Morphogenesis in Punica granatum (pomegranate)

1986 ◽  
Vol 64 (8) ◽  
pp. 1644-1653 ◽  
Author(s):  
Kanchan Jaidka ◽  
P. N. Mehra

Explants of root, hypocotyl, cotyledon, stem, shoot tip, and leaf of seedlings obtained by in vitro germination of seeds, as well as embryos excised from seeds, were utilized for the induction of callus. Murashige and Skoog basal medium supplemented with naphthaleneacetic acid (4 ppm), kinetin (2 ppm), and coconut water (15%) was found to be optimal for induction and growth of callus from all explants. Growth rate experiments were performed with callus to study the effect of different growth regulators at various concentrations. The calluses were heterogeneous in nature and consisted predominately of diploid cells, although a few polyploid cells were also observed after two and four subcultures. Plantlets, isolated roots, leaves, and shoots were differentiated in various callus cultures. The root tips and shoot tips of such plantlets revealed only diploid constitution. Embryolike structures were formed in callus on transfer to media containing naphthaleneacetic acid and 6-benzylaminopurine. Embryoid development was traced to a single cell which was invariably isolated from the rest of the callus tissue. This initial divided to form a multicelled structure which later gave rise to a globular, ovoid, or heart-shaped embryoid, or one with irregular form. The embryoids germinated into complete plantlets with root and shoot. The embryoidal initials were mostly diploid but occasional aneuploids or polyploids were observed.

2009 ◽  
Vol 2009 ◽  
pp. 1-7 ◽  
Author(s):  
Dahanayake Nilanthi ◽  
Xiao-Lu Chen ◽  
Fu-Cheng Zhao ◽  
Yue-Sheng Yang ◽  
Hong Wu

Petiole explants were obtained from in vitro grown diploid (2x=22)Echinacea purpureaplantlets. Shoots were regenerated by culturing the explants on MS basal medium containing 0.3 mg/L benzyladenine (BA), 0.01 mg/L naphthaleneacetic acid (NAA) and four concentrations (30, 60, 120, and 240 mg/L) of colchicine for 30 days, or 120 mg/L of colchicine for various durations (7, 14, 21, and 28 days). The regenerated shoots were induced to root on MS basal medium with 0.01 mg/L NAA, and then the root-tips of the regenerated shoots were sampled for count of chromosome number. It was found that a treatment duration of >7 days was necessary for induction of tetraploid (4x=44) shoots, and treatment with 120 mg/L colchicine for 28 days was the most efficient for induction of tetraploids, yielding 23.5% of tetraploids among all the regenerated shoots. Chimeras were observed in almost all the treatments. However, the ratio of tetraploid to diploid cells in a chimeric plant was usually low. In comparison with diploid plants, tetraploid plants in vitro had larger stomata and thicker roots with more root branches, and had prominently shorter inflorescence stalk when mature.


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


2013 ◽  
Vol 8 (11) ◽  
pp. 1934578X1300801 ◽  
Author(s):  
Anahi Bucchini ◽  
Laura Giamperi ◽  
Donata Ricci

This is the first report on the antioxidant and antifungal activities of callus cultures from Inula crithmoides L. (Asteraceae). Callus cultures were initiated from leaf sections, on initial culture MS basal medium supplemented with various concentrations of 2,4-D (2,4-dichlorophenoxyacetic acid), NAA (1-naphthaleneacetic acid) and IBA (indole-3-butyric acid) and a 72% survival was achieved. Significant differences between the various auxins used as phytohormones on callus growth were found. Maximum callusing was noticed on the leaf explants grown on MS basal medium supplemented with 1 mgL–1 2,4-D. Subsequently the antioxidant and antimicrobial activities of the methanol extract from calli were investigated. Antioxidant studies suggested that the methanol extracts of dark-grown and light-grown callus were able to reduce the stable free radical 2,2-diphenyl-1-picrilhydrazyl (DPPH). In the inhibition against lipid peroxidation, extracts of dark-grown callus showed the strongest effect with IC50 values better than those of the standards. The methanol extract of callus cultures had significant antifungal activity only against two of the fungi tested: Alternaria solani and Phytophthora cryptogea. Against all the other tested fungi, the I. crithmoides calli extracts showed fungistatic activity.


2018 ◽  
Vol 22 ◽  
pp. 274-281
Author(s):  
N. B. Kravets ◽  
N. V. Tulaidan ◽  
M. Z. Mosula ◽  
N. M. Drobyk

Aim. The aim of the research was to choose the conditions for microclonal propagation and obtain callus cultures from Carlina аcaulis L., Carlina cirsioides Klok and Carlina onopordifolia Besser ex Szafer, Kulcz. et Pawl plants in vitro. Methods. For microclonal propagation of С. acaulis, C. cirsioides and C. onopordіfolia we used rosettes of 2–3-month specimens and planted them on semi-solid Murashige and Skoog (MS) medium with decreased macro- and microsalts concentrations (MS/2) supplemented with kinetin (Кin) (from 1–3 mg/l) and 0.1 mg/l of 1-naphthaleneacetic acid (NAA). For induction of callus formation, we used root, stem explants from С. acaulis, C. cirsioides and C. onopordіfolia, and planted them on nutrient media MS, MS/2, and Gamborg and Eveleigh (В5) supplemented with different concentrations of cytokinins – 6-benzylaminopurine (BAP) or Кin and auxins – 2.4-dichlorophenoxyacetic acid (2.4-D) or NAA and indole-3-acetic acid (IAA). Results. MS/2 medium supplemented with growth regulators of NAA and Кin were the most efficient to provide the formation of microclones. For C. сirsioides plants, this indicator was 6.6–6.8 rosettes per graft after 6 months of cultivation and for С. acaulis and C. onopordіfolia – 4.2–5.0 and 4.8–5.2 respectively. To raise the percentage of rooting for microclones of Carlina species, it was expedient to steep them preliminarily in the solution of indole-3-butyric acid (IBA) with 1000 mg/l concentration for a minute. Optimal for obtaining callus tissue from Carlina plants was nutrient medium MS supplemented with 3 mg/l IAA, 0.5 mg/l NAA and 0.5 mg/l Kin and MS/2 with 0.1 mg/l BAP and 0.5 mg/l 2.4-D; under such conditions the percentage of callus induction exceeded 90 % for all types of explants. Conclusions. There were chosen the conditions for microclonal propagation of С. acaulis, C. cirsioides and C. onopordіfolia and worked out the schemes for enrooting obtained microclones in vitro. Capable of growing rapidly callus cultures from root and stem explants of the investigated plant species were obtained. Keywords: Carlina аcaulis L., Carlina cirsioides  Klok, Carlina onopordifolia Besser ex Szafer, Kulcz. et Pawl, in vitro, microclonal propagation, callus induction.


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


1987 ◽  
Vol 65 (1) ◽  
pp. 72-75 ◽  
Author(s):  
J. Y. Peron ◽  
E. Regnier

A method for rapid micropropagation of sea kale (Crambe maritima L.) was developed. Petiole explants placed in vitro on a medium containing 0.5 mg/L indoleacetic acid (IAA), 6.0 mg/L kinetin, and 1.5 mg/L benzylaminopurine developed callus within 15 days and shoots within 28 days. Nearly four adventitious shoots could be developed within 3 weeks by placing the initial shoot on media without IAA. To develop roots, the shoots were then transferred to the basal medium containing 0.1 to 1.0 mg/L indolbutyric or α-naphthaleneacetic acid. Rooted plantlets were obtained within 2 or 3 weeks. After an acclimatization period of 6 weeks in a greenhouse in unsterilized medium, the plantlets could be set outdoors.


1980 ◽  
Vol 22 (4) ◽  
pp. 607-614 ◽  
Author(s):  
Chi-Chang Chen ◽  
Chung-Mong Chen

Forty-six callus cultures of rice (Oryza sativa L., 2n = 24), each presumably originating from a single microspore, were established and maintained on a medium containing 2,4-D. At the end of the first transfer 24% of the cultures were nonhaploid consisting of only diploid or polyploid cells, or of cells of two ploidy levels. Nuclear fusion and endomitosis occurring during the initial stages of in vitro microspore development were postulated to account for the formation of nonhaploid callus. Seventeen cultures were studied cytologically through 19 transfers. Only in one tetraploid and one hexaploid callus did the ploidy levels of cells remain unchanged during culture. Chromosome numbers in 13 cultures fell into a geometric series. Since no diplo- and quadruplochromosomes were observed, it was inferred that endomitosis rather than endoreduplication was responsible for the changes. In spite of the tendency for chromosome doubling, the proportions of cells of different ploidy levels were fixed in the 13 cultures at later transfers. Haploid cells were eliminated from all cultures. Diploid cells became predominant in eight cultures and tetraploid cells in five, suggesting a selection for either cell type. Triploids appeared in two cultures which initially did not contain this type of cell. Limited cytological information indicated that triploid cells might have originated from tetraploid cells through reductional grouping of chromosomes accompanied by multipolar formation.


2018 ◽  
Vol 25 (03) ◽  
pp. 90-96
Author(s):  
Otgonpurev S ◽  
Munguntsooj B ◽  
Ariunjargal B ◽  
Munkhtsetseg D

Papaver nudicaule  L has a long history as a being useful for the treatment of many diseases in Asian and European countries. Aim of this study was to cultivate callus and cell suspension culture in vitro using plant phytohormones. The proliferative capacity was tested on shoot cultivated on Murashige-Skoog (MS) basal medium testing auxins: -Naphthaleneacetic acid (NAA)  in combination with cytokinine: 6-Benzylaminopurine. We determined production of alkaloids by four-week old callus and cell suspension biomass of Papaver nudicaule using thin layer chromatography comparing standard sanguianarine. This result displayed us the easier approach to isolate pure one alkaloid from the biomass that those whole- plant. Нүцгэн намуу ургамлын in vitro биомасс дахь алкалоид Хураангуй: Нүцгэн намуу (Papaver nudicaule) нь эмийн болон чимэглэлийн ач холбогдолтой ургамал юм. Уг ургамлын үрээс каллусын болон эсийн суспензийн биомассыг гарган авахдаа нафтален цууны хүчил болон бензил аденин өсөлтийн бодисын хослол бүхий тэжээлт орчинд өсгөвөрлөн өсөлтийн параметрүүдийг тогтоолоо. Ургамлын in vitro биомасст нимгэн үеийн хроматографийн аргаар сангвинарин алкалоидыг стандарт бодистой харьцуулан тодорхойлов. Сангвинарин нь ургамлын эд эрхтэний төрөлжилтөөс үл хамааран үүсдэг биологийн өндөр идэвхтэй хоёрдогч нийлэгжилтийн бүтээгдэхүүн юм. Түлхүүр үг: өсөлтийн гормоны хослол, каллусын эд, суспензийн нягт, нимгэн үеийн хроматографи


2021 ◽  
Vol 40 ◽  
pp. 01001
Author(s):  
Elen Poghosyan ◽  
Naira Sahakyan ◽  
Margarit Petrosyan ◽  
Irina Batlutskaya ◽  
Karen Trchounian

A growing demand for the ecologically pure products brings us for searching novel biotechnological approaches for plant cultivation. One of these approaches is the in vitro cultivation and further acclimatization of valuable plant species. The object of our investigation was Ajugareptance L. ornamental plant which possesses high metabolic activity. In vitro cultivation was carried out applying Murashige-Skoog nutrient medium and its modifications. Acclimatization of in vitro plants was implemented according Hazarika. In the presence of twice higher concentration of cytokinins over auxins and 0.2 mg/ml gibberellins callus culture was formed from the leaf explants. Callus tissue was formed in the presence of 0.2 mg/ml kinetin and 2 mg/ml indole-3-acetic acid which has denser structure than the first one. The shoot formation was observed on callus cultures growing on the same medium approximately after 5th passage. Callus culture growth was supported also by the adding of 2 mg/ml 2,4-dichlorophenoxyacetic acid. For the micropropagation, the already formed shoots were transferred to the nutrient medium which contains only 0.1 mg/ml 1-Naphthaleneacetic acid as a phytohormone. A. reptans culture has high regenerative ability and the micro-propagation index was 104 – 105. In vitro regenerated plants were successfully acclimatized to the soil conditions during two-week period.


Author(s):  
Rebaz Rasul Habas ◽  
Musa Turker ◽  
Fethi Ahmet Ozdemir

An efficient plant regeneration protocol was developed from in vitro germinated seeds of Petunia hybrida an ornamentally important plant in the family Solanaceae. Shoot tip and node explants of Petunia hybrida were cultured on MS basal medium supplemented with different concentrations and combinations of Benzyl amino purine (BAP), 1-Naphthaleneacetic acid (NAA), Indole-3-butyric acid (IBA) and Gibberellic acid (GA3). The highest shoot length was obtained from MS medium supplemented with 1 mg/l BAP + 1 mg/l NAA. The highest shoot number (3 shoots/explant) were obtained from MS medium supplemented with 0.6 mg/l BAP + 0.5 mg/l IBA. The isolated shoots were transferred to MS basal medium supplemented with different concentrations of GA3 ranging from 0.05, 0.2, 0.5 and 1 mg/l for shoot elongation. The highest shoot length (5.75 cm) was recorded from the MS medium supplemented with 0.2 mg/l GA3 +0.2 mg/l BAP. Rooting of regenerated shoots were achieved on MS medium supplemented with 0.1-1 mg/1 IBA and NAA. The regenerated shoots with well developed roots were successfully acclimatized and established in pots containing sterilized peat moss and grown under laboratory conditions with 70% survival rates.


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