Micropropagation

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  • 文章类型: Journal Article
    PsidiumcattleianumSabine(草莓番石榴)是一种常绿灌木,可作为水果树篱种植,并在食品和制药学科中得到了广泛的考虑。本研究旨在为卡丁草的体外繁殖设定一个有希望的方案,随着原始植物(OP)的酚类含量的分析,诱导愈伤组织(IC),和再生植株(RP)提取物,最终,评估他们的抗炎作用,抗氧化剂,和抗癌潜力。用商业漂白处理种子,HCl,和H2O2以提高发芽率并最小化污染率。将灭菌的叶外植体培养到补充有苄基腺嘌呤(BA)的Murashige和Skoog(MS)培养基上,2,4-二氯苯氧基乙酸,激动素显示出最好的愈伤组织诱导,而MS培养基补充BA,腺嘌呤硫酸盐,萘乙酸,和赤霉酸活化再生。用吲哚-3-丁酸增强MS培养基记录最大生根百分比。最后,获得的生根芽在沙子和泥炭苔藓土壤中成功适应。OP的HPLC-MS/MS图谱,RP,IC显示多种酚类代谢产物。IC提取物降低了MCF-7,HepG2和K-562癌细胞系的活力。此外,OP表现出强的抗氧化活性。P.cattleianum及其RP是酚类化合物的重要来源,有望在食品和制药行业中应用。
    Psidium cattleianum Sabine (strawberry guava) is an evergreen shrub that is grown as a fruiting hedge and has received significant consideration in the food and pharmaceutical disciplines. This study aims to set a promising protocol for in vitro propagation of P. cattleianum, along with profiling the phenolic content of the original plant (OP), induced callus (IC), and regenerated plantlets (RP) extracts, ultimately, evaluating their anti-inflammatory, antioxidant, and anticancer potential. Seeds were treated with commercial bleaching, HCl, and H2O2 to enhance the germination percentage and minimize the contamination percentage. Culturing sterilized leaf explants onto Murashige and Skoog (MS) medium supplemented with benzyl adenine (BA), 2,4-dichloro phenoxy acetic acid, and kinetin showed the best callus induction, while supplementation of MS media with BA, adenine sulfate, naphthalene acetic acid, and gibberellic acid activated regeneration. Augmentation of MS media with indol-3-butyric acid recorded the maximum rooting percentage. Finally, the obtained rooted shoots were successfully acclimatized in sand and peat moss soil. HPLC-MS/MS profiles of OP, RP, and IC showed a variety of phenolic metabolites. IC extract decreased the viability of MCF-7, HepG2, and K-562 cancer cell lines. Also, OP exhibits strong antioxidant activity. P. cattleianum and its RP are profound sources of phenolic compounds promoted for promising applications in the food and pharmaceutical industries.
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  • 文章类型: Journal Article
    本研究是在组织培养实验室进行的,园艺部门,农业学院,Damietta大学,埃及。这项研究的目的是建立适合三种进口桃砧木的微繁殖协议:冲绳(P.persica),Nemared(P.persica×P.davidiana)×P.persica),和Garnem(P.体外dulcis×P.persica)。结果表明,将外植体以20%的次氯酸钠(NaOCl)浸泡15分钟可产生最高的响应性(82.81%),生存率(96.61%),死亡率最低(3.14%),污染最低(0.24%)。Garnem基因型外植体反应最好(89.12%),生存率(90.62%),最低死亡率(0.00%),与其他基因型相比,污染最高(9.37%)。与腋芽相比,射击提示显示出最高的响应能力,生存,和死亡(100、87.40和12.59%,分别),以及最不显著的污染(0.00%)。此外,响应的百分比,幸存下来,死了,在不同的收集日期,受污染的外植体差异很大。使用的6-苄基氨基嘌呤(BAP)浓度(3至5.0mg/L)在体外增殖方面表现出相似的行为,冲绳每个外植体的生长率为3.77至6.11,4.33至8.88和3.33至7.44,Nemared,和Garnem桃子砧木,分别,表明芽增殖的数量是基因型依赖性的。此外,使用5.0mg/LBAP与0.2mg/LIBA的组合显着增加平均芽增殖(96.29%),每个外植体的枝条数(7.48),与其他处理相比,平均叶片数/外植体(16.33)。基于这些结果,在冲绳的体外繁殖过程中,不定芽的发育得到了增强,Nemared,和Garnem桃砧木通过吲哚-丁酸(IBA)和6-苄基氨基嘌呤(BAP)的协同作用。
    The present study was conducted in the Laboratory of Tissue Culture, Horticulture Department, Faculty of Agriculture, Damietta University, Egypt. The objective of this study was to establish a micropropagation protocol suitable for three imported peach rootstocks: Okinawa (P. persica), Nemared (P. persica × P. davidiana) × P. persica), and Garnem (P. dulcis × P. persica) in vitro. The results showed that soaking the explants in sodium hypochlorite (NaOCl) at 20% for 15 min produced the highest responsiveness (82.81%), survival (96.61%), with the lowest mortality (3.14%) and contamination (0.24%). Explants of the Garnem genotype had the best response (89.12%), survival (90.62%), lowest mortality (0.00%), and highest contamination (9.37%) when compared to the other genotypes. In comparison with axillary buds, the shoot tip displayed the highest responsiveness, survival, and death (100, 87.40, and 12.59%, respectively), as well as the least significant contamination (0.00%). Additionally, the percentages of responsive, survived, dead, and contaminated explants at the various collection dates varied significantly. The 6-benzylaminopurine (BAP) concentrations used (3 to 5.0 mg/L) demonstrated similar behavior in terms of in vitro proliferation, with rates of 3.77 to 6.11, 4.33 to 8.88, and 3.33 to 7.44 shoot numbers per explant for the Okinawa, Nemared, and Garnem peach rootstocks, respectively, indicating that the number of shoot proliferations is genotype-dependent. Additionally, using 5.0 mg/L BAP in combination with 0.2 mg/L IBA significantly increased average shoot proliferation (96.29%), number of shoots per explant (7.48), and average leaf number/explant (16.33) compared to the other treatments. Based on these results, adventitious bud development was enhanced during in vitro multiplication of the Okinawa, Nemared, and Garnem peach rootstocks by the synergistic interaction of indole-butyric acid (IBA) and 6-benzylaminopurine (BAP).
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  • 文章类型: Journal Article
    秘鲁安第斯山脉是几种野生黑莓物种的自然栖息地,由于缺乏技术和科学发展来支持其农业潜力,因此鲜为人知和开发。在这种情况下,进行了一项研究,以了解理化成分,生物活性化合物,抗菌活性,和四个野生黑莓的体外繁殖(Rubussp。)来自秘鲁北部高地的物种。结果表明,R.floribundus果实的总可溶性固形物(9.58±1.83°白利糖度)和可滴定酸度(1.88±0.07%柠檬酸)含量最高。R.webbaueri的果实记录了最高的总酚含量(415.06±8.69mgGAE/100gFf)。通过DPPH测定确定的抗氧化能力在物种之间差异显著,在安德科拉的果实中发现的值最高(50.27±0.11毫克TE/100克Ff)。R.webbaueri和R.andicola的果实提取物显示出更好的抗菌活性,金黄色葡萄球菌是最敏感的细菌。在体外增殖阶段,结果表明,BAP(6-苄基氨基嘌呤)在1.5mgl-1的剂量对芽数有显著影响,叶数,和射击长度。结果可能有助于遗传资源的管理。
    The Peruvian Andes are the natural habitat of several wild blackberry species that are little known and exploited due to the lack of technological and scientific development to support their agricultural potential. In this context, a study was conducted to understand the physicochemical composition, bioactive compounds, antimicrobial activity, and in vitro multiplication of four wild blackberry (Rubus sp.) species from the northern Peruvian highlands. The results indicate that fruits of R. floribundus presented the highest content of total soluble solids (9.58 ± 1.83°Brix) and titratable acidity (1.88 ± 0.07% citric acid). The fruits of R. weberbaueri recorded the highest total phenolic content (415.06 ± 8.69 mg GAE/100 g Ff). The antioxidant capacity determined by the DPPH assay varied significantly among species, with the highest value found in fruits of R. andicola (50.27 ± 0.11 mg TE/100 g Ff). The fruit extracts of R. weberbaueri and R. andicola showed better antimicrobial activity, with Staphylococcus aureus being the most sensitive bacterium. In the in vitro multiplication phase, the results show that BAP (6-Benzylaminopurine) has a significant effect at a dose of 1.5 mg l-1 on shoot number, leaf number, and shoot length. The results may help in the management of genetic resources.
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  • 文章类型: Journal Article
    珍贵的红香蕉,选择卓越的品质,成功的克隆繁殖和保存需要很强的遗传一致性。在体外红香蕉植物的生长早期确保这种均匀性至关重要,因为组织培养过程中的基因突变和染色体重排会危及克隆和种质保护。在这种情况下,分子标记在确认遗传稳定性中起着关键作用。因此,该研究旨在发现一种标记,该标记可在最初的继代培养过程中从组织培养的红香蕉中识别出组织培养的红香蕉。已鉴定出与花青素相关的标记,该标记可有效地将红香蕉与绿色变体区分开,并在各种香蕉品种中进一步验证。观赏Musa物种及其种间杂种。基于PCR的标记显示出显著的特异性,在组织培养过程中辨别红香蕉与绿叶变体。它还区分了绿色和红色的后代,削减时间和资源成本,缩短香蕉的繁殖周期。
    在线版本包含补充材料,可在10.1007/s13205-023-03868-6获得。
    The prized Red banana, selected for superior qualities, demands strong genetic uniformity for successful clonal propagation and preservation. Ensuring this uniformity early in the growth of in vitro Red banana plants is essential, as gene mutations and chromosome rearrangements during tissue culture can jeopardize both cloning and germplasm conservation. In this situation, molecular markers play a pivotal role in confirming genetic stability. Thus the study aims to discover a marker that identifies tissue-cultured Red bananas from their virescent variants during initial sub-culturing. A marker linked to anthocyanin has been identified which effectively differentiated Red bananas from virescent variants and it was further validated in various banana cultivars, ornamental Musa species and their interspecific hybrids. The PCR-based marker showed remarkable specificity, discerning Red bananas from virescent variants during tissue culture. It also distinguished green and red offspring, cutting time and resource costs, and shortening the banana breeding cycle.
    UNASSIGNED: The online version contains supplementary material available at 10.1007/s13205-023-03868-6.
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  • 文章类型: Journal Article
    用于封装和持续释放农用化学品(例如生长素)的纳米载体已经成为提供增强的生物利用度和功效以改善作物产量和营养质量的有吸引力的策略。这里,对壳聚糖作为生物聚合物纳米载体和银作为金属纳米载体对苹果砧木中微插条的体外不定根潜力的有效性进行了比较研究,第一次。合成了生长素吲哚-3-乙酸(IAA)和吲哚-3-丁酸(IBA)负载银(nAg)或壳聚糖纳米颗粒(nChi)。扫描电子显微镜和透射电子显微镜研究显示纳米颗粒的球形。IAA-nChi的平均粒径为167.5±0.1nm,而IBA-nChi的平均粒径为123.2±2.6nm。nAg-IAA和nAg-IBA颗粒的流体动力学直径测量为93.66±5nm和71.41±3nm,分别。傅里叶变换红外光谱分析证实了IAA或IBA在壳聚糖纳米颗粒中的包封。同时,在银纳米粒子上检测到IAA或IBA的特征峰。与游离的IAA或IBA应用(50.0%-33.3%)相比,装载有IAA或IBA的壳聚糖和银纳米颗粒(91.7%-62.5%)的MallingMerton106(MM106)的微插条的体外不定根明显更高,除了2.0mgL-1IBA(66.7%)。然而,在所有浓度下施用2mgL-1IBA和IBA-nChi会导致不良的大愈伤组织发育。
    Nanocarriers for encapsulation and sustained release of agrochemicals such as auxins have emerged as an attractive strategy to provide enhanced bioavailability and efficacy for improved crop yields and nutrition quality. Here, a comparative study was conducted on the effectiveness of chitosan-as a biopolymeric nanocarrier- and silver-as a metallic nanocarrier- on in vitro adventitious rooting potential of microcuttings in apple rootstocks, for the first time. Auxins indole-3-acetic acid (IAA) and indole-3-butyric acid (IBA) loaded silver (nAg) or chitosan nanoparticles (nChi) were synthesized. Scanning electron microscopy and transmission electron microscopy studies showed the spherical shape of the nanoparticles. The average particle size of IAA-nChi was 167.5 ± 0.1 nm while that of IBA-nChi was 123.2 ± 2.6 nm. The hydrodynamic diameter of the nAg-IAA and nAg-IBA particles were measured as 93.66 ± 5 nm and 71.41 ± 3 nm, respectively. Fourier transform infrared spectroscopy analyses confirmed the encapsulation of IAA or IBA in the chitosan nanoparticles. Meanwhile, the characteristic peaks of IAA or IBA were detected on silver nanoparticles. In-vitro adventitious rooting of microcuttings of Malling Merton 106 (MM 106) was significantly higher both in chitosan and silver nanoparticles loaded with IAA or IBA (91.7%-62.5%) compared to free IAA or IBA applications (50.0%-33.3%), except for 2.0 mg L-1 IBA (66.7%). However, the application of 2 mg L-1 IBA and IBA-nChi at all concentrations caused an undesirable large callus development.
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  • 文章类型: Journal Article
    右叶利比。属于马鞭草科,是塞拉多特有的。该物种是芳香的,其特征在于其叶子上存在富含单萜的腺毛。这项研究的目的是评估生长,光合色素生产,以及在不同波长和强度下体外生长的圆叶乳杆菌的化学成分。光强度由使用20、54、78、88和110μmolm-2s-1的冷白色荧光灯进行的五种处理组成。光质量包括使用不同光波长的发光二极管(LED)的六种处理,即,白色,红色,蓝色,和它们的相互作用:1R:1B,2.5R:1B,1R:2.5B45天后,生物测定参数,光合色素含量,和挥发性化合物进行了评估。20和54μmolm-2s-1的较低光照强度产生较高的生长,光合色素含量,和生物量积累。月桂烯和十五烷分别在88和110μmolm-2s-1的光强度下最高。在20和54μmolm-2s-1强度下获得最高的柠檬烯和奥西烯酮水平,分别,并且在78μmolm-2s-1强度下获得了最高的Myrcenone含量。关于光波长,红色和蓝色光谱的结合进一步刺激了植株的生长,2.5R:1B组合获得了最佳的生物特征数据和总叶绿素含量。在1R:2.5B光谱下,z-辛烯酮化合物的含量最高。单色蓝色光谱增加了月桂烯和柠檬烯的含量,但降低了月桂烯的含量,红色的光增加了。用白色光谱以及红色和蓝色组合获得最高的十五烷含量。
    Lippia rotundifolia Cham. is in the family Verbenaceae and is endemic to the Cerrado. This species is aromatic and characterized by the presence of glandular trichomes on its leaves that are rich in monoterpenes. The objective of this study was to evaluate the growth, photosynthetic pigment production, and chemical composition of L. rotundifolia grown in vitro under different light wavelengths and intensities. The light intensities consisted of five treatments using cool white fluorescent lamps at 20, 54, 78, 88, and 110 μmol m-2 s-1. The light quality consisted of six treatments using light-emitting diodes (LEDs) in different light wavelengths, namely, white, red, blue, and their interactions: 1R:1B, 2.5R:1B, and 1R:2.5B. After 45 days, the biometric parameters, photosynthetic pigment content, and volatile compounds were evaluated. The lower light intensities of 20 and 54 μmol m-2 s-1 generated higher growth, photosynthetic pigment content, and biomass accumulation. Myrcene and pentadecane were highest under light intensities of 88 and 110 μmol m-2 s-1, respectively. The highest limonene and ocimenone levels were obtained at 20 and 54 μmol m-2 s-1 intensity, respectively, and the highest myrcenone content was obtained at 78 μmol m-2 s-1 intensity. Regarding the light wavelengths, the combination of red and blue spectra further stimulated plantlet growth, and the 2.5R:1B combination obtained the best biometric data and total chlorophyll content. The z-ocimenone chemical compound contents were highest under the 1R:2.5B light spectrum. The monochromatic blue spectrum increased the myrcene and limonene content but decreased the myrcenone content, which was increased by red light. The highest pentadecane contents were obtained with the white spectrum and the red and blue combinations.
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  • 文章类型: Journal Article
    仙人掌科的几个分类群由于出于商业目的而被过度收集而受到威胁,并且该家庭的大部分被纳入《濒危动植物物种国际贸易公约》(CITES)。微繁殖可能在保持野生种群压力并促进濒危类群的异地保护方面发挥关键作用。微繁殖的限制之一是每个分类单元,有时甚至是不同基因型的植物调节剂的物种特异性要求。通过微嫁接技术,砧木直接为接穗提供必要的激素要求。在本文中,我们介绍了aselliformisEhrenberg的体外移植数据,附录ICITES列出了园艺贸易中极度濒危和追捧的物种,关于微繁殖的仙人掌-印度米勒。用无花果假单胞菌的根尖和根尖接穗进行微移植,成功率分别为97%和81%。离体转移后的存活率为85%。我们假设这种方法可以应用于其他濒危物种,仙人掌科的类群生长缓慢,从而有助于保护这个濒临灭绝的家庭。
    Several taxa of Cactaceae are endangered by overcollection for commercial purposes, and most of the family is included in the Convention on International Trade in Endangered Species of Fauna and Flora (CITES). Micropropagation may play a key role to keep the pressure off wild populations and contribute to ex situ conservation of endangered taxa. One of the limits of micropropagation is the species-specific requirement of plant regulators for each taxon and sometimes even for different genotypes. With the micrografting technique the rootstock directly provides the scion with the necessary hormonal requirements. In this paper we present data on in vitro grafting of Pelecyphora aselliformis Ehrenberg, an Appendix I CITES listed species critically endangered and sought after by the horticultural trade, on micropropagated Opuntia ficus-indica Miller. Apical and sub-apical scions of P. aselliformis were used to perform micrografting with a successful rate of 97 and 81 % respectively. Survival rate after ex vivo transfer was 85 %. We hypothesize that this method could be applied to other endangered, slow growing taxa of Cactaceae thus contributing to the conservation of this endangered family.
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  • 文章类型: Journal Article
    OBJECTIVE: To study the leaf epidermis of wild and micropropagated Dioscorea bulbifera Linn. (D. bulbifera) in order to document useful diagnostic features that may be employed for correct crude drug identification and to clear any taxonomic uncertainties in the micropropagated medicinal plant.
    METHODS: Growth responses of micropropagated D. bulbifera were observed on Murashige Skoog medium supplemented with 6-benzylamino purine (1.0 mg/L)+α-naphthaleneacetic acid (0.2 mg/L)+cysteine (20 mg/L) using nodal segments as explants. Leaves of the wild and micropropagated plants were studied microscopically.
    RESULTS: More than 80% shoot regeneration and formation of 10%-30% whitish-brown callus were observed within 3 weeks. The highest root proliferation was obtained from Murashige Skoog medium of 6-benzylamino purine (0.05 mg/L) and α-naphthaleneacetic acid (0.01 mg/L) with mean root length of (27.00±1.25) mm and elongated single shoot of mean length (38.00±11.09) mm. Leaf epidermal features that revealed similarities between the wild and micropropagated plants included amphistomatic condition, presence of mucilage, glandular unicellular trichome with multicellular head, polygonal cells with smooth walls, stomata type and shape. Slight variations included thick cuticular wall with closed stomata in wild plant compared to thin walled opened stomata in the in vitro plant. Opening of stomata accounted for larger average stomata sizes of (7.68±0.38) µm and (6.14±0.46) µm on the adaxial and abaxial surfaces, respectively of the micropropagated plant compared to the wild.
    CONCLUSIONS: The diagnostic features obtained in the study could serve as a basis for proper identification for quality control for standardization of the medicinal plant.
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