SEQUENTIAL SAMPLING AND MICROSATELLITES FOR EVALUATION OF THE GENETIC QUALITY IN CORN SEEDS LOTS

In the seed production system, genetic purity is one of the fundamental requirements for its commercialization. The present work had the goal of determined the sample size for genetic purity evaluation, in order to protect the seed consumer and the producer and to evaluate the sensitivity of microsatellite technique for discriminating hybrids from their respective relatives and for detecting mixtures when they are present in small amounts in the samples. For the sequential sampling, hybrid seeds were marked and mixed in with the seed lots, simulating the following levels of contamination: 0.25, 0.5, 1.0, 2.0, 4.0, and 6.0%. After this, groups of 40 seeds were taken in sequence, up to a maximum of 400 seeds, with the objective of determining the quantity of seeds necessary to detect the percentage of mixture mentioned above. The sensitivity of microsatellite technique was evaluated by mixing different proportions of DNA from the hybrids with their respective seed lines. For the level of mixture was higher than 1:8 (1P1:8P2; 8P1:1P2), the sensitivity of the marker in detecting different proportions of the mixture varied according to the primer used. In terms of the sequential sampling, it was verified that in order to detect mixture levels higher than 1% within the seed lot- with a risk level for both the producer and the consumer of 0.05- the size of the necessary sample was smaller than the size needed for the fixed sample size. This also made it possible to reduce costs, making it possible to use microsatellites to certify the genetic purity of corn seeds lots.

SSR genetic diversity assessment of popular pigeonpea varieties in Malawi reveals unique fingerprints

Background: Pigeonpea ( Cajanus cajan L. Millsp.) is a drought tolerant legume of the Fabaceae family and the only cultivated species in the genus Cajanus. It is mainly cultivated in the semi-arid tropics of Asia and Oceania, Africa and America. In Malawi, it is grown as a source of food and income and for soil improvement in intercropping systems. However, varietal contamination due to natural outcrossing causes significant quality reduction and yield losses. In this study, 48 polymorphic SSR markers were used to assess the diversity among all pigeonpea varieties cultivated in Malawi to determine if a genetic fingerprint could be identified to distinguish the popular varieties. Results: A total of 212 alleles were observed with an average of 5.58 alleles per marker and a maximum of 14 alleles produced by CCttc019 (Marker 40). Polymorphic information content (PIC), ranged from 0.03 to 0.89 with an average of 0.30. A neighbor-joining tree produced 4 clusters. The most commonly cultivated varieties, which include released varieties and cultivated land races, were well-spread across all the clusters observed, indicating that they generally represented the genetic diversity available in Malawi, although substantial variation was evident that can still be exploited through further breeding. Conclusion: Screening of the allelic data associated with the five most popular cultivated varieties, revealed 6 markers – CCB1, CCB7, Ccac035, CCttc003, Ccac026 and CCttc019 – which displayed unique allelic profiles for each of the five varieties. This genetic fingerprint can potentially be applied for seed certification to confirm the genetic purity of seeds that are delivered to Malawi farmers.

Assessing the taxonomic status of dingoes Canis familiaris for conservation

1. The conservation status of the dingo Canis familiaris dingo is threatened by hybridization with the domestic dog C. familiaris familiaris. A practical method that can estimate the different levels of hybridization in the field is urgently required so that animals below a specific threshold of dingo ancestry (e.g. 1/4 or 1/2 dingoes) can reliably be identified and removed from dingo populations. 2. Skull morphology has been traditionally used to assess dingo purity, but this method does not discriminate between the different levels of dingo ancestry in hybrids. Furthermore, measurements can only be reliably taken from the skulls of dead animals. 3. Methods based on the analysis of variation in DNA are able to discriminate between the different levels of hybridization, but the validity of this method has been questioned because the materials currently used as a reference for dingoes are from captive animals of unproven genetic purity. The use of pre-European materials would improve the accuracy of this method, but suitable material has not been found in sufficient quantity to develop a reliable reference population. Furthermore, current methods based on DNA are impractical for the field-based discrimination of hybrids because samples require laboratory analysis. 4. Coat colour has also been used to estimate the extent of hybridization and is possibly the most practical method to apply in the field. However, this method may not be as powerful as genetic or morphological analyses because some hybrids (e.g. Australian cattle dog × dingo) are similar to dingoes in coat colour and body form. This problem may be alleviated by using additional visual characteristics such as the presence/absence of ticking and white markings.

Isoenzimas na diferenciação de sementes de três espécies do gênero Euterpe

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Coincidence of flowering time and the productivity and quality of cauliflower hybrid seeds

A falta de sincronismo de florescimento entre as linhagens auto incompatíveis em um campo de produção de sementes híbridas de couve flor pode além de reduzir a produção de sementes comprometer a pureza genética das mesmas. Com o objetivo de estudar o efeito da coincidência de florescimento entre linhagens de couve-flor na produtividade e qualidade de sementes híbridas, foi realizado o presente experimento. Os tratamentos consistiram em seis diferentes épocas de semeadura, espaçadas a cada quinze dias, de uma linhagem de verão auto-incompatível que foi polinizada por uma linhagem de inverno que não apresenta auto-incompatibilidade. Observou-se a coincidência do florescimento das diferentes épocas de semeadura com a linhagem polinizadora. Foram avaliadas as seguintes características: área foliar média, número de flores por planta, número de síliqüas por planta, número de sementes por planta (peso e número), peso médio de 1000 sementes e foi determinado o número de sementes por síliqüa. Foi realizado ainda, o teste padrão de germinação e determinada a pureza genética das sementes para cada tratamento. A coincidência da época de florescimento entre as linhagens de couve-flor afetou diretamente a produtividade e a qualidade genética das sementes híbridas produzidas, sendo que, quanto maior foi o nível de coincidência, maior foi o número de sementes formadas por síliqüa e menor a percentagem de sementes contaminantes. Entretanto, não teve influência na qualidade fisiológica das mesmas.

Use of EP(Peroxidase) allele in soybean varietal characterization

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Assessing the taxonomic status of dingoes Canis familiaris dingo for conservation

1. The conservation status of the dingo Canis familiaris dingo is threatened by hybridization with the domestic dog C. familiaris familiaris. A practical method that can estimate the different levels of hybridization in the field is urgently required so that animals below a specific threshold of dingo ancestry (e.g. 1/4 or 1/2 dingoes) can reliably be identified and removed from dingo populations. 2. Skull morphology has been traditionally used to assess dingo purity, but this method does not discriminate between the different levels of dingo ancestry in hybrids. Furthermore, measurements can only be reliably taken from the skulls of dead animals. 3. Methods based on the analysis of variation in DNA are able to discriminate between the different levels of hybridization, but the validity of this method has been questioned because the materials currently used as a reference for dingoes are from captive animals of unproven genetic purity. The use of pre-European materials would improve the accuracy of this method, but suitable material has not been found in sufficient quantity to develop a reliable reference population. Furthermore, current methods based on DNA are impractical for the field-based discrimination of hybrids because samples require laboratory analysis. 4. Coat colour has also been used to estimate the extent of hybridization and is possibly the most practical method to apply in the field. However, this method may not be as powerful as genetic or morphological analyses because some hybrids (e.g. Australian cattle dog x dingo) are similar to dingoes in coat colour and body form. This problem may be alleviated by using additional visual characteristics such as the presence/absence of ticking and white markings.

An evaluation of genetic analyses, skull morphology and visual appearance for assessing dingo purity: implications for dingo conservation

The introgression of domestic dog genes into dingo populations threatens the genetic integrity of 'pure' dingoes. However, dingo conservation efforts are hampered by difficulties in distinguishing between dingoes and hybrids in the field. This study evaluates consistency in the status of hybridisation (i.e. dingo, hybrid or dog) assigned by genetic analyses, skull morphology and visual assessments. Of the 56 south-east Queensland animals sampled, 39 (69.6%) were assigned the same status by all three methods, 10 (17.9%) by genetic and skull methods, four (7.1%) by genetic and visual methods; and two (3.6%) by skull and visual methods. Pair-wise comparisons identified a significant relationship between genetic and skull methods, but not between either of these and visual methods. Results from surveying 13 experienced wild dog managers showed that hybrids were more easily identified by visual characters than were dingoes. A more reliable visual assessment can be developed through determining the relationship between (1) genetics and phenotype by sampling wild dog populations and (2) the expression of visual characteristics from different proportions and breeds of domestic dog genes by breeding trials. Culling obvious hybrids based on visual characteristics, such as sable and patchy coat colours, should slow the process of hybridisation.

Genetic changes, intra- and inter-specific introgression in farmed Nile tilapia (Oreochromis niloticus) in Thailand

Fourteen microsatellite loci were used to examine genetic changes of four strains in Nile tilapia (Oreochromis niloticus) derived from genetically improved farmed tilapia (GIFT) and two strains derived from a local Chitralada strain of Nile tilapia in Thailand. Reference populations, including the ninth generation of GIFT strain, the original Chitralada strain, two conspecific reference populations from Ivory Coast and Uganda, and one population each of Oreochromis mossambicus and Oreochromis aureus, were also examined. Despite minor genetic changes, three of the four GIFT-derived populations retained their purity as GIFT while genetic variation did not decline. One of the GIFT-derived populations showed high levels of introgression from the Chitralada strain. Likewise, introgression from GIFT to the Chitralada-derived populations was seen. Inter-specific introgression from O. mossambicus was observed in the GIFT reference population and one of the Chitralada-derived strains. Introgression from O. aureus was detected in one of the GIFT-derived populations with a history of intensive inter-strain crossing. However, the introgression resulted in elevated genetic variation relative to the Chitralada original strains.