Molecular genetics of rotifers: preliminary restriction napping of the mitochondrial genome of Brachionus plicatilis

Methods are presented to extract and purify mitochondrial DNA from the rotifer Brachionus plicatilis. The mtDNA obtained is of sufficient purity for digestion with restriction endonucleases. EcoR I restriction patterns are presented for 4 geographically separated clones. A restriction map based on digestion with 5 different restriction enzymes is included for one of these clones. Finally, use of mtDNA analysis for studies on the population structure and biogeography of rotifers is discussed.     

水稻Pib基因及其连锁RFLP标记在栽培稻、药用野生稻和玉米中的比较物理定位

比较遗传学研究表明,禾本科不同基因组之间存在着广泛的同线性和共线性.对水稻(Oryza sativa L.)这一模式植物与其他禾本科植物的原位杂交定位可以揭示禾本科植物基因组的共同特点和进化规律,为建立禾本科遗传大体系积累资料.实验以图位克隆法分离的水稻Pib 基因(10.3 kb)和与之连锁的RFLP标记为探针, 研究了Pib及与其连锁的RFLP标记在供试种中的同源性和物理位置. Southern杂交结果表明,Pib在玉米(Zea mays L.)基因组中有同源序列.进一步利用单色和双色荧光原位杂交技术确定了Pib在栽培稻(O.sativa ssp. indica cv. Guangluai 4)、玉米和药用野生稻(O. officinalis Wall ex Watt)染色体上的物理位置.定位结果表明,Pib基因和与之连锁的RFLP标记在这3个供试种基因组中具有同线性.     

小麦耐盐种质的筛选鉴定和耐盐基因的标记

通过对 40 0份材料的芽期、苗期鉴定 ,筛选出 11份耐盐性较强的普通小麦 (TriticumaestivumL .)、小麦和黑麦 (SecalecerealeL .)、小麦和延安赖草 (Leymuschinensis (Trin .)Tzvel.)杂交后代材料 ,其中耐盐性突出的材料有 :普通小麦品种“红蚂蚱”、“科遗 2 6”、“希望”(Hope) ;小麦与黑麦杂交后代材料 98_46、98_113、98_131;小麦与延安赖草杂交后代材料 98_16 0、98_16 1、98_16 3。耐盐性表现最突出的材料是 98_113和 98_16 0。细胞学鉴定和原位杂交及醇溶蛋白酸性聚丙烯酰胺凝胶电泳 (A_PAGE)分析和低分子量谷蛋白SDS_PAGE分析 ,证明 98_113是稳定的小麦 黑麦二体附加系 ,但具体附加的是黑麦的哪条染色体还不清楚 ;98_131是小麦 黑麦 1B/ 1R易位系。结合其他 1B/ 1R材料的耐盐表现 ,提出了黑麦 1R染色体短臂上存在耐盐基因的可能性。对 (98_16 0×BanacakaMska)F2 代分离群体苗期抗盐鉴定分析 ,表明在这一杂交组合中的耐盐性状可能由一个主效基因控制。应用SSR标记技术 ,筛选到了与 98_16 0耐盐性状连锁的SSR标记WMS6 7和WMS2 13,它们与耐盐基因的遗传距离分别为 13.9cM (centMorgan)和 31.0cM。结合小麦SSR图谱分析 ,将该主效抗性基因定位在 5BL上。     

西双版纳四种植被亚型原始林和次生林蚂蚁群落比较研究

采用样地调查法对西双版纳地区４种植被亚型原始林和次生林的蚂蚁群落作了比较研究。山地雨林和石灰岩山季雨林的次生林是原始林经过连续过度砍伐形成的树冠结构不完整的次生林;而落叶季雨林和季风常绿阔叶林的次生林是大约２０年前原始林因刀耕火种被全部砍伐,后来恢复成树冠结构完整的次生林。山地雨林和石灰岩山 雨林次生林蚂蚁特有种数目显著低于     

Recombination at the Rp1 locus of maize.

Summary The Rp1 locus of maize determines resistance to races of the maize rust fungus (Puccinia sorghi). Restriction fragment length polymorphism markers that closely flank Rp1 were mapped and used to study the genetic fine structure and role of recombination in the instability of this locus. Susceptible progeny, lacking the resistance of either parent, were obtained from test cross progeny of several Rp1 heterozygotes. These susceptible progeny usually had non-parental genotypes at flanking marker loci, thereby verifying their recombinational origin. Seven of eight Rp1 alleles (or genes) studied were clustered within about 0.2 map units of each other. Rpl ^G, however, mapped from 1–3 map units distal to other Rp1 alleles. Rp5 also mapped distally to most Rp1 alleles. Other aspects of recombination at Rp1 suggested that some alleles carry duplicated sequences, that mispairing can occur, and that unequal crossing-over may be a common phenomenon in this region; susceptible progeny from an Rp1 ^A homozygote had recombinant flanking marker genotypes, and susceptible progeny from an Rp1 ^DlRp1 ^F heterozygote showed both possible nonparental flanking marker genotypes.     

Co-localisation of host plant resistance QTLs affecting the performance and feeding behaviour of the aphid Myzus persicae in the peach tree

The architecture and action of quantitative trait loci (QTL) contributing to plant resistance mechanisms against aphids, the largest group of phloem-feeding insects, are not well understood. Comparative mapping of several components of resistance to the green peach aphid (Myzus persicae) was undertaken in Prunus davidiana, a wild species related to peach. An interspecific F(1) population of Prunus persica var. Summergrand × P. davidiana clone P1908 was scored for resistance (aphid colony development and foliar damage) and 17 aphid feeding behaviour traits monitored by means of the electrical penetration graph technique. Seven resistance QTLs were detected, individually explaining 6.1-43.1% of the phenotypic variation. Consistency was shown over several trials. Nine QTLs affecting aphid feeding behaviour were identified. All resistance QTLs except one co-located with QTLs underlying aphid feeding behaviour. A P. davidiana resistance allele at the major QTL was associated with drastic reductions in phloem sap ingestion by aphids, suggesting a phloem-based resistance mechanism. Resistance was also positively correlated with aphid salivation into sieve elements, suggesting an insect response to restore the appropriate conditions for ingestion after phloem occlusion. No significant QTL was found for traits characterising aphid mouthpart activity in plant tissues other than phloem vessels. Two QTLs with effects on aphid feeding behaviour but without effect on resistance were identified. SSR markers linked to the main QTLs involved in resistance are of potential use in marker-assisted selection for aphid resistance. Linking our results with the recent sequencing of the peach genome may help clarify the physiological resistance mechanisms.     

Physical mapping of plastid DNA variation among eleven Nicotiana species

Summary Plastid DNA of seven American and four Australian species of the genus Nicotiana was examined by restriction endonuclease analysis using the enzymes Sal I, Bgl I, Pst I, Kpn I, Xho I, Pvu II and Eco RI. These endonucleases collectively distinguish more than 120 sites on N. tabacum plastid DNA. The DNAs of all ten species exhibited restriction patterns distinguishable from those of N. tabacum for at least one of the enzymes used. All distinctive sites were physically mapped taking advantage of the restriction cleavage site map available for plastid DNA from Nicotiana tabacum (Seyer et al. 1981). This map was extended for the restriction endonucleases Pst I and Kpn I. In spite of variation in detail, the overall fragment order was found to be the same for plastid DNA from the eleven Nicotiana species. Most of the DNA changes resulted from small insertions/deletions and, possibly, inversions. They are located within seven regions scattered along the plastid chromosome. The divergence pattern of the Nicotiana plastid chromosomes was strikingly similar to that found in the genus Oenothera subsection Euoenothera (Gordon et al. 1982). The possible role of replication as a factor in the evolution of divergence patterns is discussed. The restriction patterns of plastid DNA from species within a continent resembled each other with one exception in each instance. The American species N. repanda showed patterns similar to those of most Australian species, and those of the Australian species N. debneyi resembled those of most American species.Abbreviations ims isonuclear male sterile - ptDNA plastid chloroplast DNA - Rubisco ribulosebisphosphate carboxylase/oxygenase - kbp kilobase pairs - LSU large subunit of Rubisco     

Mapping of ben genes of Pseudomonas aeruginosa

Abstract Four ben genes responsible for the conversion of benzoate to catechol in Pseudomonas aeruginosa PAO have been mapped to a 4.6 kb Kpn I fragment. ben -1 and ben -4 were known to be separate genes but now ben-1508 has been found to be different from ben-2 . The two genes were distinguished by Tn 5 mutagenesis of a cosmid clone and deletion mapping. It is likely that the four genes mapped ( ben-4, ben-2, ben-1508 and ben-1 ) correspond to the previously characterized benR (regulatory gene) and benABC (benzoate dioxygenase) respectively.     

用放射杂交板定位鸡的MC4R基因以及其在鸡和人染色体上同源区的比较分析

黑素皮质素受体(melanocortin-4 receptor,MC4R)基因的突变与猪、鼠和人等的食欲、肥胖和生长有关联性,然而对鸡的MC4R基因的功能却知之甚少。为了确定鸡的MC4R基因在染色体上的位置,使用鸡-仓鼠杂交板(ChickRH6)做了该基因的定位工作。通过扩增ChickRH6杂交板上的93个样品,然后经整合分析将mC4R基因定位在2号染色体上的标记MCW0062、BCL2和OVY附近,即2q12。这个连锁图上的5个标记基于两点分析与MC4R的LOD值都大于5。同时,以MC4R基因为标记做了鸡和人的染色体比较分析。结果显示鸡的2号染色体(GGA2)和人的18号染色体(HSA18)存在同源区,且基因BCL2和肥胖基因(obesity)位于MC4R基因附近。推测鸡的MC4R基因与人的MC4R基因可能具有相似的功能。该研究揭示了鸡和人MC4R基因的染色体分布,并用杂交放射板将鸡的MC4R基因定位在2号染色体的12区带。