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- Author or Editor: J.W. Van Sambeek x
“Ambers” is a term used to describe poorly filled, shriveled eastern black walnut (Juglans nigra L.) kernels with a dark brown or black-colored pellicle that are unmarketable. Studies were conducted to determine the incidence of ambered black walnut kernels and to ascertain when symptoms were apparent in specific tissues. The occurrence of ambered kernels was evaluated in fruit harvested from mature ‘Football’ trees growing at three sites within a commercial black walnut orchard in 2008 to 2010. Mature walnut fruit sampled from trees at Site 2 had greater odds for ambered kernels than those on trees at two other sites within the same orchard with 27% of the walnuts sampled exhibiting symptoms when examined in October. Also, black walnut fruit in 2010 had more ambered kernels than those examined in Oct. 2008 or 2009. Cropload, soil type, ambient temperatures, or precipitation was not apparently associated with a high incidence of ambered kernels. When black walnut fruit from trees at Site 2 were examined from 25 June to 6 Oct. 2011, embryos were visible in 50% of the fruit without discoloration on the first date. Stenospermocarpy (e.g., aborted or rudimentary embryos after fertilization) was observed in fruit with discolored or ambered kernels as early as 7 July. Stenospermocarpic fruit with ambered kernels had shorter embryo axis lengths (root apex to shoot apex) than fruit with non-ambered kernels on 7 July and at successive sampling dates. Cotyledon widths of ambered kernels in stenospermocarpic fruit were narrower than those of non-ambered kernels on 21 July, but symptomatic cotyledons continued to enlarge until 15 Sept. All fruit enlarged during the growing season and nut diameters varied by only 3.4 mm at harvest. Thus, visible embryo degeneration, which was associated with ambered kernels in black walnut fruit, was detected in early July when shell hardening occurs and kernel tissues are enlarging.
Hybrid poplar is traditionally established using dormant stem cuttings in tilled soils followed by chemical or mechanical weed control. In 1996, we initiated a study to evaluate the effects of site preparation and four weed control treatments on growth and morphology of three hybrid poplar clones established on a 0.2-ha tall fescue field in southern Illinois. Site preparation included application of 2000 kg/ha of 12N-12P-12K. The experiment was arranged as a split-split plot. Main plots were closely mowed tall fescue or tilled to remove the grass sod. Within each main plot, weed control treatments were applied to 1-m wide strips in rows 2.4 m apart. Weed control treatments included porous black film, solid black film, and solid white film, and a control treatment of 3.7 L/ha of glyphosate applied each spring. On 15 Apr. 1996, three 25-cm-long dormant stem cuttings from each of three clones were randomly planted 15 cm deep every 1.8 m within each row. Clonal differences existed after the first year for survival, number of stems, stem height, stem basal diameter, and stem volume, but not for number and total length of lateral branches. Nearly all tree growth measurements analyzed during the first 3 years had a highly significant interaction between type of site preparation and method of weed control. With polyethylene films, tree survival exceeded 90% on both the tilled ground and grass sod sites after 3 years; however, with the herbicide treatment survival averaged only 18% in the grass sod and 51% in tilled soil. Excluding the herbicide treatment, tree growth was better in the grass sod than in the tilled soil. Tree growth using porous black polyethylene film was usually less than that with either of the two solid polyethylene films. The best tree growth was found with a grass sod and solid white polyethylene film for weed control.
At monthly intervals for 1 year, one branch was removed from the lower crown of three 30-year-old trees of black walnut (Juglans nigra L.). The basal 1.3 m of each branch was cut into four 32-cm-long segments that were placed horizontally in shallow plastic trays filled with perlite and watered daily with tap water. Branch segments cut early in the dormant season (29 Sept., 31 Oct., or 1 Dec.) or shortly after flushing (6 June) produced few, if any, epicormic sprouts. Approximately half the branch segments cut on 3 Jan. or 3 Feb. produced one sprout that elongated slowly. Most branch segments cut in the late dormant season (2 Mar., 30 Mar., 3 May) or growing season (5 July, 4 Aug., 6 Sept.) produced one or two sprouts >20 mm long. To prepare explants for in vitro culture, the terminal 2.5 cm was harvested when sprouts exceeded 3.0 cm, trimmed of all leaves, and disinfested. Explants were placed vertically in liquid Long & Preece (LP) medium supplemented with 3% sucrose, 0.3 μM TDZ, 0.05 μM IBA, and 1 μM BA. When shoots began to elongate (4 to 6 weeks), they were then placed horizontally on agar-solidified LP medium with liquid LP overlays to induce axillary shoot proliferation. Advantages of forcing epicormic sprouts on large branch segments are: 1) they can be a source of in vitro explant material for 6 to 7 months a year, 2) aseptic cultures can be easily obtained, 3) shoots from the base of branches may show more juvenility than shoots forced from branch tips, 4) softwood shoot wilting is not a problem as with forcing shoots from branch tips, 5) the procedure does not require preparing and changing forcing solutions, and 6) branch segments should have more stored food than dormant branch tips for forcing softwood growth.
During Apr. 1999, the lower branches of mature black walnut trees were removed and cut into sections 48 cm long and placed horizontally in plastic flats filled with perlite in a shaded polyethylene-covered greenhouse. Water was applied by drip emitters and care was taken to avoid overhead water contact with the stem sections. Within 2 months, elongating, green, leafy shoots were excised, brought into the laboratory, surface disinfested and placed in vitro onto agar-solidifed Long and Preece (LP) medium with 0.3 μM thidiazuron (TDZ), 0.5 μM indolebutyric acid (IBA), and either 0.1, 1.0, or 10.0 μM benzyladenine (BA). Explants were transferred to fresh medium after 1, 3, and 5 days in vitro and every 2 weeks thereafter. After 3 months in vitro, callus was excised and and explants were all placed on LP medium with 10 μM BA and 0.5 μM IBA for 4 weeks. They were then transferred to LP with 0.3 μM TDZ, 1.0 μM BA, and 0.5 μM IBA for 2 weeks. This 4-2-week alternation of media has continued for more than 6 months. After 4 months in vitro, shoot clusters were subdivided, and individual microshoots recultured. Of the original 260 explants, 30 survived and have been subdivided into 111 cultures. These explants have produced 132 axillary shoots that are also multiplying. Adult black walnut will acclimate and proliferate in vitro, but only with careful attention to detail and regular transfers to fresh medium.
Cotyledon explants were harvested from immature walnut fruits during July and August 1991. Media consisted of either WPM with 0.1 μM 2,4-D, 5.0 μM TDZ and 1.0 g/liter casein hydrolysate or DKW with 4.4 μM BA, 0.05 μM IBA, 9.3 μM Kinetin and 250 mg/liter l-glutamine. Treatments were arranged factorially with 2 gelling agents, 7 g/liter Sigma agar or 2 g/liter Gelrite and were incubated in light or in darkness. After 4 weeks, all explants were placed on basal DKW with no growth regulators and were cultured in darkness. The best treatment tested was from seeds collected 14 weeks post-anthesis on WPM, agar, and incubation in light (22 embryos/explant, 78% embryogenesis). Use of DKW and gelrite in darkness resulted in 1 embryo/explant and 38% embryogenesis. Up to 90% shoot organogenesis also occurred on cotyledon explants from seeds collected 16 weeks post-anthesis and placed on WPM. Shoots elongated on stationary liquid DKW with 10 μM BA.
Branch tips (30 to 40 cm long) of adult black walnut were forced in a half-strength solution of Long and Preece medium (LP) salts (minus iron) plus 1 mM 8-hydroxyquinoline citrate (8-HQC). The resulting softwood shoots were surface-disinfested and cut into 1.5-cm-long nodal segments. Explants were placed on two media: Driver and Kuniyuki Walnut medium (DKW) or LP with four plant growth regulator combinations: 5 μM BA with 0.05 μM IBA, 10 μM BA, 1 nM TDZ, or 10 nM TDZ in a factorial arrangement. Gelrite was used as the gelling agent. Explants were transferred to fresh medium on days 1, 3, 5, and 7 after initiation, then weekly. Data recorded 60 days after culture initiation showed more and longer shoots and leaves, greater explant diameter, more green (living) tissue, and less exudation per explant on LP than on DKW. Greatest explant and shoot length were observed when the medium contained 10 nM TDZ. BA (10 μM) and LP were best for long-term maintenance of cultures
Genetic transformation studies are aided by use of selection agents, such as antibiotics or herbicides. To determine the level of kanamycin to be used as a selection agent, cotyledonary stage somatic embryos from J. nigra lines J26 and J28, J. nigra × J. hindsii line S11, and J. regia line SU2 were placed on gelrite solidified WPM with 1 g/liter casein hydrolysate and 250 mg/liter cefotaxime and 3% (w/v) sucrose. Dosages for inhibiting secondary embryogenesis were 40 mg/liter kanamycin for J. nigra and J. nigra × J. hindsii and 100 mg/liter for J. regia. For the bialaphos experiments, somatic embryos of J. nigra lines J26 and J28 and J. nigra × J. hindsii line S11 were cultured on gelrite solidified LP medium with 0.5 g/liter casein hydrolysate and 3% (w/v) sucrose. Between 0.1 and 1.0 mg/liter bialaphos, inhibited secondary embryogenesis.