Comparative in vitro and early nursery performance of adventitious shoots from cryopreserved cotyledons and axillary shoots from epicotyls of the same zygotic embryo of control-pollinated Pinus radiata

2005 ◽  
Vol 35 (11) ◽  
pp. 2629-2641 ◽  
Author(s):  
Cathy L Hargreaves ◽  
Lynette J Grace ◽  
Susan A van der Maas ◽  
Mike I Menzies ◽  
Satish Kumar ◽  
...  

This is the first published report comparing production and performance of adventitious shoots from cryopreserved cotyledons, with axillary shoots formed from epicotyls of the same zygotic embryo of radiata pine (Pinus radiata D. Don). Genotypes from 10 control-pollinated families of P. radiata in two treatments were compared for shoot initiation, in vitro growth, rooting, and early nursery performance. Plant growth in nursery beds was assessed by measuring height after 2 and 7 months. After 8 months in nursery beds, the physiological ages of genotypes were assessed before field planting. Genotype capture was higher from the cryopreserved cotyledons than from the epicotyls. This technique has the advantage of preserving juvenile material while field testing is done. Early shoot multiplication in both treatments was good. After four transfer cycles, epicotyl cultures showed improved elongation and higher multiplication rates. After 6 months of in vitro growth, shoots from both treatments were given auxin pulses. Shoots of adventitious origin were slower to root than epicotyl-derived shoots. Overall rooting rates were satisfactory. Plants of adventitious origin were shorter when planted into nursery beds and when transferred to field trials. Assessment of relative physiological age indicated that all plants of adventitious origin showed some increase.

HortScience ◽  
2018 ◽  
Vol 53 (10) ◽  
pp. 1482-1486 ◽  
Author(s):  
Huan Xiong ◽  
He Sun ◽  
Feng Zou ◽  
Xiaoming Fan ◽  
Genhua Niu ◽  
...  

Castanea henryi is an important woody grain tree species native to China. The objective of the current study was to find the suitable plant growth regulators (PGRs) and the optimal concentrations for direct organogenesis by using axillary shoots and cotyledonary nodes. Seeds were collected from the field, sterilized, and germinated in vitro. Axillary shoots and cotyledonary nodes of 3-week-old seedlings were used as explants. To find the suitable PGR for adventitious shoot induction, 0.5 mg·L–1 6-benzylaminopurine (6-BA), 0.1 mg·L–1 indole-3-acetic acid (IAA), 0.1 mg·L–1 2,4-dichlorophenoxyacetic acid (2,4-D), or 0.1 mg·L–1 1-naphthaleneacetic acid (NAA) was supplemented to Murashige and Skoog (MS) medium containing 0.65% agar and 3% sucrose. A high induction percentage of adventitious shoots (85.67%) was obtained from cotyledonary nodes supplemented with 0.1 mg·L–1 2,4-D. The type of explant influenced shoot proliferation rates and quality. Apical explants produced more and longer shoots than nodal segments. For shoot multiplication, 1 mg·L–1 6-BA + 0.05 mg·L–1 indole-3-butyric acid (IBA) supplemented with MS medium produced 12.33 and 6.25 shoots per explant, respectively, from apical and nodal explants. For shoot elongation and strengthening, 2 mg·L–1 6-BA + 0.05 mg·L–1 IBA supplemented with MS medium was the best combination, producing shoots with a mean length of 3.50 cm, a diameter of 0.46 cm, and about eight leaves per shoot. The greatest rooting of 76.70% and 11.33 roots per shoot was achieved when cultured in MS medium supplemented with 3.5% perlite + 1.5 mg·L–1 IBA. For acclimatization of the rooted plantlets in the greenhouse, a survival rate of 80% was achieved. This protocol—from multiplication to acclimation—is helpful to realize mass propagation of high-quality trees of chinquapin for increasing production and nut quality.


1985 ◽  
Vol 63 (12) ◽  
pp. 2172-2176 ◽  
Author(s):  
Victor M. Villalobos ◽  
Edward C. Yeung ◽  
Trevor A. Thorpe

Previous studies on shoot initiation in cultured excised cotyledons of Pinus radiata indicated that the process began early in culture on the side of the cotyledon in contact with the cytokinin-containing medium. In contrast to cotyledons cultured without cytokinin, organized structures, which have been termed promeristemoids, can be observed in cotyledons cultured with cytokinin after 5 days in culture. These structures arise from single cells in the first subepidermal cell layer at day 3. They are stable and their continued division leads to the formation of meristemoids, shoot primordia, and finally shoots with primary needles.


HortScience ◽  
1999 ◽  
Vol 34 (3) ◽  
pp. 460E-460 ◽  
Author(s):  
Virginia Miller-Roether ◽  
Paul E. Read ◽  
Erika Szendrak

The American Chestnut Foundation (ACF) has conducted a breeding program aimed at developing blight-resistant chestnut trees exhibiting the phenotype of American Chestnut (Castanea dentata). We developed a protocol for in vitro micropropagation and multiplication of candidate blight-resistant plants from the ACF breeding program. The protocol included forcing dormant shoots to budbreak, culture establishment, shoot multiplication, inducing a functional root system on the microcuttings produced by this system and establishment of autotrophic plants. Because Castanea spp. is recalcitrant to rooting, a unique bilayer method of rooting was developed. The unique bilayer consisted of a clear basal medium of 50% DKW and 50% WPM (Long and Preece), with a continuous level of 0.01 mg IBA/L and 0.2 mg BA/L. The clear basal medium was over-laid with an opaque layer. Rooting response occurred for 27 of the 31 genotypes at various frequencies. Rooted plantlets were planted in 50% peat: 50% perlite in order to become autotrophic and acclimated. Acclimated trees were planted in 10″ × 2″ Deepots® and placed in the greenhouse. These trees exhibited a very vigorous functional root system. Acclimated trees were hardened off, placed in cold storage (≈4-5 °C) for 5 months. All trees placed in cold storage broke dormancy for spring growth and ≈100 trees were sent to ACF for planting into field trials.


Author(s):  
Jiangli Shi ◽  
Zhidan Dong ◽  
Chunhui Song ◽  
Beiyang Xie ◽  
Xianbo Zheng ◽  
...  

Rootstocks play a vital role in regulating the environmental adaptability and controlling the growth and development of apple trees. M9T337, an excellent apple rootstock widely used in commercial orchards, could confer dwarf tree architectures, early fruiting and suitability for high-density planting. However, the rooting ability of M9T3337 is low when it is vegetatively propagated, and researchers have not yet established an efficient micropropagation system. The present study systematically evaluated the multiplication in adventitious shoots and the in vitro formation of adventitious roots to determine the effects of the culture media and plant growth regulators of M9T337 and a rapid micropropagation system was developed. For the shoot multiplication, the highest multiplication index of 3.93 was obtained on Murashige and Skoog (MS) media supplemented with 2.0 mg/L 6-BA, 0.1 mg/L NAA and 0.3 mg/L GA3 from 12 combinations of 6-BA and NAA. Stronger and taller adventitious shoots were grown on MS supplemented with 1.8 mg/L 6-BA and 0.5 mg/L NAA. The optimal media with 100% rooting was obtained using ½ MS supplemented with 0.3 mg/L IBA or MS supplemented with 0.6 mg/L IBA for the rooting induction, resulting in mean rooting numbers of 13.00 and 11.33, respectively. Additionally, the effect on rooting of adding 0.3 mg/L IBA or not on the 1/2 MS and MS media was compared; the results suggested that an appropriate IBA concentration was the key to successful rooting. The rooted plantlets were acclimatised in a shaded greenhouse with an 84% survival rate. The established micropropagation system could be used for the rapid propagation of M9T337 for commercial production.


Biologia ◽  
2014 ◽  
Vol 69 (5) ◽  
Author(s):  
Achuthan Hemanthakumar ◽  
Thankappan Preetha ◽  
Padmesh Pillai ◽  
Peringatulli Krishnan ◽  
Sooriamuthu Seeni

AbstractAxillary shoots were induced from shoot tip of Calamus thwaitesii suckers on Murashige and Skoog medium supplemented with 0.4 mg/L N6-benzylaminopurine and 0.1 mg/L each of thidiazuron and α-naphthaleneacetic acid (NAA). The shoots initiated were subcultured to fresh media of the same composition for shoot multiplication and multiplied shoots were transferred to half strength MS hormone-free media for shoot elongation. The elongated shoots (∼5cm) were then re-cultured to the media supplemented with 3.0 mg/L indole-3-butyric acid/4.0 mg/L NAA to raise plantlets which were subsequently analysed for genetic fidelity using inter simple sequence repeat markers. Out of 183 bands scored, 178 bands were monomorphic indicating 97.2% similarity. The observed low level of polymorphism between genotypes supports genetic consistency of these micro-clones that are likely to be genetically true to their parental origin. The clones thus obtained were hardened in the specially fabricated mist house at 29 ±2°C and 80±5% relative humidity for 3 months followed by shifting to green house for another 3 months of nursery establishment. The established plants when reintroduced to the selected forest segments of the Western Ghats, Kerala (India) showed 79.3% survival rate after 2 years of field transfer. The viable and highly reproducible in vitro cloning protocol demonstrated here for the first time can be used for the production of elite female clones for aforestation activities and sustained delivery of high quality raw materials to cane processing units for strengthening cane industry.


2020 ◽  
Vol 22 (1) ◽  
pp. 59
Author(s):  
K.K. Boguspaev ◽  
S.K. Turasheva ◽  
T.M. Seilkhanov ◽  
D.G. Faleev ◽  
M.S. Mutalkhanov ◽  
...  

Scorzonera tau-saghyz Lipsch. et G.G. Bosse is an endemic rubber producing plant, growing in mountain regions in South Kazakhstan. The rubber content in plants and the quality of biopolymer has an important impact on industrial rubber production. The results of this study showed that the amount of rubber in S. tau-saghyz roots fluctuates between 7.74% and 38.75%. The amount of synthesized and deposited rubber biopolymer particles depends on various factors such as physiological age of plant, origin, temperature, moisture and environmental conditions. We optimized the extraction method of natural rubber by using n-hexane as a solvent for direct extraction. This method allows extracting the maximum amount of rubber from 3‒4-year-old plants. NMR results show structural links of natural isoprene rubber in the root extract sample. There is a clear relationship between methyl, methine and methylene protons which corresponds to isoprene rubber structure. The samples having strongly marked singlets that are inherent for rubber functional groups confirms the stereospecific structure of rubber. Good solubility of the root extract in deuterated chloroform can characterize the low molecular weight of the polymer. NMR characterization of rubber, extracted from S. tau-saghyz roots, is reported for the first time. Regeneration in vitro provides an important opportunity for endemic preservation by rapidly increasing the number of plants. The best regeneration of adventitious shoots was obtained on MS medium containing 5.5 μM kinetin and 0.5 μM NAA. The plants were successfully acclimatized in a glasshouse with 75% of S. tau-saghyz plantlets, respectively surviving after transfer to ex vitro conditions.


1991 ◽  
Vol 116 (1) ◽  
pp. 149-155 ◽  
Author(s):  
John E. Preece ◽  
Carl A. Huetteman ◽  
W. Clark Ashby ◽  
Paul L. Roth

During 1987, we selected the six fastest-growing seedlings or clones from each of 15 provenances that represented the natural distribution range of silver maple (Acer saccharinum L.). Shoots from all 90 trees were cut into nodal segments, rooted as cuttings, and maintained as clonal stock plants in the greenhouse. Rooting was generally excellent and more than half of the clones rooted ≥90%. At the same time, explants were obtained from these field-grown trees and many were established in vitro as aseptic cultures by first pretreating with benomyl and rifampicin. Single-node explants from the greenhouse-grown clonal stock plants were also established and multiplied in vitro. There was a significant effect of clone within provenance on all in vitro growth characteristics. All clones proliferated axillary shoots, but not all at the same rates. Although statistically significant, low correlation coefficients indicated that micropropagation results were not good predictors of nursery performance of the populations from which the clones were selected, nor of the climatic conditions at the site of origin of the trees. The micropropagation system reported herein, therefore, should be applicable to a wide variety of silver maple genotypes. Chemical name used: methyl [1-[butylamino)carbonyl]-1H-benzimidazol-2-yl]carbamate(benomyl).


1986 ◽  
Vol 4 (1) ◽  
pp. 26-29
Author(s):  
Frank A. Blazich ◽  
Cheryl G. Giles ◽  
Carole M. Haemmerle

Shoot tips excised from Chapman's rhododendron (Rhododendron chapmanii A. Gray) were surface sterilized, the terminal portions were removed (decapitated) and the shoots placed in liquid Woody Plant Medium (WPM) supplemented with 8 μM (1.6 ppm) 6(γ, γ-dimethylallylamino)-purine (2iP). Within 2 to 3 months axillary shoots excised, decapitated and cultured on agarsolidified WPM supplemented with 49 μM (10 ppm) 2iP. Multiple shoot formation consisting of adventitious and axillary shoots was observed within 4 to 6 months. These shoots were transferred to WPM supplemented With 8 μM (1.6 ppm) 2iP and cultured under reduced light levels to stimulate shoot elongation. Shoots ≥ 10 mm (0.4 in.) were harvested (microcuttings) and rooted using non-in vitro procedures. Enhancement of axillary shoot multiplication was achieved by culturing decapitated axillary shoots under reduced light levels in a horizontal position on WPM supplemented WIth 8 μM (1.6 ppm) 2iP.


HortScience ◽  
1991 ◽  
Vol 26 (5) ◽  
pp. 594-596 ◽  
Author(s):  
Giovanni Iapichino ◽  
Tony H.H. Chen ◽  
Leslie H. Fuchigami

An efficient adventitious shoot production protocol has been developed for Rhododendron laetum × aurigeranum. Shoot tips taken from greenhouse-grown plants were cultured on Anderson's medium supplemented with 74 μM 2iP. Axillary shoots were excised and cultured on medium containing 23 μM IAA and 74 μM 2iP. After 6 months, brown callus developed at the cut surfaces of the shoot-tip explants. This callus produced many adventitious shoots (up to 70 per explant). Clusters of adventitious shoots were divided, subculture, and continued to proliferate shoots. An estimated 1600-fold increase in the number of shoots could be readily achieved in 6 months. In vitro rooting of adventitious shoots was accomplished in 4 weeks. Seventy-three percent of shoots rooted on 1/4 strength Anderson's medium supplemented with 28 μm IAA. Plantlet survival was 100%3 weeks after transfer to soil. Chemical names used: 1-H-indole-3-acetic acid (MA); N-(3 -methy1-2-butenyl) -1H-purine-6 amine (2iP).


2011 ◽  
Vol 72 (3) ◽  
pp. 181-190 ◽  
Author(s):  
Małgorzata Podwyszyńska ◽  
Agnieszka Marasek

The effects of TDZ and paclobutrazol on the primary regeneration on tulip flower stalk explants of six cultivars and subsequent shoot multiplication were examined. Explants, flower stalk slices, were excised from cooled and subsequently forced bulbs. The explants were incubated for two months in darkness on medium containing NAA and cytokinins, 2iP and BAP, as control, or TDZ (0.5-4 mg l<sup>-1</sup>) and paclobutrazol (0.05-0.4 mg l<sup>-1</sup>). Then, the regenerating explants were subcultured on medium with TDZ and NAA applied at low concentrations. Different regeneration capabilities were found depending on cultivar and growth regulators. The percentage of explants forming leaf-like structures ranged, on the control medium, from 80% in 'Blue Parrot' and 'Prominence' to below 30% in 'Apeldoorn' and 'Mirjoran'. TDZ, applied at optimum for each cultivar concentration, greatly increased the regeneration potential up to 70-100%. Paclobutrazol, added to the TDZ-containing medium, significantly enhanced the response of explants, resulting in high numbers of leaf-like structures formed per explant (13.7-22.8). The structures developed gradually into characteristic forms: the growing up cotyledonary leaf, the probable root primordium formed at its base, the growing downwards stolon and the shoot meristem developed finely on its tip. It is suggested that such primary regeneration may have a nature of somatic embryogenesis. Then, the adventitious shoots developed and formed clusters, which were divided into 2-3 smaller ones every two months. The growth regulators, used at initial stage, markedly influenced subsequent shoot multiplication. Thus, the most intensive shoot formation was noted with TDZ at concentrations of 0.5-2 mg l<sup>-1</sup> and paclobutrazol of 0.05-0.1 mg l<sup>-1</sup>.


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