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1.
Somatic embryogenesis and plant regeneration in two wild cotton species belong to G genome 总被引:1,自引:0,他引:1
Shu-Feng Yan Qiao Zhang Jing-Er Wang Yu-Qiang Sun M. K. Daud Shui-Jin Zhu 《In vitro cellular & developmental biology. Plant》2010,46(3):298-305
The present work describes the plant regeneration via somatic embryogenesis in two wild cotton species belonging to G genome:
Gossypium nelsonii Fryx and Gossypium australe F Muell. The role of plant hormones and carbohydrates was also evaluated for somatic embryogenesis and somatic embryo development.
Normal plants were obtained from G. nelsonii Fryx; abnormal plants and somatic embryos were obtained from G. australe F Muell. The best medium for callus induction for these G genome wild cotton species was MSB5 supplemented with 0.1 mg L−1 KT and 0.1 mg L−1 2,4-D. For embryogenic callus proliferation, the best medium used was MSB5 supplemented with 0.2 mg L−1 KT and 0.5 mg L−1 IBA. The medium MSB5 supplemented with 0.15 mg L−1 KT and 0.5 mg L−1 NAA was used successfully for root initiation and plant growth. In addition, adding CuSO4 and AgNO3 in the callus-inducing and proliferation medium resulted in a number of somatic embryos. Glucose and maltose, the carbon
sources in somatic culture, were used for callus induction, but maltose worked even better than glucose for proliferation
of embryogenic callus and development of somatic embryos. 相似文献
2.
Jing Li Yang Bo Zhao Eun Soo Seong Myong Jo Kim Won Hee Kang Na Young Kim Chang Yeon Yu Cheng Hao Li 《Plant biotechnology reports》2010,4(4):261-267
We describe culture conditions for a high-efficiency in vitro regeneration system of Papaver
nudicaule through somatic embryogenesis and secondary somatic embryogenesis. The embryogenic callus induction rate was highest when
petiole explants were cultured on Murashige and Skoog (MS) medium containing 1.0 mg l−1 α-naphthaleneacetic acid (NAA) and 0.1 mg l−1 6-benzyladenine (BA) (36.7%). When transferred to plant growth regulator (PGR)-free medium, 430 somatic embryos formed asynchronously
from 90 mg of embryogenic callus in each 100-ml flask. Early-stage somatic embryos were transferred to MS medium containing
1.0 mg l−1 BA and 1.0 mg l−1 NAA to germinate at high frequency (97.6%). One-third-strength MS medium with 1.0% sucrose and 1.0 mg l−1 GA3 had the highest frequency of plantlet conversion from somatic embryos (91.2%). Over 90% of regenerated plantlets were successfully
acclimated in the greenhouse. Secondary somatic embryos were frequently induced directly when the excised hypocotyls of the
primary somatic embryos were cultured on MS medium without PGRs. Sucrose concentration significantly affected the induction
of secondary embryos. The highest induction rate (89.5) and number of secondary somatic embryos per explant (9.3) were obtained
by 1% sucrose. Most secondary embryos (87.2–94.3%) developed into the cotyledonary stage on induction medium. All cotyledonary
secondary embryos were converted into plantlets both in liquid and on semisolid 1/3-strength MS medium with 1.0% sucrose. 相似文献
3.
Plant regeneration was achieved through direct and indirect somatic embryogenesis in Eucalyptus camaldulensis. Callus was induced from mature zygotic embryos and from cotyledon explants collected from 10, 15, 25, and 30-day-old seedlings
cultured on Murashige and Skoog (MS) basal medium supplemented with different concentrations of naphthaleneacetic acid (NAA).
Maximum callus induction from mature zygotic embryos was obtained on MS basal medium containing 1 mg l−1 NAA. The frequency of callus development varied based on the age of the cotyledon explants 10-day-old explants giving highest
percentage on MS basal medium supplemented with 1 mg l−1 NAA. Callus obtained from mature zygotic embryos gave highest frequency of somatic embryogenesis on MS basal medium containing
0.5 mg l−1 benzyladenine (BA) and 0.1 mg l−1 NAA. Separate age wise culture of the calli, obtained from cotyledons of different ages cultured separately, revealed high
somatic embryogenic potential on callus from 10-day-old cotyledons. Direct somatic embryogenesis too was obtained from hypocotyl
explants without an intervening callus phase on MS basal medium containing 0.5 mg l−1 BA. The effects of abscisic acid (ABA), sucrose, and different strengths of MS medium on somatic embryo maturation and germination
were also investigated. Number of mature somatic embryos increased with lower concentrations (0–1 mg l−1) of ABA while no significant differences were observed at higher concentrations (2–5 mg l−1) of ABA. Compared to basal medium containing lower concentrations of sucrose (1%), the MS medium supplemented with higher
levels of sucrose (4%) showed significantly lower frequency of mature somatic embryos. Basal medium without any dilution gave
the highest number of immature embryos. However, the number of mature embryos was high at higher medium dilutions. 相似文献
4.
Guang-Zhe Lin Xiao-Mei Zhao Soon-Kwan Hong Yu-Ji Lian 《Plant Cell, Tissue and Organ Culture》2011,106(1):93-103
We have developed a system for the in vitro regeneration of pasqueflowers (Pulsatilla koreana Nakai). The system was based on somatic embryogenesis and shoot organogenesis. Over a growth period of 6 weeks, multiple
shoots were initiated from leaf, petiole, and pedicel explants on Murashige and Skoog (MS) medium containing 0.5 mg l−1 indole-3-acetic acid (IAA) and zeatin (Zn), kinetin (Kin), or 6-benzyladenine (BA). We achieved 100% of adventitious shoot
induced when petiole and pedicel explants were cultured on MS, 0.5–2.0 mg l−1 Zn, and 0.5 mg l−1 IAA. Somatic embryos developed from the explants and generated shoots on MS medium containing 0.25 mg l−1 Zn and 0.5 mg l−1 IAA. Globular and heart-shaped stages of somatic embryos were observed. Histological studies have revealed the stages of
development of somatic embryos. For propagation and growth, the regenerated shoots from organogenic or embryogenic calluses
were transferred to MS medium containing either (1) 1.5 mg l−1 Zn and 0.05 mg l−1 IAA or (2) 1.0 mg l−1 BA and 0.05 mg l−1 IAA. After the length of the shoots reached 3 cm, the shoots initiated by organogenesis as well as those initiated by somatic
embryogenesis were transferred to the root induction medium. After 2 months of culture in half-strength MS with 1.5 mg l−1 α-naphthalene acetic acid (NAA), the rooting ratio was 93%. Finally, the rooted plantlets were acclimatized in a mixture
of mountain soil and perlite. 相似文献
5.
A. Othmani C. Bayoudh N. Drira M. Marrakchi M. Trifi 《Plant Cell, Tissue and Organ Culture》2009,97(1):71-79
Plant regeneration through somatic embryogenesis from young leaf explants (5–10 mm long) adjacent to the apex of 5–6 year
old offshoots of Tunisian date palm (Phœnix dactylifera L.), cultivar Boufeggous was successfully achieved. Factors affecting embryogenic callus initiation, including plant growth
regulators and explant size, were investigated. The highest induction frequencies of embryogenic calli occurred after 6–7 months
on MS medium supplemented with 10 mg l−1 2,4-D and 0.3 mg l−1 activated charcoal. The subculture of these calli onto maintenance medium resulted in the formation of proembryos. Fine chopping
and partial desiccation (6 and 12 h) of embryogenic calli with proembryos prior to transfer to MS medium supplemented with
1 mg l−1 ABA stimulated the rapid maturation of somatic embryos. Maturated somatic embryo yield per 0.5 g FW of embryogenic callus
was 51 embryos with an average maturation time of 55 days. This was increased to 422 with finely chopped callus, and 124 and
306 embryos following 6 and 12 h desiccation treatments, respectively. The average time to maturation for these 3 treatments
was 35, 43 and 38 days, respectively. Subsequent substitution of ABA in MS medium with 1 mg l−1 NAA resulted in the germination and conversion of 81% of the somatic embryos into plantlets with normal roots and shoots.
The growth of regenerated somatic plants was also monitored in the field. 相似文献
6.
Xueping Shi Xigang Dai Guofeng Liu Junwei Zhang Guogui Ning Manzhu Bao 《In vitro cellular & developmental biology. Plant》2010,46(2):117-125
An efficient protocol for secondary somatic embryogenesis in camphor tree is reported. Secondary somatic embryos (SSEs), initially
obtained from the primary embryos of a nascent embryogenic culture in 2002, were proliferated and maintained for more than
4 yr via cyclic secondary somatic embryogenesis. Throughout this period, the embryo populations retained a high level of competence
for plant regeneration. SSEs were produced on the surfaces of the cotyledons and radicular ends of maternal somatic embryos
(MSEs). Histological observations of the various stages of secondary embryo development revealed four typical stages, namely,
globular, heart-shaped, torpedo, and cotyledonary. The process of secondary embryogenesis continued in a cyclic way, with
each newly formed embryo producing a subsequent generation of secondary embryos. In order to progress developmentally beyond
proliferation cycles, cotyledonary embryos from one of embryogenic lines (L14) were cultured on Murashige and Skoog (MS) medium
with 0.1–3.0 mg l−1 abscisic acid (ABA) or 0.05–1.0 mg l−1 thidiazuron (TDZ) in darkness for 2 mo to achieve maturation. Matured embryos were then transferred to MS-based germination
medium containing either 0.1 mg l−1 TDZ, 0.2 mg l−1 indole-3-butyric acid (IBA), and 0.5 mg l−1 6-benzylaminopurine (BA) or 0.1 mg l−1 TDZ and 0.2 mg l−1 IBA and were cultured in light for germination. Over 50% of embryos matured in the presence of 0.5 mg l−1 ABA were able to germinate with shoots and poor root system. Frequencies of embryos germinating normal shoots among different
genotypes did not change significantly. A total of 93% of the shoots from the germinated embryos converted to plantlets on
half strength MS medium with 0.5 mg l−1 IBA by 3 wk. Plantlets acclimatized successfully to ex vitro conditions and developed as field-grown plants with normal appearance. 相似文献
7.
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
8.
Iyyakkannu Sivanesan Mi Young Lim Byoung Ryong Jeong 《Plant Cell, Tissue and Organ Culture》2011,107(2):365-369
A simple and efficient protocol was developed for somatic embryogenesis from leaf and petiole explants of Campanula punctata Lam. var. rubriflora Makino. Somatic embryos (SE) were obtained with greater frequency from petiole explants than from leaf explants when cultured
on Murashige and Skoog (MS) medium supplemented with 2.0 mg L−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.0 mg L−1 6-benzyladenine (BA). On this medium, a mean number of 19.5 and 31.2 SE were developed per leaf and petiole explants, respectively.
Embryos were induced both light and dark conditions but culturing the explants 2 weeks in the dark followed by 3 weeks under
light resulted in high frequency of embryo formation. Globular embryos germinated best on MS medium supplemented with 0.3%
(w/v) activated charcoal (AC) and 1.0 mg L−1 GA3. The germinated plantlets grew further on MS medium containing 0.3% AC. Plantlets were successfully acclimatized in the greenhouse
with 94% survival rate. This is the first report on induction of somatic embryogenesis in this genus and also has implications
for genetic transformation, and mass clonal propagation. 相似文献
9.
The effect of auxins, time exposure to auxin and genotypes on somatic embryogenesis from mature embryos of wheat 总被引:3,自引:0,他引:3
Mikhail Filippov Dmitry Miroshnichenko Darya Vernikovskaya Sergey Dolgov 《Plant Cell, Tissue and Organ Culture》2006,84(2):100192-100201
The effects of different factors on the embryogenesis and plant regeneration from mature embryos of Russian spring and winter
genotypes were studied. Embryogenic callus induction was achieved on MS medium supplemented with different concentrations
of 2,4-D (2,4-dichlorophenoxyacetic acid), 2,4,5-T (2,4,5-trichlorophenoxyacetic acid) or Dicamba (3,6-dichloro-o-anisic acid). Although all auxins were able to induce callus from explants with high frequency (98–100%), Dicamba was more
effective for the induction of embryogenic callus (21.8–38.3%). Maximum embryogenic callus formation and high number of regenerated
plants were observed at 12 mg l−1 of Dicamba. The time exposure to Dicamba (7, 14, 21 and 28 days) had a significant effect on efficiency of somatic embryogenesis.
When contact of explants with callus induction medium was increased from 7 to 21 days the rate of somatic embryogenesis and
number of regenerated plants per embryogenic callus gradually increased from 13.0 to 38.4% and 3.6 to 8.0%, respectively.
Supplement of additional auxins (indoleacetic acid (IAA), indolebutyric acid (IBA), and naphthaleneacetic acid (NAA)) to callus
induction medium with Dicamba had a positive effect on the rate of embryogenic callus formation, while the average number
of regenerated shoots was not affected. The best rate of somatic embryogenesis was observed at the addition of 0.5 mg l−1 IAA with Dicamba (61.0%). The optimum combination of Dicamba and IAA increased the efficiency of somatic embryogenesis and
plant regeneration from seven spring and winter wheat genotypes, thought overall morphogenic capacity was still genotype dependent. 相似文献
10.
Jonny E. Scherwinski-Pereira Rodrigo S. da Guedes Paulo César P. FerminoJr Tatiane L. Silva Frederico Henrique S. Costa 《In vitro cellular & developmental biology. Plant》2010,46(4):378-385
An efficient procedure has been developed for inducing somatic embryogenesis and regeneration of plants from tissue cultures
of oil palm (Elaeis guineensis Jacq.). Thin transverse sections (thin cell layer explants) of different position in the shoot apex and leaf sheath of oil
palm were cultivated in Murashige and Skoog (MS) (Physiol Plant 15:473–497, 1962) medium supplemented with 0–450 μM picloram and 2,4-D with 3.0% sucrose, 500 mg L−1 glutamine, and 0.3 g L−1 activated charcoal and gelled with 2.5 g L−1 Phytagel. Embryogenic calluses were evaluated 12 wk after inoculation. Picloram (450 μM) was effective in inducing embryogenic
calluses in 41.5% of the basal explants. Embryogenic calluses were maintained on a maturation medium composed of basal media,
plus 0.6 μM NAA and 12.30 μM 2iP, 0.3 g L−1 activated charcoal, and 500 mg L−1 glutamine, with subcultures at 4-wk intervals. Somatic embryos were converted to plants on MS medium with macro- and micronutrients
at half-strength, 2% sucrose, and 1.0 g L−1 activated charcoal and gelled with 2.5 g L−1 Phytagel. 相似文献
11.
A simple and efficient system was developed for rapid somatic embryogenesis from leaf explants of Merwilla plumbea, a traditional but threatened medicinal plant in South Africa. Friable embryogenic callus (FEC) was obtained from leaf explants
on embryogenic callus induction medium containing agar-solidified Murashige and Skoog (MS) salts and vitamins, 8.3 μM picloram,
2.3 μM thidiazuron (TDZ) and 20 μM glutamine. FEC was subsequently incubated in embryogenic callus proliferation medium containing
4.5 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 4.1 μM picloram for 7 days before it was transferred to liquid somatic embryo
medium (SEML) containing MS medium supplemented with 0.4 μM picloram and 0.9 μM TDZ. In SEML supplemented with 150 mg L−1 haemoglobin, 5.4–35.6 somatic embryos per settled cell volume of 500 mg FEC were obtained. These embryos were at globular
to cotyledonary developmental stages. Embryo maturation, germination and plant formation rate was 94.4% following transfer
of SEs to half-strength MS medium supplemented with 1.4 μM gibberellic acid. Plantlets transferred into soil acclimatized
in the misthouse and established successfully in the greenhouse (100%). This is the first report on induction of Merwilla plumbea somatic embryogenesis. The protocol developed offers controlled vegetative propagation by alleviating extinction threats,
ensures germplasm conservation and provides a system for physiological, biochemical, molecular and cellular studies of embryo
development. 相似文献
12.
Somatic embryogenesis and further plant regeneration were observed using zygotic embryos, young inflorescences and young leaves
ofEuterpe edulis (Palmae) as explants. Both for the cultures of zygotic embryos and inflorescences, activated charcoal in the medium was essential
for the establishment of viable cultures. Embryogenesis was induced by using a gelled basal medium with MS or Euwens salts
supplemented by high 2, 4-D levels (50–100 mg L−1). The embryogenic process was direct without a callus stage. For further development, cultures with globular or post-globular
embryos were transferred to the basal medium with 2-iP (2.5 mg L−1) and NAA (0.1 mg L−1). To convert embryos to plantlets, cultures were transferred to a third medium in which sucrose and salts were reduced to
the half-strenght of the basal medium, without growth regulators. In the case of liquid medium, with either 2, 4-D or NAA
(10–20 mg L−1). The developmental stage of each explant was critical for the induction of embryogenesis. The histological study of embryogenic
cultures revealed that in the case of zygotic embryos, somatic embryos arise directly from the surface of the cotyledonar
node, or from subepidermal tissues. In the inflorescences, a pro-embryogenic tissue is formed at the floral primordium region;
in the leaves, the first morphogenic event is cell proliferation in the vascular parenchyma. 相似文献
13.
Qi Xiang Zhang Yu Sun Heng Kang Hu Bei Chen Chun Tao Hong Hai Peng Guo Yin Hui Pan Bing Song Zheng 《In vitro cellular & developmental biology. Plant》2012,48(1):50-57
Miscanthus sinensis (Poaceae) is a typical perennial giant grass of East Asia. Due to its high photosynthetic efficiency, low input requirements,
and high biomass production, M. sinensis shows outstanding potential as a biofuel feedstock. However, the lack of an efficient tissue culture system may limit its
utilization potential. Different explants of M. sinensis were evaluated to develop an efficient tissue culture system. Shoot apices from in vitro-germinated seedling explants were tested for adventitious bud proliferation. The highest level of proliferation (multiplication
coefficient 6.69) was obtained when shoot apices were cultured on Murashige and Skoog (MS) medium supplemented with 1.0 mg L−1 6-benzyladenine (BA), 2.0 mg L−1 kinetin, 0.05 mg L−1 α-naphthalene acetic acid (NAA), 3% sucrose, and 0.8% agar. The highest rooting percentage (95.4%) was obtained when adventitious
buds were cultured on half-strength MS medium supplemented with 0.2 mg L−1 NAA, 3% sucrose, and 0.8% agar. Significant differences were found in the formation of embryogenic callus among different
explant types. The embryogenic callus derived from epicotyls had the highest regeneration capacity when cultured on a medium
supplemented with 2.0 mg L−1 2,4-dichlorophenoxyacetic acid, 0.5 mg L−1 BA, and 0.1 mg L−1 thiamine. Under these conditions, the callus induction percentage was 82%. 相似文献
14.
The halophyte Leymus chinensis (Trin.) is a perennial rhizome grass (tribe Gramineae) that is widely distributed in China, Mongolia and Siberia, where it
is produced as a forage product. In this report, we establish a highly reproducible plant regeneration system through somatic
embryogenesis. Two explants, mature seeds and leaf base segments were used; these parts displayed different responses to combinations
of growth factors that affect embryogenic callus induction, callus type optimization and plant regeneration. The highest callus
induction frequency was obtained on Murashige and Skoog (MS) medium supplemented with 2.0 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D) in the presence of 5.0 mg l−1
l-glutamic acid. The inclusion of 5.0 mg l−1
l-glutamic acid was found to significantly promote primary callus induction, embryogenic callus formation and callus status
improvement. Subculturing on maintenance medium for 1–2 months before plant regeneration was found to be essential for the
optimization of callus type and the maturation of embryogenic callus. Callus relative water content and growth rate were simultaneously
investigated during callus maintenance, and found to possibly be related to callus type. Shoots were differentiated from the
embryogenic callus on the optimal medium with MS salts containing 0.2–0.5 mg l−1 α-naphthalene acetic acid (NAA), 2.0 mg l−1 kinetin (Kn) and 2.0 g l−1 casamino acids in 71.0 and 69.2% of wild-type (WT) and Jisheng No.1 (JS) plants, respectively. Plant regeneration was variable
depending on NAA levels, and the addition of casamino acids stimulated the maturation of embryogenic callus and plant regeneration.
Transferring callus with shoots onto half-strength MS medium resulted in rooting within 1 week. The growth of regenerated
plants was also surveyed in the field. This is the first report of plant regeneration through somatic embryogenesis from mature
seeds and leaf base segments of L. chinensis. 相似文献
15.
P. Giridhar Vinod Kumar G. A. Ravishankar 《In vitro cellular & developmental biology. Plant》2004,40(6):567-571
Summary A novel protocol has been developed for inducing somatic embryogenesis from leaf cultures of Decalepis hamiltonii. Callus was obtained from leaf sections in Murashige and Skoog (MS) medium supplemented with α-naphthaleneacetic acid (NAA)+N6-benzyladenine (BA) or 2,4-dichlorophenoxyacetic acid (2,4-D)+BA. Nodular embryogenic callus developed from the cut end of
explants on media containing 2,4-D and BA, whereas compact callus developed on media containing NAA and BA. Upon subsequent
transfer of explants with primary callus onto MS media containing zeatin and/or gibberellic acid (GA3) and BA, treatment with zeatin (13.68μM) and BA (10.65 μM) resulted in the induction of the highest number of somatic embryos directly from nodular tissue. The maturation of embryos
took place along with the induction on the same medium. Embryogenic calluses with somatic embryos were subcultured onto MS
basal medium supplemented with 4.56μM zeatin+10.65 μM BA. After 4wk, more extensive differentiation of somatic embryos was observed. The mature embryos developed into complete
plantlets on growth regulator-free MS medium. A distinct feature of this study is the induction of somatic embryogenesis from
leaf explants of Decalepis hamiltonii, which has not been reported previously. By using this protocol, complete plantlets could be regenerated through indirect
somatic embryogenesis or organogenesis from leaf explants in 12–16 wk. 相似文献
16.
K. Balaraju S. Saravanan P. Agastian S. Ignacimuthu 《Acta Physiologiae Plantarum》2011,33(4):1123-1133
An improved method of direct somatic embryogenesis (SE) was developed in Swertia chirata for the first time using leaves and roots of in vitro-grown young seedlings. In the present study, 2,4-dichlorophenoxyacetic
acid (2,4-D) was assessed individually and in combination with other auxins, as well as with cytokinin for its effectiveness
to induce somatic embryos. Leaf explants with abaxial side in the medium produced maximum number of somatic embryos. This
system omits the callus stage and thus reduces the process of SE in S. chirata by 35–45 days. Embryos at different stages of development were observed. Maturation of heart stage embryos were observed
on Murashige and Skoog (MS) medium containing 1 mg L−1 2,4-D. Upon transfer to the germination medium, they were converted to cotyledonary stage and then plantlets of 33% and 68%
of them were converted to cotyledonary stage and then plantlets on MS medium supplemented with 0.05 and 0.1 mg L-1 GA3 respectively. The 2,4-D alone at 1.0 or 1.5 mg L−1 was found to be better for embryogenic tissue initiation than 2,4-D in combination with indole-3-acetic acid or α-naphthalene
acetic acid. For further embryo development, 2,4-D was combined with cytokinins such as 6-benzylaminopurine (BAP) and kinetin
or plant growth regulator free medium or medium with 50% reduced concentration of the same hormone while subculturing. Mean
germination and percentage of survival were maximum in the medium containing 1.0 mg L−1 2,4-D in combination with 0.1 mg L−1 BAP. Regenerated plantlets were morphologically and genetically identical. This method offers a vast scope for the clonal
propagation of endangered plants. 相似文献
17.
Yantree Devi Sankar-Thomas Katja Saare-Surminski Reinhard Lieberei 《Plant Cell, Tissue and Organ Culture》2008,95(2):163-173
The present study describes a protocol for plant regeneration via somatic embryogenesis in temporary immersion system (TIS)
for Camptotheca acuminata. Somatic embryos were induced by culturing hypocotyl segments from 14-day-old in vitro grown C. acuminata seedlings in TIS. Hypocotyl segments were placed in culture vessels modified with a mechanical device to support the fixation
of explants. Cultures were maintained under a 16 h photoperiod with a light intensity of 60 μmol m−2 s−1 PPF at 25 ± 1°C. After 16 weeks of incubation embryogenic calli were formed above the edge of the mechanical device in the
basal Murashige and Skoog (MS) medium containing 35 g l−1 sucrose and without hormonal supplementation. For plantlet regeneration, somatic embryos at cotyledonary stage were cultured
in three different concentrations of 6-benzylamino-purine (0.5, 1.0 and 1.5 mg l−1 BAP) and in plant growth regulator (PGR) free medium. In general, 0.5 mg l−1 BAP was found to be the most effective concentration for growth and development of Camptotheca embryos in TIS. Conversion of somatic embryos into plantlets was also successfully achieved on sterile substrates moistened
with 0.5 mg l−1 BAP. Plantlets derived from cotyledonary embryos were rooted in vitro with 0.5 mg l−1 indole-3-butyric acid (IBA) before transfer to ex vitro conditions. 相似文献
18.
Zita Demeter Gyula Surányi V. Attila Molnár Gábor Sramkó Dániel Beyer Zoltán Kónya Gábor Vasas Márta M-Hamvas Csaba Máthé 《Plant Cell, Tissue and Organ Culture》2010,100(3):349-353
Crocus heuffelianus belongs to the C. vernus (Iridaceae) species aggregate. In the Carpathian Basin and particularly in Hungary it is considered an endangered species. Therefore
our aim was to establish a tissue culture system with potential of germplasm preservation of this taxon. For in vitro culture
experiments, shoot primordia from corms were the most suitable. We induced an embryogenic callus line from those explants
on basal Murashige-Skoog (MS) medium supplemented with Gamborg’s vitamins, 2% (w/v) sucrose, 10 mg l−1 (53.7 μM) α-naphthaleneacetic acid (NAA) and 1 mg l−1 (4.44 μM) 6-benzyladenine (BA). Globular stage embryos developed on this medium and several culture conditions were used
in an attempt to obtain mature embryos and plant regeneration. Firstly a decrease of auxin/cytokinin concentration and ratio,
then secondly a decrease in the strength of culture medium and the concentration of carbon source was used, which was effective
in embryogenesis and the production of plants. Regeneration medium used in the second step was fourfold diluted MS medium
and Gamborg’s vitamins supplemented with 1% (w/v) sucrose, 0.05 mg l−1 (0.26 μM) NAA and 0.5 mg l−1 (2.22 μM) BA, with a 14/10 h photoperiod. Under these conditions we could detect all the stages of somatic embryo development
characteristic for Iridaceae. This is the first report demonstrating the production of stable tissue culture of C. heuffelianus with potential use in germplasm preservation via plant regeneration. This study could also contribute to a better understanding
of somatic embryogenesis in the Crocus genus. 相似文献
19.
Ai Hua Chen Jing Li Yang Yu Da Niu Chuan Ping Yang Gui Feng Liu Chang Yeon Yu Cheng Hao Li 《Plant Cell, Tissue and Organ Culture》2010,102(3):357-364
We developed a new protocol for highly efficient somatic embryogenesis and plantlet conversion of Schisandra chinensis. Friable embryogenic callus was induced from cotyledonary leaves and hypocotyls of germinated zygotic embryos on Murashige
and Skoog (MS) agar medium containing 2,4-dichlorophenoxyacetic acid (2,4-D). Preculture of zygotic embryos on 2,4-D-containing
medium increased embryogenic callus induction efficiency. The highest embryogenic callus induction frequency of 56.7% was
obtained from shoot apical meristem-containing hypocotyl explants from 1-week-old germinated embryos on MS medium containing
4.0 mg l−1 2,4-D. Embryogenic callus proliferation, somatic embryo (SE) formation, and subsequent plantlet conversion occurred under
optimal culture conditions. The effects of MS medium strength, sucrose, gibberellic acid (GA3), and 6-benzyladenine (BA) on SE formation and plantlet conversion were evaluated. Low MS medium strength (1/4 to 1/2) was
necessary for SE formation, and the optimal sucrose concentration was 2.0%. Supplementing medium with GA3 negatively impacted SE formation and subsequent development. BA significantly increased the number of SEs and the plantlet
conversion capacity. One-third-strength MS medium with 1.0% sucrose and 0.5 mg l−1 BA produced the highest number of SEs (309 embryos from 9 mg embryogenic callus) and the highest frequency of plantlet conversion
from germinated SEs (52.6%). When transplanted to soil, 90% of the regenerated plants developed into normal plants. 相似文献
20.
Silvia Karina Vila Hebe Yolanda Rey Luis Amado Mroginski 《Journal of Plant Growth Regulation》2007,26(3):268-277
Factors affecting somatic embryogenesis induction and conversion in paradise tree (Melia azedarach) were evaluated. Somatic embryogenesis was influenced by plant growth regulators, explant stage, carbohydrate source and
concentration, gelling agents, light, and induction times. MS medium with 4.54 μM thidiazuron (TDZ) was optimal for the induction
of embryogenic tissue. Zygotic embryos that were 1-1.5 mm long (torpedo and early cotyledonal stage) had a greater embryogenic
response than smaller or larger embryos and better conversion of somatic embryos into plants. In general, embryos that formed
in medium containing 1% or 5% carbohydrate were hyperhydrics or fused, respectively, whereas those that formed in medium with
a carbohydrate concentration of 3% had better morphology. Raffinose at 3% yielded satisfactory somatic embryo induction with
good morphology and the best values of conversion into plants. Induction and conversion of somatic embryos were superior on
medium solidified with agar A-1296. The explants maintained under 160 μmol m−2 s−1 or 1 week in darkness and later 160 μmol m−2 s−1 produced a significantly higher embryogenic index. Only 4 days of treatment on induction medium, with either raffinose or
sucrose at 3% as a carbohydrate source, were required to induce somatic embryogenesis, but longer exposure, until 18 days,
increased the yield and improved the morphology of somatic embryos. 相似文献