Chromosomal Homology and Molecular Organization of Muller''s Elements D and E in the Drosophila Repleta Species Group

Thirty-three DNA clones containing protein-coding genes have been used for in situ hybridization to the polytene chromosomes of two Drosophila repleta group species, D. repleta and D. buzzatii. Twenty-six clones gave positive results allowing the precise localization of 26 genes and the tentative identification of another nine. The results were fully consistent with the currently accepted chromosomal homologies and in no case was evidence for reciprocal translocations or pericentric inversions found. Most of the genes mapped to chromosomes 2 and 4 that are homologous, respectively, to chromosome arms 3R and 3L of D. melanogaster (Muller's elements E and D). The comparison of the molecular organization of these two elements between D. melanogaster and D. repleta (two species that belong to different subgenera and diverged some 62 million years ago) showed an extensive reorganization via paracentric inversions. Using a maximum likelihood procedure, we estimated that 130 paracentric inversions have become fixed in element E after the divergence of the two lineages. Therefore, the evolution rate for element E is approximately one inversion per million years. This value is comparable to previous estimates of the rate of evolution of chromosome X and yields an estimate of 4.5 inversions per million years for the whole Drosophila genome.     

黑腹果蝇种组五种核型的报道

通过传统的敲片、Giemsa染色的方法, 本文首次对果蝇属黑腹果蝇种组的5种果蝇 （D. constricta、D. ohnishii、D. ogumai、D. pseudobaimaii、D. tani）染色体的数目和形态进行了分析报道。分析发现：这5个种具有相同的染色体数目（2n=8）和不同的形态。D. pseudobaimaii和D. tani 为2V,1R,1D型;D. constricta染色为2V,1R,1D型且其点状染色体难以辨认;D. ohnishii和D. ogumai 具有相同形态为2V,2R。另外,还发现核型与亲缘关系之间有一定的对应性。     

Two Tests of Y Chromosomal Variation in Male Fertility of Drosophila Melanogaster

Deficiency mapping with Y autosome translocations has shown that the Y chromosome of Drosophila melanogaster carries genes that are essential to male fertility. While the qualitative behavior of these lesions provides important insight into the physiological importance of the Y chromosome, quantitative variation in effects on male fertility among extant Y chromosomes in natural populations may have a significant effect on the evolution of the Y chromosome. Here a series of 36 Y chromosome replacement lines were tested in two ways designed to detect subtle variation in effects on male fertility and total male fitness. The first test involved crossing males from the 36 lines to an excess of females in an attempt to measure differences in male mating success (virility) and male fecundity. The second test challenged males bearing each of the 36 Y chromosomes to competition in populations with males bearing a standard, phenotypically marked (BsY) chromosome. These tests indicated that the Y chromosome lines did not differ significantly in either male fertility or total fitness, but that interactions with autosomes approached significance. A deterministic population genetic model was developed allowing Y autosome interaction in fertility, and it is shown that, consistent with the experimental observations, this model cannot protect Y-linked polymorphism.     

DNA Sequence Variation at the Period Locus Reveals the History of Species and Speciation Events in the Drosophila Virilis Group

The virilis phylad of the Drosophila virilis group consists of five closely related taxa: D. virilis, D. lummei, D. novamexicana, D. americana americana and D. americana texana. DNA sequences from a 2.1-kb pair portion of the period locus were generated in four to eight individuals from each of the five taxa. We found evidence of recombination and high levels of variation within species. We found no evidence of recent natural selection. Surprisingly there was no evidence of divergence between D. a. americana and D. a. texana, and they collectively appear to have had a large historical effective population size. The ranges of these two taxa overlap in a large hybrid zone that has been delineated in the eastern U.S. on the basis of the geographic pattern of a chromosomal fusion. Also surprisingly, D. novamexicana appears to consist of two distinct groups each with low population size and no gene flow between them.     

中国特有果蝇 Drosophila curviceps 种亚组在Drosophila immigrans种组中的种系发生地位

果蝇immigrans种组中的curviceps种亚组是1992年新建立的中国特有果蝇类群。该种亚组中的物种主要分布在中国大陆和台湾。目前除了形态学水平的研究外,还没有其他证据支持建立该种亚组的合理性及其起源和种系发生地位。为了在DNA分子水平上探讨果蝇curviceps种亚组在果蝇immigrans种组中的种系发生地位,从而为今后更深入地研究中国特有果蝇,甚至为果蝇亚属的进化遗传学提供理论依据,测定了immigrans种组5个种亚组（nasuta、immigrans、hypocausta、quadrilineata、curviceps)中12个代表物种的rDNA的ITS1和部分Adh基因的序列。其中ITS1序列的长度为513－587bp,共有191个信息位点;Adh基因片段的长度在714－747bp之间,共99个信息位点。考虑到单个分子提供的信息较少,将两个分子的序列综合起来,组成一个较长的复合序列。分别根据ITS1,Adh和两个分子的复合序列排比（Alignment)结果,和最大简约法和邻接法构建分子系统树,其中根据复合序列构建的系统树与形态学研究结果最为一致。分子树显示curviceps种亚组的特种确定单独形成一个分枝,为种亚组级的分类阶元,支持了形态学将其建立为一个新种亚组。根据Kimura距离,估算了复合分子的替换速率约为每百万年1．48％,进而计算出5个种亚组的分 歧年代。结合各物种的地理分布,推测了immigrans种组的进化历史：curviceps种亚组与quadrilineata种亚组的亲缘关系最近,主要分布在中国南部的温带地区。它们之间的分歧时间大约为3．4百万年,是最年轻的两个种亚组。主要分布在苏门答腊及附近的热带地区的hypocausta种亚组的物种是最早分化出来的,与其他种亚组的分歧时间约为9．2百万年。该结果与形态学和生物地理学研究相吻合。值得一提是的,目前归属仍存在争议的物种D.neohypocausta,在分子系统树中与hypocausta种亚组的物种相距较远,而与immiagrasn种亚组的关系较近,但分枝置信度较低（＜50％）。由于还缺乏其他方面的证据,因此D.neohypocausta的归属有待今后的研究来作定论。     

Discordant Rates of Chromosome Evolution in the Drosophila Virilis Species Group

In Drosophila, the availability of polytene chromosome maps and of sets of probes covering most regions of the chromosomes allows a direct comparison of the organization of the genome in different species. In this work, we report the localization, in Drosophila virilis, D. montana, and D. novamexicana, of >100 bacteriophage P1 clones containing ~65 kilobase inserts of genomic DNA from D. virilis. Each clone hybridizes with a single euchromatic site in either chromosome 1 or chromosome 3 in D. virilis. From these data, it is possible to estimate the minimum number of inversions required to transform the map positions of the probes in one species into the map positions of the same probes in a related species. The data indicate that, in the D. virilis species group, the X chromosome has up to four times the number of inversions as are observed in chromosome 3. The first photographic polytene chromosome maps for D. montana and D. novamexicana are also presented.     

Differentiation of Muller''s Chromosomal Elements D and E in the Obscura Group of Drosophila

Twenty-two markers located on Muller's elements D or E have been mapped by in situ hybridization in six species of the obscura group of Drosophila and in D. melanogaster. The obscura species can be grouped into a Palearctic cluster (D. subobscura, D. madeirensis and D. guanche) and a Nearctic one (D. pseudoobscura, D. persimilis and D. miranda). Eleven of the probes contain known genes: E74, Acp70A, Est5, hsp28/23, hsp83, emc, hsp70, Xdh, Acph-1, Cec and rp49. The remaining probes are recombinant phages isolated from a D. subobscura genomic library. All these markers hybridize to the putative homologous chromosome or chromosomal arm of elements D and E. Thus, these elements have conserved their genic content during species divergence. Chromosomal homologies proposed previously for each element among the species of the same cluster have been compared with the present results. The distribution of markers within each element has changed considerably as inferred from pairwise comparisons of obscura species included in the two different clusters. Only chromosomal segments defined by closely linked markers have been conserved: one such segment has been detected in element D and three in element E between D. subobscura and D. pseudoobscura.     

A Chromosomal Study on 7 Species of Smilax L.

The chromosome numbers and karyotypes of 7 species of Smilax L. in Liliaceae (s. 1.) are cytotaxonomically studied in this work. Their karyotypic characters, distinction between the species and the chromosomal basis of sexual differentiation are discussed. The karyotypes of most species are first reported. The results are shown as follows (see Tables 1-4 for the chromosome parameters and the karyotype constitution; Fig. 1 for their idiograms): 1. Smilax nipponica Miq. The species is one of the herbaceous species distributed in East Asia. Two karyotypes, 2n = 26(type A) and 2n = 32 (type B), are found in the species (Plate 1: 1-7). The karyotype of No. 88032 (uncertain of -L--M--S- sexuality) is 2n = 26 = 2m + 6st + 6m + 4sm + 6sm + 2st. The karyotype has 4 pairs of L chromosomes, of which the first three pairs are subterminal, and the 4th is median. The karyotype belongs to 3B. No. 88045 (the male) and No. 88046 (the female) have 2n = 32. Their karyotypes are basically uniform, and both are -L--M-- S 2n=32= 2m+4sm+ 2st+ 2m+4sm+ 6m+ 10sm + 2st, also with 4 pairs of L chromosomes, but the 2nd pair is median, and thus different from the type A. The karyotype belongs to 3B. The first pair of chromosomes of the male are distinctly unequal in length, with the D. V. (0.93) of relative length between them obviously greater than that of the female (0.1). The pair seems to be of sex-chromosomes. Sixteen bivalents (n= 16) were observed at PMCs MI of No. 88045 (Plate 1: 4). The major difference between the karyotypes A and B are greater relative length of L chromosomes in the type A than in the type B, and the increase of chromosome number in the karyotype B mainly due to the increase of st chromosomes. Nakajima (1937)reports 2n= 30 for S. hederacea var. nipponica (=S. nipponica, Wang and Tang, 1980). 2. S. riparia A. DC. This species is also herbaceous, distributed in East Asia. Thirty chromosomes were found in root-tip cells (uncertain of sexuality). The kar -L--M--S-yotype is 2n = 30 = 8st + 6sm + 2st + 6m + 6sm + 2st (Plate 3: 1, 5), consisting mainly of sm and st chromosomes. There are 4 pairs of L chromosomes which are all subterminal and the m chromosomes appear to fall all into S category. Though the karyotype belongs to 3B, it is less symmetrical than that of S. nipponica. The species is karyologically rather different from S. nipponica, therefore. The first pair of chromosomes of this material are unequal in length, and it may be a male. The karyotype of this species is first reported. 3. S. sieboldii Miq. The species is a thorny climbing shrub, distributed in East Asia. At PMCs All, 16 chromosomes (n= 16) were found (Plate 2: 6), in accordance with Nakajima's (1933) report for a Japanese material. 4. S. china L. This species, a thorny climbing shrub, is of a wide distribution range mainly in East Asia and Southeast Asia. Two karyotypes were observed in different populations. (1) The population from Xikou has 2n = 96(6x) = 20st+L- -M- 6t + 6sm + 12st + 52(S) (Plate 3:7), of which the first three pairs of chromosomes are terminal, different from those in the other species. The arm ratios of both L and M chromosomes are larger than 2.0, which resembles those of S. davidiana. (2) PMCs MI of the population from Shangyu shew 15 chromosomes (n 15). The hexaploid of the species is recorded for the first time. Hsu (1967,1971) reported 2n = 30 from Taiwai and Nakajima (1937) recorded n = 30 from Japan, which indicates that the karyotype of the species varies not only in ploidy, but also in number. 5. S. davidiana A. DC. The somatic cells were found to have 32 chromosomes, and PMCs MI shew 16 bivalents (Plate 2: 1-5). The karyotype is 2n = 32=-L- -M- -S 8st + 4sm + 4st + 8sm + 8st. The karyotype belongs to 3B, and is less symmetrical than those in herbaceous species. The D. V. (0.20) of relative length between the two homologues of the first pair is slightly larger in the male than in the female (0.14), and it is thus difficult to determine whether they are sexual chromosomes or not. 6. S. glabra Roxb. The species is a non-thorny climbing shrub, distributed in East Asia and Southeast Asia. 32 chromosomes were found in somatic cells. The -L- -M- - Skaryotype is 2n= 32= 8st + 10st+6sm+8st (Plate 3: 2, 6),with only 3 pairs of sm chromosomes (12, 13 and 16th). The karyotype is more asymmetric than that of S. davidiana, although it is also of 3B (Table 1). The karyotype is first reported for the species. 7. S. nervo-marginata Hay. var. liukiuensis (Hay.) Wang et Tang The variety has a relatively narrow distribution range, mainly occurring in eastern China. The chromosomal number of somatic cells is 2n= 32 (Plate 3: 3-4). The karyotype is -L- -M- -S 2n = 32 = 2sm + 6st + 2sm + 2st + 2m + 6sm + 12st, evidently different from that of S. glabra. The first pair of chromosomes are submedian, and much longer than the 2nd to 4th pairs. The ratio in length of the largest chromosome to the smallest one is 4.3. The symmetric degree is of 3C, a unique type. The karyotype of the species is reported for the first time. In Smilax, the known basic numbers are 13, 15, 16 and 17. The two herbaceous species distributed in East Asia have three basic numbers: 13, 15 and 16, while the woody species studied mainly have 16, with no 13 recorded. Mangaly (1968) studied 8 herbaceous species in North America and reported 2n=26 for them except S. pseudo-china with 2n=30. Mangaly considered that a probably ancestral home of Smilax, both the herbaceous and woody, is in Southeast Asia and the eastern Himalayas, and speculated that the ancestral type of Sect. Coprosmanthus is possibly an Asian species, S. riparia. The karyotypes of the two herbaceous species in East Asia consist mostly of sm and m chromosomes, whereas those for the North American species are all of st chromosomes. Based on the general rule of karyotypic evolution, i.e. from symmetry to asymmetry, his speculation seems reasonable. Researches on sex-chromosomes of Smilax have been carried out since 1930 (Lindsay, 1930; Jensen, 1937; Nakajima, 1937; Mangaly, 1968), and they are generally considered to be the largest pair, but there is still no adequate evidence. The result of our observation on S. nipponica may confirm that the first pair of chromosomes of this species is XY type of sex-chromosomes. Chromosomes of the genus are small and medium-sized, varying between 1-6 μm, slightly larger in herbaceous species than in woody ones, larger in the karyotype of 2n=26 than in that of 2n=32. Based on karyotype constitution of the above 5 species, the karyotype in the genus is characterized by 4 pairs of L chromosomes and 2-5 pairs of M chromosomes, and mostly st and sm chromosomes, and by rather asymmetrical 3B type. The degree of symmetry in the above 5 species is from Sect. Coprosmanthus to Sect. Coilanthus, and herbaceous species towoody ones.     

Genetic Variation at Structural Loci in the Glossina morsitans Species Group

Gene diversity was investigated in four taxa of tsetse flies (Diptera: Glossinidae) including Glossina morsitans morsitans, G. m. centralis, G. swynnertoni, and G. pallidipes. Histochemical tests were performed for 35–46 isozymes. Polymorphic loci were 20% in G. morsitans morsitans, 32% in G. m. centralis, 17.6% in G. swynnertoni, and 26% in G. pallidipes. Mean heterozygosities among all loci were 6.6% in G. morsitans morsitans, 6.0% in G. m. centralis, 7.1% in G. swynnertoni, and 6.8% in G. pallidipes. Allozyme gene diversities were considerably less than those reported for many Diptera. The low gene diversities are probably related to small effective population sizes.     

Courtship Behavior Analysis in Three Sibling Species of the Drosophila virilis Group

Entomological Review - Courtship behavior was studied in three sibling species of the Drosophila virilis group: D. virilis, D. lummei, and D. littoralis. The latter species was represented by two...     

Mitochondrial DNA Polymorphism in Natural Populations of the Drosophila virilis Species Group

Restriction fragment length polymorphism (RFLP) analysis has been used to evaluate mitochondrial DNA (mtDNA) variation in 12 sibling species forming the Drosophila virilis species group. The variation thresholds corresponding to the interspecific and interstrain levels have been determined. The results indicate that interspecific hybridization has significantly contributed to the evolutionary history of the virilis species group.     

黑腹果蝇种组的核型与进化研究

观察了国内黑腹果蝇种组34种果蝇的有丝分裂中期核型,其中首次描述了一些新核型。系统地分析了黑腹果蝇种组8个种亚组之间的核型进化关系及种间亲缘关系。结果是:elegans种亚组的核型为A型;eugracilis、melanogaster和ficusphila种亚组的核型为C型;takahashii和suzukii种亚组的核型为C型和D型;montium种亚组的核型为B、C、C’、D、D’、和E型;ananassae种亚组的核型为F、G和H型。从核型分化的角度可以将黑腹果蝇种组分为5个谱系:elegans,eugracilis-melanogaster-ficusphila,takkahashii-suzukii,montium,ananassae。这与2004年Yang等的观点基本一致,正好从核型进化的角度验证了Yang通过DNA序列分析所得到的结果。差别只在于elegans种亚组,作者把它单独列为一支,认为是祖先种亚组。通过选取同一种果蝇的几个不同地域单雌系的核型分析,结果表明:同一种果蝇的核型存在地域差异。这种差异可能是由于不同生境造成,也可能是本身进化程度的差异,或是两种因素相互作用的结果。     

Chromosomal Localization and Sequence Variation of 5S rRNA Gene in Five Capsicum Species

Chromosomal localization and sequence analysis of the 5S rRNA gene were carried out in five Capsicum species. Fluorescence in situ hybridization revealed that chromosomal location of the 5S rRNA gene was conserved in a single locus at a chromosome which was assigned to chromosome 1 by the synteny relationship with tomato. In sequence analysis, the repeating units of the 5S rRNA genes in the Capsicum species were variable in size from 278 bp to 300 bp. In sequence comparison of our results to the results with other Solanaceae plants as published by others, the coding region was highly conserved, but the spacer regions varied in size and sequence. T stretch regions, just after the end of the coding sequences, were more prominant in the Capsicum species than in two other plants. High G-C rich regions, which might have similar functions as that of the GC islands in the genes transcribed by RNA PolII, were observed after the T stretch region. Although we could not observe the TATA like sequences, an AT rich segment at -27 to -18 was detected in the 5S rRNA genes of the Capsicum species. Species relationship among the Capsicum species was also studied by the sequence comparison of the 5S rRNA genes. While C. chinense, C. frutescens, and C. annuum formed one lineage, C. baccatum was revealed to be an intermediate species between the former three species and C. pubescens.     

The Genetic Basis of a Species-Specific Character in the Drosophila Virilis Species Group

The genetic basis of the species-specific dorsal abdominal stripe of Drosophila novamexicana was examined. The dorsal stripe is present in D. novamexicana and absent in all other members of the Drosophila virilis species group. Interspecific crosses between D. novamexicana and genetically marked D. virilis revealed that all four of the autosomes (except the tiny dot chromosome, which was not marked) and the sex chromosomes (the X and Y chromosome effects could not be disentangled) showed a significant effect on the width of the dorsal stripe. All the autosomes act approximately additively; only minor interactions were detected among them. No significant maternal effects were found. This means that a minimum of five loci are involved in the character difference between the two species, and this is the maximum number that this technique could discern. These results suggest that, based on the number of factors involved in the character difference, the inheritance of this character should be considered polygenic, but because chromosome 2 (the largest chromosome in the species) contributed over half of the variance toward the character difference, it is best to consider the inheritance oligogenic based on effect. The implications of these findings are discussed in light of the importance of macromutation in speciation and the sex chromosome theory of speciation.     

Chromosomal replication in Drosophila virilis

About half of the diploid genome of D. virilis is -heterochromatic (Heitz, 1934) and contains the satellite sequences found in isopycnic CsCl density gradients (Gall et al, 1971; Steinemann, 1976). The thymidine incorporation behavior of this material in the course of S phase was monitored by autoradiography. Labelled interphase nuclei show three types of labelling patterns, label exclusively confined to either eu- or -heterochromatin, and simultaneous labelling of both fractions. Using the fraction of labelled mitotic index method, the duration of the DNA-synthetic period, t_s = 11.9 ± 4.3 h and G₂ period, t_{G2 + 1/2M} = 6.9 ± 3.8 h, were determined. On the assumption that the investigated brain cells belong to an exponentially growing cell population, the cell cycle is 22.9 h long and the G₁ period lasts t_G1=4.1 h. The a-heterochromatin begins to replicate later than euchromatin and continues alone after a phase of common replication of both fractions. Noteworthy is the asynchronous termination in the proximal -heterochromatic segments of different chromosomes. Within the S phase, the first 1 h of DNA replication is exclusively confined to euchromatin, followed by 8 h of replication in both eu- and -heterochromatin and terminated by 3 h of exclusive -heterochromatin replication. Thus euchromatin has a doubling time of about 9 h and -heterochromatin of about 11 h. The -heterochromatin of D. virilis is late and slow replicating.