Micropropagation of Juvenile Eucalyptus regnans (Mountain Ash)

1991 ◽  
Vol 39 (2) ◽  
pp. 179 ◽  
Author(s):  
C Blomstedt ◽  
J Cameron ◽  
P Whiteman ◽  
SF Chandler

Node-derived shoot cultures of Eucalyptus regnans were established from in vitro grown seedlings on Murashige and Skoog basal medium supplemented with 0.5 mg L-1 (2 μm) zeatin and 0.05 mg L-1 (0.3 μm) napthaleneacetic acid. A double sterilisation method was essential to obtain clean material from seed. Microcuttings from established cultures were used to develop an efficient method for in vitro rooting. Rooting was best after a 7 day pulse on 20 mg L-1 (98 μm) indolebutyric acid. Hoagland's or Woody Plant Medium supported better rooting than MS basal medium and rooting was significantly enhanced by subculture to activated charcoal after the auxin pulse. Carbohydrate (sucrose or glucose) was essential for rooting while high light intensity was inhibitory. Optimal light conditions were a 12 h day (17 W m-2). In all, 90% of plantlets established in the nursery survived the winter.

2014 ◽  
Vol 68 (4) ◽  
pp. 275-280 ◽  
Author(s):  
Branka Vinterhalter ◽  
Vladimir Orbović ◽  
Dragan Vinterhalter

Shoot cultures of <em>Gentiana punctata</em> L. were inoculated with suspension of <em>Agrobacterium rhizogenes</em> strain A4 M70GUS. Hairy roots which appeared 2-3 weeks later were cultured on hormone-free, liquid, WPM (Lloyd and McCown 1980) basal medium for more than 5 years (60 subcultures). Growth rate of transformed roots was higher than the growth rate of nontransformed roots. Spontaneous shoot regeneration occured only in three culture vessels in subcultures No. 40 and 42. Plants had phenotype characteristics typical for <em>A. rhizogenes</em> transformed plants including: wrincled leaves, short internodes, plagiotropic roots and in general their growth rate was reduced. These plants also manifested precocious formation of flower buds without vernalization and flowering under in vitro conditions. Flowers were pale yellow, the same as in the standard phenotype.


2000 ◽  
Vol 10 (3) ◽  
pp. 397-400
Author(s):  
J.R. Fu ◽  
X.M. Huang ◽  
S.Q. Songa

AbstractThe plumules of newly-excised wampee embryos, which are more sensitive to dehydration than the roots, became more resistant to water loss when axes were allowed to sprout on woody plant medium [WPM; McCown and Lloyd (1981) Hortscience16, 453] before being dried. Pre-treatment of sprouting axes (seedlings) with sucrose incorporated in the WPM enhanced survival. Although the roots withered following further dehydration of seedlings cultured on WPM containing 60% sucrose, excised plumules were capable of generating adventitious roots when a combination of 10 mM α-napthaleneacetic acid and 10 mM indole-3-butyric acid was used during subsequent in vitro incubation.


Plants ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 755
Author(s):  
Angela Ricci ◽  
Luca Capriotti ◽  
Bruno Mezzetti ◽  
Oriano Navacchi ◽  
Silvia Sabbadini

In the present study, an efficient system for the in vitro regeneration of adventitious shoots from the peach rootstock Hansen 536 leaves has been established. Twenty regeneration media containing McCown Woody Plant Medium (WPM) as a basal salt supplemented with different concentrations and combinations of plant growth regulators (PGRs) were tested. Expanded leaves along with their petiole from 3-week-old elongated in vitro shoot cultures were used as starting explants. The highest regeneration rate (up to 53%) was obtained on WPM basal medium enriched with 15.5 μM N6-benzylaminopurine (BAP). The influences on leaf regeneration of the ethylene inhibitor silver thiosulphate (STS) and of different combinations of antibiotics added to the optimized regeneration medium were also investigated. The use of 10 μM STS or carbenicillin (238 μM) combined with cefotaxime (210 μM) significantly increased the average number of regenerating shoots per leaf compared to the control. In vitro shoots were finally elongated, rooted and successfully acclimatized in the greenhouse. The results achieved in this study advances the knowledge on factors affecting leaf organogenesis in Prunus spp., and the regeneration protocol described looks promising for the optimization of new genetic transformation procedures in Hansen 536 and other peach rootstocks and cultivars.


HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 693b-693
Author(s):  
Xiaoling Yu ◽  
Barbara M. Reed

Multiplication and elongation of shoot cultures established from mature trees of hazelnut cvs. Nonpareil and Tonda Gentile Romana were affected by changes in basal medium, carbon source and concentration, cytokinin and agar concentration. Explants on DKW medium produced significantly more shoots than those on Anderson medium or modified woody plant medium for chestnut. Explants on DKW medium with 3% glucose or fructose gave more and longer shoots than those with the other carbon sources. Cytokinins 6 benzylaminopurine (BA) and zeatin were more effective in producing shoots than kinetin and 2iP. On BA supplemented medium, the best multiplication rate was obtained with 1.5 - 2.0 mg/l. Explants grown on 0.4% agar produced more shoots than those on 0.6%, however, prolonged culture on 0.4% agar caused vitrification of lower parts of the plants. Shoot multiplication rates of these two cultivars were similar, but `Nonpareil' produced longer shoots than `Tonda Gentile Romana'.


HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 873D-873
Author(s):  
Yiqin Ruan ◽  
Mark H. Brand

Rhododendron `Montego' shoot cultures initiated from plants with and without tissue proliferation (TP and NTP) served as explant sources for all studies (Note: in vitro TP shoot cultures produce primarily dwarf shoots, some long shoots, and stem tumors). Calli induced from TP leaves and tumors and NTP leaves were cultured on woody plant (WP) medium containing NAA and 2-iP. During the first 4 weeks of culture, calli from NTP leaves had higher relative growth rates than calli from TP leaves or tumors. However, calli from TP leaves and tumors grew faster than calli from NTP leaves for all subculture periods that followed. Shoot tips (5 mm) were excised from TP dwarf shoots, TP long shoots, and NTP shoots and were cultured on WP medium with or without 15 μM 2-iP. Shoot tips from TP dwarf and long shoots multiplied on medium without 2-iP, averaging 18.4 and 1.7 shoots per shoot tip in 12 weeks, respectively. Shoot tips from NTP shoots only multiplied when maintained on 2-iP-containing medium. When placed on 2-iP-containing medium, both types of TP shoot tips produced clusters of callus-like nodules that gave rise to highly tumorized, short shoots or leafy meristems.


Author(s):  
Yelnititis Yelnititis ◽  
Sri Sunarti

In vitro culture is a promising technique for mass propagation of high-value species. Study of propagation for Acacia hybrid (A. mangium x A. auriculiformis) through this technique has been conducted using single node stem from seedlings as explants. Growth medium used was modified Murashige and Skoog (MS), basal medium Woody Plant Medium (WPM), and Gamborg (B5) supplemented. The study was conducted in two stages, namely shoot induction and shoot multiplication. The treatment tested was the Benzyl Adenine (BA) supplementation at the concentration of 0.3; 0.7; and 1.0 mgL-1 of. Observation was conducted on the frequency of shoot induction, number of shoot, shoot length and visual performance of the culture. The result showed that treatment of BA 0.7 mgL-1 on modified MS medium is the best for shoot induction, shoot multiplication and visual performace of the culture. The average of number of shoot was 2.6; 5.0 and 7.7 shoots on the first three consecutive subcultures. Changing to different basal medium on the fourth subculture showed that the treatment of BA 0.7 mgl-1 is the best condition for shoot regeneration (12.60 shoots) and shoot length (6.97 cm). The culture from this treatment showed the best visual morphological performance.Keywords:Acacia hybrid; multiplication; subculture; in vitro; BA. ABSTRAKKultur in vitro merupakan suatu teknik yang menjanjikan untuk perbanyakan massal spesies-tanaman bernilai tinggi. Penelitian perbanyakan akasia hibrid (A. mangium x A. auriculiformis) melalui kulturin vitro telah dilakukan dengan menggunakan eksplan berupa batang satu buku yang berasal dari anakan. Media tumbuh yang digunakan adalah media dasar Murashige dan Skoog (MS) yang sudah dimodifikasi, media dasar Woody Plant Medium (WPM), dan Gamborg (B5). Penelitian dilakukan dalam dua tahap yaitu induksi tunas dan perbanyakan tunas. Perlakuan yang diuji adalah penggunaan Benzyl Adenine (BA) dengan konsentrasi 0,3; 0,7 dan 1,0 mg L-1. Pengamatan dilakukan terhadap waktu induksi tunas, jumlah tunas, tinggi tunas dan penampilan biakan secara visual. Hasil penelitian menunjukkan bahwa penggunaan BA 0,7 mg L-1 pada media MS modifikasi merupakan perlakuan terbaik untuk induksi tunas, perbanyakan tunas, tinggi tunas, dan kondisi biakan secara visual. Jumlah rata-rata tunas yang dihasilkan dari perlakuan ini adalah 2,6; 5,0 dan 7,7 tunas pada subkultur pertama, kedua dan ketiga. Pada penggunaan media dasar berbeda pada subkultur keempat menunjukkan bahwa perlakuan BA 0,7 mg L-1 merupakan perlakuan terbaik dengan jumlah tunas sebanyak 12,60 tunas dan rata-rata tinggi tunas 6,97 cm. Biakan yang dihasilkan dari perlakuan tersebut mempunyai penampilan yang baik dan normal.


2014 ◽  
Vol 71 (3) ◽  
pp. 181-186
Author(s):  
Halina Wysokińska ◽  
Katarzyna Lisowska ◽  
Katarzyna Floryanowicz-Czekalska

Shoot buds isolated from in vitro shoot cultures of <em>Catalpa ovata</em> G. Don were encapsulated using 3% sodium alginate with sucrose (3%) and 50 mM calcium chloride. The morphogenic response of encapsulated buds was affected by such factors, like composition of the media and the presence of growth regulators. The highest frequency of plantlet germination from encapsulated buds (70% within 4 weeks) was obtained on Woody Plant medium (WP) (Lloyd and McCown 1980) containing indole-3-butyric acid (IBA) (1 mg/l). The process was substantially inhibited by cold-storage (4<sup>o</sup>C) of encapsulated buds. In this case, the frequency response ranged from 3% to 22% dependent on storage period (28 or 42 days) and the presence of the paraffin coat covering the alginate capsules. The plantlets developed from both unstored and stored encapsulated buds of <em>C. ovata</em> were transplanted to soil and grew in pots to phenotypically normal plants.


2008 ◽  
Vol 44 (No. 2) ◽  
pp. 66-72 ◽  
Author(s):  
L. Klčová ◽  
M. Gubišová

Plant regeneration by different techniques was evaluated in three buckwheat cultivars: indirect regeneration from cotyledons and hypocotyls, direct regeneration by nodal segment cultivation and induction of multiple shoots from seedling apices. Regenerated shoots were obtained in all procedures. The effects of BA (6-benzylaminopurine), media composition and gelling agent were tested. The regeneration efficiency of shoot apex culture was 2.65&ndash;3.33 nodal segments/explant. Cotyledon and hypocotyl segments produced 1.25&ndash;2.44 shoots per explant plated. Nodal segment cultivation yielded 4.1&ndash;4.8 new nodal segments/explant in 4 weeks. Eighty percent of shoots rooted on the basal medium. Rooting was improved (up to 95.6%) by IBA (3-indolebutyric acid) addition to the culture medium. Regenerated plantlets were transferred to the soil. The most efficient and simple micropropagation of buckwheat was nodal segment cultivation on MS medium solidified by agar with the addition of 1 mg/l BA. This method is advisable for rapid multiplication, in vitro conservation or rescue of genetic resources.


2013 ◽  
Vol 5 (3) ◽  
pp. 332-337 ◽  
Author(s):  
Mouaad Amine MAZRI

The effect of basal medium, explant size and density on shoot multiplication, growth, rooting and acclimatization of date palm cv. ‘16-bis’ was evaluated. Bud clusters of different sizes (2, 3, 4 and 5 buds per cluster) were cultured at density of 1, 2, 3 and 4 clusters on Murashige and Skoog medium (MS), woody plant medium (WPM) and Nitsch medium (NM) supplemented with 0.5 mg/L 2-naphthoxyacetic acid and 0.5 mg/L kinetin for three months (multiplication phase). Separated shoots of different sizes (


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