Functional divergence caused by ancient positive selection of a Drosophila hybrid incompatibility locus

Interspecific hybrid lethality and sterility are a consequence of divergent evolution between species and serve to maintain the discrete identities of species. The evolution of hybrid incompatibilities has been described in widely accepted models by Dobzhansky and Muller where lineage-specific functional divergence is the essential characteristic of hybrid incompatibility genes. Experimentally tractable models are required to identify and test candidate hybrid incompatibility genes. Several Drosophila melanogaster genes involved in hybrid incompatibility have been identified but none has yet been shown to have functionally diverged in accordance with the Dobzhansky-Muller model. By introducing transgenic copies of the X-linked Hybrid male rescue (Hmr) gene into D. melanogaster from its sibling species D. simulans and D. mauritiana, we demonstrate that Hmr has functionally diverged to cause F1 hybrid incompatibility between these species. Consistent with the Dobzhansky-Muller model, we find that Hmr has diverged extensively in the D. melanogaster lineage, but we also find extensive divergence in the sibling-species lineage. Together, these findings implicate over 13% of the amino acids encoded by Hmr as candidates for causing hybrid incompatibility. The exceptional level of divergence at Hmr cannot be explained by neutral processes because we use phylogenetic methods and population genetic analyses to show that the elevated amino-acid divergence in both lineages is due to positive selection in the distant past—at least one million generations ago. Our findings suggest that multiple substitutions driven by natural selection may be a general phenomenon required to generate hybrid incompatibility alleles.     

Functional conservation of the Drosophila gooseberry gene and its evolutionary alleles

The Drosophila Pax gene gooseberry (gsb) is required for development of the larval cuticle and CNS, survival to adulthood, and male fertility. These functions can be rescued in gsb mutants by two gsb evolutionary alleles, gsb-Prd and gsb-Pax3, which express the Drosophila Paired and mouse Pax3 proteins under the control of gooseberry cis-regulatory region. Therefore, both Paired and Pax3 proteins have conserved all the Gsb functions that are required for survival of embryos to fertile adults, despite the divergent primary sequences in their C-terminal halves. As gsb-Prd and gsb-Pax3 uncover a gsb function involved in male fertility, construction of evolutionary alleles may provide a powerful strategy to dissect hitherto unknown gene functions. Our results provide further evidence for the essential role of cis-regulatory regions in the functional diversification of duplicated genes during evolution.     

杜鹃花属植物杂交不亲和与败育分布研究

该研究以杜鹃花属5亚属3组17亚组38种,计109个不育组合及91个可育组合的杂交结果为依据,对4个杂交不亲和指标与败育系数及败育频度参数进行了分析。结果表明:(1)杜鹃花属不同类群间杂交的不育组合比例约为54.5%,其不亲和与败育包括不能坐果(capsul aborted,Cab型)、坐果但不能形成种子(seed aborted,Sab型)和能形成种子但不能发芽(seed not germinated,Sng型) 3种情况,其中Cab与Sab类型均可能是前合子期不亲和与后合子期不亲和的复合表征,Sng型则可以肯定为后合子期种子发育阶段败育的情况,Cab∶Sab∶Sng=81∶13∶15;而可育组合中部分败育苗无疑属于"杂种不活hybrid inviability"的败育类型。(2)有关不育类型的分布与杜鹃花属植物亲本类群及其分类与亲缘关系具有明显关联,从同一亚组内、同一亚属内到不同亚属间杂交的不育类型的分布呈Sng型→Sab型→Cab型增加的趋势,杂交双亲分类上亲缘关系越密切Sng型的频度越高,反之关系越疏远Cab型的频率越高,亚属间远交往往止于Cab型。(3)"杂种不活"为败育苗的表现形式之一,从同一亚组、同一亚属到不同亚属间杂交,其败育等级频度分布通常也从无或轻度败育到严重败育方向发展,常绿杜鹃亚属内杂交的败育情况明显低于杜鹃亚属内杂交,这与后者多倍体亲本的介入相关,而上述2亚属间杂交的苗木败育则更加严重,但映山红(R. simsii)分别与百合花杜鹃(R. liliiflorum)和毛肋杜鹃(R. augustinii)的亚属间杂交未出现败育苗分布。     

Functional conservation of a glucose-repressible amylase gene promoter from Drosophila virilis in Drosophila melanogaster

Summary Previous studies have demonstrated that the expression of the -amylase gene is repressed by dietary glucose in Drosophila melanogaster. Here, we show that the -amylase gene of a distantly related species, D. virilis, is also subject to glucose repression. Moreover, the cloned amylase gene of D. virilis is shown to be glucose repressible when it is transiently expressed in D. melanogaster larvae. This cross-species, functional conservation is mediated by a 330-bp promoter region of the D. virilis amylase gene. These results indicate that the promoter elements required for glucose repression are conserved between distantly related Drosophila species. A sequence comparison between the amylase genes of D. virilis and D. melanogaster shows that the promoter sequences diverge to a much greater degree than the coding sequences. The amylase promoters of the two species do, however, share small clusters of sequence similarity, suggesting that these conserved cis-acting elements are sufficient to control the glucose-regulated expression of the amylase gene in the genus Drosophila.Offprint requests to: D.A. Hickey     

Functional interactions between the gene tetanic and the Shaker gene complex of Drosophila

Different phenotypes associated with the tetanic (tta) mutation such as appendage contraction, maternal effect and low viability and fertility are enhanced by one extra dose of the Shaker gene complex (ShC). The tta mutation is lethal with two extra doses of ShC. In addition, tta embryos have a defective nervous system. In this paper, I analyse the interaction between tta and ShC to gain insight into their relationship. Aneuploid analysis suggests that the lethality is due to an interaction of the tta mutation with the maternal effect (ME) region of this gene complex. Mutations in the ME region of ShC partially suppress this interaction. Trans-heterozygous combinations of MEI[l(1)305] and MEIII [l(1)459] mutations causes dominant lethality in a tta background. Trans-heterozygous combinations of an MEII [l(1)1359] mutation with the cited MEI and MEIII mutations are lethal in a tta background. Double mutant combinations and gene dosage experiments, suggest that tta also interacts with the viable (V) region of ShC. These specific genetic interactions indicate that tta and the ME and V regions of ShC are functionally related. These results, together with the previous electrophysiological, molecular and biochemical studies on these mutants suggest an interaction at the protein level. Thus, in the case of the V region, the tta gene product may modulate the activity of the K⁺ channels encoded in this region. Furthermore, the extreme dosage sensitivity of the interaction between tta and ShC suggests a stoichiometric requirement for the different gene products involved, which might be physically associated and form heteromultimers.     

Functional substitution of the recE gene of Bacillus subtilis by the recA gene of Proteus mirabilis

Summary Rec mutants of Bacillus subtilis have been tested for complementation by the recA gene of Proteus mirabilis (recApm) which was introduced into B. subtilis via the plasmid pHP334. In the recE4 mutant of B. subtilis the plasmid pHP334 restored significantly the defects in RecE functions tested: UV-sensitivity, homologous recombination (transduction and transformation) and prophage induction.Although serological methods to detect the presence of RecApm protein in B. subtilis have been unsuccessful, our results strongly indicate that the recE function of B. subtilis is analogous to the recA function of P. mirabilis.Abbreviations Cm^r resistance to chloramphenicol - Em^r resistance to erythromycin - Tc^r resistance to tetracycline - SDS sodium dodecyl sulfate - UV ultraviolet - AS ammonium sulfate     

Functional conservation of hematopoietic factors in Drosophila and vertebrates

The GATA, Friend of GATA, and Runt homology domain protein families function during hematopoiesis to promote progenitor cell development and regulate lineage commitment and differentiation. The hematopoietic functions of these factors have been remarkably conserved across taxonomic groups, ranging from flies to humans. Furthermore, aspects of hematopoiesis and hemocyte function appear to be conserved. Thus, comparative studies using Drosophila and vertebrate models should enhance our understanding of blood cell development.     

Functional relationships between genes of the Shaker gene complex of Drosophila

Different mutations belonging to the HLI and HLII complementation groups of the haplolethal (HL) region of the Shaker complex (ShC) are described. The HLI complementation group includes viable (hdp), recessive lethals [l(1)1614], semidominant lethals [l(1)8384] and dominant lethals [l(1)5051,l(1)9916, l(1)13193], lack-of-function alleles that affect nervous system, cuticle and muscle development. The HLI complementation group encodes troponin I. HLII lack-of-function mutations [l(1)174 and l(l)4058] affect nervous system development. The semidominant lethal HLI mutation 1(1)8384 shows differential complementation with other mutations in the ME and HL regions of ShC. Thus, heterozygous combinations of l(1)8384 with ME mutations l(1)162 and l(1)387 are poorly viable. The same phenomenon is observed for heterozygotes of l(1)8384 with HL mutations l(1)1199, l(1)2288 and l(1)3014. These specific interactions indicate the existence of functional relationships among the genetic elements of ShC. The implications for the understanding of the functional organization of ShC are discussed.     

核桃细菌性黑斑病菌rpfG基因的功能分析

[目的] 本研究旨在揭示核桃细菌性黑斑病菌（Xanthomonas arboricola pv.juglandis,Xaj） DW3F3中rpfG基因的生物学功能,从而为核桃细菌性黑斑病防治药剂的开发提供作用靶点。[方法] 以野油菜黄单胞菌（Xanthomonas campestris pv.campestris,Xcc）8004菌株以及水稻白叶枯病菌（Xanthomonas oryzae pv.oryzae,Xoo） PXO99^A的rpfG基因为模板序列,对Xaj野生型菌株DW3F3的基因组序列进行检索。利用同源重组技术,对Xaj中rpfG基因进行敲除,并用生物化学方法对基因缺失菌株的相关毒力因子、抗逆性进行检测。[结果] 通过同源比对,在XajDW3F3的基因组中发现了与XccrpfG、XoorpfG同源的基因,并成功获得rpfG的缺失突变株ΔrpfG。与野生型相比,突变株ΔrpfG的生物被膜形成能力仅为野生型XajDW3F3的44.58%;胞外多糖产量也由野生型的8.47 mg/mL降为5.23 mg/mL;ΔrpfG的絮凝活性增加,能使菌液变澄清;运动性实验显示ΔrpfG的运动直径比野生型增加了12.38%;胞外酶的分泌也发生了不同程度的改变,突变株分泌纤维素酶的能力极显著降低,淀粉酶活性有所提高,而分泌蛋白酶的能力未发生变化;此外rpfG缺失后,Xaj对逆境（盐、酸、SDS、硫酸铜）的耐受力降低。[结论] 结果表明rpfG基因能影响核桃细菌性黑斑病菌的致病相关性状,并赋予了细菌一定的抗逆性。     

Selectively recombining B incompatibility factors of Schizophyllum commune

Summary Analysis of genetic crosses among strains of Schizophyllum commune carrying recombining B factors has revealed that not all heteroallelic pairs of B factors are able to recombine with each other. This suppression of recombination is highly specific and appears to be determined by the B factors themselves.     

胡杨WINDs基因克隆及功能初步分析*

目的: WIND(WOUND INDUCED DEDIFFERENTIATION),是属于ERF/AP2 (ETHYLENE RESPONSE FACTOR/ APETALA 2)家族的一种重要转录因子,该类基因最早被发现在拟南芥中可以与乙烯响应元件GCC-BOX和脱水响应元件DRE结合,响应干旱信号和调节乙烯水平。最近的研究发现WIND基因在植物伤口信号回应、愈伤组织形成及不定芽的产生过程中也发挥了关键作用。已有的研究阐述了WIND基因在拟南芥中控制愈伤组织形成及不定芽再生的机制,但其在木本植物中的功能尚不明确,将探究WIND基因在胡杨中与伤口信号响应及不定芽再生相关的功能,同时为在分子水平上解决胡杨再生问题提供理论依据。方法: 采用基因克隆、qRT-PCR、转基因表型分析等方法研究WIND基因在胡杨外植体伤口响应和再生不定芽过程中的作用。结果: 克隆胡杨WIND家族中的基因PeWIND1和PeWIND2,发现其编码区序列长度分别为1 050 bp和1 032 bp,编码349个和343个氨基酸,亚细胞定位均在细胞核中。组织特异性分析显示PeWIND1和PeWIND2在胡杨根、茎、叶、愈伤组织中均有表达,且在愈伤组织中表达量最高。时间表达特异性显示,在经伤口刺激后的24 h内,PeWIND1和PeWIND2基因均呈现先升高后降低的表达趋势,且均在伤口刺激后1 h达到表达量峰值。转基因植株表型统计发现,过表达PeWIND1和PeWIND2基因后转基因植株不定芽再生能力增强。结论: 在胡杨叶片有伤口刺激后,PeWIND1和PeWIND2响应伤口信号,表达量先升高后降低,PeWIND1和PeWIND2能够促进杨树茎段再生不定芽。     

茎瘤固氮根瘤菌ORS571中motA基因的功能分析

[目的] MotA是细菌的鞭毛马达蛋白,是跨膜质子通道的重要组成结构之一,在调控鞭毛运动中具有至关重要的作用。本研究探究了Azorhizobium caulinodans ORS571中鞭毛马达基因motA对菌株表型和植物互作的影响。[方法] 通过同源重组原理和三亲接合转移方法构建突变菌株∆motA,测定野生型与突变体在菌体生长、运动、固氮、胞外多糖合成、生物膜形成及根系定殖能力的差异。[结果] 与野生型相比,突变体菌体生长没有明显差异,但其运动能力完全丧失,固氮、胞外多糖合成、生物膜形成及根系定殖能力减弱。[结论] MotA鞭毛马达蛋白对A.caulinodans ORS571的运动、固氮、胞外多糖合成、生物膜形成及根系定殖能力均有调控作用。     

Functional characterization of an ammonium transporter gene from Lotus japonicus

NH₄⁺ is the main product of symbiotic nitrogen fixation and the external concentration of combined nitrogen plays a key regulatory role in all the different step of plant-rhizobia interaction. We report the cloning and characterization of the first member of the ammonium transporter family, LjAMT1;1 from a leguminous plant, Lotus japonicus. Sequence analysis reveals a close relationship to plant transporters of the AMT1 family. The wild type and two mutated versions of LjAMT1;1 were expressed and functionally characterized in yeast. LjAMT1;1 is transcribed in roots, leaves and nodules of L. japonicus plants grown under low nitrogen conditions, consistent with a role in uptake of NH₄⁺ by the plant cells.     

Direct control of antennal identity by the spineless-aristapedia gene of Drosophila

Summary Loss-of-function mutations in the spineless-aristapedia gene of Drosophila (ss ^a mutants) cause transformations of the distal antenna to distal second leg, deletions or fusions of the tarsi from all three legs, a general reduction in bristle size, and sterility. Because ss ^a mutants are pleiotropic, it has been suggested that ss ⁺ has some rather general function and that the ss ^a antennal transformation is an indirect consequence of perturbations in the expression of other genes that more directly control antennal or second leg identity. Here we test whether the ss ^a transformation results from aberrant expression of Antennapedia (Antp), a homeotic gene thought to specify directly the identity of the second thoracic segment. We find that Antp ^– ss ^a mitotic recombination clones in the distal antenna behave identically to Antp ⁺ ss ^a clones, and are transformed to second leg. This demonstrates that the ss ^a antennal transformation is independent of Antp ⁺, and suggests that ss ⁺ may itself directly define distal antennal identity. The results also reveal that Antp ⁺ is not required for the development of distal second leg structures, as these develop apparently normally in Antp ^– ss ^a antennal clones. Because Antp ^– mutations cause deletions or transformations that are restricted to proximal structures, whereas ss ^a alleles cause similar defects that are distally restricted, we suggest that ss ⁺ and Antp ⁺ may play similar, but complementary, roles in the distal and proximal portions of appendages, respectively.