黑腹果蝇黑条体突变体突变位点的鉴定

根据果蝇ebony基因的序列设计了一系列特异性引物,对1991年发现的一种黑腹果蝇的新的体色突变（Qian＆Zhang,1994）——黑条体突变型的ebony基因进行克隆和序列测定。与野生型W91910的ebony基因相比,黑条体突变型的ebony基因的编码区无明显的大片段变异,变异仅存在于数个氨基酸位点,且均不位于该基因产物的关键序列。在黑条体的ebony基因的5’端序列的克隆和序列测定中发现,与野生型w^91910及黑檀体e。的ebony基因相比,黑条体突变型的ebony基因有一个大片段的缺失,该缺失包括外显子1的206个碱基和内含子1的747个碱基,由此确定了黑条体突变体的突变类型和突变位点。     

黑腹果蝇黑条体突变型的基因定位研究

黑腹果的体色突变类型常见的有黄体（yellow,y）、黑体（black,b）和黑檀体（ebony,e）,分别位于X染色体,第二染色体和第三染色体上,.黑条体突变型是本实验室1991年9月从野外采集的黑腹果蝇野生型单雌系后代中发现的自发突变品系,为了探明黑条体突变型是原有黑体突变类型的再现还是新的突变,采用常规杂交方法和互补7实验技术对黑腹果蝇黑条体突变型的定位进行了探讨,互补测验的结果表明,黑条体与黑檀体杂交的子一代为反式排列的杂合体无互补,表现为突变型,子二代中,由于交换而产生重组类型的顺式排列的杂合体表现为野生型。因此确定黑条体突变基因（bsr）与黑檀体突变基因（e）是等位的,位于第三染色体的93D2区,但分别位于不同的位点上,属于同一顺反子的新的点突变,同时对于各体色间的相互作用及遗传传递方式的进行了讨论。     

稀有的黑檀体基因(ebony)5'UTR自发突变导致黑腹果蝇的黑条体突变

由于果蝇Drosophila群体中有很多自发突变其中包括多种体色突变,因此它是一个研究自发突变的优秀的模式体系。本研究证实我们实验室发现的一个可以引起果蝇体色突变的自发突变(bsr)是一个黑檀体(e)的等位基因,将其命名为ebsr。序列分析显示ebsr的5′端缺失了953个碱基,其中包括外显子1后端的206个碱基及相连的内含子1的747个碱基。逆转录PCR结果显示5′端的缺失导致内含子1不能从mRNA中剪接掉,由此导致该mRNA的翻译起始密码子AUG前端增加了一个3.2kb的序列。该序列导致ebsr的mRNA的5′UTR(5′-untranslated region)区较野生型基因增加近3kb的长度。通过mRNA二级结构分析发现这个增加的3kb的片段可以形成复杂的颈环结构(stem-loop)。免疫印迹结果显示该突变基因没有基因产物产生。本研究进一步证实了由于mRNA的5′UTR序列结构的改变可以影响到蛋白质的翻译。     

稀有的黑檀体基因（ebony）5′UTR自发突变导致黑腹果蝇的黑条体突变

由于果蝇Drosophila群体中有很多自发突变其中包括多种体色突变, 因此它是一个研究自发突变的优秀的模式体系。本研究证实我们实验室发现的一个可以引起果蝇体色突变的自发突变（bsr）是一个黑檀体（e）的等位基因, 将其命名为e^bsr。序列分析显示e^bsr的5′端缺失了953个碱基, 其中包括外显子1后端的206个碱基及相连的内含子1的747个碱基。逆转录PCR结果显示5′端的缺失导致内含子1不能从mRNA中剪接掉, 由此导致该mRNA的翻译起始密码子AUG前端增加了一个3.2 kb的序列。该序列导致e^bsr的mRNA的5′UTR（5′-untranslated region）区较野生型基因增加近3 kb的长度。通过mRNA二级结构分析发现这个增加的3 kb的片段可以形成复杂的颈环结构（stem-loop）。免疫印迹结果显示该突变基因没有基因产物产生。本研究进一步证实了由于mRNA的5′UTR序列结构的改变可以影响到蛋白质的翻译。     

果蝇求偶行为的影响因素及其分子基础

果蝇Drosophila melanogaster Meigen是进行行为遗传学研究的极好材料。果蝇的雄性求偶行为已经被作为行为遗传学研究的模式。文章简要介绍近年来在遗传和分子水平上对果蝇性信息素和求偶行为的研究进展,尤其是突变体在果蝇行为遗传学研究中的应用。通过对果蝇求偶行为的分析,分别介绍果蝇的性信息素及视觉、听觉、嗅觉和味觉相关基因在果蝇求偶和交配行为过程中的作用。     

黑腹果蝇的性别控制

性别的形成包括两个过程,即性别决定和性别分化。果蝇的性别控制研究包括性别决定、性别分化、性别鉴定、性别诱导和性别控制5个方面。性别决定是在两种不同发育途径之间的选择,它提供了一个研究基因调控的模式系统。果蝇的性别决定问题已经研究得相当详细［1］。性别分化是使胚胎向着雌性或雄性发育的过程,决定了性别表型。果蝇的性别分化也取得了不少研究成果。近年来,许多重要的性别调控基因已被克隆和鉴定。随着果蝇基因组全序列测定的完成,果蝇的性别控制研究将会更为深入而完善。本文对与黑腹果蝇性别决定和性别分化相关的一些问题进行综述。     

黑腹果蝇的分离变相因子

众所周知,一对基因在杂合状态中保持相对的独立性,而在配子形成时,又按原样以相同比例分离到不同的配子中去,这是生物中最基本的遗传规律———孟德尔分离定律。但实际上并不是所有基因的分离都严格遵循孟德尔分离定律,存在于黑腹果蝇中的分离变相因子（ＳｅｇｒｅｇａｔｉｏｎＤｉｓｔｏｒｔｅｒ,以下简称ＳＤ）就是一种具有减数分裂驱动（ｍｅｉｏｔｉｃｄｒｉｖｅ）性质的、违反孟德尔分离定律的特殊遗传因子,从六十年代发现至今引起了人们的广泛关注,本文从其遗传背景,结构特征及进化规律等方面分别介绍一些近期的研究结果。１…     

樱桃新害虫黑腹果蝇的生物学特性

果蝇是近几年发现危害樱桃果实的一类重要害虫,在国内外樱桃产区均有发生。天水地区危害甜樱桃的果蝇有3个种,分别是黑腹果蝇Drosophila melanogaster Meigen、铃木氏果蝇Drosophila suzukii(Matsumura)和海德氏果蝇Drosophila hydei(Sturtevant),黑腹果蝇为优势种。作者记述黑腹果蝇对甜樱桃果实的危害情况、寄主范围及其生活史、生活习性、发育历期与温度的关系等,调查发现蚂蚁是樱桃果蝇的天敌之一。     

黑腹果蝇细胞谱系分析方法进展

对动物体内单个细胞的谱系进行分析有助于追踪其在发育过程中的作用,但是体内各种组织都是由很多形态、结构、功能各不相同的细胞构成的复杂系统,这种复杂性严重阻碍了对单个细胞的研究。嵌合克隆技术(Mosaic technique)和标记技术(Labeling technique)的出现为这一研究提供了强有力的手段。文章介绍了近几年来黑腹果蝇(Drosophila melanogaster)研究中常用的7种嵌合克隆标记方法,包括FRT介导的有丝分裂重组(FRT-mediated mitotic recombination)、MARCM(Mosaic analysis with a repressible cell marker)、TSG(Twin spotgenerator)、Twin-spot MARCM、Q-MARCM(Q system-based MARCM)、Coupled MARCM和G-TRACE(Gal4technique for real-time and clonal expression)技术,详述了这些技术的原理及应用,并对不同技术进行了对比。运用这些技术研究者可以从单细胞水平进行遗传学标记和操作,特别是在神经系统等复杂系统中追踪单个细胞的发育过程。果蝇中的这些技术也将为其他模式生物追踪细胞谱系提供参考。     

黑腹果蝇先天免疫研究进展

先天免疫是昆虫适应复杂环境的关键,也是新型害虫防治的重要研究方向。昆虫通过模式识别受体识别环境中不同的病原物,激活先天免疫系统以清除病原物。昆虫的先天免疫系统主要包括体液免疫与细胞免疫,体液免疫包括免疫信号通路诱导产生抗菌肽、活性氧以及黑化作用等,细胞免疫包括血细胞的吞噬、包囊和凝结。本文将重点总结黑腹果蝇Drosophila melanogaster在模式识别受体、免疫信号通路和细胞免疫相关方面的研究进展,为进一步研究其他经济昆虫与农业害虫的免疫机制,提高生产经济效益提供参考。     

Multiple mating in wild Drosophila melanogaster revisited by microsatellite analysis

The occurrence of multiple mating in Drosophila melanogaster is of particular interest to evolutionary biologists, as seminal fluid has some toxic effects for females. Thus, it has been predicted that the number of matings per females should be low. We have tested this prediction with seven highly polymorphic microsatellite loci in inseminated females from a Viennese D. melanogaster population. In contrast to the predicted low number of matings and previous studies in natural populations, we identified the genotypes of four to six different males fertilizing the offspring of each female tested. Potential causes and consequences are discussed.     

The genetic basis of adaptive pigmentation variation in Drosophila melanogaster

In a broad survey of Drosophila melanogaster population samples, levels of abdominal pigmentation were found to be highly variable and geographically differentiated. A strong positive correlation was found between dark pigmentation and high altitude, suggesting adaptation to specific environments. DNA sequence polymorphism at the candidate gene ebony revealed a clear association with the pigmentation of homozygous third chromosome lines. The darkest lines sequenced had nearly identical haplotypes spanning 14.5 kb upstream of the protein-coding exons of ebony. Thus, natural selection may have elevated the frequency of an allele that confers dark abdominal pigmentation by influencing the regulation of ebony.     

An analysis of genetic differentiation among assortatively mating Drosophila melanogaster in Zimbabwe

African Drosophila melanogaster populations, and those from Zimbabwe in particular, have attracted much interest recently. African flies differ genetically from 'cosmopolitan' populations and were found to exhibit discriminative mating behaviour against individuals from 'cosmopolitan' populations. It has therefore been proposed that Zimbabwean and some other African populations are in an 'incipient stage of speciation'. However, whether the mating behaviour is an effective barrier against gene flow from other populations, and whether intra-population genetic differentiation has already evolved in sympatry is not known. Here, we took a population-based approach to test whether the well-characterized mating type differences have resulted in a genome-wide differentiation at the population level. Using 122 polymorphic microsatellite loci mapping to the third chromosome, we demonstrate a significant genetic differentiation between Zimbabwean flies differing in their mating behaviour. We also provide evidence to suggest that this difference is unlikely to be attributable to population structure within Zimbabwe. However, the analysis of individual microsatellite loci did not indicate more loci differentiating these two groups than expected by chance. Our data suggest that the 'Z'-'M' mating behaviour is strong enough to result in a small but significant genetic differentiation. Thus, future studies based on a larger population sample of flies characterized for their mating behaviour and using more markers are expected to provide more information on the genetic basis of the mating traits in the Zimbabwe flies.     

黑腹果蝇昼夜活动节律及行为时间分配

采用试验室单管观察记录的方法,对3,4,5日龄野生型黑腹果蝇Drosophila melanogaster Meigenw1118成虫每日活动节律进行研究。试验将果蝇活动划分为强活动(飞行和爬行)、弱活动(梳理、觅食等原地发生的运动)和静息(身体不发生移动的休息)3种类型。强活动和弱活动之和为总运动。研究结果显示,野生型黑腹果蝇w1118的昼夜活动表现为明显的双峰模态,晨峰和晚峰分别处于开、关灯前后;雌、雄果蝇总体活动无差异,关灯(18:30)前后雌蝇活动稍强于雄蝇,开灯(6:30)前后则相反;果蝇强活动的节律与总运动基本一致,而弱活动节律不明显;静息节律为单峰模式,其高峰期位于夜间1:00~5:00;雌蝇的静息活动显著多于雄蝇(P<0·05)。     

Age-dependent behavior loss in adult Drosophila melanogaster

The use of Drosophila as an organism in which to study aging has been limited by the fact that few biomarkers of aging exist in the adult. In this paper we examine behavior loss relative to longevity in wild-type populations maintained at 22°C and 29°C to determine whether behavior loss—that is, loss of ability to perform certain innate behavioral responses within a defined test interval—can be used as biomarkers of aging. We find that under controlled conditions behavior loss can be used as a landmark of aging in populations maintained at either 22°C or 29°C. The ability to perform normal geotactic and phototactic responses is lost during the reproductive phase of the adult populations, whereas motor activity is not lost until well into the death phase. We feel that the use of behavior loss, together with other parameters of longevity in Drosophila, will allow comparisons to be made between different strains or between different environmental conditions to test their effect on aging. In the companion paper we demonstrate the use of behavior loss to identify a mutation which may accelerate the aging process.     

Habitat selection by Drosophila melanogaster larvae

Drosophila melanogaster larvae are used to examine habitat choice behavior and its effect on a component of preadult fitness (pupal survivorship). We established strains of flies by collecting pupae from two microhabitats from an orchard. Strain differences in pupation site choice (on versus off fruit) persisted in a field-like laboratory assay without artificial selection. To produce heterogeneous environments, air temperature and soil water content were varied in these assays. A habitat suitability difference measure was used to determine for each environment, which microhabitat (on or off fruit) resulted in greater pupal survivorship. We found 1) that habitat choice behavior had both plastic and heritable components, 2) that strain-by-environment interactions influenced habitat choice behavior and pupal survivorship and, 3) a significant positive correlation between habitat suitability and larval habitat choice behavior.     

Simulation of gene pyramiding in Drosophila melanogaster

Gene pyramiding has been successfully practiced in plant breeding for developing new breeds or lines in which favorable genes from several different lines were integrated.But it has not been used in animal breeding,and some theoretical investigation and simulation analysis with respect to its strategies,feasibility and efficiency are needed before it can be implemented in animals.In this study,we used four different pure fines of Drosophila melanogaster,each of which is homozygous at a specific mutant gene with a visible effect on phenotype,to simulate the gene pyramiding process and analyze the duration and population size required in different pyramiding strategies.We finally got the ideal individuals,which are homozygous at the four target genes simultaneously.This study demonstrates that gene pyramiding is feasible in animal breeding and the interaction between genes may affect the final results.