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1.
《Journal of Crop Improvement》2013,27(4):432-446
Broccoli (Brassica oleracea var. italica) is an important nutritionally rich vegetable cole crop grown in the world. Environmental stress, pests, and diseases cause enormous yield losses because of a limited gene pool. Genetic manipulation is becoming an important method for broccoli improvement. The objective of present study was to evaluate the potency of thidiazuron (TDZ) as a plant growth regulator in evoking morphogenic responses in leaf and petiole explants of broccoli. An efficient, reproducible, and high frequency plant regeneration protocol has been standardized in broccoli cv. Solan green head. Leaf and petiole explants were cultured on Murashige-Skoog (MS) medium, supplemented with a wide range of TDZ concentrations. The following treatments were designed for efficient in vitro shoot regeneration: TDZ alone, TDZ with adenine, TDZ with naphthalene acetic acid (NAA), and TDZ with indole acetic acid (IAA). Among the 36 combinations of growth regulators used, the highest percentage of leaf explants producing shoot (89.25%) was recorded on MS medium containing 1.0 μM TDZ and 0.107 μM NAA. The multiple shoot regeneration response of petiole explant producing shoots (91.55%) was obtained on MS medium containing 2.0 μM TDZ and 0.107 μM NAA. Shoot multiplication and elongation were obtained on the same medium. For root regeneration in in vitro regenerated shoots, different concentrations of NAA were applied. High frequency (100%) root regeneration response with healthy and vigorous roots was observed on MS medium supplemented with 0.54 μM NAA. The regenerated plantlets with well-developed shoots and root system were transferred to pots containing cocopeat and successfully acclimatized. We recommend 1.0 μM TDZ with 0.107 μM NAA and 2.0 μM TDZ and 0.107 μM NAA combinations for adventitious shoot regeneration from leaf and petiole explants in broccoli cv. Solan green head respectively. This is the first report on high frequency organogenesis from leaf and petiole explants of broccoli cv. Solan green head using thidiazuron. 相似文献
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M. ThiyagarajanP. Venkatachalam 《Industrial Crops and Products》2012,37(1):111-117
Stevia rebaudiana is a valuable medicinal plant species and it is being used for the treatment of diabetes. Currently, there is a high demand for raw material of this medicinal herb due to ever increasing diabetes disorder among the population. In order to meet the increased demand an efficient in vitro propagation of S. rebaudiana was established. Nodal explants collected from the field were cultured on MS basal medium fortified with different concentrations of BAP (0.5-3.0 mg/l) and KIN (0.5-3.0 mg/l) individually for shoot bud induction. In vitro derived nodal buds were cultured on MS medium supplemented with different concentrations (0.5-3.0 mg/l) of BAP and KIN for multiple shoot bud regeneration. In the second experiment, in vitro derived buds were placed on MS medium supplemented with different concentrations of BAP (0.5-3.0 mg/l) in combination with 0.5 mg/l IAA or IBA or NAA for shoot bud multiplication. The highest frequency (94.50%) of multiple shoot regeneration with maximum number of shoots (15.69 shoots/explant) was noticed on MS medium supplemented with 1.0 mg/l BAP. For large scale plant production, in vitro derived nodal bud explants were cultured on MS medium fortified with 1.0 mg/l BAP, in which about 123 shoots/explant were obtained after three subcultures on the same media composition. Elongated shoots (>2 cm) dissected out from the in vitro proliferated shoot clumps were cultured on half-strength MS medium containing different concentrations of NAA (0.1-0.5 mg/l) and/or MS medium fortified with various concentrations (0.5-2.0 mg/l) of auxins (NAA, IAA and IBA) for root induction. Highest frequency of rooting (96%) was noticed on half-strength MS medium augmented with 0.4 mg/l NAA. The rooted plantlets were successfully transferred into plastic cups containing sand and soil in the ratio of 1:2 and subsequently established in the greenhouse. The present in vitro propagation protocol would facilitate an alternative method for rapid and large-scale production of this important antidiabetic medicinal plant. 相似文献
3.
为建立白头翁再生体系,以白头翁主根的切段和试管苗叶片为外植体,比较2种外植体离体培养差异,探讨不同植物生长调节剂对不定芽和愈伤组织诱导、愈伤增殖和再分化及芽苗生根的影响。结果表明:叶片可以通过愈伤组织再分化和直接产生不定芽2种途径建立再生体系,而根只诱导出了愈伤组织,增殖培养中逐渐褐化死亡;在含6-BA培养基上,叶块死亡,而根只诱导出少量愈伤组织;叶片诱导不定芽的培养基为MS+TDZ 0.3 mg/L+NAA 0.1 mg/L;愈伤组织增殖的培养基为MS+TDZ 0.2 mg/L+2,4-D 0.2 mg/L;再分化时,需要转接到TDZ和NAA组合的培养基上,其中处理组合TDZ 0.3 mg/L+NAA0.1 mg/L的再分化率达100%;生根培养基为1/2MS+NAA 0.2 mg/L+IBA 0.2 mg/L+蔗糖20 g/L。不同外植体离体培养存在差异,叶片较根易培养;TDZ对白头翁叶片培养效果较好,2,4-D对愈伤组织的诱导能力较强,而NAA适合于不定芽分化,NAA与IBA组合使用生根效果较好。 相似文献
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Salma U Rahman MS Islam S Haque N Jubair TA Haque AK Mukti IJ 《Pakistan journal of biological sciences: PJBS》2008,11(12):1638-1641
The influence of media composition on callus induction and subsequent regeneration of Rauwolfia serpentina L. Benth has been studied. High frequency (96.43%) callus induction was obtained when nodal segments from in vitro raised shoots were cultured on MS medium supplemented with 0.5 mg L(-1) BA and 2.0 mg L(-1) NAA. The callus differentiated into adventitious shoots when it was subcultured on MS medium supplemented with 2.0 mg L(-1) BA with 0.2 mg L(-1) NAA. Regenerated shoots were best rooted on half-strength MS medium with 1.0 mg L(-1) each of IBA and IAA. 相似文献
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S. L. Lam 《American Journal of Potato Research》1977,54(10):465-468
Normal plantlet formation from tuber discs in potato can be successfully inducedin vitro provided that auxin-kinetin ratio in the nutrition medium is properly adjusted. Naphthalene acetic acid (NAA) was found to be effective in adjustment of auxin level in the medium. 相似文献
7.
2个葡萄品系外植体愈伤组织诱导和植株再生 总被引：12，自引：0，他引：12
进行了无核白、红地球2个葡萄品系外植体愈伤组织诱导和植株再生的研究。通过葡萄品系无核白、红地球叶片和叶柄外植体的愈伤组织的诱导、再分化,以器官发生途径实现植株再生。结果表明:(1)愈伤组织诱导以MS+BA5mg/L+NAA0.5mg/L+蔗糖40g/L培养基最佳,诱导率为35%;同一葡萄品种的叶片与叶柄诱导愈伤组织的频率基本相同,而红地球外植体诱导愈伤组织的频率高于无核白;(2)不定芽的诱导再生以1/2MS+BA0.5mg/L+NAA0.05mg/L+CH500mg/L+蔗糖30g/L培养基最佳,平均诱导率达35.4%;叶柄来源的愈伤组织诱导不定芽的频率高于叶片愈伤组织;(3)根系的诱导以1/2MS+KT0.5mg/L+IBA1.0mg/L+AC2g/L+蔗糖15g/L培养基最佳,无核白和红地球来源的不定芽诱导生根频率基本相同,都在80%以上。 相似文献
8.
BA与NAA对药用黄花石蒜鳞片组织培养的影响 总被引：2，自引：0，他引：2
以野生药用植物黄花石蒜(Lycoris aurea Herb.)为材料,选用其鳞茎的内层鳞片、中层鳞片2个不同部位的外植体,研究MS培养基中9种不同BA与NAA的浓度配伍对黄花石蒜鳞片的组织培养效果。结果表明:MS+10mg/L BA+5mg/L NAA+0.7%琼脂+3%蔗糖,有利于内层鳞片和中层鳞片外植体不定芽、不定根的发生,产生褐变量也较少,其培养效果显著较好。在培养基中BA、NAA的浓度分别高于或分别低于10、5mg/L,其培养效果均显著较差。相关分析表明,2种外植体相同BA与NAA浓度配伍的培养效果为极显著正相关,这进一步说明,对一种外植体培养效果较好的BA与NAA的浓度配伍对另一种外植体的培养效果仍然较好。多重比较与相关分析均说明,在培养基中BA、NAA的浓度分别为10、5mg/L对内层鳞片外植体和中层鳞片外植体的培养效果均较好。 相似文献
9.
Mithilesh SinghRakhi Chaturvedi 《Industrial Crops and Products》2012,36(1):321-328
The study revealed, for the first time, accumulation of spilanthol, an antiseptic alkylamide, in in vitro cultures of Spilanthes acmella Murr., a medicinal plant of immense commercial value. To achieve this, in vitro shoots were regenerated via direct organogenesis from leaf-disc explants of Spilanthes. Shoots were induced in the presence of N6-benzylaminopurine (BAP) alone or in combination with either α-naphthalene acetic acid (NAA) or Indole-3-acetic acid (IAA) in Murashige and Skoog medium. The best treatment for shoot regeneration was MS + BAP (5.0 μM) + IAA (5.0 μM), which promoted adventitious shoot proliferation in >82% cultures with an average of 5.3 shoots per explant. Regenerated shoots rooted spontaneously with a frequency of 100% on half strength MS medium (major salts reduced to half strength) containing 50 g l−1 sucrose. The plantlets were acclimatized successfully with 90% survival rate. Additionally, ploidy stability of the regenerated plants was assessed by flow cytometry which showed that all investigated plants had the similar ploidy as that of the mother plant. For spilanthol identification, peaks eluted from HPLC were analyzed by mass spectrometry with its characteristic fragmentation pattern. For quantification studies, calibration curve was generated, which revealed a higher amount of spilanthol content (3294.36 ± 12.4 μg/g DW) in the leaves of in vitro plants compare to those of in vivo plants (2703.66 ± 9.6 μg/g DW of spilanthol). An efficient multiplication frequency, ploidy stability and enhanced spilanthol accumulation ensure the efficacy of the protocol developed for this industrially important medicinal plant. 相似文献
10.
R. Ayadi L. Hamrouni M. HananaM. Trifi M.L. Khouja 《Industrial Crops and Products》2011,33(2):474-480
The present study report a protocol for the efficient in vitro propagation of kenaf (Hibiscus cannabinus L., an industrial crop having high cellulosic fiber content) on hormone free MS medium using the shoot apex and nodal explants. Shoot tips and nodes were isolated from 15 days old seedlings cultivated on MS medium. Different combinations and concentrations of auxin/cytokinin were used and added to the MS medium to assess the shoot and root induction of theses explants. Several subcultures were drived in order to enhance the multiplication rate. Healthy and well developed in vitro propagated shoots were transferred for acclimatization under greenhouse conditions in pots filled with different substrates (sand + compost or perlite). Our results showed that shoots could elongate and root within 4-6 weeks on MS basal medium without any callus formation. However, addition of growth regulators to the MS medium leaded to a decrease in shoot and root induction rates. Indeed, the highest shoot regeneration frequency (90.5%) was obtained on MS control medium. Elongated shoots were transferred onto the same hormone free MS medium using five subcultures where the multiplication rate reached the highest value (3.66) at the fifth and last step. The in vitro rooted plantlets were acclimatized in greenhouse and successfully transplanted to natural conditions with 70% survival. 相似文献
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An improved and efficient in vitro regeneration system has been developed for Eclipta alba, a medicinally important plant, through transverse thin cell layer culture (tTCL). The transverse section of the nodal segment of field grown plants was used as tTCL explants for plant regeneration. Shoot multiplication from tTCL nodal explants was influenced by BAP and their interaction with Kin or NAA. MS medium containing 13.2 μM BAP and 4.6 μM Kin was most effective for shoot multiplication from tTCL nodal explants. Upon this medium, percent response for shoot proliferation was 100% with an average of 32.6 shoot buds per tTCL nodal explant. Regenerated shoots from tTCL nodal explants were rooted on the growth regulator free MS medium. The rooted plantlets were successfully acclimatized and established in soil with a survival frequency of 90-100%. Random amplified polymorphic DNA (RAPD) markers were used to evaluate the genetic fidelity of the micropropagated plants. RAPD profile analysis indicated that micropropagated plants were genetically similar to mother plant. 相似文献
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To reduce the time period for in vitro regeneration in annatto (Bixa orellana L.), a highly efficient two-stage plant regeneration protocol had been developed that can be used commercially. Different types of explants: nodal shoot tips, shoot tips and single nodes from in vitro grown seedlings were inoculated onto the Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of plant growth regulators. Highest number of shoot buds was obtained when nodal shoot tip explants were inoculated onto MS medium supplemented with 31.1 μM N6-benzyladenine (BA) and 14.7 μM phenylacetic acid (PAA). PAA in combination with BA exhibited a synergistic effect on shoot multiplication and elongation. Sub-culturing of the shoots onto the MS medium supplemented with BA (13.3 μM) and PAA (7.3 μM) produced elongated shoots. Elongated shoots when inoculated onto the MS medium supplemented with 4.9 μM indole-3-butyric acid (IBA) produced optimal rooting. The rooted plantlets were hardened and their field survival rate after 6 weeks time was 73%. 相似文献
15.
Salma U Rahman MS Islam S Haque N Khatun M Jubair TA Paul BC 《Pakistan journal of biological sciences: PJBS》2008,11(9):1273-1277
A protocol for mass propagation through axillary bud proliferation was established for Rauwolfia serpentina L. Benth. (Apocynaceae). MS medium supplemented with 1.5 mg L(-1) BA and 0.2 mg L(-1) NAA elicited the maximum number of shoots (4 multiple shoots) from nodal explants. These adventitious shoots were best rooted on half strength MS medium supplemented with 1.0 mg L(-1) each of IBA and IAA. The in vitro raised plants were acclimatized in glass house and successfully transplanted to field condition with almost 95% survival. 相似文献
16.
籼稻绿芽悬浮细胞原生质体再生成株 总被引：2，自引：1，他引：1
采用籼稻Hu-18绿芽为外植体,在N6附加2 mg/L 2,4-D的培养基上诱导愈伤组织。加20 d后转人AA 培养基进行悬浮培养。继代培养45 d左右形成了分裂旺盛的胚性细胞悬浮系。即从绿芽诱导至胚性细胞悬浮系的建立仅用了65 d左右。继代后前6 d,细胞干重几乎每2 d增加1倍,而培养液的渗透压及pH 值迅速下降。取继代后4 d的悬浮细胞游离原生质体,产率为8.74X106/g鲜重。纯化后的原生质体在KPR培养基中进行琼脂糖包埋培养,原生质体植扳率为12.0％ 。将20 d后的小愈伤组织(0.1 mm)转入 N6附加0.5 mg/L 2,4- D、1 mg/ L BA、1 mg/L KT、0.3 mg/L ZT的培养基上增殖,待长成直径2～3 mm 左右时,用N6附加1 mg/L BA,1 mg/L KT,0.3 mg/L ZT的培养基进行分化培养, 5 d后同时出现芽根生长,最终再生成绿色植株,绿苗分化率为3.5株/10000个原生质体。 相似文献
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G. SivanandhanM. Arun S. MayavanM. Rajesh T.S. MariashibuM. Manickavasagam N. SelvarajA. Ganapathi 《Industrial Crops and Products》2012,37(1):124-129
Calli were obtained from leaf, cotyledon and internode explants of in vitro-grown plants of Indian cultivar of Withania somnifera in MS medium supplemented with 2, 4-D (2.0 mg l−1) and Kinetin (0.2 mg l−1). The brown, semi-friable callus (500 mg FW) derived from leaf explants produced higher number of primary adventitious roots (9 roots/callus) in half strength MS medium fortified with IBA (0.5 mg l−1) and NAA (0.1 mg l−1). The primary adventitious roots with an inoculum mass of 15 g FW were cultured for 6 weeks in the same medium for secondary adventitious root proliferation. Elicitation of abiotic elicitor, aluminium chloride at 10 mg l−1 at the end of 4 weeks culture with 4 h exposure time enhanced withanolides productivity. Under similar culture conditions, the biotic elicitor, chitosan at 100 mg l−1 stimulated higher production of all withanolides when compared to aluminium chloride treatment. This is the first report on the use of callus-derived adventitious root culture for the enhanced production of withanolides upon chitosan elicitation. 相似文献
18.
《Industrial Crops and Products》2007,25(2):169-177
Scutellaria species have been used in many traditional medical systems and is well known among the Native American tribes as a strong emmenagogue and as a female medicinal herb. The inoculation of arbuscular mycorrhiza fungi (AMF) into the roots of micropropagated plantlets could help not only mass propagate these species, but also help grow in marginal, phosphorus deficient soils. Leaves, shoot apices, and nodal segments from wild as well as from greenhouse-grown plants were used to initiate cultures in Murashige and Skoog (MS) medium supplemented with cytokinins benzyladenine (BA), kinetin, thidiazuron (TDZ), naphthalene acetic acid (NAA), and indole butyric acid (IBA) Among all the explants tested for shoot bud induction, only shoot tips and nodal explants were responsive. Explants swelled and became rough on the surface at the end of 3-week incubation with many green shoot buds. Two to 3 weeks after transfering to rooting media with or without IBA, all shoots developed roots. In vitro raised plants were acclimatized in a mist chamber and transferred to 6-l containers in the greenhouse to study the role of AMF on plant growth and development. Inoculation with AMF, showed positive effects on plant growth, particularly root development compared with the control plants. Among the five AMF strains tested, S3004 increased plant height and fresh weights of shoot, root, and seed. 相似文献
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S. Kumar R. Tiwari A. Chandra A. Sharma R. K. Bhatnagar 《Grass and Forage Science》2013,68(3):459-468
In vitro direct plant regeneration of lucerne was achieved by simultaneous application of thidiazuron (TDZ) and 6‐benzyladenine (BA) in Murashige and Skoog (MS) medium. Seedlings were germinated and grown for 6 d on growth regulator–containing MS medium. The shoot tip, consisting of the apical meristem along with parts of the cotyledonary leaves and hypocotyl, was then cultured on a medium containing the growth regulator(s). Adventitious budding of the shoot tip was promoted synergistically by treatment with TDZ and BA, and a maximum of thirty‐five shoots per explant was obtained on a medium supplemented with 2 mg L?1 TDZ and 1 mg L?1 BA. Plant regeneration frequency varied from 67 to 93%, and five Indian lucerne cultivars responded well to the regeneration protocol. The Agrobacterium‐mediated transformation frequency from co‐cultivated explants was 13% following multiple shoot induction. Southern analysis of the T0 plants and T1 progenies confirmed stable inheritance of the hpt marker gene. Agrobacterium infection of the explant caused a significant reduction in the plant regeneration frequency (23%) and the number of shoots induced (11) when compared with uninfected explants. A single shoot tip provided sufficient material to regenerate and establish twenty‐seven lucerne plants, whereas only nine plants could be regenerated from an Agrobacterium co‐cultivated explant. This transformation protocol could represent a valuable improvement over existing ones for lucerne. 相似文献