Micropropagation Techniques Research Articles

The impacts of AMPEP K+ (Ascophyllum marine plant extract, enhanced with potassium) on the growth rate, carrageenan quality, and percentage incidence of the damaging epiphyte Neosiphonia apiculata on four strains of the commercially important carrageenophyte Kappaphycus, as developed by micropropagation techniques

Four strains of Kappaphycus spp. (viz. K. alvarezii—tambalang brown and green, K. striatus—sacol green) and K. malesianus (aring-aring) were grown in Semporna, Sabah, for three consecutive growth cycles from August 2018 to January 2019, each for a duration of 45 days. The study was established in order to determine the impacts of a commercially produced phyco-stimulant: Ascophyllum marine plant extract powder, enhanced with potassium (i.e., AMPEP K+) on micropropagated Kappaphycus strains on the mean daily growth rate (DGR), quality characteristics of extracted, refined carrageenan from the harvested biomass (i.e., yield, viscosity, and gel strength), and the percentage incidence of the damaging, endophytic Neosiphonia apiculata, while in open water cultivation. Results showed that the AMPEP K+-treated thalli were significantly different (P < 0.01), in terms of the three major assessment criteria used in this study. Amongst the four cultivars, and under the conditions tested, K. striatus was the most resistant to the incidence of N. apiculata, particularly when treated with AMPEP K+. The use of the brown seaweed-derived extract, as a phyco-stimulant, applied to a commercially cultivated, red seaweed for the promotion of thallus growth, reduction of biotic stress caused by endophytes, and important improvements to commercially valuable traits, such as carrageenan quality, are encouraging. It is recommended that these protocols should be adopted in crop management to assist the Malaysian seaweed industry.

In vitro grafting – twenty-first century’s technique for fruit tree propagation

ABSTRACT In vitro grafting is a novel micropropagation technique, which has been mainly used for research purposes such as eliminating endogenous pathogens and studying shoot-root signalling or scion-rootstock compatibility. Through this technique, an in vitro grown scion is grafted on/into a micro-rootstock under aseptic conditions. Enhanced juvenility, health and growth potential of in vitro grafts, compared to traditional field/bench grafts, make this technique a great fit for mass propagation of fruit trees under current global challenges. However, to date, in vitro grafting has not been employed for commercial propagation of any crop trees.

Micronutrient Delivery System for Induction of Organogenesis in Banana in vitro Cultures

Micropropagation techniques represent one of the technologies, which allows the large-scale production of banana and the culture medium composition is one of the major factors affecting in vitro propagation of plants. The objective of this study was to evaluate the efficiency of a new micronutrient delivery system based on ionic Cu and Zn for inducing in vitro organogenesis in two banana cultivars in vitro cultures, Grand Naine and Pysang Ceylon. The first experiment evaluated different concentrations of BAM-FX® (0.16, 0.32, 0.64, 1.28, and 2.56 µl ml-1) added to the MS culture medium. The concentration of 0.16 µl ml-1 BAM-FX® provided the best results for in vitro shoot and root growth and development. Therefore, a second experiment was performed to evaluate the potential of combining BAM-FX® with a reduced concentration of MS medium (¼, ½, and ¾ strength), or the use of BAM-FX® alone without MS medium. Results indicate that MS at ¾ strength combined with 0.16 µl ml-1 BAM-FX® provided proper in vitro shoot and root growth and development. The use of BAM-FX® in vitro requires additional studies to verify the feasibility of this product for efficient micropropagation of banana, as well as for other species.

Intraspecific functional trait structure of restoration‐relevant species: Implications for restoration seed sourcing

Abstract Recent research has highlighted the existence of significant intraspecific trait variation within and among populations of plant species. This inherent variation within species means there is a wealth of trait diversity from which to source germplasm for use in ecological restoration. However, it remains unclear how to source materials from this pool of trait diversity in order to achieve desired outcomes and support ecosystem function in a restoration context. We provide a framework to study the structure of trait variation across populations and genotypes in an effort to bridge functional trait research with the sourcing of native plant materials for restoration. We investigated the structure of intraspecific functional trait variation in three forb species used in restoration on the Colorado Plateau to (a) understand the structure of functional trait variation within and among populations, and (b) determine if individual and multivariate functional traits differ between populations while accounting for trait variation within and among genotypes. We found considerable functional trait variation at all sampling levels, with variation within populations often surpassing variation among populations. Still, we observed population‐level differences in trait values in 8 of the 12 species‐by‐trait combinations and populations largely segregated in multivariate trait space. Synthesis and applications. Using micropropagation techniques, we uncovered population‐level differences in functional trait variation, suggesting that mixing populations to create restoration germplasm following a regional admixture provenancing approach could lead to increased functional diversity in restorations. However, the substantial trait variation identified within some populations of our study species also suggests a similar potential when utilizing genotypically diverse material from even a single population. Further research on these and other species is needed to understand the structure of intraspecific functional trait variation and how it impacts ecosystem function. The approach outlined in this study can assist researchers in assessing the underlying trait variation present in various restoration materials and provide managers with more detailed information that can help make germplasm sourcing decisions.

Phenolic composition and biological activities of the in vitro cultured endangered Eryngium viviparum J. Gay

Eryngium viviparum is an endangered species that requires management efforts based on complementary ex situ conservation strategies, such as in vitro culture. This study was carried out to evaluate the phenolic composition and the antioxidant, cytotoxic, and antimicrobial properties of E. viviparum aerial and root parts obtained by this micropropagation technique. The HPLC-DAD-ESI/MSn analysis showed that phenolic compounds were more abundant in the root (102 ± 4 mg g−1 extract) than in the aerial part (40.6 ± 0.8 mg g−1 extract) of the plant. The major compound was trans rosmarinic acid, followed by trans 3-O-caffeoylquinic acid. The root extract also showed higher antioxidant activity, with a result close to that of trolox in the thiobarbituric acid reactive substances (TBARS) formation inhibition assay, and a moderate toxicity to lung (NCI-H460), breast (MCF-7) and liver (HepG2) tumour cells. It was also more effective than ketoconazole against Penicillium ochrochloron. In turn, the aerial part extract inhibited Salmonella typhimurium more effectively than ampicillin. This study highlights E. viviparum as an unexplored source of bioactive compounds with potential application in the food, pharmaceutical, and other industrial sectors. Consequently, it promotes the interest of conserving this endangered species.

Optimization of Protoplast Isolation and Micropropagation Techniques in Rose

Rose is the dominant flower in the ornamental industry because of its multiple aesthetic values. Optimization of plant growth regulators in media was achieved for one of the important species of rose. The nodal explants were tested on a full strength MS media with different hormone concentrations of 6-benzylaminopurine (BAP) and Kinetin (Kn). Maximum response was obtained for a combined hormone concentration of 6-benzylaminopurine (3.5mg/L) and kinetin (1.5mg/L) with shoot regeneration from nodes, internodes and apical meristematic tissue. 6-benzylaminopurine alone showed shoot regeneration only from internodes and nodes, while kinetin alone showed shoot regeneration only from apical meristematic tissue. Protoplast of Rosa sp. and Rosa sinensis were isolated enzymatically and fused using polyethylene. The protoplasts were observed to fuse on different planes in an irregular pattern.

Nutritional Qualities of Ginger Mutant Lines Grown in the Humid Tropical Agroecology of Nigeria

The present study was aimed to determine the variations in nutritional qualities of 15 mutant lines and two landraces of ginger (Zingiber officinale). Fifteen (15) gamma (γ)-ray induced mutants lines and two landraces of ginger were planted in 2017 early cropping season in the Teaching and Research Farm, Department of Crop Science, Faculty of Agriculture, Forestry and Wildlife Resources Management, University of Calabar, Calabar, Nigeria. To evaluate the nutritional qualities of these seventeen ginger genotypes at maturity, proximate analysis was carried out in the Biochemistry Laboratory of the National Root Crop Research Institute Umudike, Abia State, Nigeria. Using standard and official protocols of the Association of Official Analytical Chemists (AOAC). Results showed that the ginger lines varied significantly (P &lt; 0.01) in all their proximate attributes. The moisture content ranged from 10.13% (UG1) to 12.95% (UG2). Mean dry matter was 88.89%; UG1 and UG2 had the highest (89.89%) and lowest (87.05%) dry matter content, respectively. Mean crude protein was 7.74%; UG2-9-01 and UG2-11-03 had the highest (8.25%) and lowest (7.29%) crude protein respectively. UG1-5-38 and UG1-5-22 had the highest (8.12%) and lowest (6.41%) crude fibre content respectively. The oleoresin content ranged from (6.25%) in UG2-9-01 to (9.09%) in UG1-11-07. UG1-5-04 and UG1-5-22 had the highest (2.88%) and lowest (2.22%) ash content respectively. UG2-9-01 had the highest carbohydrate content of (65.10%). While UG1-5-52 had the lowest (61.27%) The result showed that the ginger lines used in this study had high mean carbohydrate (62.85%) and protein (7.74%) contents as such can be used as supplementary sources of these nutrients for human and livestock. UG1-7-24, UG1-11-07 and UG1-5-18 with high oleoresin contents of 9.11%, 9.09% and 9.05% respectively are recommended to ginger breeders as useful genotypes for improving other ginger lines through micropropagation techniques especially when breeding for oleoresin quality, which is an important quality of ginger. In conclusion, further evaluation and testing of these ginger lines is recommended.

A Review on Micropropagation Culture Method

Micropropagation is a vegetative propagation conducted under controlled and aseptic conditions in the microenvironment of the culture vessel, which have the all growth requirements of a plant in the natural conditions. Recently different techniques of propagation have been developed which could facilitate large scale production of plants and for the improvement of the species. An overview on the in vitro propagation via meristem culture, callus culture and protoplast culture etc. are presented here. Today micropropagation techniques are applied in order to produce large numbers of new high-quality plants in a relatively short time and space, in low cost and can also be preserved.&#x0D;

The endangered Saharan cypress (Cupressus dupreziana): do not let it get into Charon's boat.

The current state-of-the-art creates a sound basis for the preservation of unique species Cupressus dupreziana provided that targeted effort and care is devoted to the accomplishment of multiplication protocols. This review is to summarize known data on Cupressus dupreziana-specific characteristics, including abiotic stress resistance, and natural reproduction, and estimates the possibilities of ex situ conservation with an emphasis on the cultivation in vitro of this endangered species. As there is only limited information about the cultivation of C. dupreziana in vitro, we have included relevant data on the related species Cupressus sempervirens, where micropropagation techniques are well established, along with other information on species with similar fates and life strategies-Saharan olive Olea europaea subsp. laperrinei and myrtle Myrthus nivellei. The aim of this work is to enhance general understanding and to promote an interest in this relict plant species to contribute to more wide-ranging studies and to increase its chance of preservation. Besides others, reducing the number of species threatened with extinction is an essential and immediate task as high genetic variability of ecosystems is crucial for their stability under changing climatic conditions.

A Mutation Study on Gerbera jamesonii: An Important Ornamental Plant

The demand for variations in ornamental plants is always on high for the development of different types of color and other morphological changes etc. In this context, the mutation is a very much helpful and promising approach among the floriculturists and very well recognized for the development of novel varieties. Most of the researchers used the micropropagation techniques for large scale propagation of ornamental plants. Micropropagation not only enhances the rate of propagation but also produce true to type plants in a relatively short time and space. In this study, we use a combination of mutation and micropropagation strategies in Gerbera jamesonii plant.

Оптимизация методики клонального микроразмножения Stevia rebaudiana Bertoni

Оптимизация методики клонального микроразмножения Stevia rebaudiana Bertoni

A general introduction of propagation and micropropagation techniques of Macadamia Nut (Macadamia spp.)

A general introduction of propagation and micropropagation techniques of Macadamia Nut (Macadamia spp.)

English

Dendrobium primulinum Lindl. (D. Primulinum L.) is one of the important epiphytic orchid species for horticultural and commercial use. It is listed as a rare and critically endangered species of orchid. The present study is intended to conserve the orchid species through the micropropagation technique. The in vitro seed germination and seedling growth was carried out by taking a mature pod of D. Primulinum L. in Murashige and Skoog (MS) media, and the MS media supplemented with a varied concentration (0.5, 1.0, 1.5 and 2 mg/L) of 6-benzylaminopurine (BAP), and a fixed concentration (0.5 mg/L) of α-naphthalene acetic acid (NAA). Sequential phases of seed germination, protocorm formation, and seedling development in the presence of growth regulators were determined in the study. The significance of hormonal effects was determined by using One-Way Analysis of Variance (ANOVA, p≤0.05). The seed germination started after two weeks of culture in the media supplemented with BAP. The maximum seedling growth was obtained in the media supplemented with 1.5 mg/L of BAP. Although the hormone-free basal medium revealed an ideal condition for seed germination and spherules formation, the presence of an appropriate concentration of growth regulators such as 0.5 mg/L NAA or a combined 0.5 mg/L BAP + 0.5 mg/L NAA expressed a synergistic effect to enhance the protocorm formation and seedling development. Key words: In vitro, Dendrobium primulinum, 6-benzylaminopurine (BAP), α-Naphthalene acetic acid (NAA).

An efficient protocol for commercial micropropagation of malanga (Colocasia esculenta L. Schott) using temporary immersion

Temporary immersion systems allow semi-automatization and reduce production costs during micropropagation on a commercial scale. The objective of this study was to evaluate temporary immersion (TI) as a new technique for commercial micropropagation of malanga (Colocasia esculenta). The effect of the semi-solid and liquid medium using the Temporary Immersion Bioreactor (TIB) was compared. For the TIB culture system, the immersion frequency and volume of culture medium per explant on shoot development were evaluated. At 30 d of culture, the number of shoots per explant, shoot length and number of leaves per shoot were scored. In addition, the chlorophyll content during multiplication and survival during acclimatization were evaluated in the different culture systems. The results showed that the highest shoot production, shoot length and number of leaves per shoot were obtained in TI with an immersion frequency of 2 min every 4 h and with a volume of culture medium of 25 and 50 mL per explant. An increase in chlorophyll content in TI was observed with an immersion frequency every 4 h and with a culture medium volume of 50 and 100 mL per explant. Regarding the percentage of survival during acclimatization, no significant differences were observed between culture systems, immersion frequencies and volume of culture medium per explant. In conclusion, the evaluated immersion system is an alternative for commercial micropropagation of C. esculenta.

Shoot proliferation and in vitro organogenesis from shoot apex and cotyledonary explants of royal poinciana (Delonix regia), an ornamental leguminous tree

Cytokinin supplementation was essential for the induction of shoot proliferation and in vitro organogenesis from shoot apices and cotyledons of D. regia, respectively. Micropropagation techniques have been established for woody species to facilitate their mass clonal propagation and to better understand their processes of regeneration. The goal of this work was to evaluate the potential of shoot apex and cotyledonary explants of Delonix regia, commonly known as royal poinciana, to be induced to undergo shoot proliferation and in vitro organogenesis, respectively. Shoot apices and cotyledons obtained from in vitro-germinated seedlings were inoculated onto Murashige and Skoog (MS) medium supplemented with different concentrations (0.125, 0.250, 0.5, 1.0 and 2.0 mg L−1) of 6-benzyladenine (BA) and a control without BA. After 30 days of cultivation, multiple shoot formation and adventitious shoots were observed in shoot apices and cotyledonary segments of D. regia, respectively. Both explant sources exhibited a dose-dependent behavior with increased fresh mass and shoot number with increasing BA concentrations. The greatest number of shoots was observed in explants cultured on BA-supplemented (2.0 mg L−1) medium; however, the leaflets of shoots grown in MS medium with higher BA concentrations were smaller. Histological analysis revealed that the formation of adventitious shoots occurred through the reprogramming of mesophyll cells in the edge regions of the cotyledonary explants. The results obtained here provide new information for optimizing and establishing efficient regeneration systems and micropropagation of royal poinciana, an important ornamental tree species.

Evaluation of induction of somatic embryogenesis from cotyledonary leaves of Banana Passion fruit (Passiflora mollissima) L.H Bailey

Passiflora mollissima L.H Bailey is an endemic species from the Andes mountain of Colombia in South America with important edible fruits and medicinal and economical properties. The development of improved micropropagation techniques is necessary to provide rapid and efficient clonal propagation of elite genotypes with high resistance and uniform production, as well as a system that can be used for genetic transformation. For this reason, the investigation focused on the evaluation of induction of somatic embryogenesis in P. mollissima from cotyledonary leaves, the effect of growth regulator concentrations and the orientation of explants on embryo production. Histological analyses of somatic embryogenesis were performed every 10 days after induction over 38 days of exposure to the medium. Results showed somatic embryo formation on Murashige and Skoog, (1962) culture medium supplemented with 4.5 μM 2,4-diclorophenoxyacetic acid plus 4.5 μM 6-benzyladenine. The results obtained are applicable to knowledge of non-zygotic embryogenesis in passionflower of the Andean region, for the purpose of improvement and commercial use. It is noteworthy that this is the first study in the induction and obtaining of embryos in P. mollissima.

Genomic alterations in coding region of tissue culture plants of Coffea arabica obtained through somatic embryogenesis revealed by molecular markers

In coffee, the micropropagation technique can be efficiently used in mass multiplication of superior F1 hybrids which is difficult using the conventional method. However, the in vitro regeneration protocol is genotype-specific. Therefore the development of an efficient regeneration protocol is critical to the success of large scale propagation of any coffee hybrids. In the present study, regeneration of two improved rust tolerant arabica hybrids S.4202 and S.4932 was achieved through somatic embryogenesis. The MS media supplemented with different concentrations of 2, 4-D, IAA and Kn were tested for callus induction using foliar explants. The optimum somatic embryogenesis in S.4202 and S.4932 was obtained when explants were incubated in callus induction media containing 0.4 to 0.6 mg/l 2, 4-D. The in vitro regenerated plants were planted in the field and were found to be phenotypically normal. The genetic fidelity of in vitro regenerated plants of both the genotypes along with their mother plant was tested using 20 SRAP and 12 SCoT markers. The average genetic similarity between the tissue culture-derived plants and mother plants of S.4202 was 0.977 using SRAP and 0.987 using SCoT markers. Similarly, the average genetic similarity between the mother plant and the in vitro derived plants of S.4932 was 0.966 and 0.993 using SRAP and SCoT markers respectively. The differential fragments amplified by SRAP primers in the regenerated plants were sequenced and in silico analysis of the fragments revealed the occurrence of somaclonal variation in the coding region with functional attribution to zinc finger protein. Plant regeneration through somatic embryogenesis was achieved in two improved cultivars of Coffea arabica. The genetic fidelity of in vitro regenerated plants was tested using both SRAP and SCoT molecular markers. The differential fragments amplified in in vitro regenerated plants using SRAP primers were sequenced and in silico analysis revealed the genomic alterations in the coding region.

TÉCNICA DE ESQUEJES EN AGRICULTURA: UNA ALTERNATIVA A LA VANGUARDIA

&lt;p&gt;The multiple effects in agriculture due to plant morphogenic conditions lead to the concern to search viable alternatives for the propagation and feasible development of crops. Therefore, it has had to resort to new techniques, such is the case of micropropagation techniques such as cuttings, which are based on the promotion and multiplication of plants, quickly, efficiently and in large quantities; due this reason it has been considered one of the most successful biotechnological methods in agriculture. The technique of cuttings results in a new plant identical to the plant from which it precedes; this quick and simple technique can be performed on most plants. One way of guaranteeing root development properly during micropropagation techniques is the use of certain growth regulators, of which the most used are the auxins, such as indole-3-acetic acids (AIA), Naphthaleneacetic acid (ANA) and indolbutyric acid (AIB). These auxins promote the initiation of roots and have been attributed the ability to stimulate cell divisions in the radicle. In addition to the auxins, the roots of the cuttings can be favored using treatments based on hormones, microorganisms and bioactive compounds. The morphogenic characteristics of the plants and the application of treatments, among other factors, contribute to the successful establishment of a crop.&lt;/p&gt;

Micropropagation of clonal lines of thorny artichoke (Cynara scolymus L.)

The aim of the study was to evaluate the in vitro propagation ability of 10 clonal lines of thorny globe artichoke ( Cynara scolymus L.). The study methodology comprised five stages of evaluation. The stages evaluated were initiation, multiplication, rooting, acclimatization, and transplant to the field. The study began with the initiation of dissected shoot tips of 10 clonal lines in test tubes containing the Murashige and Skoog (MS) medium. Best results were obtained when explants were cultured on an induction medium containing MS + naphthalene acetic acid (NAA) 1.0 mg l−1 + benzyl aminopurine (BA) 1.0 mg l−1, highlighting clonal lines L-250, L-132, and L-62. Because of high rates of vitrification and phenolization in the initial stage, clonal lines L-24, L-127, and L-142 were discarded from the study. Therefore, only seven clonal lines were included for evaluation in the multiplication stage. Once the microplants were obtained under laboratory condition in the culture medium, they were immediately transferred to a proliferation medium containing MS + BA 1.0 mg l−1. Only in three clonal lines (L-132, L-200, and L-250), a high multiplication rate (3.5 shoots/explant) was achieved with axillary bud formation. Of the seven clonal lines evaluated, clonal line L-250 achieved the highest rates in the variables shoot height (3.38 cm), number of leaves (13.4), and number of shoots/explant (4.4). In the rooting stage, clonal line L-250 obtained a significant improvement by transferring plantlets to direct acclimatization after 20 days of in vitro root induction in a medium containing MS + NAA 1.0 mg l−1. Similarly, in the acclimatization stage, the clonal line L-250 showed a significant result. Then, in the transplantation stage, the plants were transplanted to the field with 100% rooting; 30 days after the transplantation, the clonal line L-250 obtained 100% survival in the field than the control treatments (offspring from two locations were used – Mito and Alayo). As the rooting period is reduced by approximately 20 days by inducing direct root formation under greenhouse conditions, the micropropagation technique is optimized with the protocol used in this study.

Productivity of Rhizomes and Starch Quantification in Cultures of Different Vegetative Propagules of Arrowroot

Arrowroot rhizomes are rich in carbohydrate and are commonly grown by family farmers who have an important source of income in this activity and play a prominent role in the conservation of the species. There are few studies on the phytotechnical aspects of culture. The objective of this work was to evaluate the productive capacity of the &amp;lsquo;common&amp;rsquo; arrowroot using different sizes and forms of propagation, aiming at the production of rhizomes and arrowroot starch, in different agricultural crops. The experimental design was the randomized block in a 4 &amp;times; 2 factorial scheme, with 6 replicates. Four types of rhizome propagation (rhizomes-seeds of small size with a weight of 20 to 30 g; rhizomes-seeds of average weight between 30.01 to 45 g; rhizomes-seeds of large size weighing between 45.01 a 60 g, and seedlings produced in tissue culture), in two agricultural years (2015/2016 and 2016/2017). The variables total production, number of rhizomes, extraction yield and total starch production were evaluated. The type of propagule used interfered in the yield of rhizomes (9.85 to 34.75 t ha-1) and in the production of arrowroot starch (1.76 to 7.68 t ha-1). The vegetative propagation with pieces of rhizomes-seeds between 20 and 60 g was more viable than the micropropagation technique. Although the soil and climate conditions showed differences between the agricultural crops studied, they did not significantly influence the yield of rhizomes and arrowroot starch, by the type of propagule used.