Nick Gawel

Restriction fragment length polymorphism (RFLP)-based phylogenetic analysis of Musa.

SummaryRandom genomic probes were used to detect RFLPs in 19 Musa species and subspecies. A total of 89 phylogenetically informative alleles were scored and analyzed cladistically and phenetically. Results were in general agreement with morphology-based phylogenetic analyses, with the following exceptions: our data unambiguously places M. boman in section Australimusa, and indicates M. beccarii is very closely related to M. acuminata. Additionally, no support was found for the separation of section Rhodochlamys from section Musa. A comparison of morphology-based and RFLP-based phylogenetic analyses is presented.

CHLOROPLAST DNA RESTRICTION FRAGMENT LENGTH POLYMORPHISMS (RFLPS) IN MUSA SPECIES

SummaryTaxonomic and phylogenetic determinations within the genus Musa are established using a numerical, morphology-based scoring system. However, within this system, the classification and relationships of some types are disputed. The application of chloroplast DNA (cpDNA) restriction fragment length polymorphism (RFLP) analysis to Musa taxonomy provided valuable, supplemental information about the classification of, and relationships between, Musa species and subspecies. Whole-cell DNA was extracted from lyophilized Musa leaf-blade tissue and digested with various restriction enzymes, Southern blotted onto nylon membranes, and probed using radioactively labeled heterologous orchid cpDNA fragments. Phylogenies were inferred from cpDNA RFLP patterns using PAUP software. The relationships between most species examined were as expected; however, some species (M. beccarii and M. basjoo) did not conform to the conventional morphology-based phylogeny.

Genetic control of somatic embryogenesis in cotton petiole callus cultures

SummaryThree commercial varieties (Acala SJ-5, Coker 312 and Paymaster 303) and three exotic accessions (T1, T25 and T169) of cotton (Gossypium hirsutum L.) were tested for ability to undergo somatic embryogenesis. Sections of split petiole were cultured on 3 media and evaluated for embryogenesis after 180 days. Embryogenic T25 and Coker 312 plants were selected and crossed in a diallel with non-embryogenic Acala SJ-5, Paymaster 303, T1 and T169 plants. F1, F2 and BC1 populations were generated and tested for embryogenesis on a medium of MS salts and vitamins (1962) plus (per liter) 4.0 mg NAA, 1.0 mg Kn, 30 g glucose, 100 mg myo-inositol, 2.0 g Gelrite and 0.75 g MgCl2. Segregation for both occurrence and magnitude of embryogenesis was observed, suggesting the action of more than one gene.

Somatic embryogenesis in two Gossypium hirsutum genotypes on semi-solid versus liquid proliferation media

A comparison of semi-solid vs. liquid embryo proliferation media was made using two Gossypium hirsutum L. genotypes (Coker 312 and T25) and two callus initiation media. Sections of petioles from mature, flowering plants were cultured on two modified Murashige and Skoog media. Medium 1 included 4.0 mg l-1 NAA and 1.0 mg l-1 kinetin; medium 2 contained 0.1 mg l-1 2,4-D and 0.1 mg l-1 kinetin. After six weeks, callus was removed from each explant and divided in half. One callus portion was placed in liquid proliferation medium and the other on semi-solid (0.2% Gelrite) proliferation medium. Composition of proliferation medium was identical to that of initiation medium, except no growth regulators were added. Embryos were counted after eight weeks. The percentage of explants forming callus was influenced by genotype/initiation medium combination. Analysis of variance procedures revealed significant variability for callus initiation media, proliferation media (semi-solid or liquid), and an initiation medium x genotype interaction. Paired t-tests indicated that more embryos were produced in liquid proliferation medium (227.3 embryos/culture) than on semi-solid proliferation medium (134.6 embryos/culture).

Detecting genetic diversity in diploid bananas using PCR and primers from a highly repetitive DNA sequence

SummaryThe polymerase chain reaction (PCR) was used to detect polymorphisms among 29 diploid clones of Musa acuminata Colla. from Papua New Guinea. Primer sequences were derived from a 520 bp highly repetitive DNA sequence isolated from M. acuminata ssp. malaccensis. Primers, used individually, detected a total of 48 polymorphisms that were scored as unit characters and used to generate a Jaccards similarity index. Principal coordinate analysis (PCO) was used to cluster clones and the unweighted paired-group method of analysis (UPGMA) was used to compute genetic distance among the materials examined. The abundance of diversity within the PNG diploids examined reflects the extreme genetic variability within the M. acuminata gene pool. PCR, utilizing primers from a highly repetitive sequence, is a rapid means of detecting genetic diversity in M. acuminata.

Cytoplasmic genetic diversity in bananas and plantains

SummaryConcerns 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.

Abscisic acid-induced growth inhibition of sweet potato (Ipomoea batatas L.) in vitro

The inhibitory effects of abscisic acid (ABA) on in vitro growth and development of axillary buds from nodal segments of sweet potato (Ipomoea batatas L.) was investigated. ABA at concentrations of 0.01, 0.1, 1.0 or 10.0 mg 1-1 inhibited axillary bud and root development and subsequent plantlet growth. ABA at 10 mg 1-1 completely inhibited axillary shoot development but did not affect the viability of cv. Jewel explants over a culture period of 365 days. Transfer of nodal segments cultured for 90, 180 or 365 days from basal medium containing 10 mg 1-1 ABA to growth regulator-free media resulted in rapid and normal plantlet development. Gibberellic acid at 0.1, 1.0 or 10.0 mg 1-1 in the presence of ABA at 0.1, 1.0 or 10.0 mg 1-1 did not counteract the ABA-induced growth inhibition. Although ABA totally inhibited the growth of 6 sweet potato plant introductions at a concentration of 10.0 mg 1-1, the efficacy of ABA as a suppressant of shoot growth varied with genotype.

Chemical and environmental growth regulation of sweetpotato (Ipomoea batatas (L.) Lam.) in vitro

The need for conservation of biotic diversity is well recognized. However, improved techniques for the efficient, cost effective-preservation of plant germplasm are needed. The conservation and distribution of plant germplasm in vitro is gaining acceptance. However, increased usage is dependent upon the ability of curators to minimize culture maintenance requirements. This report examines the effect of various levels of sucrose, photoperiod, temperature, sorbitol and mannitol on minimal growth storage of Ipomoea batatas (L.) Lam. Growth was reduced 50% with a temperature reduction of from 21.1 to 15.6°C. Sucrose concentrations of 15 and 20 g l-1 resulted in reduced plant stature with few adverse effects on plantlet viability or morphology. Reduction of photoperiod from 16 to 4 h produced smaller, slightly chlorotic, but otherwise normal plants. The addition of sorbitol or mannitol to culture media generally produced undesirable effects on gross plant morphology and loss of apical dominance. Genotype x growth retarding treatment interactions were observed for all variables examined.