plant regeneration were difficult to transform using Agrobacterium . To increase the transformation efficiency in castor, we are developing protocols for plant regeneration through adventitious shoot formation by testing various tissues from mature seed
Yeh-Jin Ahn and Grace Qianhong Chen
Robert L. Geneve
Adventitious root formation in debladed petiole cuttings of English ivy proceeds via a direct rooting pattern for the easy-to-roof juvenile phase, while the difficult-to-root mature phase roots through the indirect pattern, Juvenile petiole cuttings treated with NAA (100 μM) plus the polyamine biosynthesis inhibitor, DFMA (1 mM), formed an increased number of roots per cutting initiated through the indirect rooting pattern. The increased formation and the change in rooting pattern were reversed by the addition of putrescine (1 mM). Delaying auxin application to petiole cuttings for 15 days, also induced juvenile petioles to root by the indirect pattern. This could be reversed by rebounding the base of the cutting prior to auxin application at day 15. The data support the use of the terms “pre-competent root forming cells” (PCRFC) and `induced competent root forming cells' (ICRFC) to describe the target cells for the initial events of root formation for the direct and indirect patterns, respectively,
R.N. Trigiano, K.M. Kaveriappa, S.E. Schlarbaum, M.T. Windham, and W. Witte
DNA amplification fingerprinting (DAF) was Used to characterize both parents (different cultivars) in breeding experiments with Cornus florida. Putative hybrids were fingerprinted and true crosses identified by finding unique male parent products in amplification profiles. Both manual and honey bee mediated pollinations successfully produced hybrid seed. Axillary buds from seedlings were used to initiate proliferating shoot cultures on woody plant medium with 4.5 μm BA. Initiation and development of adventitious roots were dependent on IBA (4.1 μm), sucrose (0–2%), and agar (0.2–0.6%) concentrations. About 40–50% of the microshoots produced roots and were acclimatized to greenhouse conditions. Cultures have been maintained without loss of regeneration potential for over 2 years. Clonal material can be reentered into the breeding program or used to evaluate horticultural characteristics in different environments and locales.
Thomas E. Marler and Louann C. Guzman
Growth and physiological responses of Intsia bijuga trees to flooding were determined in a series of five container experiments to assess the relative tolerance of this species to flooding. The first measurable response to flooding was reduced leaf gas exchange, which began within 5 to 6 days of the onset of flooding. Development of hypertrophied lenticels at the water line and paraheliotropic leaflet movement were evident by 17 days of flooding. Emergence of adventitious roots on the stem above the water line began after about 30 days of flooding. Leaflet abscission was greatly accelerated by flooding. After more than 3 months of flooding, regrowth of roots, stems, and leaves began within two weeks of draining the medium. The data and observations support a relative ranking of moderate flood tolerance for Intsia bijuga.
James R. McKenna and Ellen G. Sutter
The use of auxin-impregnated toothpicks stimulated adventitious root formation in genotypes of Juglans `Paradox' that had been backcrossed to J. regia. These genotypes were selected as potential rootstocks because of improved tolerance to cherry leaf roll virus and Phytophthora spp. Other auxin applications including quick dips and talc formulations had little or no effect. The use of toothpicks lowered the concentration of IBA necessary for root initiation compared to previously reported results using quick dips. Toothpicks were inserted transversely into holes drilled 1 to 2 cm above the base of cuttings. Callus and roots always formed at the location of the toothpicks rather than at the base of the cutting. Roots were formed using this method in simple layering, hardwood, and semi-hardwood cuttings. Of all the cuttings that rooted, 90% rooted with toothpicks whereas only 10% rooted using a quick dip. This method may have potential for increasing the efficiency of rooting other difficult-to-root plants.
Louis Anella and Thomas H. Whitlow
Changes in photosynthesis and root morphology during flooding were studied in container-grown 2- to 3-year-old Acer rubrum L. saplings. The seed was collected from opposite ends of a hydrologic gradient: two upland xeric sites [the George Washington National Forest in Page County, Va. (38°25'N, 78°35'W), and the Robinson Forest in Perry County, Ky. (37 °9'N, 83°7'W)] and a floodplain hydric site [the Presquile National Wildlife Refuge along the James River in Chesterfield County, Va. (37°21'N, 77°16'W)]. Each xeric site, containing various half-sib families (maternal parent known, paternal parent from one or more sources), was compared to a different hydric site half-sib family. After 1 week of flooding, the xeric-site trees had lower net photosynthesis than the hydric-site trees and remained significantly lower for the duration of the study. After an initial drop, hydric-site plants showed a recovery in net photosynthesis, indicating a greater ability to acclimate to a flooding stress. Seventy-one percent of the hydric-site plants developed adventitious roots and all retained their leaves. Xeric-site plants did-not develop adventitious roots and 61% were defoliated after 60 days of continuous flooding. The results indicate ecotypic differentiation in physiological response between Acer rubrum populations collected from opposite ends of a hydrologic gradient.
Joseph J. King and Dennis P. Stimart
In an attempt to analyze genetically the interaction of endogenous auxin concentration and adventitious root formation, an EMS mutagenized M2 population of Arabidopsis thaliana was screened for mutants with altered abilities to form adventitious roots. A selected recessive nuclear mutant, rooty (rty), is characterized by extreme proliferation of roots, inhibition of shoot development and other morphological alterations suggestive of auxin or ethylene effects. The rty phenotype occurs in wild type seedlings grown on auxin containing medium and relatively normal growth is stimulated in rty seedlings growing on cytokinin containing medium. Analysis by GC-MS found that endogenous IAA concentrations in rty are 2 to 17 times higher than in wild type depending on tissue type and IAA form. Dose response experiments with IAA and NAA indicated that rty does not express increased sensitivity to auxin. These data suggest that the rty phenotype is due to elevated endogenous auxin. A genetic map location for rty and possible roles for the wild type RTY gene product in regulating auxin concentration will be presented.
S. Snapp, W. Kirk, B. Román-Avilés, and J. Kelly
Fusarium root rot is a major limiting factor in snap bean (Phaseolus vulgaris L.) production. The level of genetic resistance in commercial bean cultivars is minimal and disease is frequently exacerbated by environmental factors. We investigated the contribution of vigorous, adventitious roots to enhancing root rot tolerance in snap bean. Seedling root system architecture was evaluated in 17 recombinant inbred lines (RILs) from a cross of a resistant snap bean line (FR266) and a susceptible dry bean cultivar (Montcalm). The RILs varied in tolerance to Fusarium root rot. Although overall length and branching density (as measured by fractal dimension and meristem numbers) of root systems were not related to root rot resistance, the lateral root number at the root: shoot interface was positively correlated with genotype tolerance (R 2 = 0.6*). Root diameter was also positively correlated with tolerance; this is consistent with the hypothesis that larger adventitious and basal roots are beneficial under disease stress. A field-based study of commercial snap bean cultivars compared raised and flat-bed systems of production, in a soil inoculated with Fusarium solani f. sp. phaseoli. Substantially greater yields (40% to 90%) were observed in raised beds. Root vigor was relatively high (root length density >0.2 cm·cm−3) and root rot scores were lower with raised than with flat-beds, in 2001, but not in 2000. Overall, this is suggestive that integrated crop management practices can improve lateral root vigor and reduce root rot severity.
S. Yusnita, Robert L. Geneve, and Sharon T. Kester
In vitro shoot multiplication of white Eastern redbud was successful using two-node mature explants from the initial spring flush on a woody plant medium (WPM) supplemented with benzylaminopurine (BAP). Optimal shoot proliferation was obtained at 10-15 μM BAP. Treatment with thidiazuron produced fasciated (stunted) adventitious shoots which failed to elongate. Successive subcultures increased the ability of explants to form shoots. However, shoot tip necrosis became a problem after 7-8 subcultures. Shoot tip necrosis is being studied by comparing shoot multiplication on bacto-agar vs. gelrite, increasing the Ca concentration in WPM and by trying to reduce the phenolic exudate by the explants with PVP or activated charcoal. Microshoots >3 cm long were rooted by pulse treatment on half strength WPM containing 300 μM IBA or NAA before being moved to hormone free WPM. There was a different morphology between IBA and NAA induced roots, although the number of roots were comparable. IBA treated microcuttings developed branched, fine roots, whereas NAA treated plants produced unbranched, coarse roots. Rooted microshoots were successfully acclimated to greenhouse condition.
Wilfredo Colon, Mike Kane, Dewayne Ingram, and Hilton Biggs
Stage 2 micropropagules were transferred into woody plant medium supplemented with either 0, 0.1, 1, 10, 100 mg/L ABA (Abscisic acid) and with or without 1 mg/L IBA (Indole-3-butyric acid), Significant decreases in total dry weight and shoot length were observed at 1, 10 and 100 mg/L of ABA regardless of IBA concentration, Leaf area was significantly reduced in all treatments by increasing ABA levels. In the absence of IBA no callus formed but lateral roots developed. Another experiment using ABA levels of 0, 0.1, 0.5 and 0, 1 mg/L IBA was conducted. Total number of roots decreased with increasing ABA levels. Adventitious roots which formed on the stem and roots originating from root primordia were observed in all ABA levels with IBA, Callus did not form in the treatments lacking IBA. Scanning electron microscopy was used to document morphological differences due to ABA, Abscisic acid levels in leaf tissue were assayed using immunological techniques.