Occurrence of triploids in oil palm and their origin.

BACKGROUND AND AIMS: Oil palms showing exceptional vigour and dubbed as 'giant palms' were identified in some progeny during breeding. A panel of phenotypical traits were studied to characterize these trees. The hypothesis that gigantism and other anomalies might be linked to polyploidy was investigated. METHODS: Twenty sib pairs of palms from different crosses, each comprising a giant and a normal oil palm, were studied by flow cytometry with rice 'Nipponbare' as standard reference. In parallel, palms were assessed in the field using 11 phenotypic traits. A principal component analysis (PCA) was conducted to define relationships between these phenotypical traits, and a linear discriminant analysis (LDA) to predict ploidy level and giant classification. Finally, a co-dominant molecular marker study was implemented to highlight the sexual process leading to the formation of 2n gametes. KEY RESULTS: The first group of oil palms presented an oil palm/rice peak ratio of around 4.8 corresponding to diploid oil palms, whereas the second group presented a ratio of around 7, classifying these plants as triploid. The PCA enabled the classification of the plants in three classes: 21 were normal diploid palms; ten were giant diploid palms; while 11 were giant triploid palms. The LDA revealed three predictors for ploidy classification: phyllotaxy, petiole size and circumference of the plant, but surprisingly not height. The molecular study revealed that triploid palms arose from 2n gametes resulting from the second division restitution of meiosis in parents. CONCLUSIONS: This study confirms and details the process of sexual polyploidization in oil palm. It also identifies three phenotypical traits to assess the ploidy level of the giant oil palms in the field. In practical terms, our results provide a cheap scientific method to identify polyploid palms in the field.

Origin of C. latifolia and C. aurantiifolia triploid limes: the preferential disomic inheritance of doubled-diploid 'Mexican' lime is consistent with an interploid hybridization hypothesis.

Background and Aims: Two main types of triploid limes are produced worldwide. The 'Tahiti' lime type (Citrus latifolia) is predominant, while the 'Tanepao' type (C. aurantiifolia) is produced to a lesser extent. Both types result from natural interspecific hybridization involving a diploid gamete of C. aurantiifolia 'Mexican' lime type (itself a direct interspecific C. micrantha × C. medica hybrid). The meiotic behaviour of a doubled-diploid 'Mexican' lime, the interspecific micrantha/medica recombination and the resulting diploid gamete structures were analysed to investigate the possibility that 'Tahiti' and 'Tanepao' varieties are derived from natural interploid hybridization. Methods: A population of 85 tetraploid hybrids was established between a doubled-diploid clementine and a doubled-diploid 'Mexican' lime and used to infer the genotypes of 'Mexican' lime diploid gametes. Meiotic behaviour was studied through combined segregation analysis of 35 simple sequenbce repeat (SSR) and single nucleotide polymorphismn (SNP) markers covering the nine citrus chromosomes and cytogenetic studies. It was supplemented by pollen viability assessment. Key Results: Pollen viability of the doubled-diploid Mexican lime (64 %) was much higher than that of the diploid. On average, 65 % of the chromosomes paired as bivalents and 31.4 % as tetravalents. Parental heterozygosity restitution ranged from 83 to 99 %. Disomic inheritance with high preferential pairing values was deduced for three chromosomes. Intermediate inheritances, with disomic trend, were found for five chromosomes, and an intermediate inheritance was observed for one chromosome. The average effective interspecific recombination rate was low (1.2 cM Mb-1). Conclusion: The doubled-diploid 'Mexican' lime had predominantly disomic segregation, producing interspecific diploid gamete structures with high C. medica/C. micrantha heterozygosity, compatible with the phylogenomic structures of triploid C. latifolia and C. aurantiifolia varieties. This disomic trend limits effective interspecific recombination and diversity of the diploid gamete population. Interploid reconstruction breeding using doubled-diploid lime as one parent is a promising approach for triploid lime diversification.

Immunological and physiological parameters of Biomphalaria alexandrina snails exposed to Azadirachta indica plant.

BACKGROUND: Plant molluscicides could be appropriate for snail control measures against schistosomiasis in endemic areas. OBJECTIVES: The present study was undertaken to evaluate the immunological and physiological responses of Biomphalaria (B.) alexandrina snails to the effect of methanol extract of Azadirachta (A.) indica plant. MATERIALS AND METHODS: Haemolymph samples were collected from snails treated with LC25 from methanol extract for 1 month and untreated snails. The collected haemolymph samples from treated and untreated snails with tested plant were used for flow cytometric analysis of cell cycle. RESULTS: The obtained results indicated that hemolymph samples from B. alexandrina snails contained two morphologically distinct types of hemocytes, designated as Hyalinocyte and Granulocyte cells. In addition, the number of both snail's hyalinocytes and granulocytes and the mortality rate was significantly increased with treatment with A. indica extract. Phagocytosis in group treated with tested plant was highly significant increased than control one indicating a highly increase response of snail against the treatment. The lipid peroxide and glucose levels in hemolymph of treated snails were elevated while the protein and glycogen contents showed a decrease in soft tissues when compared with the control group. In addition, the activity level of some enzymes representing glycolytic enzymes as hexokinase (HK), pyruvate kinase (PK), phosphofructokinase (PFK), lactate dehydrogenase (LDH),and glucose phosphate isomerase (GPI); glycogenolytic enzymes as glycogen phosphorylase, glucose-6-phosphatase (G-6-Pase); gluconeogenic enzymes as fructose-1-6 diphosphatase (F-D-P ase), phosphoenolpyruvate carboxykinase (PEPCK) was also significantly reduced in response to treatment. CONCLUSIONS: It was concluded that the application of methanol extracts of A. indica plant may be helpful in snail control as it interferes with the snails'immunology and physiology.

Foundation characteristics of edible Musa triploids revealed from allelic distribution of SSR markers.

BACKGROUND AND AIMS: The production of triploid banana and plantain (Musa spp.) cultivars with improved characteristics (e.g. greater disease resistance or higher yield), while still preserving the main features of current popular cultivars (e.g. taste and cooking quality), remains a major challenge for Musa breeders. In this regard, breeders require a sound knowledge of the lineage of the current sterile triploid cultivars, to select diploid parents that are able to transmit desirable traits, together with a breeding strategy ensuring final triploidization and sterility. Highly polymorphic single sequence repeats (SSRs) are valuable markers for investigating phylogenetic relationships. METHODS: Here, the allelic distribution of each of 22 SSR loci across 561 Musa accessions is analysed. KEY RESULTS AND CONCLUSIONS: We determine the closest diploid progenitors of the triploid 'Cavendish' and 'Gros Michel' subgroups, valuable information for breeding programmes. Nevertheless, in establishing the likely monoclonal origin of the main edible triploid banana subgroups (i.e. 'Cavendish', 'Plantain' and 'Mutika-Lujugira'), we postulated that the huge phenotypic diversity observed within these subgroups did not result from gamete recombination, but rather from epigenetic regulations. This emphasizes the need to investigate the regulatory mechanisms of genome expression on a unique model in the plant kingdom. We also propose experimental standards to compare additional and independent genotyping data for reference.

Production of haploids from anther culture of banana [Musa balbisiana (BB)].

We report here, for the first time, the production of haploid plants of banana Musa balbisiana (BB). Callus was induced from anthers in which the majority of the microspores were at the uninucleate stage. The frequency of callus induction was 77%. Callus proliferation usually preceded embryo formation. About 8% of the anthers developed androgenic embryos. Of the 147 plantlets obtained, 41 were haploids (n=x=11). The frequency of haploid production depended on genotypes used: 18 haploid plants were produced from genotype Pisang klutuk, 12 from Pisang batu, seven from Pisang klutuk wulung and four from Tani. The frequency of regeneration was 1.1%, which was based on the total number of anthers cultured. Diploid plants (2n=2x=22) were also observed in the regenerated plants. The haploid banana plants that were developed will be important material for the improvement of banana through breeding programmes.

Effect of double infection with Schistosoma mansoni and Echinostoma liei on some physiological parameters of Biomphalaria alexandrina.

The survival rate and fecundity of B. alexandrina were greatly influenced when exposed to either S. mansoni or E. liei miracidia. The snails exhibited much lower survival rate and fecundity when double exposed to both S. mansoni and E. liei miracidia than single exposure and control snails. The results indicated a disruption in the snail metabolism due to exposure to S. mansoni and E. liei miracidia and this effect was more pronounced in case of double exposure to the two parasites. Protein concentrations in hemolymph and tissues significantly reduced in all exposed snail groups than in the control group. A significant elevation in the levels of aspartate aminotransferase (ASAT) and alanine aminotransferase (Asat) enzymes was recorded in hemolymph and tissues of exposed snail than unexposed (control snails). The ASAT/ALAT ratios in tissue and hemolymph of single-exposed and unexposed B. alexandrina did not exceed 1, while it increased up to 1.18 in hemolymph of double exposed snails. There were significant increases in the levels of acid and alkaline phosphatases enzymes in exposed snails.

Maternal inheritance of chloroplast genome and paternal inheritance of mitochondrial genome in bananas (Musa acuminata).

Restriction fragment length polymorphisms (RFLPs) were used as markers to determine the transmission of cytoplasmic DNA in diploid banana crosses. Progenies from two controlled crosses were studied with heterologous cytoplasmic probes. This analysis provided evidence for a strong bias towards maternal transmission of chloroplast DNA and paternal transmission of mitochondrial DNA in Musa acuminata. These results suggest the existence of two separate mechanisms of organelle transmission and selection, but no model to explain this can be proposed at the present time. Knowledge of the organelle mode of inheritance constitutes an important point for phylogeny analyses in bananas and may offer a powerful tool to confirm hybrid origins.

A molecular marker-based linkage map of diploid bananas (Musa acuminata).

A partial molecular linkage map of the Musa acuminata diploid genome is presented. This map is based on 58 RFLP, four isozyme and 28 RAPD markers segregating in an F2 population of 92 individuals. A total of 90 loci was detected, 77 of which were placed on 15 linkage groups while 13 segregated independently. Segregation distortions were shown by 36% of all loci, mostly favoring the male parent. Chromosome structural rearrangements were believed to be one of the main causes of these distortions. The use of genetic linkage data to further the genetic and evolutionary knowledge of the genus Musa, as well as to help improve the design of breeding strategies, is discussed.