Both kinetin and BA promoted in vitro shoot formation from hypocotyl explants of Lupinus texensis Hook. placed on Murashige and Skoog (MS) medium. With either cytokinin, shoot formation was best at ≈4.5 μm. Adventitious root formation was observed only on tissue culture-derived shoots placed in MS media containing 5.4 to 54 μM NAA. IAA and IBA, at concentrations ranging from 5 to 55 μm, failed to stimulate rooting. Even at the optimal concentration of NAA, only 14% of the shoots produced roots. Thus, although hypocotyl explants readily produced shoots, adventitious root formation on these shoots occurred with relatively low frequency. Chemical names used: 6-benzylaminopnrine (BA); indole-3-acetic acid (IAA); indole-3-butyric acid (IBA); 6-furfurylaminopurine (kinetin); 1-naphthaleneacetic acid (NAA).
The regenerative capacity of mature pecan [Carya illinoinensis (Wangenh.) K. Koch] embryonic tissues was demonstrated after pretreating mature nuts to eliminate associated endogenous contaminants. Cultured cotyledon segments were induced to form adventitious roots in a medium with 50 μm NAA. A regeneration medium with 20 μm BA and 5 μm IBA stimulated prolific axillary shoot production from the embryonic axis without causing cotyledon abscission. Cotyledon retention was essential for shoot initiation and long-term development. Eighty-five percent of the shoots emerging from embryonic axes formed at the cotyledonary nodes. Thirty percent of the microshoots rooted on an auxin-free medium after preculture in a medium with 20 μm IBA. TDZ (25 μm) stimulated callus production from the cotyledonary nodes and radicles. Adventitious buds emerged on the callus surface and internally in callus. Chemical names used: a -naphthaleneacetic acid (NAA); 6-benzylaminopurine (BA); indole-3-butyric acid (IBA); N-phenyl-N'-1,2,3-thidiazol-5-ylurea (TDZ).
Multiple shoots were obtained from shoot tips (2 to 3 mm) derived from mature plants (5 to 6 years old) of Citrus reticulata Blanco cv. Khasi mandarin and C. limon Burm.f. cv. Assam lemon when cultured on Murashige and Skoog (MS) medium, supplemented with (mg·liter-1) 1.0 BAP, 0.5 kinetin, and 0.5 NAA. Root induction was observed when 7-week-old single shoots (≈ 2 cm long) of both Citrus species were cultured on MS medium supplemented with (mg·liter-1) 0.25 BAP, 0.5 NAA, and 0.5 IBA. These plantlets were successfully established in the soil. Chemical names used: naphthalene acetic acid (NAA), indole 3-butyric acid (IBA), and benzylamino purine (BAP).
Hardwood and softwood stem cuttings of 5-year-old Atlantic white cedar [Chamaecyparis thyoides (L.) B.S.P.] were cut to 12-cm (short) or 24-cm (long) lengths, treated with 0 to 15 g IBA/liter in 50% isopropyl alcohol, and rooted in a raised greenhouse bench under intermittent mist. When hardwood cuttings were collected in February, short cuttings survived and rooted better than long cuttings. Survival and percent rooting for softwood cuttings collected in late August was virtually 10070 regardless of cutting length. Long cuttings produced more roots and longer roots with hardwood and softwood material. IBA was unnecessary for rooting, but it markedly increased the number of roots. Chemical name used: 1H-indole-3-butyric acid (IBA).
Softwood shoots of apple (Malus domestica Borkh.) rootstocks M.9 and MM.106 were banded with Velcro for up to 20 days before cuttings were propagated. Percent rooting and the number of roots per cutting were significantly improved by banding for 10 to 20 days, with and without IBA application. As the duration of stem banding increased from 0 to 20 days, percent rooting and the number of roots of both M.9 and MM.106 cuttings increased linearly or curvilinearly. Stem banding also stimulated budbreak of cuttings. In M.9, banding resulted in a higher survival rate and increased new shoot growth of transplanted cuttings after 4 months. Percent budbreak and new shoot growth were highly correlated with the number of roots per cutting in both cultivars. The effects of stem banding on budbreak and subsequent growth of the cuttings were largely due to the enhanced rooting of cuttings. Chemical names used: 1H-indole3-butyric acid (IBA).
Two cold-tolerant species (Eucalyptus macarthurii Deane et Maiden and E. smithii R.T. Baker), a cold-tolerant hybrid (E. macarthurii×E. grandis Hill ex Maiden), and E. saligna Sm. were propagated in vitro from nodal explants collected from field-grown seedlings and from clonal hedges. Shoot growth was initiated on modified Murashige and Skoog (MS) medium containing BA at 0.1 mg·liter-1. Modified MS medium with BA (0.2 mg·liter-1) and NAA (0.01 mg·liter-1) was most effective in promoting shoot proliferation. Root initiation was achieved on half-strength modified MS medium with 2 mg IBA/liter. Rooted plants were hardened and established in the field. Chemical names used: N-(phenylmethyl)-1EZ-purin-6-amine (BA); 2-(1-naphthyl)acetic acid (NAA); 1H-indole-3-butyric acid (IBA).
Scion wood of `Desirable' pecan [Carya illinoinensis (Wangenh.) K. Koch] was grafted onto the lateral roots of 70-year-old `Van Deman' seedling rootstocks for evaluation as an alternative to planting nursery-grown trees for orchard cultivar conversion. Grafting treatments included application of IBA, method of grafting, position of graft, and grafting time. Survival was higher for grafts treated with IBA than those without IBA, for modified bark grafts positioned beneath the soil line than for either modified hark grafts positioned above the soil line or inlay grafts, and for grafts made 6 to 8 weeks after budbreak than later in the season. Techniques developed in this study demonstrate that cultivar conversion of > 75% is possible. Chemical name used: lH -indole-3-butyric acid (IBA).
The relative stabilities of IAA and IBA under various tissue culture procedures were determined. IBA was significantly more stable than IAA to autoclaving. IBA was also found to be more stable than IAA in liquid Murashige and Skoog medium (MS) under growth chamber conditions. The stabilities of IBA and IAA were similar in agar-solidified MS. Light provided by cool-white fluorescent bulbs promoted degradation of IAA and IBA in both liquid and agar media. Activated charcoal in concentrations as high as 5% was found to adsorb more than 97% of IAA and IBA in liquid MS. These results have important implications for the preparation, storage, and handling of IBA and IAA in plant tissue culture. Chemical names used: indole-3-acetic acid (IAA); indole-3-butyric acid (IBA).
Axillary shoots of cacao (Theobroma cacao L.), induced in vitro with cytokinins (BA or TDZ), elongated and produced leaves only in the presence of cotyledons and/or roots. Detached axillary shoots, which do not grow in `vitro under conventional tissue culture protocols, rooted with auxin and developed normally in vivo. Detached axillary shoots from cotyledonary nodes and single-node cuttings from mature plants were induced to elongate and produce normal leaves in the presence of 20,000 ppm CO2 and a photosynthetic photon flux density (PPFD) of 150 to 200 μmol·s-1·m-2. Subculture nodal cuttings continued to elongate and produce leaves under elevated CO2 and light levels, and some formed roots. Subculture of microcuttings under CO2 enrichment could be the basis for a rapid system of micropropagation for cacao. Chemical names used: N -(phenylmethyl) -1 H -purin-6-amine (BA); 1 H -indole-3-butyric `acid (IBA); α -naphthaleneacetic acid (NAA); thidiazuron (TDZ).
Plantlets were recovered from axillary bud cultures of muscadine grape (Vitis rotundifolia, `Summit'). Nodal segments 0.5 to 1.0 cm long were cultured in Murashige and Skoog (MS) basal medium supplemented with 5, 10, 20, or 40 μm BA. Best total shoot production was obtained with 10 μm BA; with higher BA levels, shoots were unexpanded and exhibited high mortalities. MS medium supplemented with IBA enhanced rooting by increasing rooting percentage and number per plantlet. Shoots previously proliferated on medium with 5 μm BA rooted significantly better than those multiplied on 10 μM BA. Shoot vigor during rooting was greater in shoots proliferated on 5 vs. 10 μm BA. Root development was not significantly affected by liquid vs. agar-solidifted medium or shoot length. Chemical names used: N-(phenylmethyl) -1H-purin-6-amine (BA), 1H-indole-3-butyric acid (IBA).