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1.
Wildgermplasmconsistsofanimportantbasisforgeneticimprovementofcultivatedrice.OryzameyerianaisoneofthethreewildOryzaspeciesfoundinChina[1].AccessionsoriginatedinYunnanProvinceexhibitstrongresistancetobacterialleafblight[2].O.meyerianagrowsonthehillsideunde… 相似文献
2.
Guangcun He Lihui Shu Lanjie Liao Xiaohui Yin Lahong Sheng Xiaoling Wang 《中国科学C辑(英文版)》1998,41(4):393-399
Oryza meyeriana Baill is one of the three wild rice species found in Chiia.O. mcyeriana possesses valuable characteristics but is reluctant in cell culturein vitro. In a series of experiments, callus with no regeneration ability was induced from young panicle ofO. meyeriana. The callus was subcultured and propagated. Embryogenic cell clones were obtained after cryopreswation. Suspension cultures were established and protoplasts were isolated and regenerated into plants. Results of artificial inoculation ofXanthomonas campestris pv.Oryzae showed that the strong resistance did not change in the regenerated plants. The development of protoplast-to-plant system is an important progress towards utilization ofO. meyeriana via cellular engineering. The experiments demonstrated that cryopreservation of plant calli was a new way to obtain embryogenic cell line. 相似文献
3.
A simple and efficient procedure for cryopreservation of embryogenic cells of aromatic Indica rice varieties 总被引:2,自引:0,他引:2
Embryogenic suspension cells of two commercially cultivated aromatic Indica rice varieties, Basmati 385 and Pusa Basmati 1, were cryopreserved using a simple one-step freezing procedure that does not require a controlled-rate freezer. The procedure involves osmotic pre-conditioning of cells with mannitol, addition of a cryoprotectant solution consisting of sucrose, dimethyl sulfoxide, glycerol, proline, and modified R2 medium, cooling to –25°C for 2 h in a freezer, and then storage in liquid nitrogen. After rapid thawing at 45°C, these cultures showed post-thaw cell viability of 5.6 to 10.5% and formed actively dividing, readyto-use cell suspensions in 20–35 d when cultured directly into liquid medium. Plants were regenerated from cell clumps as well as from colonies formed by protoplasts that were isolated from suspension cells re-established from cryopreserved cells, with frequencies higher (54–98%) than, or comparable to, those obtained from three to four-month-old original non-frozen cell cultures. Cell viability and regeneration frequencies of post-thawed Pusa Basmati 1 cultures were similar to those obtained from the suspension cells cryopreserved using the conventional slow-freezing procedure which involves pre-freezing cells to –40°C at the rate of –0.2°C per min prior to immersion in liquid nitrogen. In Basmati 385, however, cells frozen at ––25°C showed lower post-thaw cell viability than those preserved using the slow-freezing procedure, but these cells produced cell suspensions that had greater shoot morphogenetic potential. The study indicates the beneficial effect of this simple freezing procedure, not only for preserving desirable cultured cells but also for an enrichment of embryogenic cells.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- DMSO
dimethylsulfoxide
- LN
liquid nitrogen
- MS
Murashige and Skoog (1962) medium
- NAA
-napthaleneacetic acid
- pcv
packed cell volume
- TTC
2,3,5-triphenyltetrazolium chloride 相似文献
4.
Alejandro Vazquez-Tello Makoto Hidaka Takeshi Uozumi 《Plant Cell, Tissue and Organ Culture》1995,40(2):169-177
Embryogenic cell suspensions of Lavatera thuringiaca L. were established from leaf petiole and shoot regeneration was achieved when cells were plated on medium without growth regulators. We tested three methods for protoplast culture, isolated from a one-year old embryogenic cell suspension, to determine the best conditions for L. thuringiaca protoplast culture and shoot regeneration. The highest protoplast plating efficiency was obtained with the agaroseembedded method, reaching 30%, while the nursing culture method gave 5% when the protoplasts were plated over Whatman paper No. 2. However, the same nursing culture failed to produce protoplast-derived microcalluses when the protoplasts were plated on a nitrocellulose filter. The liquid thin layer method gave the lowest plating efficiency with only 0.5%. Shoot regeneration from protoplast-derived microcalluses was achieved in two steps; first, globular embryo development was favored in medium low in auxin (2,4-d and BA at 0.01 and 0.05 mg 1-1, respectively), second, the globular embryos further differentiate into shoots in medium without growth regulators or in medium containing GA3 (0.5 to 1.0 mg 1-1).Abbreviations 2,4-d
2,4-dichlorophenoxyacetic acid
- BA
benzyladenine
- GA3
gibberellic acid
- NAA
-naphthaleneacetic acid
- IBA
indole-3-butyric acid 相似文献
5.
Compact calli initiated from young inflorescences of Oryza sativa L. (rice) on the Linsmaier and Skoog's (LS) medium containing 1 to 2.5mg/l of 2,4-dichlorophen-oxyacetic acid (2,4-D) were
used for regeneration studies. After smooth and compact nodules appeared, these calli were transferred to the regeneration
medium containing indole-3-acetic acid (IAA) and either kinetin or 6-benzylaminopurine (BAP). Somatic embryos developed in
ten days and were examined by histological studies. Some of the embryos showed scutellum-like structures and a coleoptile-coleorhiza
bipolar organization. Regenerated plants had the normal chromosome number of 2n=24. 相似文献
6.
Yan CQ Qian KX Yan QS Zhang XQ Xue GP Huangfu WG Wu YF Zhao YZ Xue ZY Huang J Xu GZ Wu P 《Plant cell reports》2004,22(8):569-575
Bacterial blight is one of the major diseases affecting rice productivity. To improve the resistance of cultivated rice to bacterial blight, we introduced a bacterial blight resistance trait from Oryza meyeriana, a wild rice species, into an elite japonica rice cultivar (Dalixiang) using asymmetric somatic hybridization. One hundred and thirty-two independent lines were regenerated. The hybrid plants possessed several morphological features of the donor species, O. meyeriana. Random amplified polymorphic DNA analysis revealed that hybrid plants exhibited banding patterns derived from their parental genotypes. For the majority of the hybrids, resistance to bacterial blight pathogens was intermediate to that observed for O. meyeriana and O. sativa (cv. Dalixiang). Four of the hybrid lines exhibited a high bacterial blight resistance, but it was less than that observed for O. meyeriana. These results demonstrate that O. meyeriana can be used as a good genetic source for improving bacterial blight resistance in commercial rice cultivars through asymmetric somatic hybridization.Abbreviations
2,4-D:
2,4-Dichlorophenoxyacetic acid
-
IOA:
Iodoacetamide
-
NAA:
-Naphthaleneacetic acid
-
PEG:
Polyethylene glycol
Communicated by P. Lakshmanan 相似文献
7.
GAO Lizhi GE Song HONG Deyuan ZHANG Jiongwei LUO Qingyan TAO Guoda XU Zaifu 《中国科学:生命科学英文版》1999,42(1):102-108
In order to determine genetic diversity ofOryza meyeriana (Zoll. et Mor. ex Steud.) Baill., 12 enzyme systems encoded by 17 loci were electrophoretically analyzed in 164 individuals
of seven populations from Simao Prefecture, Yunnan Province, China. In comparison with those seed plants with the same life
history and breeding systems, as well as the other species in the genusOryza, the species shows rather low levels of genetic diversity (A = 1.1,P = 8.0 %, Ho = 0.004 and He = 0.015) within populations and high genetic differentiation among populations. Fst was up to 0. 649, suggesting that 64. 9% of total genetic variability exists among populations. Considering high genetic
differentiation among populations from a limited geographic region, most of the populations of the species are worth being
protected, and therefore, great natural protection regions should theoretically be established in which a great deal of populations
should be involved for developingin situ conservation management. Meanwhile, some priory localities forin situ conservation ofO. meyerzana in Yunnan Province, were proposed.
Project supported by the Grant of the President of the Chinese Academy of Sciences. 相似文献
8.
Motoshige Kawata Satoshi Harada Baltazar Antonio Kiyoharu Oono 《Plant Cell, Tissue and Organ Culture》1992,28(1):1-10
Protoplasts were isolated from callus derived from a single homozygous seed of Oryza sativa L. var. Norin 8. Thirty protoclones were randomly selected and these showed variation in regeneration frequency ranging from 0–87% with an average of 52%. The potential for regeneration of each protoclone as reflected in the regeneration frequency was analyzed five times over a period of 250 days and showed that the protoclones can be classified into three types, namely: protoclones with high regeneration frequency; protoclones with low regeneration frequency, both of which maintained their respective levels of regeneration potential; and protoclones with gradually decreasing regeneration frequency. Secondary protoclones established from protoplasts isolated from some of these protoclones and regenerated 2–3 times for a period of 120 days also showed further reduction in regeneration frequency. The polypeptide composition analyzed by two dimensional gel electrophoresis suggests the presence of specific polypeptides related to regeneration potential. Analysis of ploidy level based on plant morphology and pollen size suggests the predominance of tetraploids among the regenerated plants. 相似文献
9.
Plant regeneration from 2-month-old callus cultures derived from immature leaves of 7-day-old aseptic seedlings and mature embryos of the African wild rice Oryza longistaminata was achieved at 20% and 100% frequency, respectively. The morphogenic potential of the embryo-derived calluses dropped from 100% at the third subculture to 12.5 % at the 12th subculture. Five-month-old morphogenic calluses were used to establish a fast-growing suspension culture which, when plated onto semisolid medium, still retained its ability to regenerate plantlets 9 months after initiation. Histological analyses demonstrated that late plant regeneration from established callus and suspension cultures occured through organogenesis, although some embryogenesis events may have taken place during initiation of these cultures.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- NAA
-naphthalenacetic acid
- BAP
6 benzylaminopurine
- PAS
Periodic acid shiff 相似文献
10.
Christian Schöpke Ludwig E. Müller Hans-Willy Kohlenbach 《Plant Cell, Tissue and Organ Culture》1987,8(3):243-248
Somatic embryogenesis was achieved in protoplast cultures of coffee. Protoplasts were isolated from cell suspension-derived somatic embryos of Coffea canephora. After repeated subculture in a medium supplemented with 0.5 mg/l of each of kinetin, 2,4-dichlorophenoxyacetic acid (2,4-D), and naphthaleneacetic acid (NAA), microcalli developed. Transfer of these microcalli to a medium lacking growth regulators resulted in the formation of globular embryos. Upon subculture without growth regulators they grew to well-differentiated embryos, Eventually some of them developed to plantlets which were transferred to the greenhouse for further observation. 相似文献
11.
F. Pupilli S. Arcioni F. Damiani M. Pezzotti 《Plant Cell, Tissue and Organ Culture》1990,23(3):193-199
Plant regeneration from leaf- and cotyledon-derived calli and from protoplast-derived tissue has been obtained in Lotus pedunculatus. Callus induction was achieved with 2,4-D and plant regeneration required the following two media sequences: bud formation was stimulated by IAA and BA and shoot growth by kinetin. Root formation occurred in the presence of IAA. Cotyledon protoplasts showed a low plating efficiency and plant regeneration was achieved via an intervening callus phase.Abbreviations BA
6-benzyladenine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- 2iP
N6--2-isopentenyl-adenine
- NAA
-naphthaleneacetic acid 相似文献
12.
An improved procedure for plant regeneration from indica and japonica rice protoplasts 总被引:1,自引:0,他引:1
R. K. Jain G. S. Khehra S. -H. Lee N. W. Blackball R. Marchant M. R. Davey J. B. Power E. C. Cocking S. S. Gosal 《Plant cell reports》1995,14(8):515-519
Plant regeneration from protoplasts of two commercially cultivated Indian indica rice varieties, Pusa Basmati 1 and Java, has been accomplished by plating embryogenic cell suspension-derived protoplasts on the surface of filter membranes overlying agarose-embedded feeder cells of Lolium multltiflorum and Oryza ridleyi, combined with the use of a maltose-containing shoot regeneration medium. Embryogenic cell suspension cultures of Pusa Basmati 1 and Jaya were initiated from mature seed scutellum-derived calli in liquid R2 medium modified by the addition of 560 mg l–1 of proline and 1.0 % (w/v) maltose. In both varieties, protoplast plating efficiencies up to 0.4 % were obtained, depending on the nature of the feeder cells. L. multiflorum feeder cells induced a 6-fold higher plating efficiency than feeder cells of O. ridleyi. In combination, O. ridleyi and L. multiflorum feedercells further enhanced protoplast plating efficiency. Protoplast-derived cell colonies were not obtained from protoplasts of either indica varieties in the absence of feeder cells. MS-based medium containing kinetin (2.0 mg l–1) and -naphthaleneacetic acid (0.5 mg 1–1), together with sucrose and maltose both at 1.5 % (w/v), induced green shoot regeneration in 44 % of protoplast-derived tissues, depending on the feeder cells used for protoplast culture. In both varieties, tissues obtained using O. ridleyi feeder cells were more morphogenic than tissues obtained using L. multiflorum feeder cells, either alone or in combination with cells of O. ridleyi. In the japonica rice variety Taipei 309, this new procedure resulted in a 30-fold increase in plant regeneration from protoplasts compared to previous published procedures.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- FDA
fluorescein diacetate
- GPFs
growth promoting factors
- NAA
-naphthaleneacetic acid
On leave from Department of Genetics, Haryana Agricultural University, Hisar, IndiaOn leave from Biotechnology Centre, Punjab Agricultural University, Ludhiana, India 相似文献
13.
Tang K. Sun X. An D. Power J. B. Cocking E. C. Davey M. R. 《Plant Cell, Tissue and Organ Culture》2001,66(2):149-153
An efficient and rapid procedure has been developed to establish embryogenic cell suspension cultures of two Japonica Chinese
commercial rice cultivars, Zhonghua 8 and Eryi 105. Embryogenic cell suspensions of both varieties were established from 0.5–1.0
g fresh weight of embryogenic callus in AA medium within 2.5 months of the initiation of callus from sterilised seeds. The
previously reported subculture of callus on semi-solid medium for 4–8 weeks prior to transfer into liquid medium was unnecessary
and caused delay in the establishment of embryogenic cell suspensions. Protoplasts were isolated reproducibly from cell suspensions
up to 18 months after their initiation, with protoplast plating efficiencies attaining 0.15–0.37%. Reproducible plant regeneration
from 14–26% of the protoplast-derived tissues was achieved without the requirement for nurse cells.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
14.
Somatic embryogenesis and plant regeneration from embryogenic suspension cultures of perennial ryegrass 总被引:1,自引:0,他引:1
Ousama M. Faizzaghmout William A. Torello 《In vitro cellular & developmental biology. Plant》1990,26(4):419-424
Summary Embryogenic callus induced from mature caryopses of perennial ryegrass (Lolium perenne L.) were placed in liquid half-strength Murashige and Skoog (MS) basal medium and supplemented with 6.0 mg/l 2,4-dichlorophenoxy
acetic acid (2,4-D), 3 g/l (w/v) casein hydrolysate (CH), and B5 vitamins, to initiate fast-growing highly embryogenic cell
suspension cultures. Newly initiated suspension cultures contained a high level of large non-embryogenic cells (NE) with relatively
few embryogenic (E) cells. Cell types were separated by discontinuous Percolls gradients or by filtering the newly initiated
cultures through 31-μm nylon mesh. The growth conditions of the E cell were optimized by testing various media components
including 2,4-D and sucrose, and subculture diluton ratio. Optimal shoot formation occurred after pretreatment of the embryogenic
cells on solidified callus maintenance medium supplemented with 60 mg/l cefotaxime for 4 weeks prior to transfer to regeneration
medium Regeneration media consisted of half-strength MS basal medium supplemented with B5 vitamins, 0.5 mg/l fluridone, and
0.5 mg/l BA. Most plants regenerated were albino with only a few green plants.
Journal Paper number MAES 2959 of the Massachusetts Agricultural Experiment Station. 相似文献
15.
Rice (Oryza sativa L.) plants of the indica cultivar IR54 were regenerated from protoplasts. Conditions were developed for isolating and purifying protoplasts from suspension cultures with protoplast yields ranging from 1·106 to 15·106 viable protoplasts/1 g fresh weight. Protoplast viability after purification was generally over 90%. Protoplasts were cultured in a slightly modified Kao medium in a Petri plate by placing them onto a Millipore filter positioned on top of a feeder (nurse) culture containing cells from a suspension culture of the japonica rice, Calrose 76. Plating efficiencies of protoplasts ranged from 0.5 to 3.0%; it was zero in the absence of the nurse culture. Protoplast preparations usually contained no contaminating cells, and when present, the number of cells never exceeded 0.1% of the protoplasts. After three weeks the Millipore filter with callus colonies were transferred off feeder cells and onto a Linsmaier and Skoog-type medium for an additional three weeks. Selected callus colonies that had embryo-like structures were then transferred to regeneration medium containing cytokinins, and regeneration frequencies up to 80% were obtained. Small shoots emerged and were transferred to jars for root development prior to transferring to pots of soil and growing the plants to maturity in growth chambers. Of the cytokinins evaluated, N6-benzylaminopurine was the most effective in promoting shoot formation; however, kinetin was also somewhat effective. Regeneration medium could be either an N6 or Murashige and Skoog basal medium. Of 76 plants grown to maturity, 62 were fertile, and the plant heights averaged about three-fourths the height of seed-grown plants.Two other suspension cultures of IR54, one developed from the protoplast callus of the initial IR54 line, and the other developed from callus produced by mature seeds, have yielded protoplasts capable of regenerating plants when using cells of the Calrose 76 suspension as a nurse culture. In addition, protoplasts obtained from three-week-old primary callus of immature embryos of IR54 were capable of regenerating plants when using the same culture conditions.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- pcy
packed cell volume
- BAP
N6-benzylaminopurine
- FDA
fluorescein diacetate
- FW
fresh weight
- IAA
indole-3-acetic acid
Media AA
Muller and Grafe (1978)
- CPW
Frearson et al. (1973)
- Kao*
Kao (1977)
- LS
Linsmaier and Skoog (1965)
- MS
Murashige and Skoog (1962)
- N6
Chu et al. (1975)
- PCM
Ludwig et al. (1985) 相似文献
16.
Ana Sofia Duoue Ana Sofia Pires Dulce Metelo Dos Santos Pedro Fevereiro 《In vitro cellular & developmental biology. Plant》2006,42(3):270-273
Summary Plants were suecessfully régenerated via somatic embryos from 3-yr-old cell suspension cultures of Medicago truncatula Gaertin. cv. Jemalong line M9-10a. The cultures were originally initiated from callus induced in well-expanded leaflets of
30 d in vitro-grown plants, Suspension cultures were established in stirred-liquid Murashige and Skoog (MS) basal salts and vitamins supplemented
with 2.3 μM 2.4-dichlorophenoxyacetic acid (2,4-D) and 2.3 μM kinetin (Kin) and subeultured weekly. Somatic embryogenesis induction step was conducted in liquid MS medium containing 0.45
μM 2,4-D and 0.91 μM zeatin (Zea), during 1,2, and 3wk after subculture. Induced and non-induced cultures were transferred to solid embryo proliferation
medium [EPM-MS basal salts and vitamins solidified with 0.2% (w/v) gelrite]. Somatic embryos developed until the late torpedo/dicotyledonary
stages. We found that the best condition for the development of somatic embryos was achieved when suspension cultures were
not subjected to the induction step. Induction of 1 and 2 wk led to a decrease in the recovery of somatic embryos and the
3-wk treatment resulted in no differentiation of somatic embryos. Plant regeneration was obtained in all conditions (except
for 3wk induction) when embryos were transferred to an embryo conversion medium [ECM, similar to EPM but solidified with 0.7%
(w/v) agar]. Embryo conversion rates were 54.5±1.6, 52.5±18.5, and 41.6±8.4% for 0, 1, and 2 wk induction treatments, respectively.
These plants were successfully transferred to the greenhouse where they matured and produced seeds. 相似文献
17.
A. Amante-Bordeos L. A. Sitch R. Nelson R. D. Dalmacio N. P. Oliva H. Aswidinnoor H. Leung 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1992,84(3-4):345-354
Summary
Oryza minuta J. S. Presl ex C. B. Presl is a tetraploid wild rice with resistance to several insects and diseases, including blast (caused by Pyricularia grisea) and bacterial blight (caused by Xanthomonas oryzae pv. oryzae). To transfer resistance from the wild species into the genome of cultivated rice (Oryza sativa L.), backcross progeny (BC1, BC2, and BC3) were produced from interspecific hybrids of O. sativa cv IR31917-45-3-2 (2n=24, AA genome) and O. minuta Acc. 101141 (2n=48, BBCC genomes) by backcrossing to the O. sativa parent followed by embryo rescue. The chromosome numbers ranged from 44 to 47 in the BC1 progeny and from 24 to 37 in the BC2 progeny. All F1 hybrids were resistant to both blast and bacterial blight. One BC1 plant was moderately susceptible to blast while the rest were resistant. Thirteen of the 16 BC2 progeny tested were resistant to blast; 1 blast-resistant BC2, plant 75-1, had 24 chromosomes. A 3 resistant: 1 susceptible segregation ratio, consistent with the action of a major, dominant gene, was observed in the BC2F2 and BC2F3 generations. Five of the BC1 plants tested were resistant to bacterial blight. Ten of the 21 BC2 progeny tested were resistant to Philippine races 2, 3, and 6 of the bacterial blight pathogen. One resistant BC2, plant 78-1, had 24 chromosomes. The segregation of reactions of the BC2F2, BC2F3, and BC2F4 progenies of plant 78-1 suggested that the same or closely linked gene(s) conferred resistance to races 2, 3, 5, and 6 of the bacterial blight pathogen from the Philippines. 相似文献
18.
Summary To increase plantlet regeneration frequency, rice callus was dehydrated in a Petri dish with a single layer of filter paper prior to transfer to the regeneration medium. With a 24 h dehydration treatment, the regeneration frequency was increased to 47 %, while the regeneration frequency of the untreated control was less than 5 %. This relatively simple method provides an alternative method for improving the regeneration frequency of rice callus. 相似文献
19.
Identification of fungal (Magnaporthe grisea) stress-induced genes in wild rice (Oryza minuta) 总被引:4,自引:0,他引:4
Shim KS Cho SK Jeung JU Jung KW You MK Ok SH Chung YS Kang KH Hwang HG Choi HC Moon HP Shin JS 《Plant cell reports》2004,22(8):599-607
To identify fungal stress-related genes in wild rice, Oryza minuta, we constructed a subtracted library using suppression subtractive hybridization in combination with mirror orientation selection. DNA chips containing 960 randomly selected cDNA clones were applied by reverse Northern analysis to eliminate false positive clones from the library and to prescreen differentially expressed genes. In total, 377 cDNA clones were selected on the basis of their signal intensities and expression ratios. Sequence analyses of these 377 cDNA fragments revealed that 180 of them (47.7%) represented unique genes. Of these180 cDNAs, 89 clones (49.6%) showed significant homologies to previously known genes, while the remaining 91 did not match any known sequences. The putative functions of the 180 unique ESTs were categorized by aligning them with MIPS data. They were classified into seven different groups using microarray data-derived expression patterns and verified by Northern blotting.Abbreviations
ER:
Endoplasmic reticulum
-
EST:
Expressed sequence tag
-
MIPS:
Munich Information Center for Protein Sequences
-
MOS:
Mirror orientation selection
-
NCBI:
National Center for Biotechnology Information
-
omfi:
Oryza minuta fungal-stress induced
-
PCD:
Programmed cell death
-
PDI:
Proteins disulfide isomerase
-
SSH:
Suppression subtractive hybridization
Communicated by I.S. ChungK.S. Shim and S.K. Cho contributed equally to this work. 相似文献
20.
Chun-Nong Huang Jun-Hui Wang Qiu-Sheng Yan Xue-Qin Zhang Qin-Feng Yan 《Plant cell reports》1995,14(11):730-734
Summary Rice cells were precultured for 10 d in medium containing 60 g/L sucrose and subsequently for 1 d in medium supplemented with 0. 4 M sorbitol. After loading with 25%PVS2 at 22°C for 10 min and dehydration in 100%PVS2 at 0°C for 7. 5 min,they were plunged into liquid nitrogen directly. Survival was 45. 0 ±5.1% (based on the reduction of triphenyl tetrazolium chloride)following warming and unloading. For regrowth, cells were plated on semi-solid medium replenished with 40%(w/v) starch for 2d prior to reculture. Cell suspensions were reestablished and plants were regenerated from recovered cells. Twenty eight plants set seeds in the greenhouse.Abbreviations PVS
plant vitrification solution
- P
preculture
- LN
liquid nitrogen
- TTC
triphenyl tetrazolium chloride
- 2,4-D
2,4-dichlorophenoxyacetic acid
- DMSO
dimethyl sulfoxide
- EG
ethylene glycol
- BSA
bovine serum albumin 相似文献