RAPD profiling in detecting genetic variation in endemic Coelonema (Brassicaceae) of Qinghai-Tibet Plateau of China

Random amplified polymorphic DNA ( RAPD) markers were used to measure genetic diversity of Coelonema draboides ( Brassicaceae), a genus endemic to the Qilian Mountains of the Qinghai-Tibet Plateau. We sampled 90 individuals in 30 populations of Coelonema draboides from Datong and Huzhu counties of Qinghai Province in P. R. China. A total of 186 amplified bands were scored from the 14 RAPD primers, with a mean of 13.3 amplified bands per primer, and 87% ( 161 bands) polymorphic bands (PPB) was found. Analysis of molecular variance (AMOVA) shows that a large proportion of genetic variation (84.2%) resides among individuals within populations, while only 15.8% resides among populations. The species shows higher genetic diversity between individuals than other endemic and endangered plants. The RAPDs provide a useful tool for assessing genetic diversity of rare, endemic species and for resolving relationships among populations. The results show that the genetic diversity of this species is high, possibly allowing it to adapt more easily to environmental variations. The main factor responsible for the high level of differentiation within populations and the low level of diversity among populations is probably the outcrossing and long-lived nature of this species. Some long-distance dispersal, even among far separated populations, is also a crucial determinant for the pattern of genetic variation in the species. This distributive pattern of genetic variation of C. draboides populations provides important baseline data for conservation and collection strategies for the species. It is suggested that only populations in different habitats should be studied and protected, not all populations, so as to retain as much genetic diversity as possible.

Variabilidade genética em acessos e cultivares de quatro espécies de Brachiaria estimada por marcadores RAPD.

A importância do gênero Brachiaria para o desenvolvimento da pecuária nas regiões tropicais e subtropicais é notória. Brachiaria brizantha, Brachiaria decumbens, Brachiaria ruziziensis e Brachiaria humidicola estão entre as principais espécies utilizadas como forrageiras e fazem parte do programa de melhoramento da Embrapa Gado de Corte. Neste trabalho, a técnica de Random Amplified Polymorphic DNA (RAPD) foi utilizada para estimar a variabilidade genética em 14 genótipos dessas quatro espécies, incluindo cultivares e acessos utilizados como genitores em cruzamentos inter e intra-específicos. Foram utilizados 47 primers que amplificaram 396 bandas, escoradas como "1" para presença e "0" para ausência, e uma matriz de similaridade genética foi construída usando o coeficiente de Jaccard. Os valores de similaridade genética variaram de 0,49 a 0,87, dos quais os acessos mais similares (C48 e B105) pertencem à mesma espécie, B. brizantha. As cultivares B. brizantha cv. Marandu e B. humidicola cv. BRS Tupi foram as mais divergentes. As análises de agrupamento obtidas pelos métodos Unweighted Pair-group Method with Arithmetical Average (UPGMA) e de Tocher foram similares, dividindo os acessos em quatro grupos que correspondem às espécies estudadas. O dendrograma obtido com o método UPGMA mostrou que B. brizantha, B. decumbens e B. ruziziensis são geneticamente mais próximas entre si do que B. humidicola. Além disso, este trabalho possibilitou a identificação de primers que podem ser utilizados com a finalidade de identificação precoce de híbridos quando esses genótipos forem utilizados como genitores.

A biotecnologia nos programas de melhoramento de forrageiras tropicais da Embrapa Gado de Corte.

Neste trabalho procurou-se apresentar e discutir, de forma ampla, o uso da biotecnologia e seu potencial para os programas de melhoramento de forrageiras tropicais realizados na Embrapa Gado de Corte. O uso da biotecnologia nesses programas é uma atividade recente, mas importantes resultados vêm sendo gerados a fim de auxiliar o processo de obtenção de novas cultivares forrageiras. A maioria dos trabalhos apresentados utiliza marcadores Random Amplification of Polymorphic DNA (RAPD) para aplicações em curto prazo: estudos de diversidade em acessos de bancos de germoplasma, identificação de híbridos e estimação da taxa de cruzamento. Aplicações em médio e longo prazos do uso de marcadores, como mapeamento genético e seleção auxiliada por marcadores moleculares (SAMM), ainda necessitam de maiores investimentos, tanto na busca de novos marcadores, quanto no desenvolvimento de populações adequadas para esses estudos. Em 2007, teve início uma nova linha de pesquisa nessa unidade, a prospecção de genes com características econômicas. Genes para a tolerância ao alumínio são o foco dessa pesquisa que pretende explorar a sintenia entre os genomas de gramíneas, visando ao desenvolvimento de cultivares de braquiária mais tolerantes ao alumínio. A Embrapa Gado de Corte vem investindo em pessoal e aquisição de equipamentos para avançar não só na produção de cultivares de forrageiras mais adaptadas às necessidades de um mercado cada vez mais exigente, como também no crescimento institucional do setor de biotecnologia.

Type-omega DPLs

Type-omega DPLs (Denotational Proof Languages) are languages for proof presentation and search that offer strong soundness guarantees. LCF-type systems such as HOL offer similar guarantees, but their soundness relies heavily on static type systems. By contrast, DPLs ensure soundness dynamically, through their evaluation semantics; no type system is necessary. This is possible owing to a novel two-tier syntax that separates deductions from computations, and to the abstraction of assumption bases, which is factored into the semantics of the language and allows for sound evaluation. Every type-omega DPL properly contains a type-alpha DPL, which can be used to present proofs in a lucid and detailed form, exclusively in terms of primitive inference rules. Derived inference rules are expressed as user-defined methods, which are "proof recipes" that take arguments and dynamically perform appropriate deductions. Methods arise naturally via parametric abstraction over type-alpha proofs. In that light, the evaluation of a method call can be viewed as a computation that carries out a type-alpha deduction. The type-alpha proof "unwound" by such a method call is called the "certificate" of the call. Certificates can be checked by exceptionally simple type-alpha interpreters, and thus they are useful whenever we wish to minimize our trusted base. Methods are statically closed over lexical environments, but dynamically scoped over assumption bases. They can take other methods as arguments, they can iterate, and they can branch conditionally. These capabilities, in tandem with the bifurcated syntax of type-omega DPLs and their dynamic assumption-base semantics, allow the user to define methods in a style that is disciplined enough to ensure soundness yet fluid enough to permit succinct and perspicuous expression of arbitrarily sophisticated derived inference rules. We demonstrate every major feature of type-omega DPLs by defining and studying NDL-omega, a higher-order, lexically scoped, call-by-value type-omega DPL for classical zero-order natural deduction---a simple choice that allows us to focus on type-omega syntax and semantics rather than on the subtleties of the underlying logic. We start by illustrating how type-alpha DPLs naturally lead to type-omega DPLs by way of abstraction; present the formal syntax and semantics of NDL-omega; prove several results about it, including soundness; give numerous examples of methods; point out connections to the lambda-phi calculus, a very general framework for type-omega DPLs; introduce a notion of computational and deductive cost; define several instrumented interpreters for computing such costs and for generating certificates; explore the use of type-omega DPLs as general programming languages; show that DPLs do not have to be type-less by formulating a static Hindley-Milner polymorphic type system for NDL-omega; discuss some idiosyncrasies of type-omega DPLs such as the potential divergence of proof checking; and compare type-omega DPLs to other approaches to proof presentation and discovery. Finally, a complete implementation of NDL-omega in SML-NJ is given for users who want to run the examples and experiment with the language.

Avaliação inicial do RAPD como forma de acessar a variabilidade genética na espécie rica em linalol Croton cajuçara Benth. (sacaca).

O objetivo deste trabalho foi avaliar a utilidade dos métodos praticados no Laboratório de Biotecnologia Vegetal da Embrapa Amazônia Ocidental, incluindo o teste da reprodutividade das bandas, para acessar a variabilidade genética entre plantas de sacaca por meio de marcadores moleculares do tipo RAPD ("Random Amplified Polymorphic DNA").

Construction of a Festuca pratensis BAC library for map-based cloning in IFestulolium substitution lines

Iain S. Donnison, Donal M. O Sullivan, Ann Thomas, Peter Canter, Beverley Moore, Ian Armstead, Howard Thomas, Keith J. Edwards and Ian P. King (2005). Construction of a Festuca pratensis BAC library for map-based cloning in Festulolium substitution lines. Theoretical and Applied Genetics, 110 (5) pp.846-851 Sponsorship: BBSRC;BBSRC RAE2008

An association mapping approach to identify flowering time genes in natural populations of Lolium perenne (L.)

Sk?t, L., Humphreys, M. O., Armstead, I. P., Heywood, S., Sk?t, K. P., Sanderson, R., Thomas, I. D., Chorlton, K. H., & Sackville Hamilton, N. R. (2005). An association mapping approach to identify flowering time genes in natural populations of Lolium perenne (L.). Molecular Breeding, 15(3), 233-245. Sponsorship: BBSRC RAE2008

Typability and Type Checking in the Second-Order Λ-Calculus Are Equivalent and Undecidable (Preliminary Draft)

We consider the problems of typability[1] and type checking[2] in the Girard/Reynolds second-order polymorphic typed λ-calculus, for which we use the short name "System F" and which we use in the "Curry style" where types are assigned to pure λ -terms. These problems have been considered and proven to be decidable or undecidable for various restrictions and extensions of System F and other related systems, and lower-bound complexity results for System F have been achieved, but they have remained "embarrassing open problems"[3] for System F itself. We first prove that type checking in System F is undecidable by a reduction from semi-unification. We then prove typability in System F is undecidable by a reduction from type checking. Since the reverse reduction is already known, this implies the two problems are equivalent. The second reduction uses a novel method of constructing λ-terms such that in all type derivations, specific bound variables must always be assigned a specific type. Using this technique, we can require that specific subterms must be typable using a specific, fixed type assignment in order for the entire term to be typable at all. Any desired type assignment may be simulated. We develop this method, which we call "constants for free", for both the λK and λI calculi.

A Direct Algorithm for the Type Interference in the Rank 2 Fragment of the Second--Order λ-Calculus

We study the problem of type inference for a family of polymorphic type disciplines containing the power of Core-ML. This family comprises all levels of the stratification of the second-order lambda-calculus by "rank" of types. We show that typability is an undecidable problem at every rank k ≥ 3 of this stratification. While it was already known that typability is decidable at rank ≤ 2, no direct and easy-to-implement algorithm was available. To design such an algorithm, we develop a new notion of reduction and show how to use it to reduce the problem of typability at rank 2 to the problem of acyclic semi-unification. A by-product of our analysis is the publication of a simple solution procedure for acyclic semi-unification.

System F with Constraint Types

System F is a type system that can be seen as both a proof system for second-order propositional logic and as a polymorphic programming language. In this work we explore several extensions of System F by types which express subtyping constraints. These systems include terms which represent proofs of subtyping relationships between types. Given a proof that one type is a subtype of another, one may use a coercion term constructor to coerce terms from the first type to the second. The ability to manipulate type constraints as first-class entities gives these systems a lot of expressive power, including the ability to encode generalized algebraic data types and intensional type analysis. The main contributions of this work are in the formulation of constraint types and a proof of strong normalization for an extension of System F with constraint types.

What Are Polymorphically-Typed Ambients?

Abstract: The Ambient Calculus was developed by Cardelli and Gordon as a formal framework to study issues of mobility and migrant code. We consider an Ambient Calculus where ambients transport and exchange programs rather that just inert data. We propose different senses in which such a calculus can be said to be polymorphically typed, and design accordingly a polymorphic type system for it. Our type system assigns types to embedded programs and what we call behaviors to processes; a denotational semantics of behaviors is then proposed, here called trace semantics, underlying much of the remaining analysis. We state and prove a Subject Reduction property for our polymorphically typed calculus. Based on techniques borrowed from finite automata theory, type-checking of fully type-annotated processes is shown to be decidable; the time complexity of our decision procedure is exponential (this is a worst-case in theory, arguably not encountered in practice). Our polymorphically-typed calculus is a conservative extension of the typed Ambient Calculus originally proposed by Cardelli and Gordon.

A Formal Semantics for Weak References

A weak reference is a reference to an object that is not followed by the pointer tracer when garbage collection is called. That is, a weak reference cannot prevent the object it references from being garbage collected. Weak references remain a troublesome programming feature largely because there is not an accepted, precise semantics that describes their behavior (in fact, we are not aware of any formalization of their semantics). The trouble is that weak references allow reachable objects to be garbage collected, therefore allowing garbage collection to influence the result of a program. Despite this difficulty, weak references continue to be used in practice for reasons related to efficient storage management, and are included in many popular programming languages (Standard ML, Haskell, OCaml, and Java). We give a formal semantics for a calculus called λweak that includes weak references and is derived from Morrisett, Felleisen, and Harper’s λgc. λgc formalizes the notion of garbage collection by means of a rewrite rule. Such a formalization is required to precisely characterize the semantics of weak references. However, the inclusion of a garbage-collection rewrite-rule in a language with weak references introduces non-deterministic evaluation, even if the parameter-passing mechanism is deterministic (call-by-value in our case). This raises the question of confluence for our rewrite system. We discuss natural restrictions under which our rewrite system is confluent, thus guaranteeing uniqueness of program result. We define conditions that allow other garbage collection algorithms to co-exist with our semantics of weak references. We also introduce a polymorphic type system to prove the absence of erroneous program behavior (i.e., the absence of “stuck evaluation”) and a corresponding type inference algorithm. We prove the type system sound and the inference algorithm sound and complete.

Optimised crystal morphologies for active pharmaceutical ingredients and related studies

The majority of active pharmaceutical ingredients (APIs) are crystalline solids in their pure forms. Crystalline solids have definable morphologies, i.e. shape and size. Crystal morphology is determined by both the internal structure of the crystals and external factors during growth from solution. The morphology of a crystal batch can affect key processes during manufacturing. Companies generally accept whatever morphology the manufacturing process provides and deal with any subsequent problems by costly trouble‒shooting. Rational design of optimised morphologies for crystalline pharmaceutical solids would be a very significant technical and commercial advance. Chapter one introduces the concept of crystal nucleation and growth. The phenomenon of polymorphism alongside the causes and impact is discussed. A summary of the scope of instrumentation used in the investigation of crystal polymorphism and morphology, including crystal size distribution (CSD), is also included. Chapter two examines the research carried out during an exploration of the optimum crystallisation parameters of phenacetin. Following a morphological study, the impact this induces on particle density and flow properties is examined. The impact of impurities on the crystallisation properties of phenacetin is investigated. Significantly, the location of impurities within individual crystals is also studied. The third chapter describes an industrial collaboration looking at the resolution and polymorphic study of trometamol and lysine salts of ketoprofen and 2‒phenylpropionic acid (2‒PPA). Chapter four incorporates a solid state study on three separate compounds: 2‒chloro‒4‒nitroaniline, 4‒hydroxy‒N‒phenylbenzenesulfonamide and N‒acetyl‒D‒glucosamine‒6‒O‒sulfate. 2‒Chloro‒4‒nitroaniline and 4‒hydroxy‒N‒phenylbenzenesulfonamide both produced interesting, extreme morphologies which warranted further investigation as part of a collaborative study. Following a summarisation of results in chapter five, chapter six contains the full experimental details, incorporating spectral and other analytical data for all compounds synthesised during the course of the research.

Interstitial insertion of varying amounts of ABL-containing genetic material into chromosome 22 in Ph-negative CML.

We studied the cells from three selected patients with Ph-chromosome-negative chronic myeloid leukemia (CML) by Southern blotting, polymerase chain reaction, and in situ hybridization of informative probes to metaphase chromosomes. All three patients had rearrangement of M-BCR sequences in the BCR gene and expression of one or other of the mRNA species characteristic of Ph-positive CML. Leukemic metaphases studied after trypsin-Giemsa banding were indistinguishable from normal. The ABL probe localized both to chromosome 9 and 22 in each case. A probe containing 3' M-BCR sequences localized only to chromosome 22, and not to chromosome 9 as would be expected in Ph-positive CML. Two new probes that recognize different polymorphic regions distal to the ABL gene on chromosome 9 in normal subjects localized exclusively to chromosome 9 in two patients and to both chromosomes 9 and 22 in one patient. These results show that Ph-negative CML with BCR rearrangement is associated with insertion of a variable quantity of chromosome 9 derived material into chromosome 22q11; there is no evidence for reciprocal translocation of material from chromosome 22 to chromosome 9.

Partitioning of reproduction among queens in the Argentine ant, Linepithema humile

We investigated how queens share parentage (skew) in the Argentine ant, Linepithema humile, a social insect with multiple queens (polygyny). Overall, maternity of 546 male and female sexuals that mated successfully was determined with microsatellites in 26 colonies consisting of two queens and workers. The first main finding was that queens all contributed to sexual production. However, there was a significant departure from equal contribution to male and female sexual production in a notable proportion of colonies. Overall, reproductive skew for sexual (male and female) production was relatively low but higher than reproductive skew for egg production. The second interesting result was that there was a trade-off in the relative contribution of queens to male and female production. The queens contributing more to male production contributed significantly less to female sexual production. Finally, there was no significant association between colony productivity and the degree of reproductive skew. The relatively low reproductive skew is in line with predictions of the so-called concession models of reproductive skew because, in the Argentine ant, relatedness between queens is low and ecological constraints on dispersal nonexistent or weak. © 2001 The Association for the Study of Animal Behaviour.