Adelheid R. Kuehnle
University of Hawaii
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Featured researches published by Adelheid R. Kuehnle.
Plant Cell Reports | 1992
Adelheid R. Kuehnle; Nellie Sugii
Transformed dendrobium orchids (Dendrobium x Jaquelyn Thomas hybrids) were recovered from protocorms bombarded by particles coated with the plasmid pGA482GG/cpPRV4, which contains the plant expressible Nos-NPT II and papaya ringspot virus (PRV) coat protein (CP) genes. Approximately 280 protocorms from four crosses were bombarded and potentially transformed tissues were identified by growth and green color on half-strength Murashige and Skoog medium supplemented with 2% sucrose and 50–100 mg 1−1 kanamycin sulfate. Kanamycin concentrations that prevented growth of nontransformed tissues could not be used for long-term selection because such levels suppressed the regeneration of potentially transformed tissues. PCR and restriction analysis 21 months after treatment found 13 of 13 plants from two crosses, which appeared kanamycin-tolerant, to contain the Nos-NPT II gene, while only one of these plants carried the vector-linked PRV CP-gene. These results support use of particle bombardment for transformation of this important ornamental monocot.
Plant Cell Reports | 1992
Adelheid R. Kuehnle; Fure-Chyi Chen; Nellie Sugii
SummaryA method for the production of somatic embryos and subsequent plant regeneration for Anthurium andraeanum Linden ex André (Monocotyledonae) hybrids is described. Whole leaf blade explants, derived from plantlets grown in vitro, formed translucent embryogénic calli at their basal ends within one month of culture in the dark. Secondary somatic embryos formed frequently and without an intervening callus on surfaces of primary embryos. Embryogenesis was induced with three genotypes using a modified half-strength Murashige and Skoog (MS) medium supplemented with 1.0 to 4.0 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.33 to 1.0 mg l−1 kinetin. A combination of 2% sucrose with 1% glucose in the medium favored embryogenesis over 3% sucrose alone. Whole leaf blades on medium solidified with 0.18% Gelrite produced more somatic embryos than leaves on medium with 0.7% Bacto-agar. Within two to three months after culture initiation, embryos were transferred to modified MS medium containing 0.2 mg l−1 6-benzyladenine (BA) and 2% sucrose and placed in the light for conversion into plantlets. Rooted plantlets were recovered and transferred into pots with tree fern fiber medium and grown in the greenhouse.
Plant Cell Tissue and Organ Culture | 1997
Fure-Chyi Chen; Adelheid R. Kuehnle; Nellie Sugii
Root explants from in vitro-grown plants of Anthurium andraeanum ‘Alii’ and Anthurium interspecific hybrid ‘UH1060’ readily produced multiple shoots under weak light on a modified Murashige & Skoog medium containing 2.2 μM BA. Regenerated ‘UH1060’ plants grew normally and flowered within 16 months after transfer to the greenhouse. Cocultivation of root cuttings with Agrobacterium tumefaciens LBA4404 carrying the binary vector pCa2Att resulted in kanamycin-resistant ‘Anuenue’ shoots transgenic for neo and att and recovered more than one year after culture on selection media. Transformation efficiency (number of explants with transgenic shoots per total explants) was 1.3%. Other cultivars (‘UH1003’, ‘UH1060’, ‘Rudolph’, and ‘Mauna Kea’) failed to produce shoots under the transformation condition employed.
In Vitro Cellular & Developmental Biology – Plant | 1995
Guo-Ling Nan; Adelheid R. Kuehnle
SummaryFive parameters were examined for their effect on transformation ofDendrobium tissues by microprojectile bombardment. The superpromoter in pBI426 produced at least 1.5 times as many transient transformants as the single cauliflower mosaic virus 35S promoter in pBI121 (37 to 69% vs. 0 to 44%) with dark and frequent GUS (β-glucuronidase) staining. Tissue, genotype, and type of microparticle significantly affected transient GUS activity. Higher expression was seen in protocormlike bodies and in hybrid UH44 compared to etiolated shoots and protocorms and to hybrids M61 and K1329-39. Microparticles of 1.6-µm Bio-Rad gold were more effective than 1.0-µm ASI gold. Transient GUS activity did not differ among protocormlike bodies bombarded using helium propellant pressures of 650, 900, or 1100 psi. Transgenic plants were recovered fromDendrobium UH800 protocormlike bodies bombarded with pBI426-coated, 1.1-µm tungsten particles using an early-model gunpowder-driven apparatus with an estimated stable transformation rate of 11.7%. One transgenic plant ofDendrobium UH44 was recovered from etiolated shoot explants bombarded with pBI121-coated, 1.1-µm tungsten particles using the Dupont PDS-1000 with a stable transformation rate of 0.17%. Positive selection results showed 100 to 200 mg·liter−1 kanamycin to be appropriate for regeneration of transgenic plants from protocormlike bodies, protocorms, and etiolated shoot explants over a 3- to 9.5-mo. period.
American Journal of Botany | 1998
Tracie K. Matsumoto; Adelheid R. Kuehnle; David T. Webb
Morphological, anatomical, and histochemical aspects of zygotic embryogenesis by Anthurium andraeanum Lind. were investigated from 4 to 24 wk postpollination. Anatomical features were correlated with morphology of the spadix and capacity of embryos to germinate in vitro. Development from a single-cell zygote to fully mature seed takes 24 wk. The suspensor was two ranked and obvious during the early stages of embryogeny. It was apparent by week 8, substantial until week 14, and diminished rapidly until its absence by week 22. Differentiation of the shoot apex, cotyledon, and protoderm occurs at 14 wk. The embryo starts to derive nutrition from the endosperm at this time, and germination of cultured ovules reached 56%. By 20 wk the shoot apex had visible leaf primordia and the root apex was clearly defined. The cotyledon was well developed and surrounded the shoot tip. The storage of protein and starch was at its greatest in the endosperm and embryo. Furthermore, 100% germination of cultured ovules and embryos occurred at 20 wk and thereafter. Fully mature embryos at 24 wk are green and contain protoxylem elements.
In Vitro Cellular & Developmental Biology – Plant | 1997
Koon-Hui Wang; Adelheid R. Kuehnle; Brent S. Sipes
SummaryA reliable method to screen Anthurium for burrowing nematode resistance and tolerance in vitro was developed using 17 genetically distinct Anthurium cultivars. Based on nonparametric data analysis, tolerance and resistance were found to be independent traits to be evaluated separately. An effective parameter for tolerance evaluation was ranking of relative leaf retention, whereas an effective parameter for resistance evaluation was the ranking of nematode reproduction, log(Rf+1). A comparison of the ranking of leaf retention with ranking of nematode reproduction clustered the cultivar responses to burrowing nematode infection into four groups: intolerant and resistant, moderately tolerant but susceptible, intolerant and susceptible, and tolerant and susceptible. ‘Ozaki’ was identified as an intolerant reference, ‘Nitta’ as a susceptible reference. ‘Blushing Bride’ was the most tolerant cultivar among those screened, but it may not be an ideal tolerant reference due to its low vigor. Future screening for burrowing nematode-tolerant and-resistant cultivars in Anthurium should include ‘Ozaki’ and ‘Nitta’ as internal controls. Evaluation of resistance should be based on a resistance index obtained by log(Rf of hybrid tested +1) divided by log(Rf of ‘Nitta’ +1); tolerance should be based on ranking of relative leaf retention.
Plant Cell Tissue and Organ Culture | 1992
Adelheid R. Kuehnle; Elizabeth D. Earle
Several factors affecting the success of selection in plant populations were examined for their relevance to in vitro selection. Three in vitro selection schemes and two growth assessment procedures were evaluated for effectiveness in selecting for a mitochondrial trait in maize: resistance to the insecticidal compound methomyl. Regenerable maize callus was derived from immature embryos of the three-way hybrid P39/IL766A2 x W182BN containing Texas male sterile cytoplasm (cms-T). Either low, gradually increasing, or high selection pressures were used to grow callus over a period of 3–5 months. There was no significant difference in recovery of resistant plants using these 3 methods. Growth of callus on medium containing methomyl was assessed by increase in fresh weight during the final month of selection or by increase in number of callus pieces over the course of selection. These quantitative measures of growth were unreliable indicators for gain in resistance within the callus population. A procedure for recovery of methomyl resistant and male-fertile cms-T plants is suggested.
Archive | 1997
Haruyuki Kamemoto; Adelheid R. Kuehnle
Archive | 1999
Haruyuki Kamemoto; Teresita D. Amore; Adelheid R. Kuehnle
Hortscience | 2004
Adelheid R. Kuehnle; T. Fujii; Fure-Chyi Chen; Anne M. Alvarez; Nellie Sugii; R. Fukui; Stacie L. Aragon; Jesse M. Jaynes