Patricia Lebrun

An analysis of genetic diversity in coconut (Cocos nucifera) populations from across the geographic range using sequence-tagged microsatellites (SSRs) and AFLPs.

Abstract Genetic diversity in 31 individuals from 14 coconut populations across the entire geographic range (2–3 individuals per population) was assessed using sequence-tagged microsatellites (or simple sequence repeats, SSRs) and amplified fragment length polymorphism (AFLP). From the 39 SSR primer sets tested, only two gave patterns that could not be scored and used in the data analysis. The remainder included five SSRs that gave double-locus profiles in which one locus could still be scored separately. The 37 SSRs revealed between 2 and 16 alleles per locus and a total of 339 alleles in the 14 populations. Gene diversity (D = 1-Σpi2) ranged from 0.47 to 0.90. Two of the four Dwarf populations were homozygous at all 37 loci, which is consistent with their autogamous (self-fertilising) reproduction. One Dwarf population was heterozygous at one locus but the other (Niu Leka Dwarf), which is known to be cross-pollinating, showed high levels of heterozygosity. Generally, diversity was higher in populations from the South Pacific and South East Asia. Three SSR loci (CNZ46, CN2A5, CN11E6) gave distinct genotypes for all but two populations. The East African populations had higher heterozygosities than those from West Africa, and the populations from Tonga and Fiji generally had distinct alleles from those of the South Pacific. AFLP analysis with 12 primer combinations gave a total of 1106 bands, of which 303 were polymorphic (27%). Similarity matrices were constructed from the two data sets using the proportion of shared alleles for SSRs and a Jaccard coefficient for AFLPs. In each case cluster and principal co-ordinates analyses were performed, with the resultant dendrograms and plots revealing similar relationships among the populations for both approaches. There was generally a good separation of populations, and phenetic relationships were in agreement with those previously shown by RFLPs. The use of SSRs and AFLPs in genetic-diversity analysis for the establishment of germplasm collections is discussed.

Genetic diversity among wild and cultivated populations of Hevea brasiliensis assessed by nuclear RFLP analysis

Restriction fragment length polymorphism was assessed in wild and cultivated populations of Hevea brasiliensis using random probes from an Hevea nuclear library. One-hundred-and-sixty-four individuals were surveyed, and the results discussed in the light of previous work performed on isozyme variation. Both studies show that germplasm collections have led to an effective enrichment of the genetic resources available for Hevea breeding, and that cultivated clones have conserved a relatively high level of polymorphism, despite their narrow genetic base and their high level of inbreeding. An equivalent level of polymorphism is revealed by random nuclear probes and isozymes. However, the genetic structuring of the diversity appears more striking using RFLP markers. Wild accessions can be divided into three genetic groups according to their geographical origin. The present results are an essential guide to the incorporation of wild material in breeding schemes.

DNA fingerprints in Hevea brasiliensis (rubber tree) using human minisatellite probes

The suitability of DNA fingerprints as genetic markers in Hevea brasiliensis was tested. Using the human minisatellite probe 33.6, the 73 cultivated (Wickham) clones studied exhibited individual-specific, reproducible and somatically stable hybridization patterns, with stably inherited fragments. This probe has been found to be a powerful new tool for clonal identification as an alternative to isozymatic markers.

The Panama Tall and the Maypan hybrid coconut in Jamaica: did genetic contamination cause a loss of resistance to Lethal Yellowing?

We applied Bayesian population assignment methods to assess the trueness to type of four populations of the coconut cultivar Panama Tall (PNT) located in Jamaica and found that two of them presented a high percentage of off-types, while genetic contamination was low in the two others. The PNT is the pollen parent of the MAYPAN hybrid, which used to be planted in Jamaica to control an epidemic disease: Lethal Yellowing. The main source of contamination was the susceptible Jamaica Tall, thus increasing the susceptibility in the resulting MAYPAN progeny. The incidence of genetic contamination seems however to be insufficient to be the only cause of the latest outbreak of the disease. Neither the MAYPAN nor its parents can be said resistant in the present context of Jamaica.

QTL analysis of fruit components in the progeny of a Rennell Island Tall coconut (Cocos nucifera L.) individual

We investigated the genetic factors controlling fruit components in coconut by performing QTL analyses for fruit component weights and ratios in a segregating progeny of a Rennell Island Tall genotype. The underlying linkage map of this population was already established in a previous study, as well as QTL analyses for fruit production, which were used to complement our results. The addition of 53 new markers (mainly SSRs) led to minor amendments in the map. A total of 52 putative QTLs were identified for the 11 traits under study. Thirty-four of them were grouped in six small clusters, which probably correspond to single pleiotropic genes. Some additional QTLs located apart from these clusters also had relatively large effects on the individual traits. The QTLs for fruit component weight, endosperm humidity and fruit production were found at different locations in the genome, suggesting that efficient marker-assisted selection for yield can be achieved by selecting QTLs for the individual components. The detected QTLs descend from a genotype belonging to the “Pacific” coconut group. Based on the known molecular and phenotypic differences between “Pacific” and “Indo-Atlantic” coconuts, we suggest that a large fraction of coconut genetic diversity is still to be investigated by studying populations derived from crosses between these groups.

Population structures of Brazilian tall coconut (Cocos nucifera L.) by microsatellite markers

Coconut palms of the Tall group were introduced to Brazil from the Cape Verde Islands in 1553. The present study sought to evaluate the genetic diversity among and within Brazilian Tall coconut populations. Samples were collected of 195 trees from 10 populations. Genetic diversity was accessed by investigating 13 simple sequence repeats (SSR) loci. This provided a total of 68 alleles, ranging from 2 to 13 alleles per locus, with an average of 5.23. The mean values of gene diversity (He ) and observed heterozygosity (Ho ) were 0.459 and 0.443, respectively. The genetic differentiation among populations was estimated at θ^P=0.1600and the estimated apparent outcrossing rate was ta = 0.92. Estimates of genetic distances between the populations varied from 0.034 to 0.390. Genetic distance and the corresponding clustering analysis indicate the formation of two groups. The first consists of the Baía Formosa, Georgino Avelino, and São José do Mipibu populations and the second consists of the Japoatã, Pacatuba, and Praia do Forte populations. The correlation matrix between genetic and geographic distances was positive and significant at a 1% probability. Taken together, our results suggest a spatial structuring of the genetic variability among the populations. Geographically closer populations exhibited greater similarities.

Recent lethal yellowing outbreak: why is the Malayan Yellow Dwarf Coconut no longer resistant in Jamaica?

In Jamaica, the Maypan, a hybrid of Malayan Yellow Dwarf (MYD) and Panama Tall coconut, previously considered highly resistant, is currently being devastated by an epidemic outbreak of lethal yellowing disease. There are several possible causes for this change. In this study, we checked that affected planting material in Jamaica is genetically the same as the material shown to be resistant. We compared the deoxyribonucleic acid (DNA) of MYD sampled in four locations in Jamaica with a reference DNA of the same cultivar collected in five different countries. The results of our analyses showed more variation at 34 simple sequence repeat loci in Jamaica than in the rest of the world providing clear evidence for the presence of about 16% of alleles that do not match the usual typical MYD genotype. These alleles appear to have already been present in the introduced germplasm. This rules out a possible cause of the new outbreak: The observed heterogeneity may have caused some loss of resistance but is insufficient to explain a massive outbreak of the disease.

Use of RFLP markers to study the diversity of the coconut palm

The geographical spread and diversity of the coconut palm in terms of growth habit, and fruit shape and colour, have led to the identification of a large number of varieties. For instance, the international list drawn up for IPGRI contains over 300 names of cultivars corresponding to traditionally grown populations or those improved by mass selection. Although some of them undoubtedly represent poorly defined or even extinct populations, almost 60 have been exchanged internationally by research stations over the past 50 years. This reflects breeders interest in collecting and using coconut germplasm. As a result, 17 countries in the COGENT network that have contributed to the CGRD database now have over 900 accessions, and have undertaken to provide a priority list of descriptors.

Genetic diversity in Brazilian tall coconut populations by microsatellite markers

A diversidade genetica em dez populacoes de coqueiro gigante do Brasil foi determinada utilizando 13 marcadores microssatelites. Foi detectado um total de 68 alelos, com media de 5,23 alelos por loco. A media da heterozigosidade esperada e observada foi 0,459 e 0,443 respectivamente. O numero de locos por populacao variou de 36 a 48, com media de 40,9 locos. Foram encontrados 12 alelos exclusivos e destes, um foi considerado como alelo raro esporadico e oito como alelos raros localizados. As metodologias de associacao dos individuos de acordo com a populacao de referencia e com base na analise fatorial de correspondencia forneceram resultados semelhantes e permitiram a formacao de dois grupos, o primeiro formado pelas populacoes de Baia Formosa, Georgino Avelino e Sao Jose do Mipibu e o segundo pelas populacoes de Japoata, Pacatuba e Praia do Forte. Esses resultados revelam um alto nivel de diversidade genetica nas populacoes brasileiras.