In Vitro Reproductive Development of a Diploid Wild Species, Arachis duranensis1

1994 ◽  
Vol 21 (2) ◽  
pp. 139-143
Author(s):  
Q. L. Feng ◽  
H. E. Pattee ◽  
H. T. Stalker

Abstract Embryo abortion at an early stage of reproductive development is a major impediment for introgressing germplasm from wild to cultivated species of Arachis by interspecific hybridization. Ovule and embryo culture techniques have been used to rescue aborting hybrid embryos, but increased efficiency and recovery of very young tissues are still needed. The objective of this study was to induce growth and differentiation of A. duranensis proembryos. Seven-, 10-, and 14-d-old peg tips were cultured on a modified basal medium containing MS and B5 media combinations with 16 combination treatments using three growth regulators—1-naphthaleneacetic acid, gibberellic acid, and 6-benzylaminopurine—each at four levels. The results showed that seeds could be obtained in vitro by peg tip culture of four- to 16-celled proembryos. The favorable concentration ranges of growth regulators for pod formation and embryo development were 0.5-2.0 mg/L NAA, 0.05-0.5 mg/L GA3, and 0.05-0.2 mg/L 6-BAP. Over all three selected ages of pegs, the three best combinations of growth regulators resulted in 4.8, 4.7, and 3.5% pod formation, respectively.

2021 ◽  
Author(s):  
Yuan-yuan Meng ◽  
Shi-jie Song ◽  
Sven Landrein

Abstract Passiflora xishuangbannaensis (Passifloraceae) is endemic to a few sites of Mengyang nature reserve in Yunnan, Xishuangbanna and less than 40 individuals have been recorded. Nine Passiflora species are endemic to Yunnan with most species occurring in South America, making P. xishuangbannaensis highly significant and emblematic to the conservation work in the region. This study is designed to provide the first protocol for in vitro organogenesis and plant regeneration for ex situ conservation and reintroduction for an Asian Passiflora species. Using internodes, petioles and tendrils we optimize calli formation and root elongation using several plant growth regulators, individually or in combination. We also assess the genetic stability of regenerated cells. The maximum callus induction and shoot bud differentiation were both achieved on half Murashige and Skoog basal medium supplemented with 4.44 µM 6-Benzylaminopurine and 1.08 µM 1-Naphthaleneacetic acid. The best rooting was achieved from 30 days old, regenerated shoots on half Murashige and Skoog basal medium supplemented with 1.08 µM 1-Naphthaleneacetic acid. Micropropagated plants were subjected to inter simple sequence repeat markers analyses. Collectively, 86 bands were generated from 6 primers of which 12 bands were polymorphic, showing genetic variation between the regenerated plantlets and the original plant. Response to plant growth regulators was more specific than most other studies using South American species, which could be explained by the morphological and physiological differences between South American and Asian Passiflora species


HortScience ◽  
2000 ◽  
Vol 35 (5) ◽  
pp. 948-952 ◽  
Author(s):  
Luping Qu ◽  
James Polashock ◽  
Nicholi Vorsa

A very efficient adventitious regeneration (shoot organogenesis) system for cranberry (Vaccinium macrocarpon Ait.) leaves was developed. A basal medium consisting of Anderson's rhododendron salts and Murashige and Skoog's (MS) organics, supplemented with 10.0 μm thidiazuron (TDZ) and 5.0 μm 2ip, was effective for adventitious regeneration from leaves for the five cranberry cultivars tested: `Early Black', `Pilgrim', `Stevens', `Ben Lear', and `No. 35'. Parameters examined included: 1) varying combinations of three plant growth regulators (TDZ, 2ip, and NAA); 2) explant orientation (adaxial vs. abaxial side in contact with the medium); and 3) leaf position relative to the apical meristem from the source plant. Cultivars varied in regeneration frequency, but cultivar × growth regulator interaction was nonsignificant. With optimal treatment conditions, regeneration occurred on more than 95% of the explants, with `Early Black' and `Pilgrim' producing as many as 100 shoot meristems per explant. At all concentrations tested, NAA (as low as 0.1 μm) increased callus formation and significantly reduced regeneration. Emerging adventitious shoots were always observed on the adaxial side of the leaves regardless of explant orientation on the medium. Regeneration was much greater when the abaxial side was in contact with the medium, and was not related to leaf position on the source plants. Elongation of adventitious shoots began ≈2 weeks after transfer to the basal medium without growth regulators. Cuttings of elongated shoots rooted 100% both in vitro in the basal medium and ex vitro in shredded sphagnum moss. The high regeneration efficiency achieved by using this system will be very useful in the application of techniques, such as Agrobacterium- and particle bombardment-mediated transformation. Chemical names used: 1-phenyl-3-(1,2,3-thiadiazol-5-yl) urea (thidiazuron, TDZ); N6-(γ-γ-dimethyallylamino) purine (2ip); α-naphthaleneacetic acid (NAA).


1993 ◽  
Vol 73 (3) ◽  
pp. 871-878 ◽  
Author(s):  
Hélène Desilets ◽  
Yves Desjardins ◽  
Richard R. Bélanger

Different culture media were compared at the initiation and multiplication steps to develop a rapid production system for geranium (Pelargonium × hortorum) in vitro. Different salt dilutions of the Murashige and Skoog (MS) (1962) mineral medium were used in combination with different concentrations of 1-naphthaleneacetic acid (NAA) and 6-benzyladenine (BA) in order to optimize initiation of shoots of four geranium cultivars. The use of a MS basal medium with half-strength macrosalts supplemented with 0.11 μM NAA and 0.89 μM BA gave the best results in initiation. More than 40% of the apices initiated on this medium produced multiple shoots within a month. Subsequently, the effect of different concentrations of growth regulators was quantified by the mean of "shoot doubling time" evaluation. The shortest time recorded was 10.5 d for a theoretical production of 1 × 109 plantlets/apex/year. This is the first quantitative evaluation of geranium production in vitro. Geranium plantlets rooted easily on a half-strength MS medium without growth regulators. Acclimatization of geranium plantlets was characterized by high survival rates (94%) and the plants thus produced were phenotypically comparable to seed-derived plants. Key words: Geranium, micropropagation, shoot doubling time, in vitro


2001 ◽  
Vol 11 (2) ◽  
pp. 206-208 ◽  
Author(s):  
M.A. Norton ◽  
R.M. Skirvin

A method has been developed for micropropagation of the difficult-to-root winegrape cultivar `Norton' (Vitis aestivalis). Plants were established in vitro from axillary bud cuttings of field-grown plants. Four levels of 6-benzylaminopurine (BA) and three levels of naphthaleneacetic acid (NAA) were tested in a factorial arrangement for their effectiveness in promoting multiplication of shoots from single-node explants. Three levels of NAA and two concentrations of Murashige and Skoog (MS) basal medium were tested for their effectiveness in promoting rooting of shoot tips. The greatest number of shoots per axillary bud in combination with the greatest shoot length were produced with 4 μmol·L-1 [0.90 mg·L-1 (ppm)] BA. NAA had no effect on shoot multiplication. NAA was not required for in vitro rooting. All rooted plants survived the transition to soil.


1995 ◽  
Vol 22 (2) ◽  
pp. 135-141
Author(s):  
Q. L. Feng ◽  
H. E. Pattee ◽  
H. T. Stalker

Abstract Research on in vitro embryo culture in Arachis has the objective of rescuing interspecific hybrid embryos which abort before they reach maturity. This study explored effects of the three exogenous plant growth regulators 1-naphthaleneacetic acid (NAA), gibberellic acid (GA3), and 6-benzylaminopurine (6-BAP); sucrose; and medium pH on in vitro fruit and embryo development of A. hypogaea L. by culturing 10-d-old peg tips. Results indicated that medium containing 0.5 to 1.0 mg L-1 NAA was optimal for in vitro pod formation and embryo development. GA3 did not have a significant influence and 6-BAP had negative effects on both in vitro fruit and embryo development. High concentrations of 6-BAP and NAA induced callus which inhibited ovary enlargement and embryo development. Sixty g L-1 sucrose was the best concentration for ovary enlargement and embryo development. Acidic medium was needed for in vitro reproductive development with pH 4.5–6.5 the most favorable. A pod formation frequency of 81%, a seed production rate of 90% (from pods recovered in vitro), and plant recovery of 33% were obtained for a medium containing 1.0 mg L-1 NAA and 0.5 mg L-1 GA plus 60 g L-1 sucrose at pH 5.8. In vitro-recovered cotyledonary embryos between 4 and 10 mm long germinated precociously into seedlings at relatively higher frequencies than morphologically mature embryos which produced more vigorous plants.


1970 ◽  
Vol 19 (1) ◽  
pp. 89-99
Author(s):  
K. Choudhary ◽  
M. Singh ◽  
M. S. Rathore ◽  
N. S. Shekhawat

This long term study demonstrates for the first time that it is possible to propagate embryogenic Vigna trilobata and to subsequently initiate the differentiation of embryos into complete plantlets. Initiation of callus was possible on 2,4-D. Somatic embryos differentiated on modified MS basal nutrient medium with 1.0 mg/l  of 2,4-D and 0.5 mg/l  of Kn. Sustained cell division resulted in globular and heart shape stages of somatic embryos. Transfer of embryos on to a fresh modified MS basal medium with 0.5 mg/l of Kn and 0.5 mg/l of GA3 helped them to attain maturation and germination. However, the propagation of cells, as well as the differentiation of embryos, were inhibited by a continuous application of these growth regulators. For this reason, a long period on medium lacking these growth regulators was necessary before the differentiation of embryos occurred again. The consequences for improving the propagation of embryogenic cultures in Vigna species are discussed. Key words: Pasture  legume, Vigna trilobata, Globular, Heart shape, somatic embryogenesis D.O.I. 10.3329/ptcb.v19i1.4990 Plant Tissue Cult. & Biotech. 19(1): 89-99, 2009 (June)


Agronomy ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 320
Author(s):  
Nisar Ahmad Zahid ◽  
Hawa Z.E. Jaafar ◽  
Mansor Hakiman

Ginger (Zingiber officinale Roscoe) var. Bentong is a monocotyledon plant that belongs to the Zingiberaceae family. Bentong ginger is the most popular cultivar of ginger in Malaysia, which is conventionally propagated by its rhizome. As its rhizomes are the economic part of the plant, the allocation of a large amount of rhizomes as planting materials increases agricultural input cost. Simultaneously, the rhizomes’ availability as planting materials is restricted due to the high demand for fresh rhizomes in the market. Moreover, ginger propagation using its rhizome is accompanied by several types of soil-borne diseases. Plant tissue culture techniques have been applied to produce disease-free planting materials of ginger to overcome these problems. Hence, the in vitro-induced microrhizomes are considered as alternative disease-free planting materials for ginger cultivation. On the other hand, Bentong ginger has not been studied for its microrhizome induction. Therefore, this study was conducted to optimize sucrose and plant growth regulators (PGRs) for its microrhizome induction. Microrhizomes were successfully induced in Murashige and Skoog (MS) medium supplemented with a high sucrose concentration (>45 g L−1). In addition, zeatin at 5–10 µM was found more effective for microrhizome induction than 6-benzylaminopurine (BAP) at a similar concentration. The addition of 7.5 µM 1-naphthaleneacetic acid (NAA) further enhanced microrhizome formation and reduced sucrose’s required dose that needs to be supplied for efficient microrhizome formation. MS medium supplemented with 60 g L−1 sucrose, 10 µM zeatin and 7.5 µM NAA was the optimum combination for the microrhizome induction of Bentong ginger. The in vitro-induced microrhizomes sprouted indoors in moist sand and all the sprouted microrhizomes were successfully established in field conditions. In conclusion, in vitro microrhizomes can be used as disease-free planting materials for the commercial cultivation of Bentong ginger.


Author(s):  
Ileana MICLEA ◽  
Rita BERNAT

The aim of the current research was to find the best plant growth regulators for the multiplication of Sarracenia purpurea. Murashige and Skoog medium (MS) was prepared with macronutrients and micronutrients at 1/3 strength, full strength vitamins, supplemented with 30 g/l sucrose and 5 g/l phytagel and autoclaved. After cooling 0.5 mg\l α-naphthaleneacetic acid (NAA), 5 mg\l 6-benzyladenine (BA) or 0.5 mg\l NAA + 3 mg\l BA were added. Young S. purpurea plants were selected and transferred to media with or without plant growth regulators and cultured for 12 weeks. At the end of this time frame number of roots, root length (cm) and number of shoots were evaluated and differences were analysed by the analysis of variance and interpreted using the Tuckey test. The largest number of roots grew in medium supplemented with 0.5 mg\l NAA but the the absence of plant growth regulators increased their length. The best conditions for shoot multiplication were provided by supplementing 1/3MS with 5 mg\l BA.


2008 ◽  
Vol 34 (No. 2) ◽  
pp. 77-83 ◽  
Author(s):  
S. Kumar ◽  
V. Awasthi ◽  
K. Kanwar J

The influence of growth regulators and nitrogenous compounds on in vitro bulblet formation and growth was studied in two hybrids of <i>Lilium</i>. Bulbscales isolated from pre-cooled bulbs of hybrids Rosato and Marco Polo were used. The basal portion with plate (5 &times; 6 mm) of inner bulbscales was cultured on Murashige and Skoog (MS) medium containing 0.5 or 1 mg/dm<sup>3</sup> naphthaleneacetic acid (NAA) and/or benzyladenine (BA). The presence of NAA (0.5 mg per dm<sup>3</sup>) showed higher explant regeneration, producing about three bulblets per explant as compared to control. About four bulblets per explant were produced at both concentrations of BA. The bulblets with significantly higher fresh weight were obtained on medium containing NAA. Approximately a three-fold increase of bulblet fresh weight was observed with all the concentrations of TDZ in both cultivars. The bulblets cultured with nitrogenous compounds after attaining the size of 14&minus;16 cm flowered during the second year of the growing period without any phenotypic variations.


2003 ◽  
Vol 83 (4) ◽  
pp. 873-876 ◽  
Author(s):  
A. N. Aziz ◽  
R. J. Sauvé ◽  
S. Zhou

Daylily (Hemerocallis sp. ‘Stella de Oro’) callus cultures initiated from ovules were bombarded with gold particles coated with plasmid harboring Basta® resistance gene. Resulting putative transgenic calli were selected after 3 wk on semi-solid Murashige and Skoog’s (MS) basal medium supplemented with 10 mg L-1 1-naphthaleneacetic acid, 2 mg L-1 6-benzylaminopurine and 3 mg L-1 phosphinothricin (PPT). Surviving calli regenerated shoots after 2 mo on semi-solid MS medium supplemented with 2 mg L-1 thiadiazuron and 1 mg L-1 PPT. Polymerase chain reaction and Southern blotting were used to confirm independent transformation events. Key words: Basta® resistance, in vitro, Hemerocallis


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