水稻T-DNA插入启动子捕获系的初步筛选

利用农杆菌介导法将含报道基因β-glucuronidase(GUS)无启动子的转化载体pCAMBIA1300GUSAHyg导入籼稻明恢86获得水稻启动子捕获系.水稻启动子捕获系潮酶素抗感分离比χ2分析表明:39个水稻启动子捕获系中有28个为单T-DNA位点插入.2 653个水稻启动子捕获系报告基因GUS表达检测结果表明...     

灰葡萄孢蚕豆分离物的遗传多样性

【目的】灰葡萄孢是引起蚕豆赤斑病、花腐病和荚腐病的重要病原菌,其为包含两个称作类群I和类群II的复合种,具有高度的遗传变异。然而,危害蚕豆的灰葡萄孢复合种地位及遗传特征尚未被研究。论文旨在对中国6个不同地理来源蚕豆灰葡萄孢复合种的地位进行鉴定,研究其遗传多样性及群体间的遗传分化,为探明病害流行规律及制定防治策略提供依据。【方法】在PDA培养基上观察分离物形态特征。用Flipper和Boty特异性引物检测分离物的转座子基因型。葡萄孢复合种所属类群通过Bc-hch基因的PCR扩增酶切多态性鉴定,SSR标记分析解析其遗传多样性和遗传分化。【结果】100个灰葡萄孢分离物均为灰葡萄孢类群II,共产生菌核型、菌丝型和分生孢子型3种类型菌落形态,其中76个分离物为菌核型。92个分离物为转座子基因型,分别含有Boty、Flipper和Boty+Flipper,其中仅含Boty分离物最多,占检测分离物的61.0%。江苏分离物只含有单一Flipper,重庆、四川、甘肃和河北分离物只含有单一Boty。基于SSR分析,100个分离物被鉴定为92个多位点基因型,6个群体的平均基因多样性指数和平均基因型多样性指数分别为0.5140和0.9982;分子方差分析（AMOVA）发现群体内遗传变异占总变异的86.70%。标准关联指数rD值范围为0.0312-0.1261,平均0.0491;遗传固定指数（FST）在0.0085-0.2650,平均为0.1330。UPGMA聚类将100个分离物划分为10个遗传组,大部分遗传组包含不同地理来源的分离物。贝叶斯（Bayesian）推理分析将92个多位点基因型分为两个遗传类群,其中重庆和四川群体聚为一类,青海群体单独聚为一类,而江苏、河北和甘肃群体由两个不同遗传类群组成。【结论】源于蚕豆的灰葡萄孢群体由灰葡萄孢类群II组成,分离物的菌落形态以菌核型为主,绝大多数分离物为转座子基因型,但转座子基因型分布明显存在地域差异。调查的6个群体都具有高水平的遗传多样性,遗传变异主要来源于群体内,生殖方式以有性繁殖为主,大部分群体间存在中等到大的遗传分化。     

水稻抗稻瘟病基因Pi-d2基因标签的建立与应用

建立抗稻瘟病基因的分子标记对培育抗稻瘟病品种有重要意义。研究利用71个广西地方菌株检测地谷对稻瘟病菌的抗性,并利用地谷中Pi-d2基因与广恢998等位基因在基因组序列上一个碱基的差异,设计出以抗病基因本身序列为引物的基因标签M-Pid2,通过分析地谷与广恢998的F2群体的基因型与对稻瘟病抗感分离的吻合度来验证该标签的准确性,用M-Pid2扩增62份水稻种质验证该标签的特异性。结果表明,地谷能抗71个地方稻瘟病菌株中的59个菌株,抗性频率达83.1%。用M-Pid2扩增地谷与广恢998的F2群体的基因型与其对稻瘟病的抗感分离完全吻合;用M-Pid2扩增62份水稻种质,仅在地谷中扩增出629bp特异条带。从而确认抗稻瘟病基因Pi-d2为地谷的主效抗稻瘟病基因,且在广西有育种利用价值;标记M-Pid2能准确用于抗稻瘟病基因Pi-d2的辅助选择。     

瓜类细菌性果斑病菌突变体文库的构建及群体感应信号分子突变株的筛选

利用三亲杂交的方法将带卡那抗性的Mini-Tn5转座子随机插入瓜类细菌性果斑病菌xjl12基因组DNA中,构建插入不同位点的突变体文库,通过信号分子高效检测菌株JZAI,筛选群体感应信号分子失活突变株.挑取2株野生株和筛选出的4株突变株,进行Southern杂交,结果表明,转座子Mini-Tn5以单拷贝随机诱变瓜类细菌性果斑病菌获得成功;对筛出的4株突变株进行了致病性和烟草过敏反应测试,发现突变株明显降低了致病性,且2株突变株(5-17,2-57)在烟草上无过敏反应.瓜类细菌性果斑病菌突变体文库的成功构建及筛选到不同突变位点的群体感应信号分子失活突变株,对从基因水平研究瓜类细菌性果斑病菌致病分子机理及群体感应相关基因奠定了基础.     

Geolocation of Atlantic cod (Gadus morhua) movements in the Gulf of Maine using tidal information

Information derived from archival tags (digital storage tags, DSTs) were used to backtrack the migration of 11 tagged Atlantic cod (Gadus morhua) during 2001 in Massachusetts Bay, the Gulf of Maine, and Georges Bank. The DST tags continuously recorded time, temperature and depth. To geolocate fish positions during its time at large, we first extracted the tidal signal from the pressure recordings on the DST tags, and then compared the resulting data to data predicted with a Massachusetts Bay tidal model that provided us with geographical coordinates at a given date and time. Using least‐squares criteria within an estimated geographical region of confidence that was constrained by biological and statistical information (e.g. swimming speed, known release and recapture location, and bottom depth) we were able to geolocate the fish. The resultant geolocated migration tracks indicate a large degree of movement of Atlantic cod in the region and an elevated importance of the Great South Channel (GSC) migration corridor between Massachusetts Bay and the western Georges Bank and Nantucket Shoals region. This observation contrasts strongly with inferences of limited movements by Atlantic cod based on conventional tag recapture methods (mean of 1200 km traveled versus 44 km traveled as measured by conventional tagging and geolocation, respectively). This study demonstrates that geolocation methodologies applied to archival tag studies hold great promise of becoming an important new tool for fisheries managers to quantify the movements of fishes. It also points out the need for greater collaboration between fisheries scientists and oceanographers, and particularly for the development of improved tidal models to cover stock regions more accurately and with higher precision.     

Stock structure and seasonal distribution patterns of American lobster, Homarus americanus, inferred through movement analyses

Movement influences the annual distribution patterns of a species and is an important determinant of stock structure. In situations where monitoring programs have quantified movement or distribution patterns by sampling during particular times of the year, seasonal changes in abundance as well as the degree of connectivity among adjacent stocks can be underestimated. Here, a summer abundance trawl survey was combined with a 1-year mark-recapture tagging study to infer seasonal changes in distribution within and among American lobster (Homarus americanus) stocks. Within the study area, lobsters were concentrated in central Northumberland Strait (Canada) during August, yet their observed dispersal behaviour implied that density declined in the central portion and increased in the northern portion of Northumberland Strait during winter. Stock mixing among management zones was not observed and individual tendencies to move were predicted to decline precipitously in early December. These movement patterns are consistent with the hypothesis of seasonal limitation by hard-substrate habitat availability causing population redistribution. Such information can ultimately be useful when assessing changes in abundance or exploitation rates, and for guiding management efforts.     

转座子在害虫生物防治中的应用

转座子是一类散布在基因组中序列重复的DNA片段,它们可以不依靠同源重组而能在基因组中移动。综述了转座子的特征、分类、种类和分布,介绍了利用转座子获得的转基因昆虫和改良杀虫剂在害虫生物防治中的应用,并对转座子在害虫生物防治中的研究和应用前景进行展望。     

家蚕一种hAT家族转座酶基因的分析

[目的]对家蚕中一个潜在的hAT家族转座酶编码基因BmhAT进行分析.[方法]通过生物信息学方法,分析BmhAT转座酶的结构域及序列同源性.采用Real-time PCR方法,对BmhAT在野蚕及家蚕不同品种中的拷贝数进行分析以及在mRNA水平上测定BmhAT在家蚕不同组织中的相对表达量.[结果]BmhAT与许多hAT家族转座酶在序列上有很高的同源性,并具有hAT家族转座酶的保守结构域;BmhAT在家蚕基因组中存在约3-6个拷贝;在家蚕后部丝腺和脂肪体中的表达量分别是参照基因actin 3的0.4倍和0.6倍.[结论]BmhAT可能是家蚕中一个有活性的转座酶基因,编码家蚕一种新的hAT家族转座酶.     

鱼类标志放流技术的研究现状

标志放流（tagging）技术是一种研究鱼类洄游和鱼类资源的方法，先在鱼体做上标志，然后通过重新捕获标志鱼，根据标志放流记录和重捕记录，绘制鱼类标志放流和重捕的分布图，以推测鱼类游动的方向、路线、范围和速度等；若进一步结合鱼体长度、重量和年龄资料，则可研究鱼类的生长和死亡规律，以及检验增殖放流的效果等。此外，根据鱼类标志放流的结果还可估算鱼类种群数量的变动。我国的鱼类标志技术研究起步较晚，有关报道较少，因此，开展这方面的工作非常迫切。本文对鱼类标志方法的研究进展予以综述，以期促进我国鱼类标志技术的应用与发展。     

Development of a large population of activation-tagged mutants in an elite indica rice variety

In this study, we have generated more than 12,000 activation-tagged mutants in a high-yielding indica rice variety, 'BPT 5204', employing maize Ac/Ds system. Different transgenic plants obtained were analysed based on expression patterns of green fluorescence protein (GFP), red fluorescence protein (RFP), herbicide (Basta) tolerance and molecular analyses. T₁ seeds of pSQ5 and pSQ5-bar transgenics, when germinated separately on hygromycin (50 mg/L) and phosphinothricin (5 mg/L) containing medium, revealed a segregation of 3 tolerant : 1 susceptible plants. The germinal transposition frequency of Ds element in different T₂ progeny of rice plants was found to be about 18.0%. Different stable tagged mutants exhibited marked increases in plant height, number of tillers, leaf size, panicle size, seed size and number of grains per plant. The overall results indicate that the genes associated with these traits are upregulated by the enhancer element in activation-tagged mutants. As such, the various tagged mutant lines appear promising and serve as a valuable genetic resource for identification of key genes determining different agronomic traits of rice.