molestus mosquito from different geographical localities favors the hypothesis on the common ancestry of urban mosquitoes.     

低温胁迫下二倍体杂交种高山松光系统Ⅱ功能稳定性研究

以人工气候室内的高山松(Pinus densata)及其亲本油松(P.tabulaeformis)和云南松(P.yunnanensis)实生苗为材料,通过测定低温胁迫和恢复后其幼苗的快速叶绿素荧光诱导动力学(JIP-test)参数,分析高山松及其亲本的光系统Ⅱ(PSⅡ)功能稳定性以及对低温胁迫的响应差异,以探讨高山松适应高海拔低温环境的光合生理生态机制。结果表明:经过33 d的低温(12℃/5℃,昼/夜)处理后,高山松PSⅡ功能参数单位截面积吸收的能量(ABS/CSo)、捕获的能量(TRo/CSo)、电子传递的能量(ETo/CSo)、耗散的能量(DIo/CSo)、有活性反应中心的数量(RC/CSo),以及原初光化学反应的最大量子产额(φPo)、吸收光能用于QA-以后的电子传递的能量比例(φEo)、捕获光能用于QA-以后的电子传递的能量比例(Ψo)、光能利用能力参数(PIabs)、2 ms时相对可变荧光(VJ)、荧光上升的初始斜率(Mo)、荧光曲线与Fm所围面积(Area)、介于F0和Fm之间的相对面积(Sm)和QA氧化还原次数(N)均与处理前无显著差异;油松和云南松的φPo、φEo、Ψo、PIabs、Area与对照差异显著,云南松的VJ、Mo、Sm、N、ETo/CSo、DIo/CSo、ABS/CSo也与对照存在显著差异;温度恢复之后(25℃/18℃,昼/夜),这些荧光参数的恢复程度表现为高山松>油松>云南松。可见,低温胁迫显著影响了油松和云南松的光能传递和光能利用能力以及受体库大小,且云南松遭受伤害的程度高于油松,而高山松的PSⅡ功能及其稳定性在遭受低温胁迫时明显高于亲本油松和云南松,且胁迫解除后恢复程度高,从而使得高山松能够在高海拔低温环境中累积代谢所需的有机物,保证其正常的生长和发育。     

New evidence from mitochondrial DNA of a progenitor-derivative species relationship between black spruce and red spruce (Pinaceae)

Mitochondrial DNA (mtDNA) markers were used to assess the genetic diversity in allopatric populations of black spruce (Picea mariana [Mill.] BSP) and red spruce (P. rubens Sarg.). Patterns of mitochondrial haplotypes (mitotypes) were strikingly different between the two species. All mtDNA markers surveyed were polymorphic in black spruce, revealing four different mitotypes and high levels of mtDNA diversity (P(p) = 100%, A = 2.0, H = 0.496). In contrast, populations of red spruce had only two mitotypes and harbored low levels of ggenetic diversity (P(p) = 13.2%, A = 1.1, H = 0.120). When the southernmost allopatric populations of red spruce were considered, only one mitotype was detected. As previously reported for nuclear gene loci, the diversity observed for mtDNA in red spruce was a subset of that found in black spruce. Comparison of present and previously published data supports the hypothesis of a recent progenitor-derivative relationship between these species, red spruce presumably being derived by allopatric speciation of an isolated population of black spruce during the Pleistocene.     

Substoichiometrically different mitotypes coexist in mitochondrial genomes of Brassica napus L

Cytoplasmic male sterility (CMS) has been identified in numerous plant species. Brassica napus CMS plants, such as Polima (pol), MI, and Shaan 2A, have been identified independently by different researchers with different materials in conventional breeding processes. How this kind of CMS emerges is unclear. Here, we report the mitochondrial genome sequence of the prevalent mitotype in the most widely used pol-CMS line, which has a length of 223,412 bp and encodes 34 proteins, 3 ribosomal RNAs, and 18 tRNAs, including two near identical copies of trnH. Of these 55 genes, 48 were found to be identical to their equivalents in the "nap" cytoplasm. The nap mitotype carries only one copy of trnH, and the sequences of five of the six remaining genes are highly similar to their equivalents in the pol mitotype. Forty-four open reading frames (ORFs) with unknown function were detected, including two unique to the pol mitotype (orf122 and orf132). At least five rearrangement events are required to account for the structural differences between the pol and nap sequences. The CMS-related orf224 neighboring region (～5 kb) rearranged twice. PCR profiling based on mitotype-specific primer pairs showed that both mitotypes are present in B. napus cultivars. Quantitative PCR showed that the pol cytoplasm consists mainly of the pol mitotype, and the nap mitotype is the main genome of nap cytoplasm. Large variation in the copy number ratio of mitotypes was found, even among cultivars sharing the same cytoplasm. The coexistence of mitochondrial mitotypes and substoichiometric shifting can explain the emergence of CMS in B. napus.     

Polymorphism of mitochondrial DNA and distribution of cytoplasmic symbionts in the populations of two-spot ladybird beetle Adalia bipunctata

In geographically distant populations of ladybird beetle Adalia bipunctata from Eurasia mitotypes and infection with symbiotic bacteria Spiroplasma and Rickettsia were determined. All populations examined demonstrated mtDNA polymorphism and striking differences in prevalence of bacteria (from about 50% of individuals infected with Spiroplasma in St.-Petersburg population and 50% of the Rickettsia prevalence in Kem' population to complete absence of bacteria in the population from Archangelsk). In the populations studied a total of 14 mitotypes were discovered, including two mitotypes that were remarkably different from the others in nucleotide composition. Mitotype 10, which was the most different from all the others, was found in all populations from Germany to Transbaikalia, excluding the population from Tashkent. Linkage disequilibrium between mitotype 10 and the Rickettsia infection was confirmed. Infection with the Spiroplasma bacteria was typical of the individuals with haplotype 1 and relative to it. The results obtained supported the conclusion on the association between infection with Spiroplasma and Rickettsia and certain mitotype of A. bipunctata, which was the consequence of either absence or rare horizontal transfer of symbionts and ancientness of the first contact between the bacteria and A. bipunctata ladybird beetles.     

Comparative restriction enzyme analysis of mitochondrial DNA in several populations of lake resident chars of the genus Salvelinus

Restriction enzyme analysis was employed in studying the mitochondrial DNA (mtDNA) ATPase6/ND4L region in several Northeast Asian populations of resident lake char of the genus Salvelinus. On evidence of mitotypes, genetic similarity was assumed for populations of neiva (Ueginskoe Lake), lake resident char from the northern coast of the Sea of Okhotsk (Mak-Mak Lake and Elekchanskie lakes), and dolly varden. Mitotype AAAA proved to be common for these populations. Lake char populations of the Juliet and Maxi lakes (the basin of the Kolyma river) had mitotype DBAA, which is similar to mitotype DBAB observed earlier in Taranetz char S. taranetzi from Chukotka. The divergence between mitotypes AAA and DBAA was estimated at 0.3%. Different origins were assumed for the lake resident char populations from the basins of the Sea of Okhotsk and of the Kolyma River, the former originating in the Pacific and the latter, in the Arctic basins.     

The Pines and Pine Forests of Xizang

Six species of pines are distributed in Xizang. They are: Pinus griffithii McClell., P. armandi Franch. and P. gerardiana Wall. of haploxylon pines and P. densata Mast., P. yttnnanensis French. and P. roxburghii Sarg. of diploxylon pines. According to the relation of these pines with water-temperature conditions, 4 ecological types may be divided: the warm-temp erate and wet type (P. griffithii), the warm-temperate and dry type (P. yunnanensis, P. roxburghii), the temperate-cold and moist type (P. armandi) and the temperate-cold and dry type (P. densata). The composition and structure of every pine community reflect the ecological environments of the given pine in the region. The main pine comnmnity in Xizang are P. griffithii forest and P. densata forest. The P. griffithii forest is distributed on the southern side of Himalayas, while the P. densata forest on the northern side of Himalayas and the southern part of Hengtuan mountains. This indicates that the Himalaya range is a clear boundary and there is difference in water-temperature condition between southern and northern parts. They belong different vegetation regions. The different distribution of other several pine forests reflects the difference of environmental conditions within these two regions. These facts have significance in the investigation of the regularity of vegetation distribution and vegetation division in Xizang. Besides, the vertical distribution of pines cannot be used as a marker to divide the altitudinal belts due to the wide range of adaptation of pines, though there must be regularity of vertical distribution too.     

Polymorphism of mitochondrial DNA and infection with symbiotic cytoplasmic bacterium <Emphasis Type="Italic">Wolbachia pipientis</Emphasis> in mosquitoes of the <Emphasis Type="Italic">Culex pipiens</Emphasis> (Diptera,Culicidae) complex from Russia

A total of 208 mosquitoes of the Culex pipiens complex from 15 basement and terrestrial populations collected in different regions of the European part of Russia and Siberia were examined by genetic methods. Among these, two major mitotypes, M and P, were identified. These mitotypes differed by six substitutions in the 246-bp mitochondrial DNA cytochrome oxidase I gene fragment examined. Urban basement mosquitoes C. pipiens (form molestus) were characterized by the presence of mitotype M and infection with the endosymbiotic bacteria of the genus Wolbachia. Mosquitoes of the C. pipiens complex inhabiting opened biotopes harbored mitotype P, or its variety, mitotype P1, and were not infected with Wolbachia. Thus, in natural conditions marked linkage disequilibrium between cytoplasmic elements, mitochondrial DNA and Wolbachia, can be observed. Similarity of mitotypes in form molestus mosquito from different geographical localities favors the hypothesis on the common ancestry of urban mosquitoes.Translated from Genetika, Vol. 41, No. 3, 2005, pp. 320–325.Original Russian Text Copyright © 2005 by Shaikevich, Vinogradova, Platonov, Karan, Zakharov.     

Comparative Restriction Enzyme Analysis of Mitochondrial DNA in Several Populations of Lake Resident Chars of the Genus Salvelinus

Restriction enzyme analysis was employed in studying the mitochondrial DNA (mtDNA) ATPase6/ND4L region in several Northeast Asian populations of resident lake char of the genus Salvelinus. On evidence of mitotypes, genetic similarity was assumed for populations of neiva (Ueginskoe Lake), lake resident char from the northern coast of the Sea of Okhotsk (Mak-Mak Lake and Elekchanskie lakes), and dolly varden. Mitotype AAAA proved to be common for these populations. Lake char populations of the Juliet and Maxi lakes (the basin of the Kolyma River) had mitotype DBAA, which is similar to mitotype DBAB observed earlier in Taranetz char from Chukotka. The divergence between mitotypes AAAA and DBAA was estimated at 0.3%. Different origins were assumed for the lake resident char populations from the basins of the Sea of Okhotsk and of the Kolyma River, the former originating in the Pacific and the latter, in the Arctic basins.     

Origin of honeybees (Apis mellifera L.) from the Yucatan peninsula inferred from mitochondrial DNA analysis

Honeybees (Apis mellifera L.) sampled at sites in Europe, Africa and South America were analysed using a mitochondrial DNA restriction fragment length polymorphism (RFLP) marker. These samples were used to provide baseline information for a detailed analysis of the process of Africanization of bees from the neotropical Yucatan peninsula of Mexico. Radical changes in mitochondrial haplotype (mitotype) frequencies were found to have occurred in the 13-year period studied. Prior to the arrival of Africanized bees (1986) the original inhabitants of the Yucatan peninsula appear to have been essentially of southeastern European origin with a smaller proportion having northwestern European ancestry. Three years after the migration of Africanized bees into the area (1989), only very low levels of maternal gene flow from Africanized populations into the resident European populations had occurred. By 1998, however, there was a sizeable increase in the proportion of African mitotypes in domestic populations (61%) with feral populations having 87% of mitotypes classified as African derived. The results suggest that the early stages of Africanization did not involve a rapid replacement of European with African mitotypes and that earlier studies probably overestimated the prevalence of African mitotypes.