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1.
Shelake Rahul Mahadev Angappan Kathithachalam Murugan Marimuthu 《In vitro cellular & developmental biology. Plant》2011,47(5):611-617
In vitro propagation has played a key role for obtaining large numbers of virus free, homogenous plants, and for breeding of plantains
and bananas (Musa spp.). Explant sources utilized for banana micropropagation include suckers, shoot tips, and floral buds. The present study
employed male floral meristems as explant material for micropropagation of hill banana ecotypes (AAB) ‘Virupakshi’ and ‘Sirumalai.’
Immature male floral buds were collected from healthy plants from hill banana growing areas. Exposure of explants to ethyl
alcohol (70%, v/v) for 30 s, then mercuric chloride (0.1%, w/v) for 30 s, followed by three independent rinses of 5 min each in autoclaved, double-distilled water satisfactorily reduced
the contamination. Male floral bud explants were cultured on Murashige and Skoog (MS) basal medium supplemented with different
combinations of 6-benzylaminopurine (BAP), coconut water, naphthaleneacetic acid, gibberellic acid, and additional supplements.
MS medium supplemented with 5 mg l−1 BAP and coconut water (15%) was the most efficient media for shoot initiation and multiple shoot formation (15 shoots from
a single part of a floral bud). The best response for shoot elongation was obtained using the combination of basal MS, 5 mg l−1 BAP, 1 mg l−1 naphthaleneacetic acid and 1.5 mg l−1 gibberellic acid. Regenerated shoots were rooted in basal MS medium within 15–20 d. The rooted plantlets were transferred
to a soil mixture and maintained at a temperature of 25 ± 2°C for 10 d and then at room temperature (30–32°C) for 2 wk, before
transferring to a greenhouse. The regenerated plantlets showed 100% survival. 相似文献
2.
Su-Juan Zhao Zhong-Chun Zhang Xiang Gao Gulsum Tohsun Bao-Sheng Qiu 《Plant Cell, Tissue and Organ Culture》2009,99(1):9-16
An efficient micropropagation system for mining ecotype Sedum alfredii Hance, a newly identified Zn/Cd hyperaccumulator, was developed. Frequency of callus induction reached up to 70% from leaves
incubated on Murashige and Skoog (MS) medium supplemented with 1.0 mg l−1 2,4-dichlorophenoxy acetic acid (2,4-D) and 0.5 mg l−1 6-benzyladenine (BA), and 83% from internodal stem segments grown on MS medium with 0.1 mg l−1 2,4-D and 0.1 mg l−1 BA. Callus proliferated rapidly on MS medium containing 0.2 mg l−1 2,4-D and 0.05 mg l−1 thidiazuron. The highest number of adventitious buds per callus (17.3) and frequency of shoot regeneration (93%) were obtained
when calli were grown on MS medium supplemented with 2.0 mg l−1 BA and 0.3 mg l−1 α-naphthalene acetic acid (NAA). Elongation of shoots was achieved when these were incubated on MS medium containing 3.0 mg l−1 gibberellic acid. Induction of roots was highest (21.4 roots per shoot) when shoots were transferred to MS medium containing
2.0 mg l−1 indole 3-butyric acid rather than either indole 3-acetic acid or NAA. When these in vitro plants were acclimatized and transferred
to the greenhouse, and grown in hydroponic solutions containing 200 μM cadmium (Cd), they exhibited high efficiency of Cd
transport, from roots to shoots, and hyperaccumulation of Cd. 相似文献
3.
Bilal Haider Abbasi Mubarak Ali Khan Tariq Mahmood Mushtaq Ahmad Muhammad Fayyaz Chaudhary Mir Ajab Khan 《Plant Cell, Tissue and Organ Culture》2010,101(3):371-376
The morphogenic potential and free-radical scavenging activity of the medicinal plant, Silybum marianum L. (milk thistle) were investigated. Callus development and shoot organogenesis were induced from leaf explants of wild-grown
plants incubated on media supplemented with different plant growth regulators (PGRs). The highest frequency of callus induction
was observed on explants incubated on Murashige and Skoog (MS) medium supplemented with 5.0 mg l−1 6-benzyladenine (BA) after 20 days of culture. Subsequent transfer of callogenic explants onto MS medium supplemented with
2.0 mg l−1 gibberellic acid (GA3) and 1.0 mg l−1 α-naphthaleneacetic acid (NAA) resulted in 25.5 ± 2.0 shoots per culture flask after 30 days following culture. Moreover,
when shoots were transferred to an elongation medium, the longest shoots were observed on MS medium supplemented with 0.5 mg l−1 BA and 1.0 mg l−1 NAA, and these shoots were rooted on a PGR-free MS basal medium. Assay of antioxidant activity of in vitro and in vivo grown
tissues revealed that significantly higher antioxidant activity was observed in callus than all other regenerated tissues
and wild-grown plants. 相似文献
4.
Soha E. Mostafa Nabila S. Karam Rida A. Shibli Feras Q. Alali 《Plant Cell, Tissue and Organ Culture》2010,103(1):111-121
A protocol for micropropagation of Arbutus andrachne from seeds was developed. Results indicated that none of the seeds cultured on Murashige and Skoog (MS) medium, with or without
plant growth regulators (PGRs), germinated. Seeds soaked in 250 mg l−1 gibberellic acid (GA3) at 4°C for 3 days, then cultured on water-agar medium containing 2.0 mg l−1 GA3 exhibited 80–100% germination and developed into usable seedlings. Shoot proliferation was tested on MS or B5 medium containing
different concentrations of cytokinin. No shoot proliferation was observed on PGR-free medium. Proliferation was more successful
on MS than on B5 medium. On both media, the most successful proliferation was obtained using zeatin as a cytokinin type. Rooting
was tested on MS medium containing different concentrations of auxin. Rooting failed on PGR-free medium and on medium containing
indole-3-acetic acid (IAA), 0.25 or 0.5 mg l−1 indole-3-butyric acid (IBA), or 0.25, 0.5 or 2.0 mg l−1 α-naphthaleneacetic acid (NAA). Rooting (40%) was most successful with 1.0 mg l−1 NAA. Rooted plantlets exhibited 80% survival in all mixtures of peatmoss and perlite, and acclimatized plants were successfully
grown in the greenhouse. Quantitative analysis of arbutin performed on in vivo and in vitro leaves using high-performance
liquid chromatography (HPLC) revealed that in vivo leaves contained higher arbutin content (0.3–0.81% w/w) than in vitro leaves
(0.09% w/w). The highest yield of arbutin in vivo was detected in leaves collected in August, and the lowest yield in leaves
collected in December. 相似文献
5.
Sandra Gonçalves Laura Fernandes Anabela Romano 《Plant Cell, Tissue and Organ Culture》2010,101(3):359-363
A novel protocol suitable for the micropropagation of the endangered species Tuberaria major using seedlings as explants is reported. Using this protocol, we studied the effects of explant type (apical shoots and nodal
segments) and cytokinins [6-benzyladenine (BA), kinetin, and zeatin (ZEA)] on shoot proliferation. Explant type significantly
influenced the proliferation frequency and mean number of shoots, with nodal segments showing a higher proliferation capacity.
The mean number of shoots was significantly higher when the explants were cultured in half-strength (1/2) MS medium supplemented
with 0.2 mg l−1 BA (6.83 ± 0.77 shoots) or ZEA (6.55 ± 0.71 shoots). The shoots showed a great rooting capacity that was significantly influenced
by the concentration of MS macronutrients but not by the concentration of auxins. The highest rooting frequencies (97–100%)
were obtained in 1/2 MS medium with or without plant growth regulators. The plants obtained were easily acclimatized to ex
vitro conditions, with 97% surviving after 6 weeks. The micropropagated plants were successfully reintroduced into their natural
habitat and exhibited normal development. In conclusion, our culture protocol, with efficient seed germination, subsequent
multiplication of nodal explants using ZEA at 0.2 mg l−1, and successful ex vitro establishment of well-rooted plantlets on 1/2 MS medium, provides a simple and reliable methodology
for the large-scale propagation of T. major, thereby contributing to germplasm preservation of this endangered species. 相似文献
6.
7.
A simple protocol for direct shoot organogenesis and plant regeneration in Lessertia frutescens using hypocotyl and cotyledon segments is reported. l-canavanine content in the derived shoots is also quantified. Media containing different concentrations and combinations of
the cytokinins kinetin (K) and benzyladenine (BA) were tested for shoot induction potential. The best shoot regeneration rate
(83%) was obtained from hypocotyl segments cultured in Murashige and Skoog (MS) medium supplemented with 1 mg l−1 K; these hypocotyls also produced the largest number of shoots per explant (3.5) and the longest shoots per explant (13.3
mm). The best shoot regeneration rate (46%) using cotyledons as explant material was obtained in MS medium supplemented with
1 mg l−1 K and 1 mg l−1 BA or with 5 mg l−1 K and 0.5 mg l−1 BA. The highest number of cotyledon-derived shoots (1.5) was obtained in MS medium containing 2 mg l−1 K and 0.5 mg l−1 BA, and the longest cotyledon-derived shoots (6.1 mm) were obtained in MS medium containing 1 mg l−1 K and 0.5 mg l−1 BA. Shoots derived from hypocotyls cultured on media containing 1 mg l−1 K contained the highest quantity of l-canavanine (1.42 mg g−1) relative to the control (0.52 mg g−1). Shoots derived from cotyledons cultured on media containing 2 mg l−1 K contained the highest quantity of l-canavanine (2.07 mg g−1) compared to the control. Scanning electron microscopy revealed that shoots regenerated directly from the wounded epidermal
tissue, although callus formation was observed in most cultures. Young shoot clusters proliferated into healthy adventitious
shoots that were subsequently transferred directly onto rooting medium (MS medium containing 4 mg l−1 indole-3-butyric acid), eliminating the need for an additional multiplication or elongation phase. The in vitro plants were
successfully acclimatized in a growth chamber, achieving an 85% survival rate. 相似文献
8.
In vitro regeneration protocol for Anethum graveolens (Apiaceae) was developed using leaf explants. MS basal medium used in experiments was augmented with various hormones for
caulogenic and rhizogenic response. The optimum callus induction (100%) was obtained by leaf explants on MS media fortified
with BA (0.5 mg l−1) singly and in combination with NAA (0.1 and 0.2 mg l−1). BA at 0.5 mg l−1, KN at 1.0 mg l−1 and NAA at 0.1 mg l−1 induced highest number of multiple shoots (10.0 ± 0.25) per explant and they also showed in vitro flowering within 3 weeks
of culture. Influence of adenine sulfate on regeneration frequency of callus was evaluated. The highest frequency of rooting
(100%) with 6.0 ± 0.25 roots per explants was obtained in one-fourth strength MS medium supplemented with 1/4 MS + IBA 0.5 mg l−1 within 4 weeks of transfer to the rooting medium. In vitro flowering (35%) was obtained with MS fortified with BA alone and
also in combination with KN and NAA (5.3 ± 0.42 flowers per explants). In vitro flowering response was tested with different
carbohydrates (fructose, glucose, mannose and sorbitol) and optimized. Hardening was successfully attained under controlled
conditions inside the plant tissue culture room. The proposed method could effectively be applied for the conservation and
clonal propagation to meet the pharmaceutical demands of this medicinally important species. 相似文献
9.
Junli Wang Jue Wang Kun Liu Xuan Xiao Weizhen Gong Yuan Lu Mingfei Liu Dongting Xu 《In vitro cellular & developmental biology. Plant》2010,46(5):445-450
An efficient micropropagation system for Hylotelephium tatarinowii (Maxim.) H. Ohba, a rare medicinal plant, has been developed. Callus induced from leaf explants placed onto Murashige and
Skoog (MS) medium with supplementation of plant growth regulators. When the concentration of 2,4-dicholorophenoxy acetic acid
was as high as 2.0 mg l−1 in combination with 0.5 mg l−1 6-benzylaminopurine (6-BAP), the callus induction rate reached 92.1%. Adventitious shoots were observed on callus exposed
to 1.0 mg l−1 6-BAP, with 81.5% frequency of shoot regeneration after 30 d. Flower buds appeared after subculture. Regenerated shoots could
flower normally in vitro. Up to 100% of the regenerated shoots formed complete plantlets on half-strength MS medium without any growth regulator, with
an average of 5.9 roots per shoot explant. Quantitative analysis of flavonoids and rutin showed that the phytochemical profile
of callus and regenerated plants was similar to that of wild plants. 相似文献
10.
The present study describes the potential of in vitro grown adventitious roots of Hypericum perforatum L. commonly known as St. John’s wort at low nutrient and auxin levels in the liquid medium for micropropagation. Roots were
regenerated from shoot-derived callus on MS medium containing 4.0 mg l−1 Indole-3 acetic acid (IAA). IAA and Indole-3 butyric acid (IBA) were equally effective for the induction of roots from shoot
cultures. Half strength MS medium containing 1.0 mg l−1 IAA was most found suitable for culturing roots in liquid medium. A total biomass of 4.13 ± 0.67 g comprising 226 ± 34.4
shoots and shoot buds along with roots was obtained per culture starting with 200 mg roots inoculum. Pretreatment with kinetin
(2.0 mg l−1) enhanced the shoot multiplication. Shoots proliferated profusely from excised roots in static liquid medium supported with
glass bead matrix. Growtek™ vessel was found suitable and cost effective system for high throughput plantlet production. In vitro grown roots regardless
of their source of origin were an excellent and easy to handle source of explant for aseptic production of plantlets without
loosing the morphogenetic potential over the generations. The plants exhibited 84–99% similarity among themselves through
RAPD. The in vitro shoots produced can either be multiplied or rooted perpetually, and alternatively they can also be explored
for the in vitro production of hypericin and hyperforin. 相似文献
11.
Kailash Choudhary M. Singh M. S. Rathore N. S. Shekhawat 《Plant biotechnology reports》2009,3(3):205-211
An efficient in vitro regeneration protocol for moth bean [Vigna aconitifolia (Jacq.) Marechal] via somatic embryogenesis has been developed. Embryogenic callus cultures were established from the cotyledonary
node as explant on semi-solid Murashige and Skoog (MS) medium supplemented with 0.75 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.5 mg l−1 6-benzylaminopurine (BA) and with various additives (50 mg l−1 ascorbic acid and 25 mg l−1 each of adenine sulphate, citric acid and l-arginine). Numerous somatic embryos differentiated on MS basal nutrient medium supplemented with 0.25 mg l−1 2,4-D and 0.5 mg l−1 of kinetin (Kin). Sustained cell division resulted in the formation of cell aggregates, which progressed to the globular-
and heart-shaped somatic embryos and then, if they differentiated properly, to the torpedo shape and cotyledonary stages.
The transfer of embryos onto fresh MS basal medium containing 0.2 mg l−1 BA and 2.0 mg l−1 gibberellic acid enabled the embryos to achieve complete maturation and germination. More than 80% of somatic embryos were
converted into true-to-type fertile plants. In vitro-regenerated plantlets with well-developed roots were successfully hardened
in a greenhouse and established in soil. 相似文献
12.
A. V. Raghu Kuzhiyumparambil Unnikrishnan S. P. Geetha Gerald Martin Indira Balachandran 《In vitro cellular & developmental biology. Plant》2011,47(4):506-515
Embelia ribes, an important vulnerable medicinal liana, was regenerated through organogenesis and embryogenesis using leaf explants. Leaf
explants produced organogenic calluses on MS medium supplemented with 1.0 mg l−1 2,4-dichlorophenoxy acetic acid (2,4-D) and 0.5 mg l−1 6-benzylaminopurine. Shoot regeneration was obtained from organogenic calluses on MS medium containing different concentrations
of thidiazuron (TDZ) and indole-3-acetic acid (IAA). The frequency of shoot bud organogenesis was highest (23.9 shoots/explant)
in MS medium containing 0.5 mg l−1 TDZ and 0.1 mg l−1 IAA. The best result for induction of embryogenic callus was noticed in the combination of 2.0 mg l−1 TDZ and 0.5 mg l−1 2,4-D. This callus, maintained in the same medium, showed the highest differentiation of embryos (56.5%) after 6 wk of culture.
Embryos were transferred to MS medium supplemented with different concentrations of TDZ, and this facilitated conversion of
embryos into plants. After 6 wk of subculture, MS medium with 0.05 mg l−1 TDZ favored the highest percentage (52.2%) embryo conversion. As per the present protocol, 52.2% of the embryos underwent
conversion, and a mean number of 29.5 shoots per culture was obtained. Shoots developed from both types of calluses were rooted
on half-strength MS basal medium supplemented with 1.0 mg l−1 indole-3-butyric acid. HPLC-UV assay demonstrated the highest embelin content (5.33% w/w) in the embryogenic callus cultures. Embelin was isolated from embryogenic callus and was identified using IR and 1H NMR studies. 相似文献
13.
Li-Hua Zhu Xiao-Qin Wu Hong-Ye Qu Jing Ji Jian-ren Ye 《Plant Cell, Tissue and Organ Culture》2010,102(1):121-128
A protocol was developed for the micropropagation of Pinus massoniana and mycorrhiza formation on rooted microshoots. Seedling explants were first cultured on Gresshoff and Doy (GD) medium supplemented
with 6-benzyladenine (BA) alone or in combination with α-napthaleneacetic acid (NAA) to stimulate the formation of intercotyledonary
axillary buds. The frequency of axillary bud induction was up to 97% on medium supplemented with 4.0 mg l−1 BA and 0. 2 mg l−1 NAA, and the average number of buds per explant reached up to 5.5 on medium with 4.0 mg l−1 BA and 0.1 mg l−1 NAA. Axillary buds elongated rapidly after being transferred to half-strength GD medium containing activated charcoal (0.1%
w/v). Shoot proliferation was achieved by cutting elongated shoots into stem segments and subculturing on GD medium containing
2 mg l−1 BA and 0.2 mg l−1 NAA. Root primordia were induced in 82% of shoots when transferred to half-strength GD medium containing 0.2 mg l−1 NAA. Root elongation was achieved in a hormone-free GD agar medium or a perlite substrate. Rooted plantlets were inoculated
with the mycelium of ectomycorrhizal fungus Pisolithus tinctorius and the formation of ectomycorrhiza-like structures was achieved in vitro. 相似文献
14.
Jatropha curcas L. (Physic nut) is a commercially important non-edible oil seed crop known for its use as an alternate source of biodiesel.
In order to investigate the morphogenic potential of immature embryo, explants from four developmental stages were cultured
on medium supplemented with combinations of auxins and cytokinins. It was found that the size of embryo is critical for the
establishment of callus. Immature embryos (1.1–1.5 cm) obtained from the fruits 6 weeks after pollination showed a good response
of morphogenic callus induction (85.7%) and subsequent plant regeneration (70%) with the maximum number of plantlets (4.7/explant)
on Murashige and Skoog’s (MS) medium supplemented with IBA (0.5 mg l−1) and BA (1.0 mg l−1). The above medium when supplemented with growth adjuvants such as 100 mg l−1 casein hydrolysate + 200 mg l−1
l-glutamine + 8.0 mg l−1 CuSO4 resulted in an even higher frequency of callus induction (100%). Plant regeneration (90%) with the maximum number of plantlets
(10/explant) was achieved on MS medium supplemented with 500 mg l−1 polyvinyl pyrrolidone + 30 mg l−1 citric acid + 1 mg l−1 BA + 0.5 mg l−1 Kn + 0.25 mg l−1 IBA. It was observed that plantlet regeneration could occur either through organogenesis of morphogenic callus or via multiplication
of pre-existing meristem in immature embryos. The age of immature embryos and addition of a combination of growth adjuvants
to the culture medium appear to be critical for obtaining high regeneration rates. Well-developed shoots rooted on half-strength
MS medium supplemented with 0.5 mg l−1 IBA and 342 mg l−1 trehalose. The rooted plants after acclimatization were successfully transferred to the field in different agro-climatic
zones in India. This protocol has been successfully evaluated on five elite lines of J. curcas. 相似文献
15.
Meiru Li Hongqing Li Xiaoying Hu Xiaoping Pan Guojiang Wu 《Plant Cell, Tissue and Organ Culture》2010,102(3):321-327
Zoysia tenuifolia Willd. ex Trin. is one of the most popularly cultivated turfgrass. This is the first report of successful plant regeneration
and genetic transformation protocols for Z. tenuifolia using Agrobacterium tumefaciens. Initial calli was induced from stem nodes incubated on a Murashige and Skoog (1962) (MS) medium supplemented with 2 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 1 mg l−1 6-benzyladenine (BA), with a frequency of 53%. Compact calli were selected and subcultured monthly on the fresh medium. Sixty-nine
percent of the calli could be induced to regenerate plantlets when the calli incubated on a MS medium supplemented with 0.2 mg l−1 BA under darkness. For genetic transformation, calli were incubated with A. tumefaciens strain EHA105 harboring the binary vector pCAMBIA 1301 which contains the hpt gene as a selectable marker for hygromycin resistance and an intron-containing β-glucuronidase gene (gus-int) as a reporter gene. Following co-cultivation, about 12% of the callus explants produced hygromycin resistant calli on
MS medium supplemented with 2 mg l−1 2,4-D, 1 mg l−1 BA, 50 mg l−1 hygromycin, 500 mg l−1 cefotaxime after 8 weeks. Shoots were regenerated following transfer of the resistant calli to shoot induction medium containing
0.2 mg l−1 BA, 50 mg l−1 hygromycin, and 250 mg l−1 cefotaxime, and about 46% of the resistant calli differentiated into shoots. Finally, all the resistant shoots were rooted
on 1/2 MS media supplemented with 50 mg l−1 hygromycin, 250 mg l−1 cefotaxime. The transgenic nature of the transformants was demonstrated by the detection of β-glucuronidase activity in the primary transformants and by PCR and Southern hybridization analysis. About 5% of the total
inoculated callus explants produced transgenic plants after approximately 5 months. The procedure described will be useful
for both, the introduction of desired genes into Z. tenuifolia and the molecular analysis of gene function. 相似文献
16.
An efficient regeneration protocol for genetic transformation was developed from the leaves of an 11-year-old Phtinia × fraseri “Red Robin” tree. A high frequency of adventitious buds (88.63 ± 1.38%) and the highest maximum mean number of adventitious
buds per explant (4.65 ± 0.48) were obtained in light conditions on Murashige–Skoog (MS) medium containing 2 mg l−1 benzyladenine (BA) and 0.2 mg l−1 α-Naphthaleneacetic acid (NAA). After preculturing for 6 days, over 95% of the shoots successfully rooted on 1/2 MS medium
supplemented with 0.3 mg l−1 indole-3-butyric acid (IBA) within 3 weeks. For genetic transformation, two crucial parameters (30 mg l−1 kanamycin for selection and 30-min suspension time) were optimized. Taken together, a reliable transformation system with
an efficiency of more than 5% was established. This genetic transformation protocol can be utilized for further genetic manipulation
of the Photinia tree. 相似文献
17.
Meiru Li Hongqing Li Xiaoying Hu Xiaoping Pan Guojiang Wu 《Plant Cell, Tissue and Organ Culture》2011,106(3):363-371
Saussurea involucrata is a valuable traditional Chinese medicinal herb. This is the first report of a successful genetic transformation protocol
for S. involucrata using Agrobacterium tumefaciens. Leaf explants were incubated with A. tumefaciens strain EHA105 harboring the binary vector pCAMBIA 1301, which contains the hpt gene as a selectable marker for hygromycin resistance and an intron-containing β-glucuronidase gene as a reporter gene. Following
co-cultivation, about 23.7% of the explants produced hygromycin-resistant calli on MS basal medium (Murashige and Skoog in
Physiol Plant 15: 473–497, 1962) supplemented with 1 mg l−1 benzyladenine (BA), 0.1 mg l−1 α-naphthaleneacetic acid (NAA), 0.1 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D), 20 mg l−1 hygromycin, and 500 mg l−1 cefotaxime. Shoots were regenerated following transfer of the resistant calli to shoot induction medium containing 1.5 mg l−1 BA, 0.1 mg l−1 NAA, 0.25 mg l−1 gibberellic acid (GA3), 20 mg l−1 hygromycin, and 250 mg l−1 cefotaxime, and about 67.5% of the resistant calli differentiated into shoots. Finally, 80% of the hygromycin-resistant shoots
rooted on MS media supplemented with 0.2 mg l−1 NAA, 20 mg l−1 hygromycin, and 250 mg l−1 cefotaxime. The transgenic nature of the transformants was demonstrated by detection of β-glucuronidase activity in the primary
transformants and by Southern blot hybridization analysis. About 16% of the total inoculated leaf explants produced transgenic
plants after approximately 5 months. Using this optimized transformation system, a rice ortholog of the Arabidopsis FLOWERING LOCUS T gene, Hd3a, was transferred into S. involucrata. Introduction of this gene caused an early-flowering phenotype in S. involucrata. 相似文献
18.
This study demonstrates the morphogenic potential of pulvinus, an important organ situated at the base of the petiole or rachis
of leguminous plants. Plant regeneration via pulvinus-derived calli of Caesalpinia bonduc has been achieved. Organogenic calli have been derived from the explant 45 days after culture on Murashige and Skoog (MS)
medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) alone or in combination with 6-benzylaminopurine (BA). Optimum
callus induction (100%) occurred when the pulvini were cultured on MS medium fortified with 6 mg l−1 2,4-D and 1 mg l−1 BA. The highest shoot induction was obtained when the calli were transferred to MS medium supplemented with 5 mg l−1 BA and 1 mg l−1 indole-3-acetic acid (IAA). On this medium, 87% cultures responded with an average number of 4.2 shoots per culture. The
maximum root induction from the regenerated shoots was observed on half strength MS medium containing 6 mg l−1 indole-3-butyric acid (IBA). Here 100% shoots rooted with a mean number of 6.3 roots per shoot. The regenerated plantlets
were acclimatized and subsequently showed normal growth. This efficient protocol will be helpful for propagating elite clones
on a mass scale and could be utilized for genetic transformation study. 相似文献
19.
In vitro regeneration and morphogenesis studies in common bean 总被引:1,自引:0,他引:1
Kingdom Kwapata Robab Sabzikar Mariam B. Sticklen James D. Kelly 《Plant Cell, Tissue and Organ Culture》2010,100(1):97-105
An efficient protocol for high frequency in vitro regeneration of multiple shoots and somatic embryos from the embryonic axis
of common bean (Phaseolus vulgaris) was developed. Ten common bean cultivars representing a wide range of diversity among current commercial market classes
were used for in vitro regeneration evaluation in our study. These cultivars were tested on 63 different media formulations
consisting of combinations of cytokinins, namely benzyladenine (BA) and thidiazuron (TDZ) at concentration levels of 0.0,
1.0, 2.5, 5.0 and 10.0 mg l−1 and auxin, namely naphthalene acetic acid (NAA) and indole-3-acetic acid (IAA) at concentration levels of 0.0, 0.05, 0.1
and 1.0 mg l−1. P. vulgaris cv. Olathe pinto bean performed the best producing over 20 multiple shoots per explant while cv. Condor black bean was the
poorest with nine multiple shoots per explant. The optimum media for regeneration of multiple shoots was 4.4 mg l−1 Murashige and Skoog (MS) containing 2.5 mg l−1 BA and 0.1 mg l−1 IAA supplemented with 30 mg l−1 silver nitrate. Adventitious shoots and somatic embryos were regenerated on 4.4 mg l−1 MS medium containing 1 mg l−1 TDZ and 0.05 mg l−1 NAA supplemented with 30 mg l−1 silver nitrate or activated charcoal. Efficient and effective rooting of plantlets was achieved by dipping the cut end base
of in vitro regenerated shoots in 1.0 mg l−1 indole-3-butyric acid (IBA) solution and culturing on media containing 4.4 mg l−1 MS supplemented by 0.1 mg l−1 IAA, NAA or IBA. 相似文献
20.
Rakesh Yadav Pooja Arora Dharmendar Kumar Dinesh Katyal Neeraj Dilbaghi Ashok Chaudhury 《Plant biotechnology reports》2009,3(3):175-182
Simple, reproducible, high frequency, improved plant regeneration protocol in Eastern Cottonwood (Populus deltoides) clones, WIMCO199 and L34, has been reported. Initially, aseptic cultures established from axillary buds of nodal segments
from mature plus trees on MS liquid medium supplemented with 0.25 mg l−1 KIN and 0.25 mg l−1 IAA. Nodal and internodal segments were found to be extra-prolific over shoot apices during course of aseptic culture establishment,
while 0.25 mg l−1 KIN concentration played a stimulatory role in high frequency plant regeneration. Diverse explants, such as various leaf
segments, internodes, and roots from in vitro raised cultures, were employed. Direct plant regeneration was at high frequency
of 92% in internodes, 88% in leaf segments, and 43% in root segments. This led to the formation of multiple shoot clusters
on established culture media with rapid proliferation rates. Many-fold enhanced shoot elongation and growth of the clusters
could be achieved on liquid MS medium supplemented with borosilicate glass beads, which offer physical support for proliferating
shoots leading to faster growth in comparison to semi-solid agar or direct liquid medium. SEM examination of initial cultures
confirmed direct plant regeneration events without intervening calli. In vitro regenerated plants induced roots on half-strength
MS medium with 0.15 mg l−1 IAA. Rooted 5- to 6-week-old in vitro regenerated plants were transferred into a transgenic greenhouse in pots containing
1:1 mixture of vermicompost and soil at 27 ± 2°C for hardening and acclimatization. 14- to 15-week-old well-established hardened
plants were transplanted to the field and grown to maturity. The mature in vitro raised poplar trees exhibited a high survival
rate of 85%; 4-year-old healthy trees attained an average height of 8 m and an average trunk diameter of 25 cm and have performed
well under field conditions. The regeneration protocol presented here will be very useful for undertaking genetic manipulation,
providing a value addition to Eastern Cottonwood propagation in future. 相似文献