To improve the efficiency of iris plant regeneration, we tested the influence of several combinations of Kin and NAA in culture media on the induction of morphogenesis and the subsequent development of plantlets. The highest rates of regeneration (67%) were consistently observed in induction media containing 0.5 μm NAA and either 2.5 or 12.5 μm Kin. Developing medium containing 1.25 μm BA was optimal for high regeneration rates and a high percentage of plantlets simultaneously developing shoots and roots. Rooted plantlets were easily acclimatized and transplanted to various soil mixtures, then grown in the greenhouse. Under optimal conditions as many as 8000 plantlets could be regenerated from 1 g of cells in ≈4 months. Chemical names used: kinetin (Kin); 1-naphthaleneacetic acid (NAA); N6-benzyladenine (BA).
Yuexin Wang, Zoran Jeknić, Richard C. Ernst, and Tony H.H. Chen
Samir C. Debnath
The effects of TDZ (0, 0.1, 1, 5 and 10 μm) and explant orientation on adventitious shoot regeneration of `Erntedank' lingonberry were studied. Moderate concentration (1 to 5 μm) of TDZ supported bud and shoot regeneration, but strongly inhibited shoot elongation. TDZ initiated cultures were transferred to medium containing 1-2 μm zeatin and produced usable shoots after one additional subculture. Adventitious bud and shoot regeneration was greatly influenced by explant orientation. Elongated shoots were rooted on a 2 peat: 1 perlite (v/v) medium, and the plantlets were acclimatized and eventually established in the greenhouse with 80% to 90% survival rate.
James A. Kapaun and Zong-Ming Cheng
Plants were regenerated from leaf tissue of greenhouse-grown seedlings of Siberian elm (Ulmus pumila L.). Shoot regeneration was induced on Murashige and Skoog (MS) medium containing 5 to 10 μm of BA. Up to 55% of the leaf explants formed shoots with an average of 2.4 shoots per explant. Addition of 2.5 or 5 μm of IBA failed to enhance regeneration. Thidiazuron at 0.5 or 1.0 μm also induced shoot regeneration, but the shoots failed to elongate as well as shoots regenerated from media containing BA. Incubation in darkness for 7, 14, or 21 d had little effect in promoting shoot regeneration, except that incubation for 21 d increased shoot regeneration on the medium with 5.0 μm BA. Genotypes differed in shoot regeneration potential, with regeneration frequencies ranging from 13% to 55%. Regenerated shoots were micropropagated on Driver and Kuniyuki Walnut medium. Ninety percent of microcuttings rooted directly in potting soil. This regeneration system will be valuable for genetic transformation and cell selection of Siberian elm. Chemical names used: 6-benzylaminopurine (BA); indole-3-butyric acid (IBA); N-phenyl-N′ -1,2,3-thiadiazol-5-ylurea (thidiazuron, TDZ).
Constance A. Parks and Thomas H. Boyle
The effects of stratification, BA, thiourea, and GA3 were examined on germination of Liatris spicata (L.) Willd. seed. Seeds were germinated at 20 °C and numbers of germinated seed were counted daily for 21 days. The final germination percentage (G) for seeds stored dry at 4 °C for 0 to 10 weeks ranged from 52% to 64%, while stratification at 4 °C for 10 weeks increased G to 98% and decreased the days to 50% of final germination (T50) and the days between 10% and 90% germination (T90-T10). Aqueous solutions of BA at 10 or 100 mg·L-1 applied to blotter paper increased G and decreased T50 but did not affect T90-T10. In a separate experiment, dry seeds were treated for 3 minutes in BA at 0 to 1126 mg·L-1 dissolved in acetone. G values increased quadratically, whereas T50 and T90-T10 values decreased quadratically in response to BA concentration. A 3-minute preplant acetone permeation of seeds with BA at 225 or 1127 mg·L-1 yielded G and T50 values similar to those obtained with 10 weeks of stratification. Seeds immersed in thiourea at 0.76 or 7.61 mg·L-1 for 24 hours prior to sowing had higher G and lower T50 values than controls (0 mg·L-1 thiourea), but T90-T10 values were similar for all treatments. Seeds treated with GA3 at 1, 10, or 100 mg·L-1 in H2O did not differ from controls (0 mg·L-1 GA3) in G, T50, or T90-T10. Infusion of BA via acetone may be a practical means of breaking seed dormancy and accelerating germination in L. spicata. Chemical names used: N-(phenylmethyl)-1H-purine-6-amine [benzyladenine (BA)]; gibberellic acid (GA3); thiocarbamide (thiourea).
Juan Guillermo Cruz-Castillo
Anil P. Ranwala and William B. Miller
The effects of Promalin® [PROM; 100 mg·L–1 each of GA4+7 and benzyladenine (BA)] sprays on leaf chlorosis and plant height during greenhouse production of ancymidol-treated (two 0.5-mg drenches per plant) Easter lilies (Lilium longiflorum Thunb. `Nellie White') were investigated. Spraying with PROM at early stages of growth [36 or 55 days after planting (DAP)] completely prevented leaf chlorosis until the puffy bud stage, and plants developed less severe postharvest leaf chlorosis after cold storage at 4 °C for 2 weeks. When PROM was sprayed on plants in which leaf chlorosis had already begun (80 DAP), further leaf chlorosis was prevented during the remaining greenhouse phase and during the postharvest phase. PROM caused significant stem elongation (23% to 52% taller than controls) when applied 36 or 55 DAP, but not when applied at 80 DAP or later. The development of flower buds was not affected by PROM treatments. Although PROM sprays applied at 55 DAP or later increased postharvest flower longevity, earlier applications did not. Chemical names used: N-(phenylmethyl)-1H-purine 6-amine (benzyladenine, BA); α-cyclopropyl-α-(p-methoxyphenyl)-5-pyrimidinemethanol (ancymidol).
Virginia M. Keith and Mark H. Brand
Significant occurrences of phenotypic variation have been noted in micropropagated Rhododendron. Studies were undertaken to determine what aspects of micropropagation lead to variation. Rhododendron `Molly Fordham' was used to evaluate growth parameters and the incidence of variation in plants that originated from 3 month and 54 month old cultures. Plants from 3-month-old cultures were significantly wider than plants from 54-month-old cultures. Rhododendron `Aglo', `Molly Fordham', and `Scintillation' were used to evaluate growth and the incidence of variation in plants grown from microcutting bases and rerooted microcutting tips (retips). Three-month-old retips were significantly taller and wider than bases of the same age, but possessed fewer branches. The influence of in vitro N6-[2-isopentenyl]adenine (2-iP) concentration on the growth and phenotype of regenerated plants of `Aglo', `Molly Fordham', and `Scintillation' was examined. Data taken 3 months post-acclimation indicate that growth and the incidence of variation in response to 2-iP concentration is cultivar dependent.
Ryutaro Tao and Akira Sugiura
Callus cultures were initiated in the dark from leaf primordia, stem internodes, and young leaves of adult Japanese persimmon (Diospyros kaki L.) to induce adventitious buds. A high frequency of regeneration occurred on Murashige and Skoog medium (MS) with half the normal NH4NO3 and KNO3 concentration (1/2N) and containing 10 μm zeatin or 1 μm 4PU-30 in combination with 0.1 μm IAA, or MS(1/2N) medium containing 0.03 to 0.1 μ m IAA or 0.01 to 0.03 μm NAA combined with 10 μm zeatin. No significant differences in the capacity of regeneration were observed among the calli from different explant sources. Only eight of 16 cultivars formed adventitious buds on MS(1/2N) medium containing 10 μm zeatin and 0.1 μm IAA, with the percentage of explants forming adventitious buds ranging from 2% to 72%. Chemical names used: indole3-acetic acid (IAA); 1-naphthaleneacetic acid (NAA); N-phenyl-N'-(2-chloro-4-pyridyl)urea (4PU-30).
O. Pérez-Tornero, F. Ortín-Párraga, J. Egea, and L. Burgos
Apricot (Prunus armeniaca L. cv.'Helena') shoots grown on a proliferation medium containing 3% sucrose, 0.4 mg·L–1 benzyladenine (BA), and 0.04 mg·L–1 indolebutyric acid (IBA) and solidified with 0.6% agar were stored at three different temperatures in the dark for up to 24 weeks. All shoots remained viable for 24 weeks when stored at 3 °C, while at 14 °C the percentage of survival decreased quickly after 12 weeks of storage. At 7 °C, percentage of survival started to decline after 18 weeks of storage. Shoots stored at 3 °C had the highest regeneration rates and shoot lengths following transfer to standard proliferation conditions. This temperature also had a beneficial effect on shoot proliferation during the first 12 to 18 weeks of the experiment.
Xabier Barandiaran, Nieves Martín, María Fernanda Rodríguez-Conde, Antonio Di Pietro, and Jesus Martín
The influence of different callus induction media on the regeneration process in garlic was tested. The auxin 2,4 dichlorophenoxyacetic acid frequently used in garlic tissue culture was found to be detrimental when used at the levels described in the literature. However, combinations of growth regulators commonly used for dicot tissue culture produced high levels of callus induction and regeneration that could be used efficiently in a transformation program.