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1.
提高榨菜离体培养植株再生频率 总被引:11,自引:0,他引:11
采用榨菜“浙桐1号”品种为材料,以MS为基本培养基,通过对不同植物生长调节剂的组合和不同外植体等主要因素的筛选,大幅度提高了榨菜离体培养植株再生频率。结果表明,2mg/L6.BA 0.2mg/L2,4-D的组合较为适宜,其不定芽再生频率可达50%,且外植体以下胚轴为好:而CPPU和2,4-D的适宜组合为1.5mg/L 0.2mg/L,其不定芽再生频率高达66.67%,最适外植体为带柄子叶。同时,研究结果显示,添加0.25~1mg/L的GA,对榨菜已分化的不定芽的伸长有抑制作用;子叶柄和下胚轴外植体的分化具有极性现象。 相似文献
2.
Phenylacetic acid improves bud elongation and in vitro plant regeneration efficiency in Capsicum annuum L. 总被引:1,自引:0,他引:1
A highly efficient three-stage protocol for the regeneration of chilli pepper (Capsicum annuum L.) from cotyledon explants was developed. This protocol used PAA in both the shoot-bud induction medium and the medium for
elongation of the shoot buds. A superior medium for the induction of buds from the cotyledons was MS medium supplemented with
BA (5 or 7 mg/l) + PAA (2 mg/l). Buds were elongated during the second stage on medium containing BA (2 or 5 mg/l) + PAA (2 mg/l).
On this medium most of the buds elongated, and their number also increased due to the formation of new buds; bud elongation
was achieved in 100% of the cultures provided the buds were induced in the primary stage on a medium supplemented with BA+PAA.
The shoots that elongated in the second-stage rooted at 100% frequency on a medium supplemented with NAA (1 mg/l). The complete
plantlets with well-developed root and shoot systems were transferred to field conditions where they grew to maturity, flowered
and fruited normally. While shoot-bud induction from the cultured cotyledons was also observed on media supplemented with
BA (5 or 7 mg/l) alone or in combination with IAA (0.2–2 mg/l), buds induced on these media were often distorted, with most
not developing into normal shoots in the second-stage subculturing; a rosette of buds was seen in the second stage subculturing.
On the other hand, PAA in combination with BA in the primary induction medium and second-stage medium promoted normal development
and the elongation of shoot buds.
Received: 28 July 1998 / Revision received: 22 December 1998 / Accepted: 19 February 1999 相似文献
3.
为了提高辣椒子叶不定芽的伸长率和遗传转化效率,本研究以保加利亚尖椒子叶为外植体,通过正交试验分别对影响保加利亚尖椒子叶不定芽伸长的激素组合以及遗传转化参数进行了优化。结果表明:诱导不定芽伸长的最佳激素组合为0.2mg/LIAA+1.0mg/LGA3+0.1mg/LPBU,不定芽伸长率最高为60%;以5mg/L潮霉素为选择压,预培养时间为4d、共培养时间为2d、侵染时间为20min时,诱导的抗性不定芽比率最高。本研究建立的辣椒再生及遗传转化体系为辣椒转基因研究奠定一定的基础。 相似文献
4.
The effect of copper sulphate on differentiation and elongation of shoot buds from cotyledonary explants of Capsicum annuum L. cv X-235 was investigated. Shoot buds were induced on medium supplemented with 22.2 μM BAP and 14.7 μM PAA. Elongation
of shoot buds was obtained on MS medium containing 13.3 μM BAP + 0.58 μM GA3. Both shoot induction and elongation media were supplemented with different levels of CuSO4 (0–5 μM). The levels of CuSO4 in the induction as well as elongation medium highly influenced the shoot bud formation and their subsequent elongation.
Highest number of shoot buds per explant was obtained when the concentration of CuSO4 was increased 30 times to the normal MS level. Shoot buds formation frequency i.e., the number of shoots formed per explant
was increased two fold as compared to those formed on control. Elongation both in terms of percentage and length of shoots
was better than that on control. Healthy elongated shoots were rooted on MS medium supplemented with 5.7 μM IAA. Rooted plantlets
were transferred to field conditions. 相似文献
5.
Andrzej Borychowski Katarzyna Niemirowicz-Szczytt Magdalena Jędraszko 《Acta Physiologiae Plantarum》2002,24(3):257-264
Morphogenetic potential of hypocotyl and cotyledon explants of the three Polish Capsicum annuum L. cultivars (Kujawianka, Passat and Zorza) was studied to develop a reliable plant regeneration protocol. Out of 8 and 15
combinations of growth regulators used in the first and second series respectively, the best medium contained BAP (5 mg·l−1) and IAA (1 mg·l−1). Media containing thidiazuron (TDZ) and IAA proved to be worse than those with BAP and IAA. Additionally, it was indicated
that hypocotyl explants placed upside-down developed more adventious buds. ‘Passat’ was the most responsive variety (approximately
40 % of both types of explants produced buds).
In the second part of experiment the aim was to stimulate shoot induction in the conditions most adapted to Agrobacterium transformation. ‘Bryza’ replaced cv ‘Kujawianka’ and proved to be better than ‘Passat’, previously distinguished as a highly
responsive cultivar. The experiments confirmed a significant effect of the hypocotyl explant length and induction period on
shoot regeneration. Finally, the optimum concentration of carbenicillin and kanamycin was determined. 相似文献
6.
Yeh-Jin Ahn Louisa Vang Thomas A. McKeon Grace Q. Chen 《In vitro cellular & developmental biology. Plant》2007,43(1):9-15
An efficient plant regeneration protocol was established for castor (Ricinus communis L.). Hypocotyl tissue from zygotic embryo axis produced adventitious shoots when treated with either thidiazuron (TDZ, 1 μM)
or 6-benzylaminopurine (BA, 20 μM). TDZ resulted in more than a threefold higher rate of shoot induction (a maximum of 24.2
shoots per explant) than BA (6.8 shoots). Our results also showed that the pretreatment of explants in the dark increased
the number of shoots regenerated per explant by 82% and 36% with TDZ and BA, respectively. The elongation of hypocotyl tissue
in the dark appears to be the primary cause of the increase. Comparable rates of rooting were achieved on the media supplemented
with either indole-3-butyric acid (IBA, 84.3%) or 1-naphthaleneacetic acid (NAA, 87.4%) at 5 μM. However, IBA was more efficient
in promoting root and shoot development, resulting in a higher rate of establishment (93.5%) in the soil, compared to the
rate with NAA (39.5%). Histological analysis showed the adventitious induction of the shoot buds originated from the cortex
of the hypocotyl tissue. 相似文献
7.
Improvement of regeneration in pepper: a recalcitrant species 总被引:2,自引:0,他引:2
Najet Gammoudi Tania San Pedro Ali Ferchichi Carmina Gisbert 《In vitro cellular & developmental biology. Plant》2018,54(2):145-153
Organogenesis is influenced by factors like genotype, type of explant, culture medium components, and incubation conditions. The influence of ethylene, which can be produced in the culture process, can also be a limiting factor in recalcitrant species like pepper. In this work, bud induction was achieved from cotyledons and hypocotyls—from eight pepper cultivars—on Murashige and Skoog (MS) medium supplemented with 22.2 μM 6-benzyladenine (6BA) and 5.71 μM indole-3-acetic acid (IAA), in media with or without silver nitrate (SN) (58.86 μM), a suppressor of ethylene action. In the SN-supplemented medium, the frequencies of explants with buds and with callus formation were lower in both kinds of explant, but higher numbers of developed shoots were isolated from explants cultured on SN. Bud elongation was better in medium with gibberellic acid (GA3) (2.88 μM) than in medium free of growth regulators or supplemented with 1-aminocyclopropane-1-carboxylic acid (ACC) at 34.5 μM. However, isolation of shoots was difficult and few plants were recovered. The effect of adding SN following bud induction (at 7 d) and that of dark incubation (the first 7 d of culture) was also assessed in order to improve the previous results. When SN was added after bud induction, similar percentages of bud induction were found for cotyledons (average frequency 89.37% without SN and 94.37% with SN) whereas they doubled in hypocotyls (50% without SN and 87.7% with SN). In addition, in both kinds of explant, the number of developed plants able to be transferred to soil (developed and rooted) was greatly increased by SN. Dark incubation does not seem to improve organogenesis in pepper, and hypocotyl explants clearly represent a better explant choice—with respect to cotyledonary explants—for the pepper cultivars assayed. 相似文献
8.
A high frequency shoot regeneration system for ornamental kale [Brassica oleracea L. var. acephala (D.C.) Alef.] was firstly established from seedling cotyledon and hypocotyl explants. The ability of cotyledon and hypocotyl
to produce adventitious shoots varied depending upon genotype, seedling age and culture medium. The maximum shoot regeneration
frequency was obtained when the explants from cv. Nagoya 4-d-old seedlings were cultured on Murashige and Skoog (MS) medium
supplemented with 3 mg dm−3 6-benzylaminopurine (BA) and 0.1 mg dm−3 naphthaleneacetic acid (NAA). The frequency of shoot regeneration was 65.0 % for cotyledons, 76.1 % for hypocotyls; and the
number of shoots per explant was 4.3 for cotyledons, 8.2 for hypocotyls. Hypocotyl explants were found to be more responsive
for regeneration when compared with cotyledons. Among the 4 cultivars tested, Nagoya showed the best shoot regeneration response.
The addition of 3.0 mg dm−3 AgNO3 was beneficial to shoot regeneration. Roots were formed on the base of the shoots when cultured on half-strength MS medium. 相似文献
9.
P. G. Golegaonkar A. S. Kantharajah 《In vitro cellular & developmental biology. Plant》2006,42(4):341-344
Summary
In vitro plantlet regeneration was obtained from cultured cotyledon and young leaf explants of five Indian chile pepper cultivars
(Capsicum annuum L. evs. Gujarat-1, Gujarat-2, Guntur-4, Selection-49, and Jwala). Adventitious shoot buds (ASB) were regenerated directly
from cotyledon and young leaf explants in all the five cultivars on media containing benzyladenine (BA) alone or in combination
with 1-naphthaleneacetic acid (NAA). Regeneration frequency was highly influenced by cultivar explant type, media combination
and their interactions, except the interaction between cultivar and explant, for number of ASB per explant. Percent contribution
of individual source suggested that selection of explant type followed by medium combination and cultivars was essential for
obtaining high-frequency ASB induction. Across different cultivars the young leaf explant was found to be the most responsive
explant, while Murashige and Skoog (MS) medium containing BA alone (17.8, 26.6, and 35.5 μM) was found to be the best medium for the production of maximum number of ASB. Between the two explants, shoot elongation
was observed with ASB obtained from young leaf explants on MS medium containing BA (2.2 and 4.4 μM) and gibberellie acid (GA3) (1.4, 2.9, 4.3 and 5.8 μM). The MS medium fortified with 4.4 μM BA+2.9μM GA3 was optimum for shoot elongation. Elongated shoots were rooted on liquid MS medium supplemented with 2.9 μM indole-3-acetic acid (IAA) and successfully established ex vitro. 相似文献
10.
Summary Thirteen soybean genotypes representing maturity groups IV−VI were compared for organogenic responses on three media cultured
under two lighting conditions with hypocotyl sections excised from 7-d-old seedlings. All soybean lines responsed by producing
adventitious shoots on the acropetal end of the hypocotyl explants, confirming genotype-independence of shoot initiation.
Media containing 6-benzyladenine (BA; 5.0–10 μM) induced the greatest numbers of shoots. Histological studies confirmed the adventitious nature of arising shoots by indicative
formation of meristematic zones and shoot primordia from parenchymatous tissues of central pith and plumular trace regions
of the hypocotyl. Incompletely excised cotyledonary buds also contributed to shoot initiation. Degrees of responses were media-dependent
and varied with regard to genotype. Centennial, Epps, and Lyon gave the greatest individual responses. Between cultivars (across
all treatments), the regeneration potential (percentage of explants producing meristem-like structures or shoot primordia)
4 wk after initiation ranged from 47 to 75%. Four wk later, regenerative ability (number of shoots produced per responding
explant) and regeneration efficiency (number of shoots produced per explant plated) yielded 1.4–7.1 and 1.0–5.0 shoots, respectively.
The optimized protocol included initiation on a medium containing 5.0 μM BA for 4 wk, then transfer onto a shoot elongation medium (0.36 μM BA) for 4 wk. For 11 genotypes tested, 66–100% of excised shoots produced roots after 4 wk on media containing 12.5–29.2
μM indole-3-butyric acid. Of 109 regenerants transplanted to soil, 94% survived and no sterility has been observed on those
mature enough to flower. 相似文献
11.
Durga Singh Nagar Suman Kumar Jha Jigar Jani 《Physiology and Molecular Biology of Plants》2015,21(2):287-292
A reproducible protocol developed for in vitro regeneration of Milletia pinnata using hypocotyl segments. Multiple shoots were induced from hypocotyl explants through direct adventitious shoot bud regeneration. The proximal end of hypocotyls was responsive for shoot bud induction. Silver nitrate and adenine sulphate had a positive effect on shoot bud induction and elongation. The maximum response and number of shoot bud produced in media supplemented with 8.88 μM BAP with 108.6 μM adenine sulphate and 11.84 μM silver nitrate. Elongated shoots were harvested and successful rooting of microshoots achieved on MS media supplemented with 9.84 μM IBA, with 81.1 % rooting. Remaining shoot buds sub-cultured for further multiplication and elongation. Each subculture produced eight to nine elongated microshoots up to four subcultures. The rooted microshoots were successfully hardened and transferred to field. 相似文献
12.
In this work we report a new method forin vitro chili pepper (Capsicum annuum L.) plant regeneration based on shoot formation from wounded hypocotyls. Chili pepper seeds were surface sterilized and germinated on agar (0.8%) at 25 ± 2°C in the dark. Five factors that may influence shoot regeneration were studied: age of seedlings, hypocotyl wounding site, time elapsed between wounding the hypocotyls and decapitation of seedlings, culture media and cultivars. In order to study the influence of the first three factors on shoot regeneration, the apical, middle or basal hypocotyl regions of seedlings of cv. Mulato Bajio at different stages of development (9, 15, 16, 21 and 28 d old) were wounded with a syringe needle, and the seedlings were cultured on MS semisolid medium without growth regulators at 25 ± 2°C under a 16/8 h light/dark photoperiod (daylight fluorescent lamps; 35 mol m-2 s--1) until decapitation. The seedlings were decapitated (3 mm below the cotyledons) at different times after wounding (0, 2, 4, 10, 12 and 14 d), and each explant was evaluated for bud and shoot formation ( 5 mm in length) at the wounded site after 30 d of incubation. In general, seedlings at the stage of curved hypocotyl (9 d old) wounded in the apical region of hypocotyl were the best explants for shoot regeneration when inoculated on culture medium without growth regulators. Decapitation after wounding also influenced the shoot regeneration efficiency, with 10–14 d being the best period. Up to 90% shoot regeneration in cv. Mulato Bajio was obtained under these conditions. Statistically significant differences were observed for shoot formation among 21 cultivars tested. Regeneration of whole plants was achieved by rooting the shoots with indole-3-butyric acid pulses of 60 mg L–1 for 3 h and then subculturing on MS medium without growth regulators. 相似文献
13.
S. K. Yadav M. Gopala Krishna M. Maheswari M. Vanaia B. Venkateswarlu 《Journal of plant biochemistry and biotechnology.》2010,19(2):267-270
An efficient protocol for shoot bud induction and proliferation employing half cotyledonary node with intact cotyledon explants derived from two-day-old seedlings of mung bean pre-conditioned on 6- benzylaminopurine (BAP) has been achieved. Explants were cultured for four weeks each on MS B5 + 12.5 μM BAP and MS B5 + 5 μM BAP +0.05 μM α-naphthaleneacetic acid (NAA ), respectively, as shoot bud induction and shoot elongation and proliferation media, gave the best regeneration response. The removal of the pre-existing buds from explants at 12 days in shoot bud induction medium led to enhanced regeneration response. Light microscopic observations on 14-day-old explants confirmed direct organogenesis route of regeneration. Elongated shoots (>2 cm) excised from the regenerating cultures were successfully rooted on half MS B5 medium containing 2.46 μM indolebutyric acid (IBA). About 90% of the rooted plantlets, efficiently hardened in pots having soil and farm yard manure, flowered and produced pods with viable seeds upon reaching maturity. 相似文献
14.
15.
Vinod Kumar Ashwani Sharma Bellur Chayapathy Narasimha Prasad Harishchandra Bhaskar Gururaj Parvatam Giridhar Gokare Aswathanarayana Ravishankar 《Acta Physiologiae Plantarum》2007,29(1):11-18
Direct shoot bud induction and plant regeneration was achieved in Capsicum frutescens var. KTOC. Aseptically grown seedling explants devoid of roots, apical meristem and cotyledons were inoculated in an inverted
position in medium comprising of Murashige and Skoog (Physiol Plant 15:472–497, 1962) basal medium supplemented with 2-(N-morpholine) ethanesulphonic acid buffer along with 2.28 μM indole-3-acetic acid, 10 μM silver nitrate and either of 13.31–89.77 μM
benzyl adenine (BA), 9.29–23.23 μM kinetin, 0.91–9.12 μM zeatin, 2.46–9.84 μM 2-isopentenyl adenine. Profuse shoot bud induction
was observed only in explants grown on a media supplemented with BA (26.63 μM) as a cytokinin source and 19.4 ± 4.2 shoot
buds per explant was obtained in inverted mode under continuous light. Incorporation of polyamine inhibitors in the culture
medium completely inhibited shoothoot bud induction. Incorporation of exogenous polyamines improved the induction of shoot
buds under 24 h photoperiod. These buds were elongated in MS medium containing 2.8 μM gibberellic acid. Transfer of these
shoots to hormone-free MS medium resulted in rooting and rooted plants were transferred to fields. This protocol can be efficiently
used for mass propagation and presumably also for regeneration of genetically transformed C. frutescens. 相似文献
16.
亚麻组织培养高频不定芽诱导体系 总被引:1,自引:0,他引:1
对适合南方地区冬季种植的纤用亚麻品种组织培养过程中基本培养基、激素配比、外植体材料的基因型和苗龄以及再生不定芽的生根条件进行了比较研究。结果表明, 适合于亚麻白花品种组织培养的最佳培养基为YB1, 不定芽诱导率可达98.50%。在此培养基上, 白花、黑亚4号、K6531、K7697、HI026、HI045、I039和阿丽亚那下胚轴不定芽的诱导率分别为98.50%、98.50%、56.50%、42.47%、54.40%、0、27.13%和97.30%, 平均出芽数为11.43、9.33、2.17、0.77、 1.10、0、0.90和10.68。苗龄为7-10天的下胚轴最适于诱导不定芽, 随苗龄增加, 不定芽的诱导率呈下降趋势。RB5培养基最适于不定芽的生根,生根率达100%, 平均生根数为15.3。实验还确定了亚麻对卡那霉素、氨苄青霉素和头孢霉素的抗性浓度阈值。 相似文献
17.
亚麻组织培养高频不定芽诱导体系 总被引:6,自引:0,他引:6
对适合南方地区冬季种植的纤用亚麻品种组织培养过程中基本培养基、激素配比、外植体材料的基因型和苗龄以及再生不定芽的生根条件进行了比较研究。结果表明,适合于亚麻白花品种组织培养的最佳培养基为YB1,不定芽诱导率可达98.50%。在此培养基上,白花、黑亚4号、K6531、K7697、HI026、HI045、I039和阿丽亚那下胚轴不定芽的诱导率分别为98.50%、98.50%、56.50%、42.47%、54.40%、0、27.13%和97.30%,平均出芽数为11.43、9.33、2.17、0.77、1.10、0、0.90和10.68。苗龄为7-10天的下胚轴最适于诱导不定芽,随苗龄增加,不定芽的诱导率呈下降趋势。RB5培养基最适于不定芽的生根,生根率达100%,平均生根数为15.3。实验还确定了亚麻对卡那霉素、氨苄青霉素和头孢霉素的抗性浓度阈值。 相似文献
18.
Explants from 13-d old pepper (Capsicum annuum, L. cv. Early California Wonder) seedlings were cultured in Murashige and Skoog (MS) medium supplemented with different levels
of 1-naphthalene acetic acid (NAA) and 6-benzylamino purine (BAP). Multiple shoot-buds proliferated from the cut surfaces
of cotyledon, shoot-tip and hypocotyl explants in one month. The best NAA to BAP combinations (mg/l: mg/l) for multiple shoot-bud
regeneration of the above three explant types were 0.1 ∶ 5.0, 0.0 ∶ 5.0, and 0.1 ∶ 10.0, respectively. Root explants did not
express any new morphogenetic response in all hormonal combinations tested. Regenerated shoot-buds were excised from the explants
and cultured in 1/2X or 1X MS medium supplemented with different levels of Indole-3-acetic acid (IAA) or NAA. When cultured
in full strength MS medium with 0.5 mg/l IAA or 0.4 mg/l NAA, 70% of the buds rooted in one month. Plantlets were established
successfully ex vitro under greenhouse mist and grown to maturity. 相似文献
19.
Callus was produced on cotyledon, shoot tip, hypocotyl and root explants of twoCorchorus species on several media. Cytokinin was necessary for callus production on cotyledon explants. BothC.olitorius genotypes produced most callus on media with zeatin and either NAA or IAA, and theC.capsularis genotype produced most callus on media with IAA and either zeatin or BA. High frequencies of regenerated shoots were obtained
from shoot tip explants of both species, from the apical meristem and from callus. Media with 2.0 mg 1−1 BA were superior for both species, and media with zeatin were equally good forC.capsularis only. More regeneration was obtained for all genotypes after subculture of callus on media with 2.0 mg 1−1 zeatin. Cotyledon callus produced less regeneration, also with differences between genotypes; explants of both genotypes
ofC.olitorius produced regeneration on a medium with NAA and zeatin, and theC.capsularis genotype produced regeneration on a medium with IAA and BA. Limited regeneration from root explant callus was obtained forC.capsularis only on medium with BA and IAA. Regeneration was not obtained from hypocotyl callus. Further regeneration of shoots of both
species was obtained from secondary callus after subculture, and from nodal segments of regenerated shoots and of seedling
shoots cultured on basic MS medium without growth hormones. Roots were produced on about 80% of all shoots after transference
to medium with 0.2 mg 1−1 IBA, and rooted plantlets survived and flowered normally after transference to compost. 相似文献
20.
J. Ponsamuel D. V. Huhman B. G. Cassidy D. Post-Beittenmiller 《Plant cell reports》1998,17(5):373-378
Shoot buds were induced from plumular explants of peanut (Arachis hypogaea L., cv `Okrun') preconditioned on medium containing 2,4-dichlorophenoxyacetic acid and kinetin and then transferred to regeneration
medium containing benzylaminopurine and β-naphthoxyacetic acid. Buds differentiated 25 days following transfer to regeneration medium. Each explant produced 30 to
40 buds, but only 4 shoots. The remaining buds were dormant and did not produce shoots when maintained on regeneration medium.
Shoots were regenerated continuously, however, when explants were subsequently transferred to shoot conversion medium containing
1 μM brassin, benzylaminopurine and β-naphthoxyacetic acid, respectively. Approximately 5 shoots were harvested every 30 days after transfer to shoot conversion
medium for up to 7 months. No further shoot production was observed from explants maintained on regeneration medium without
brassin. Regenerated shoots could be rooted and produced viable seeds. This procedure provides an efficient and reliable system
for regeneration and transformation studies using cv `Okrun'.
Received: 9 April 1997 / Revision received: 27 August 1997 / Accepted: 20 September 1997 相似文献