Musa Germplasm Research Articles

Apparent Male Fertility in Musa Germplasm

Current efforts to produce improved genotypes of plantain and banana (Musa spp.) depend on crossing female-fertile clones with accessions that produce viable pollen. Musa accessions (168) were screened for production of viable pollen based on staining with acetocarmine glycerol jelly. Diploid hybrids and landraces produced significantly more pollen than triploids and tetraploids, suggesting more successful crosses when using diploid accessions as male parents. There was a positive correlation between the amount of pollen produced and the level of viability in both hybrids (r = 0.65, P≤ 0.01) and landraces (r = 0.61, P≤ 0.01). This finding suggests that closely associated genetic factors determine these characteristics in Musa, while environmental conditions also may influence the quality and quantity of pollen produced. Pollen production at anthesis was absent in 28 accessions. Of the 140 accessions with pollen, 67 were sufficiently fertile for use as male parents in the breeding program.

Genotype-by-environment Interaction in Musa Germplasm Revealed by Multi-site Evaluation in Sub-saharan Africa

Two multilocational trials, one comprising 18 Musa clones in three locations and another of 20 genotypes across 11 locations, were set up in 1991 and 1992, respectively, to assess the genotype-by-environment interaction (GxE) for important traits and to select stable high-yielding and black sigatoka (BS)-resistant genotypes. Combined ANOVAs showed significant differences among environments and among genotypes for all traits. GxE affected all growth and yield parameters, except fruit girth. Host response to BS disease also showed significant GxE, but there was no cross-order season-by-year interaction. Hence, genotypic response to BS can be assessed in 1 year during the rainy season, when disease pressure is highest. Genotype-by-location effects were more important than the nonsignificant genotype-by-year effects, supporting the need for multilocational trials. Stability analysis showed that full-sib plantain hybrids (TMPx1) exhibited different host responses to BS as well as different interaction patterns, suggesting that selection for stable BS resistance is possible. The BS-resistant TMPx genotypes had higher yields than the plantain landraces, but showed differences in yield stability. TMPx 1658-4, 2796-5, 5511-2, and 6930-1 have been selected as stable high-yielding hybrids, while the initial best selections (TMPx 548-4 and 548-9) were top yielders only in good environments. [Vuylsteke, D., R. Swennen, and R. Ortiz. 1993. Registration of 14 improved tropical Musa plantain hybrids with black sigatoka resistance. HortScience 28:957–959.]

DNA fingerprinting of Musa cultivars with oligodeoxyribonucleotide probes specific for simple repeat motifs

Using synthetic oligodeoxyribonucleotide probes against restriction-digested genomic DNA, we have established DNA fingerprinting of Musa cultivars. Of all the enzymes used, Eco RI and Hin dIII were found to be most informative, giving rise to individual specific band patterns with oligonucleotide probes of 15- to 18-base residues. Of the several probes and enzyme combinations used, the 15mer GACA probe with Eco RI and Hin dIII digests revealed a maximal level of polymorphism, and the probability of obtaining an identical band pattern between any two random genotypes was calculated to be 1.50 × 10 −9 and 1.59 × 10 −9, respectively. Oligonucleotide probes longer than 22 residues were also used but did not hybridize. The present approach is useful for cultivar identification and for overall genome analysis to establish relatedness among the various accessions of the Musa germ-plasm originating from different geographic locations. The relevance of using synthetic oligonucleotide probes based on simple repeat motifs for achieving DNA fingerprinting pattern is discussed.

Banana weevil resistance and corm hardness in Musa germplasm

Banana weevil (Cosmopolites sordidus Germar) is a major limiting factor in the cultivation of plantains and bananas (Musa spp. L.) in sub-Saharan Africa. The larvae damage the crop by tunneling in the corm. Chemical control is feasible but not sustainable, whereas host plant resistance is safe and has long term benefits. Banana weevil damage and infestation levels and corm hardness were assessed in Musa germplasm in order to determine the genetic control and potential mechanisms of resistance to this insect pest. Susceptibility/resistance to weevil was measured by the percentage coefficient of infestation (PCI) and damage in cross sections (CS) of the corm. All plantains were equally susceptible to the banana weevil, while a wild banana accession and some cooking and dessert banana cultivars showed high levels of resistance. Differential genotypic responses were observed in euploid plantain-banana hybrids. Segregation results suggest that host plant response to weevil in Musa is controlled by gene(s) exhibiting partial dominance towards the resistant parent and modifier genes with additive and dosage effects for susceptibility in the plantain parent. In natural banana germplasm, resistant clones showed increased corm hardness, as measured by a penetrometer in longitudinal and cross sections of outer and central corm tissues. This might suggest a non-preference mechanism for weevil resistance. However, the lack of correlation between corm hardness with PCI and CS scores in the segregating progenies suggested that other mechanisms may be more important in conferring resistance to banana weevil.

678 PB 186 RAPD GENETIC MARKERS FOR CHARACTERIZATION OF MUSA GENETIC RESOURCES

Fifty-seven accessions of Musa including cultivated clones Of 6 genomic groups (AA, AB, AAA, AAB, ABB, ABBB), M. balbisiana (BB), M. acuminata ssp. banksii (AA), M. acuminata ssp. malaccensis (AA) and M. velutina were examined for random amplified polymorphic DNA (RAPD) genetic markers using PCR with sixty 10-mer random primers. Forty-nine of 60 tested primers gave reproducible DNA amplification patterns. The number of bands resolved per amplification was primer dependent and varied from 1 to a maximum of 24. The size range of the amolification products also differed with the select& primer sequence/genotype and ranged from 0.29 to 3.0 kb. RAPD data were used to generate Jaccard's similarity coefficients which were analyzed phenetically. Phenetic analysis separated clones into distinct groupings that were in agreement with clusterings revealed when data were subsequently analyzed by principal coordinate analysis (PCO). In both the phenetic and the PCO analyses, previously unclassified cultivars grouped with cultivars previously classified for their genomic group based on morphological keys. The implications of RAPD analysis for Musa germplasm classification, clonal identification, and management are discussed.

The use of RAPD for identifying and classifying Musa germplasm.

Using the technique of random amplified polymorphic DNA (RAPD), we have identified 116 amplification products in Musa germplasm using nine primers. This has enabled us to identify RAPD markers that are specific to each of nine genotypes of Musa representing AA, AAA, AAB, ABB, and BB genomes. The pattern of variation observed following the application of multivariate analyses to the RAPDs banding data is very similar to the pattern of variation defined using morphological characters and used to assign Musa material into the different genome classes.

RFLP characterization of Indian Musa germplasm for clonal identification and classification

Nineteen single-copy clones isolated from a PstI genomic library (cv. Maiden Plantain), and eight Vigna chloroplast DNA clones were used to probe total genomic DNA digests of 57 genotypes of Musa from India. The 19 genomic clones detected a total of 107 polymorphisms among the 57 genotypes. Principal coordinates and phenetic analyses of these data placed cultivars and species into distinct groups that were in general agreement with a previously published RAPD-based classification of these same plant materials. The 107 polymorphisms were sufficient to differentiate each clone from every other clone. Heterologous Vigna chloroplast DNA probes were used to characterize the cytoplasm of Musa cultivars and species. PCO analysis of these RFLPs were detected both within and between the generally recognized genome groups, indicating multiple hybridization pathways in the origin of hybrid clones. Data presented demonstrate that RFLPs are sufficiently abundant to classify Musa germplasm and that genetic relationships among Musa cultivars, based upon RFLP data, are in general agreement with relationships determined by analysis of morphology and RAPDs.

Cytoplasmic genetic diversity in bananas and plantains

Concerns over yield declines in bananas and plantains due to the spread of Black Sigatoka disease in Musa have drawn attention to the collection of Musa germplasm and its use in conventional and biotechnological improvement programs. This report demonstrates the use of chloroplast DNA (cpDNA) restriction fragment length polymorphisms (RFLPs) for differentiating cytoplasms of various Musa clones. DNA was extracted from lyophilized leaf blade tissue and digested with either Eco R1, Hind III, Bam H1 or Pst I. Southern blots onto nylon membranes were probed with radioactively labeled heterologous orchid and lettuce cpDNA fragments. Among the 14 Musa clones examined, a single balbisiana and four acuminata-type cytoplasms were differentiated. The ability to distinguish between cytoplasms and to place plants within a cytoplasmic grouping demonstrates the usefulness of RFLP technology in evaluating diversity and determining the ancestry of Musa clones.