Dispersal of Transgenes through Maize Seed Systems in Mexico

Objectives

Current models of transgene dispersal focus on gene flow via pollen while neglecting seed, a vital vehicle for gene flow in centers of crop origin and diversity. We analyze the dispersal of maize transgenes via seeds in Mexico, the crop''s cradle.

Methods

We use immunoassays (ELISA) to screen for the activity of recombinant proteins in a nationwide sample of farmer seed stocks. We estimate critical parameters of seed population dynamics using household survey data and combine these estimates with analytical results to examine presumed sources and mechanisms of dispersal.

Results

Recombinant proteins Cry1Ab/Ac and CP4/EPSPS were found in 3.1% and 1.8% of samples, respectively. They are most abundant in southeast Mexico but also present in the west-central region. Diffusion of seed and grain imported from the United States might explain the frequency and distribution of transgenes in west-central Mexico but not in the southeast.

Conclusions

Understanding the potential for transgene survival and dispersal should help design methods to regulate the diffusion of germplasm into local seed stocks. Further research is needed on the interactions between formal and informal seed systems and grain markets in centers of crop origin and diversification.     

Habitat Deterioration, Habitat Destruction, and Metapopulation Persistence in a Heterogenous Landscape

Levins's unstructured metapopulation model predicts that the equilibrium fraction of empty habitat patches is a constant function of the fractionhof suitable patches in the landscape and that this constant equals the threshold value for metapopulation persistence. Levins's model thus suggests that the minimum amount of suitable habitat necessary for metapopulation persistence can be estimated from the fraction of empty patches at steady state. In this paper we construct several more realistic structured metapopulation models that include variation in patch quality and the rescue effect. These models predict both positive and negative correlations between the fractions of suitable patches and empty patches. The type of correlation depends in an intricate manner on the strength of the rescue effect and on the quality distribution of the patches to be destroyed. Empty patches can be considered as the resource limiting metapopulation growth. Our results demonstrate that the correlation between the fractions of suitable patches and empty patches is positive if and only if the average value of the resource decreases as the number of patches increases.     

Evolution of the Genes Encoding Seed Storage Proteins in Sugarcane and Maize

Prolamins, the seed storage proteins of maize, sorghum and coix were also found in sugarcane. Prolamins are grouped into structurally distinct classes termed the α-, β-, γ- and δ-prolamins. Orthologues for almost all of the α-, β-, γ- and δ-prolamins classes were identified in sugarcane. In maize, there are two molecular weight classes of α-prolamins, the 22 and 19 kD α-zeins. Sugarcane also possesses both the 22 kD and the 19 kD α-prolamins, which we denote as caneins, whereas sorghum and coix contain only the 22 kD α-prolamin (α-kafirin and α-coixin, respectively). Amino acid sequence alignments of the 22 and 19 kD α-prolamins from these plants revealed that both the 19 kD α-zein and the 19 kD α-canein lack around 20 amino acids at the sixth α-helix domain. We postulate that the 19 kD α-prolamins originated from a deletion of the sixth α-helix of a 22 kD counterpart in the saccharum lineage. Saccharum and sorghum diverged around five to nine million years ago (Mya), when only the 22 kD α-prolamins existed. The 19 kD α-canein must therefore have emerged after this time. Sorghum possesses a 19 kD α-prolamin similar to that of sugarcane and maize, but it contains the sixth α-helix domain lacking in the 19 kD α-zein and the 19 kD α-canein. This sorghum α-prolamin that we called 19 kD-like α-kafirin must be the ancestor of the 19 kD α-canein. The 19 kD-like α-kafirin could also be the ancestor of the 19 kD α-zein but it is also possible that the genes encoding the 19 kD α-zein and the 19 kD α-canein have evolved separately in these close groups.     

两捕食-食饵自治扩散系统的持续生存

本文得到了自治Lotka-Voltrerra两捕食-食饵扩散系统持续生存及捕者灭绝的条件.     

Insights into Persistence Mechanisms of a Zoonotic Virus in Bat Colonies Using a Multispecies Metapopulation Model

Rabies is a worldwide zoonosis resulting from Lyssavirus infection. In Europe, Eptesicus serotinus is the most frequently reported bat species infected with Lyssavirus, and thus considered to be the reservoir of European bat Lyssavirus type 1 (EBLV-1). To date, the role of other bat species in EBLV-1 epidemiology and persistence remains unknown. Here, we built an EBLV-1−transmission model based on local observations of a three-cave and four-bat species (Myotis capaccinii, Myotis myotis, Miniopterus schreibersii, Rhinolophus ferrumequinum) system in the Balearic Islands, for which a 1995–2011 serological dataset indicated the continuous presence of EBLV-1. Eptesicus serotinus was never observed in the system during the 16-year follow-up and therefore was not included in the model. We used the model to explore virus persistence mechanisms and to assess the importance of each bat species in the transmission dynamics. We found that EBLV-1 could not be sustained if transmission between M. schreibersii and other bat species was eliminated, suggesting that this species serves as a regional reservoir. Global sensitivity analysis using Sobol''s method revealed that following the rate of autumn−winter infectious contacts, M. schreibersii''s incubation- and immune-period durations, but not the infectious period length, were the most relevant factors driving virus persistence.     

Traditional Maize Storage Methods of Mayan Farmers in Yucatan, Mexico: Implications for Seed Selection and Crop Diversity

For small-scale farmers who maintain genetically diverse crop populations, aspects of the storage of their seeds and harvest may be just as important for successful farming as those related to productivity. The community of Yaxcaba, Yucatan, Mexico was studied to understand how the conditions under which Maya farmers store their maize harvests influence their seed selection practices and the diversity of maize varieties grown. Most farmers select their maize seed based primarily on ear characteristics and secondarily on grain characters. Farmers incorporate storability concerns by selecting for an archetypal healthy ear and by conducting selection in several steps between harvest and planting of the subsequent crop generation. Most farmers store their maize harvest in the husk, initially in the field and then in rustic granaries constructed of logs and palm thatch, in which farmers conserve separately their different seed lots. All local landrace populations show morphological adaptation (principally husk characteristics) for local storage conditions, indicating that storability has been an important selection pressure on traditional maize. Storability also appears to be a key factor working against the straightforward adoption of improved maize seed in Yaxcaba. Local farmers value many qualities of improved maize varieties, but their principal complaint is that improved maize cannot be conserved reliably under local storage conditions. Nearly all farmers who utilize improved seed stock in Yaxcaba grow locally adapted or ‘creolized’ versions of improved varieties, displaying characteristics of local maize landraces that facilitate their storage.     

Detection of Transgenes in Local Maize Varieties of Small-Scale Farmers in Eastern Cape,South Africa

Small-scale subsistence farmers in South Africa have been introduced to genetically modified (GM) crops for more than a decade. Little is known about i) the extent of transgene introgression into locally recycled seed, ii) what short and long-term ecological and socioeconomic impacts such mixing of seeds might have, iii) how the farmers perceive GM crops, and iv) to what degree approval conditions are followed and controlled. This study conducted in the Eastern Cape, South Africa, aims primarily at addressing the first of these issues. We analysed for transgenes in 796 individual maize plants (leaves) and 20 seed batches collected in a village where GM insect resistant maize was previously promoted and grown as part of an governmental agricultural development program over a seven year period (2001–2008). Additionally, we surveyed the varieties of maize grown and the farmers’ practices of recycling and sharing of seed in the same community (26 farmers were interviewed). Recycling and sharing of seeds were common in the community and may contribute to spread and persistence of transgenes in maize on a local or regional level. By analysing DNA we found that the commonly used transgene promoter p35s occurred in one of the 796 leaf samples (0.0013%) and in five of the 20 seed samples (25%). Three of the 20 seed samples (15%) included herbicide tolerant maize (NK603) intentionally grown by the farmers from seed bought from local seed retailers or acquired through a currently running agricultural development program. The two remaining positive seed samples (10%) included genes for insect resistance (from MON810). In both cases the farmers were unaware of the transgenes present. In conclusion, we demonstrate that transgenes are mixed into seed storages of small-scale farming communities where recycling and sharing of seeds are common, i.e. spread beyond the control of the formal seed system.     

Metapopulation Dynamics Enable Persistence of Influenza A,Including A/H5N1, in Poultry

Highly pathogenic influenza A/H5N1 has persistently but sporadically caused human illness and death since 1997. Yet it is still unclear how this pathogen is able to persist globally. While wild birds seem to be a genetic reservoir for influenza A, they do not seem to be the main source of human illness. Here, we highlight the role that domestic poultry may play in maintaining A/H5N1 globally, using theoretical models of spatial population structure in poultry populations. We find that a metapopulation of moderately sized poultry flocks can sustain the pathogen in a finite poultry population for over two years. Our results suggest that it is possible that moderately intensive backyard farms could sustain the pathogen indefinitely in real systems. This fits a pattern that has been observed from many empirical systems. Rather than just employing standard culling procedures to control the disease, our model suggests ways that poultry production systems may be modified.     

Low Rate of Between-Population Seed Dispersal Restricts Genetic Connectivity and Metapopulation Dynamics in a Clonal Shrub

Clonal species normally have low seed production, low recruitment rates and long lifespans, and it is expected that the rates of long-distance dispersal (LDD) of seeds will be low as well. Banksia candolleana is a clonal shrub in Mediterranean-type, fire-prone sclerophyll shrublands of southwestern Australia, whose reproductive biology and population dynamics contrast with those of co-occurring nonclonal congeneric species, all of which are restricted to a mosaic of sand dunes set within a matrix of inhospitable swales. Using microsatellite markers, we genotyped 499 plants in all 15 populations of B. candolleana within a 12-km² area, assessed population genetic differentiation, and quantified the effective rate of interpopulation seed dispersal through genetic assignment of individuals to populations. We measured life history, reproductive and demographic attributes, and compared these with two co-occurring Banksia species, a non-clonal resprouter and a nonsprouter. B. candolleana has much higher levels of population genetic differentiation, and one-third the rate of interpopulation seed migration, as the other two species (2.2% vs 5.5−6.8% of genotyped plants inferred to be immigrants), though distances reached by LDD are comparable (0.3−2.3 km). The low rate of interpopulation dispersal was supported by an analysis of the age structure of three populations that suggests a mean interdune migration rate of <800 m in 200 years, and 60% of suitable dunes remain uninhabited. Thus, B. candolleana has poor properties for promoting long-distance dispersal. It is unclear if these are idiosyncratic to this species or whether such properties are to be expected of clonal species in general where LDD is less critical for species survival.     

Occurrence and Persistence of Erythromycin Resistance Genes (erm) and Tetracycline Resistance Genes (tet) in Waste Treatment Systems on Swine Farms

Animal manure from modern animal agriculture constitutes the single largest source of antibiotic resistance (AR) owing to the use of large quantities of antibiotics. After animal manure enters the environment, the AR disseminates into the environment and can pose a potentially serious threat to the health and well-being of both humans and animals. In this study, we evaluated the efficiency of three different on-farm waste treatment systems in reducing AR. Three classes of erythromycin resistance genes (erm) genes (B, F, and X) conferring resistances to macrolide–lincosamides–streptogramin B (MLS_B) and one class of tetracycline resistance genes (tet) gene (G) conferring resistance to tetracyclines were used as models. Real-time polymerase chain reaction assays were used to determine the reservoir sizes of these AR genes present in the entire microbiome. These classes of AR genes varied considerably in abundance, with erm(B) being more predominant than erm(F), erm(X), and tet(G). These AR genes also varied in persistence in different waste treatment systems. Aerobic biofiltration reduced erm(X) more effectively than other AR genes, while mesophilic anaerobic digestion and lagoon storage did not appreciably reduce any of these AR genes. Unlike chemical pollutants, some AR genes could increase after reduction in a preceding stage of the treatment processes. Season might also affect the persistence of AR. These results indicate that AR arising from swine-feeding operations can survive typical swine waste treatment processes and thus treatments that are more effective in destructing AR on farms are required.     

Porcine E. coli: Virulence-Associated Genes,Resistance Genes and Adhesion and Probiotic Activity Tested by a New Screening Method

We established an automated screening method to characterize adhesion of Escherichia coli to intestinal porcine epithelial cells (IPEC-J2) and their probiotic activity against infection by enteropathogenic E. coli (EPEC). 104 intestinal E. coli isolates from domestic pigs were tested by PCR for the occurrence of virulence-associated genes, genes coding for resistances to antimicrobial agents and metals, and for phylogenetic origin by PCR. Adhesion rates and probiotic activity were examined for correlation with the presence of these genes. Finally, data were compared with those from 93 E. coli isolates from wild boars.Isolates from domestic pigs carried a broad variety of all tested genes and showed great diversity in gene patterns. Adhesions varied with a maximum of 18.3 or 24.2 mean bacteria adherence per epithelial cell after 2 or 6 hours respectively. Most isolates from domestic pigs and wild boars showed low adherence, with no correlation between adhesion/probiotic activity and E. coli genes or gene clusters. The gene sfa/foc, encoding for a subunit of F1C fimbriae did show a positive correlative association with adherence and probiotic activity; however E. coli isolates from wild boars with the sfa/foc gene showed less adhesion and probiotic activity than E. coli with the sfa/foc gene isolated from domestic pigs after 6 hour incubation.In conclusion, screening porcine E. coli for virulence associated genes genes, adhesion to intestinal epithelial cells, and probiotic activity revealed a single important adhesion factor, several probiotic candidates, and showed important differences between E. coli of domestic pigs and wild boars.     

Old Crop,New Society: Persistence and Change of Tartary Buckwheat Farming in Yunnan,China

Traditional agricultural systems are important for crop genetic resources conservation. Many scholars have addressed the problem of traditional cultivar replacement by modern varieties, but few have investigated the entire loss of traditional crops from farming systems. Our prior research suggested that tartary buckwheat (Fagopyrum tataricum (L.) Gaertn.) agriculture is rapidly decreasing in Yunnan China, the center of the crop’s origin and an important repository of on-farm genetic diversity. Using interdisciplinary methodology to determine whether the crop is indeed in decline and evaluate reasons behind planting trends, we found a combination of interacting agroecological, socio-cultural, and institutional factors influence tartary buckwheat planting in Yunnan. Farmer ethnicity, presence of a commercial market for the crop, and government agricultural policy were particularly important. Low commercial value of the crop compared to other alternatives, reduction of available farmland, and labor shortages were major reasons cited for crop abandonment. Despite an overall reduction in tartary buckwheat planting across a wide spectrum of farms, we also found the crop retains subsistence, rotational and cultural value for many Yunnan farmers. We conclude that although Yunnan farms are increasingly managed commercially rather than for subsistence, tartary buckwheat will continue to occupy a niche in the agricultural landscape.     

玉米叶绿体atpB及atpE基因在大肠杆菌中的表达

本实验构建了在大肠杆菌中能表达玉米叶绿体ＣＦ１完整β亚基、缺失Ｎ端２３个氨基酸残基的β亚基以及完整ε亚基的表达质粒。ＳＤＳ－聚丙烯酰胺凝胶电泳及Ｗｅｓｔｅｒｎｂｌｏｔ实验表明,在分别含这些表达质粒的细菌中β、ε亚基蛋白的合成量在诱导３ｈ后即达到饱和,其含量各占细菌体总蛋白的１０％左右。这些表达产物在细菌体中形成不溶性的包含体。可通过对菌体蛋白的分级分高,将表达产物溶于５ｍｏｌ／Ｌ的尿素溶液中。