Conventional Propagation Methods Research Articles

Secondary somatic embryogenesis in Cork oak: influence of plant growth regulators

Cork oak (Quercus suber L.) is one of the most important Mediterranean forest tree species, and the Maâmora forest-Morocco is considered to be the world largest cork oak stand. Clonal propagation of Quercus suber via somatic embryogenesis is an alternative to conventional tree propagation methods. However, complete maturation of somatic embryos is considered the major bottleneck for mass propagation of Quercus suber during somatic embryogenesis. This study focuses on influences of cytokinins on secondary somatic embryogenesis. Cytokinins showed a negative effect on the induction of secondary somatic embryogenesis. For the various concentrations tested, the number of secondary somatic embryos was significantly reduced except in the case with low dose of 2iP, which showed a slight increase. Abscisic acid (ABA) may be beneficial at low concentrations between 0.3 and 0.5 mg/l. The best number of secondary embryos was given at 0.5 mg/l ABA with an average number of 5 embryos from each primary embryo, which represents a slight increase below 10% compared to control medium. Also, we found that the number of responsive embryos presenting secondary somatic embryos based on their cotyledons decreased once adding ABA. Phenylurea derivatives TDZ and DPU do not promoted the process of secondary somatic embryogenesis. DPU showed no significant effect with a similar response to that of the control medium and TDZ showed an inhibitory effect. Gibberellic acid (GA3) partially blocked the process of somatic embryos multiplication, but promoted remarkably the germination of cotyledonary embryos without requesting a particular protocol. Our results provide a new insight into embryo development, establishing the basis for further research toward improvement of secondary somatic embryogenesis in Moroccan cork oak.

Clonal Propagation of Walnuts (Juglans spp.): A Review on Evolution from Traditional Techniques to Application of Biotechnology.

Walnuts (Juglans sp.) are allogamous species. Seed-derived plants are not always superior to the selected parent. Clonal propagation of selected stock plants is an essential requirement for the clonal fidelity of the descendants and to maintain their genetic structure. Selection of the desired plant is realized only after reaching maturity, and characterizing and evaluating the performance of adult trees require a long time. Clonal propagation methods ensure proper transmission of characters to descendants and can be used effectively in breeding programs. The commercialization of a cultivar or rootstock depends on the success of vegetative propagation. Walnuts, like other tree species, are recalcitrant to conventional vegetative propagation methods and even non-conventional in vitro culture (micropropagation). Elucidation of factors determining the success of cloning of desired plants would contribute to understanding current limitations for most genotypes of Juglans. We outline the role of grafting and cuttings and stool layering, as well as in vitro culture on walnut multiplication. These techniques are, in practice, entirely different; nevertheless, they are affected by common factors. The incompatibility of stock-scion and the reduced ability of stem cuttings to root are the main bottlenecks for grafting and cutting, respectively. Genotype, age, and physiological status, reinvigoration or rejuvenation-treatment of donor plant, period of harvesting and processing of explants critically affect the results of methods followed. The in vitro culture technology is the most suitable for walnut cloning. This also has constraints that affect commercial propagation of most desired genotypes. We describe comprehensive results and synthesis in this review on the asexual reproduction of walnuts, providing a better comprehension of the limiting factors and the ways to overcome them, with direct implications on commercial propagation and the releasing of outstanding genotypes.

Long-Term Field Evaluation of Conventional vs. Micropropagated Plants of Chrysanthemum cinerariifolium

Pyrethrum is a perennial herbaceous plant endemic to the eastern coast of the Adriatic Sea, and introduced in large areas of nearly all continents, where it is cultivated for the industrial extraction of pyrethrins. Pyrethrins are a group of six closely related monoterpene esters, widely used as natural insecticides. The world production of natural pyrethrins is lower than the market demand, and a wider introduction of this crop within the Mediterranean agrosystems could be an appealing opportunity for farmers and manufacturers. The availability of adequate amounts of selected plant material to bring into cultivation is, however, one of the major issues. Therefore, the in vitro propagation of elite pyrethrum genotypes could be a suitable alternative to conventional propagation methods. In this paper, we present the results of a 9-year field comparison between pyrethrum plants coming from an in vitro propagation protocol and plants obtained by cutting from the same mother plants. Furthermore, since plantlets derived from in vitro regeneration may experience ploidy changes, we evaluated the stability of the ploidy level of pyrethrum-micropropagated plants by flow cytometry (FCM) analysis. FCM screening revealed no differences among the morphotypes and between them and the mother plant. Likewise, the field evaluation of plants gave no significant differences between flower yields in both groups. Hence, micropropagation was confirmed as an easy, efficient and reproducible method to obtain large quantities of selected pyrethrum genotypes.

Block paralleled BPM simulation of an optical waveguide with cross-sectional random refractive index distribution

Image transport mediated by transverse Anderson localization requires an optical waveguide to have high contrasted indices and subwavelength feature size in random distribution. Super-fine mesh therefore needs to be adopted in numerical solution techniques. Existing numerical simulation approaches such as the conventional beam propagation method (BPM), however, shows low efficiency and is often unstable with fine mesh and high contrasted random index distribution. In this work, we propose a block algorithm based on a 3D finite-difference full-vector BPM, which is appealing as a reliable tool in numerical simulation of optical waveguides with random refractive index distribution.

Plants in vitro propagation with its applications in food, pharmaceuticals and cosmetic industries; current scenario and future approaches.

Plant tissue culture technique employed for the identification and isolation of bioactive phytocompounds has numerous industrial applications. It provides potential benefits for different industries which include food, pharmaceutical and cosmetics. Various agronomic crops i.e., cereals, fruits, vegetables, ornamental plants and forest trees are currently being used for in vitro propagation. Plant tissue culture coupled with biotechnological approaches leads towards sustainable agricultural development providing solutions to major food security issues. Plants are the rich source of phytochemicals with medicinal properties rendering them useful for the industrial production of pharmaceuticals and nutraceuticals. Furthermore, there are numerous plant compounds with application in the cosmetics industry. In addition to having moisturizing, anti‐ageing, anti‐wrinkle effects; plant-derived compounds also possess pharmacological properties such as antiviral, antimicrobial, antifungal, anticancer, antioxidant, anti-inflammatory, and anti-allergy characteristics. The in vitro propagation of industrially significant flora is gaining attention because of its several advantages over conventional plant propagation methods. One of the major advantages of this technique is the quick availability of food throughout the year, irrespective of the growing season, thus opening new opportunities to the producers and farmers. The sterile or endangered flora can also be conserved by plant micro propagation methods. Hence, plant tissue culture is an extremely efficient and cost-effective technique for biosynthetic studies and bio-production, biotransformation, or bioconversion of plant-derived compounds. However, there are certain limitations of in-vitro plant regeneration system including difficulties with continuous operation, product removal, and aseptic conditions. For sustainable industrial applications of in-vitro regenerated plants on a large scale, these constraints need to be addressed in future studies.

Fairness in graph-based semi-supervised learning

Machine learning is widely deployed in society, unleashing its power in a wide range of applications owing to the advent of big data. One emerging problem faced by machine learning is the discrimination from data, and such discrimination is reflected in the eventual decisions made by the algorithms. Recent study has proved that increasing the size of training (labeled) data will promote the fairness criteria with model performance being maintained. In this work, we aim to explore a more general case where quantities of unlabeled data are provided, indeed leading to a new form of learning paradigm, namely fair semi-supervised learning. Taking the popularity of graph-based approaches in semi-supervised learning, we study this problem both on conventional label propagation method and graph neural networks, where various fairness criteria can be flexibly integrated. Our developed algorithms are proved to be non-trivial extensions to the existing supervised models with fairness constraints. Extensive experiments on real-world datasets exhibit that our methods achieve a better trade-off between classification accuracy and fairness than the compared baselines.

Induction, Proliferation, Regeneration and Kinsenoside and Flavonoid Content Analysis of the Anoectochilus roxburghii (Wall.) Lindl Protocorm-like Body.

Anoectochilus roxburghii (Wall.) Lindl has been used in Chinese herbal medicine for treating various ailments. However, its wild resources are endangered, and artificial cultivation of the plant is limited by the low regeneration rate of conventional propagation methods. The lack of A. roxburghii resources is detrimental to the commercial production of the plant and kinsenoside, which is unique to Anoectochilus species. To develop highly efficient methods for A. roxburghii micropropagation and find alternative resources for kinsenoside production, we created an induction, proliferation, and regeneration of PLBs (IPR-PLB) protocol for A. roxburghii. We also analyzed the kinsenoside and flavonoid contents during the induction and proliferation of PLBs. The best media of IPR-PLB for PLB induction and proliferation (secondary PLB induction and proliferation), shoot formation, and rooting medium were Murashige and Skoog (MS) + 3 mg/L 6-benzylaminopurine (6-BA) + 0.5 mg/L naphthaleneacetic acid (NAA) + 0.8 mg/L zeatin (ZT) + 0.2 mg/L 2,4-dichlorophenoxyacetic acid (2, 4-D), MS + 3 mg/L 6-BA + 0.5 mg/L NAA, and MS + 0.5 mg/L NAA, respectively. On these optimized media, the PLB induction rate was 89 ± 2.08%, secondary PLB induction rate was 120 ± 5%, secondary PLB proliferation rate was 400 ± 10% and 350 ± 10 % in terms of the quantity and biomass at approximately 1 month, shoot induction rate was 10.5 shoots/PLB mass, and root induction rate was 98%. All plantlets survived after acclimation. Darkness or weak light were essential for PLB proliferation, and light was crucial for PLB differentiation on these optimized media. The kinsenoside contents of PLBs and secondary PLBs were 10.38 ± 0.08 and 12.30 ± 0.08 mg/g fresh weight (FW), respectively. Moreover, the peak kinsenoside content during the proliferation of secondary PLBs was 34.27 ± 0.79 mg/g FW, which was slightly lower than that of the whole plant (38.68 ± 3.12 mg/g FW). Two flavonoids exhibited tissue- or temporal-specific accumulation patterns, and astragalin accumulated exclusively during the first 2 weeks of cultivation. The IPR-PLB protocol for A. roxburghii may facilitate the efficient micropropagation of A. roxburghii plants. Furthermore, the PLBs are a good alternative resource for kinsenoside production.

Biology and biotechnological aspect of sweet potato (Ipomoea batatas L.): a commercially important tuber crop

This review highlights the economic importance of sweet potato and discusses new varieties, agronomic and cultivation practices, pest and disease control efforts, plant tissue culture protocols, and unexplored research areas involving this plant. Abstract Sweet potato is widely consumed in many countries around the world, including India, South Africa and China. Due to its valuable nutritional composition and highly beneficial bioactive compounds, sweet potato is considered a major tuber crop in India. Based on the volume of production, this plant ranks seventh in the world among all food crops. Sweet potato is considered a "Superfood" by the 'Centre for Science in the Public Interest' (CSPI), USA. This plant is mostly propagated through vegetative propagation using vine cuttings or tubers. However, this process is costly, labour-intensive, and comparatively slow. Conventional propagation methods are not able to supply sufficient disease-free planting materials to farmers to sustain steady tuber production. Therefore, there is an urgent need to use various biotechnological approaches, such as cell, tissue, and organ culture, for the large-scale production of healthy and disease-free planting material for commercial purposes throughout the year. In the last five decades, a number of tissue culture protocols have been developed for the production of in vitro plants through meristem culture, direct adventitious organogenesis, callus culture and somatic embryogenesis. Moreover, little research has been done on synthetic seed technology for the in vitro conservation and propagation of sweet potato. The current review comprehensively describes the biology, i.e., plant phenotypic description, vegetative growth, agronomy and cultivation, pests and diseases, varieties, and conventional methods of propagation, as well as biotechnological implementation, of this tuber crop. Furthermore, the explored and unexplored areas of research in sweet potato using biotechnological approaches have been reviewed.

POTENSI DAN ANALISA BIAYA PENYEDIAAN BIBIT TEBU (Saccharum officinarum L.) UNGGUL SECARA IN VITRO DI PT. GUNUNG MADU PLANTATIONS

<p><em>Sugarcane productivity included cane yield and sugar extracted rate (rendemen) fluctuated annually. The main causes of low sugarcane productivity is quality of planting material with no high sugar extracted rate (sugar rendemen) potential, resistance to pests and diseases, stability production, tolerance to climate changed and early age of harvest. For increasing of the yield productivity and producing a superior sugarcane planting material with competitiveness cost, the vegetative propagation through in vitro culture technique via shoot tip culture was carried out. The objective of this research is to produce a superior sugarcane planting in vitro and analyze the costs required. The research was carried out at the Tissue Culture Laboratory, PT. Gunung Madu Plantations (GMP) in March-May 2021. The research stages started from the preparation of explant source plants, sterilization of explants and tools, manufacture of stock solutions and planting media, subculture, and analysis of the costs needed to provide superior sugarcane seeds. The results showed that the basal swelling of explants, increasing the number of shoots, root elongation, and the method of providing superior sugarcane seeds from apical shoot explants in vitro were relatively more competitive with the price of plantlets per culture tube contained 10 clones costly were Rp. 20,328, compared with conventional propagation methods. with the price of sugar cane seedlings material were Rp 1,603/stem.</em></p>

J-Matrix time propagation of atomic hydrogen in attosecond fields

The J-Matrix approach for scattering is extended to the time-dependent Schrödinger equation (TDSE) for one electron atoms in external few cycle attosecond fields. To this purpose, the wave function is expanded in square integrable (L^2) Sturmian functions and an equation system for the transition amplitudes is established. Outside the interaction zone, boundary conditions are imposed at the border in the L^2 function space. These boundary conditions correspond to outgoing waves (Siegert states) and minimize reflections at the L^2 boundary grid. Outgoing wave behaviour in the asymptotic region is achieved by employing Pollaczek functions. The method enables the treatment of light - atom interactions within arbitrary external fields. Using a partial wave decomposition, the coupled differential equation system is solved by a Runge-Kutta method. As a proof of the method ionization processes of atomic hydrogen in half and few cycle attosecond fields are examined. The electron energy spectrum is calculated and the numerical implementation will be presented. Different forms of the interaction operator are considered and the convergence behaviour is discussed. Results are compared to other studies which use independent approaches like finite difference methods. Remarkable agreement is achieved even with strong field strengths of the electromagnetic field. It is demonstrated that expanding in L^2 functions and imposing boundary conditions at the limit in the L^2 function space can be an advantageous alternative to conventional propagation methods using complex absorbing potentials or complex scaling.

Development of an efficient in vitro regeneration system in Costus speciosus - an important herbal insulin plant

Costus speciosus is being rapidly eliminated in its natural habitats in India and driven to a nearly threatened degree of extinction because of non-selective collection and over-exploitation. The commercial cultivation of C. speciosus is impeded owing to its poor seed viability, reduced germination rate, and weak rooting potential of vegetative cuttings. As costus could not be multiplied rapidly by conventional propagation methods, in-vitro regeneration technology was experimented with to standardize explants in addition to the combinations of plant growth regulators (PGRs) to fortify the growing media that helps to produce the enormous number of plants in a shorter duration. Several kinds of explants viz., leaves and nodes were cultured into the Murashige and Skoog (MS) medium reinforced with different combinations and doses of PGRs. A higher frequency (51.67 ± 1.7%) of the regenerated adventitious shoot was obtained from nodal segments cultured onto MS medium fortified with 0.5 mg/l Thidiazuron (TDZ) and 2.0 mg/l of 6-benzyl amino purine (BAP). TDZ in combination with BAP and Indole-3-acetic acid (IAA) exhibited a remarkable effect on shoot multiplication and elongation. Further, MS medium with 0.5 mg/l TDZ, 1.0 mg/l 2, 4-dichlorophenoxyacetic acid, and 0.5 mg/l IAA yielded a great range of callus initiation (86.66 ± 1.6%) from leaf explants. Leaf-derived callus results in the greatest shoot regeneration (56.66 ± 1.7%) response on TDZ (0.5 mg/l) and BAP (1.0 mg/l) reinforced MS medium. Elongated shoots from node and callus cultures when inoculated with the MS medium reinforced along with 1.0 mg/l of IAA and 0.5 mg/l of Indole-3-butyric acid (IBA) produced optimum rooting (88.33 ± 1.66%). The rooted young plantlets were effectively acclimatized to the environmental climate and their greenhouse survival rate was 85%.

Global fast terminal sliding mode based radial basis function neural network for accurate fault estimation in nonlinear systems

The problem of quick and accurate fault estimation in nonlinear systems is addressed in this article by combining the technique of radial basis function neural network (RBFNN) and global fast terminal sliding mode control (GFTSMC) concept. A new strategy to update the neural network weights, by using the global fast terminal sliding surface instead of conventional error back propagation method, is introduced to achieve real time, quick and accurate fault estimation which is critical for fault tolerant control system design. The combination of online learning ability of RBFNN, to approximate any nonlinear function, and finite time convergence property of GFTSMC ensures quick detection and accurate estimation of faults in real time. The effectiveness of the proposed strategy is demonstrated through simulations using a nonlinear model of a commercial aircraft and considering a wide range of sensors and actuators faults. The simulation results show that the proposed method is capable of quick and accurate online fault estimation in nonlinear systems and shows improved performance as compared to conventional RBFNN and other techniques existing in literature.

Somatic embryogenesis mediated micropropagation of polyembryonic cultivars of mango Vellaikolumban and Olour using nucellus tissue

Abstract Somatic embryogenesis mediated micropropagation from nucellus tissue was attempted in polyembryonic mango at the ICAR-Indian Institute of Horticultural Research, Bengaluru during 2016–2019. Studies were conducted to optimize the different stages of somatic embryogenesis namely induction, proliferation, conversion, maturation, germination of somatic embryos and ex vitro establishment of plantlets using nucellus tissue. For culture initiation, various developmental stages of fruits ranging from less than 20 days post-pollination to more than 60 days-post pollination of cultivars Vellaikolumban and Olour were utilized. Fruits of 30–40 days post-pollination in Vellaikolumban and 40–50 days post-pollination in Olour gave better percentage callusing with fewer days for callus initiation and callus formation. Embryonic calli developed on induction medium containing Rugini olive (RO) constituents, 6% sucrose, and 5 ppm each of 2, 4-Dichlorophenoxyacetic acid (2, 4-D) and Giberellic acid (GA3). Upon shifting to proliferation medium (RO with 6% sucrose, 5 ppm each of 2, 4-D and GA3, 400 mgL−1of glutamine and 20% v/v coconut water) pro-globular and globular embryos were formed within 4–5 weeks. Further development of somatic embryos through early heart, late heart and cotyledonary stages occurred in conversion medium (half strength B5 salts, full RO organics, 5 ppm of Indole-3-Acetic Acid (IAA), 20% coconut water and 200 mgL−1 of casein hydrolysate). Maturation of embryos was obtained by using 0.01 mg L−1of Abscisic acid (ABA) and 100 mg L−1of Polyethylene glycol (PEG). Germination of embryos with shoot and root initiation was observed in the presence of 5 ppm of zeatin. Our results show that somatic embryogenesis can be used as an alternative method to conventional propagation methods for rapid multiplication of uniform planting material in polyembryonic mango cultivars.

Micropropagation from Juvenile Material of <i>Annona senegalensis</i> Pers.

Annona senegalensis is a savannah species present in the wild in tropical and sub-tropical African regions. It is used by the local populations for its many economic, pharmacological, nutritional and ecological interests. However, the strong pressure exerted on this species as well as the increasing degradation of the ecosystems, in which it evolves, seriously compromise its sustainability and development. Conventional methods of vegetative propagation are not sufficient to ensure its durable regeneration and the renewal of endemic populations in Senegal. In this context, it was undertaken in vitro propagation, from 30 days old juvenile seedlings. Thus, axillary, cotyledonary and apical nodes were in vitro cultured in Murashige and Skoog (1962) medium (MS), with different concentrations of cytokinins and/or auxins. BAP used alone at 2 mg•L−1 proved to be more beneficial for micropropagation of different types of explants compared to Kinetin used alone or in combination with BAP, especially for axillary and cotyledonary nodes. The BAP-NAA combination allowed to have a better elongation of newly formed shoots. For shoots of apical and cotyledonary nodes origin, a rhizogenic induction of 5 days, with IBA at 25 mg•L−1, resulted in a better rooting rate with, respectively, 75% (for 2.22 roots) and 66.67% (for 4.17 roots). For newly formed shoots of axillary origin, a 24-hour rooting induction with IBA at 50 mg•L−1 gave a rooting rate of 58.33% (for 2.4 roots). After 30 days of acclimatization, the survival rate was 75% for the young plants from the apices and 83.33% for those originating from the cotyledonary and axillary nodes. This protocol can therefore be adopted for the in vitro clonal propagation of A. senegalensis juvenile material.

In vitro Propagation of Banana (Musa paradisiaca L.) Plant Using Shoot Tip Explant

Banana is a fruit crop which has high demand in Ethiopia, but its production is constrained by lack of disease free planting material with conventional propagation methods. For shoot initiation, shoot tip explants were cultured on MS medium supplemented with 0.5, 1.0, 1.5 and 2.0 mg/L BAP. Similarly, MS medium supplemented with BAP at 1.0, 1.5, 2.0 mg/L in combination with IBA at 0.25 and 0.50 mg/L were used for shoot multiplication. Half- strength MS medium augmented with IBA at 1.0, 2.0, 3.0 and 4.0 mg/l were used for root induction. MS medium without PGRs were used as controls. Finally, hardening of the in vitro derived plantlets was carried out in green house both in the primary and secondary acclimatization stages. Results showed that the highest shoot initiation percent (93.40%), highest mean number of shoots per explant (4.67) and lesser day for shoot induction (11.00) were observed in explant cultured on MS + 1.0 mg/L BAP. With shoot multiplication, highest shooting percent (92.60%), maximum number of shoots (7.67) and highest shoot length (5.27 cm) were recorded on MS + 1.5 mg/L BAP + 0.5 mg/L IBA. The highest rooting percent (93.40%), maximum root number per shoot (7.67) and highest root length (11.00 cm) were found on a half strength MS medium + 2.0 mg/L IBA. The survival rate of plantlets were 96.00% in coco peat substrate in primary acclimatization and 97.92% in forest soil, sand and manure substrates mixed at 3:2:1 ratio in secondary acclimatization. Overall, the result showed that the PGRs type, concentrations and combinations used are effective for mass propagation of banana variety studied in this experiment.

An efficient micropropagation protocol for Ficus carica cv. Golden Orphan suitable for mass propagation

Fig ( Ficus carica L.) is from the Moraceae family and is rich in essential nutrients, natural fiber and vitamins. The Golden Orphan cultivar is a yellow fig that is sweet, firm and can thrive in tropical climates. Current conventional propagation methods for the multiplication of plant stocks were found to be less efficient in producing healthy plant stocks. Micropropagation offers an alternative in the production of plant stocks resembling the quality of mother plants. The current study aims to fine tune the micropropagation protocol of the Golden Orphan cultivar for efficient production of plant stocks. Sterile nodal bud explants were subjected to different media treatments, carbon source and combinations of auxin and cytokinin. Rooting of explants was induced in auxin and rooted explants were acclimatized in cocopeat. The optimal basal medium for shoot elongation was Murashige and Skoog (MS) Medium, whereas 20 g/L sucrose resulted in the highest shoot length (2.00 ± 0.22 cm) facilitating the production of nodal segments for subsequent multiplication. The highest shoot induction (4.15 ± 0.43 shoots/explant) was observed in the treatment of 0.8 mg/L 6-Benzylaminopurine (BAP), whereas the combination of 0.8 mg/L BAP and 0.5 mg/L indole-3-acetic acid (IAA) resulted in the highest shoot length (2.23 ± 0.09 cm). Treatment of 0.4 mg/L indole-3-butyric acid (IBA) produced the optimal percentage of rooting (83.33%) whereas acclimatization with cocopeat resulted in maximum survival rate. The present study has successfully optimised the crucial aspects of micropropagation for the Golden Orphan cultivar suitable for rapid plant stock production. • The establishment of micropropagation protocol for Ficus carica cv. Golden Orphan. • High multiplication rate in basal medium, minimal amount of sucrose and cytokinin. • Maximum acclimatization efficiency in cocopeat. • Acclimatized plants successfully produced fruits that resembles mother plants.

SUBSTITUTION OF SYNTHETIC HORMONES WITH ORGANIC MATERIALS ON THE GROWTH OF ORCHID PLANTS (Phalaenopsis amabilis) AS A GROWTH REGULATORY SUBSTANCE IN VITRO

In Indonesia, the agribusiness sector has begun to expand due to the increase in entrepreneurship of orchid plants. Several genera and species of orchid plants are promising to be traded as ornamental plants, one of which is the moon orchid (Phalaenopsis amabilis). However, the limited number of seeds or plants produced by conventional propagation methods takes a long time to get new tillers. Therefore, tissue culture techniques can be used to grow and reproduce moon orchid plants. In the success of in vitro culture, the balance of growth regulators is an essential factor. On the other hand, synthetic hormones such as 1-naphthalenaecetic acid (NAA) and 6-benzylaminopurine (BAP) are relatively expensive. The study in this research is to substitute the synthetic hormones NAA and BAP with organic ingredients that are cheap and easy to find. Such as Purple Sweet Potatoes and Beans and find out the right concentration of organic matter. This study used a completely randomized design with two factorials (concentration of purple sweet potato and green beans) with three repetitions. PLB (Protocorm Like Bodies) moon orchid (Phalaenopsis amabilis) was treated with: positive control (2 ppm), negative control (without the addition of organic or synthetic hormones), and variations in the concentration of purple sweet potato and green beans organic matter. The result was that C4D0 (500g/L purple sweet potato extract and 0 g/L green bean extract) was an excellent formulation, although there was no significant interaction between purple sweet potato and green bean extracts against moon orchid.

A Review on Chemical Composition, Biosynthesis of Steviol Glycosides, Application, Cultivation, and Phytochemical Screening of Stevia rebaudiana (Bert.) Bertoni

Aims: Stevia rebaudiana which is commonly known as sweet leaf herb is used to extract non caloric sweet steviol glycosides. Thus based on available reports about its chemical composition, biosynthesis of steviol glycosides, application, cultivation, and phytochemical screening an attempt has been made to review to Stevia rebaudiana in context of its medicinal and pharmaceutical importance.
 Methodology: Literature search have been done in the web using google scholar, PubMed as search platform. More than two-thirds of the references are within 15 years.
 Results: According to the literature search S. rebaudiana leaves contain non caloric steviol glycosides and pharmaceutically important other phytochemicals. Miccropropagation is the best propagation method to overcome difficulties in conventional propagation methods.
 Conclusion: It is concluded that in vitro cultures has potential to extract important phytochemicals used in various pharmaceutical, food (flavoring agents, food additives) and perfume industries.

Effective Propagation of Selaginella tamariscina through Optimized Medium Composition

Selaginella tamariscina is a medicinal plant that contains a variety of plant secondary metabolites; however, it is currently being collected indiscriminately from its native habitats. Hence, we have developed an efficient propagation method for S. tamariscina. Explants grown in vitro were cultured in Murashige and Skoog medium of various strengths (1/16–2x), and the highest number of sporophytes (65.7) were obtained with 1/4x MS medium. Culturing explants at various lengths (3–12 mm) for 12 weeks indicated 12 mm as the most appropriate size for sporophyte propagation. We then evaluated various concentrations of individual components, sucrose (0–5%), total nitrogen (7.5–30 mM), nitrogen ratio (3:0–0:3), and agar (0.6–0.8%), in the 1/4x MS medium for explant growth for 12 weeks. The maximum number of sporophytes were formed in media containing 3% sucrose, 15 mM nitrogen, and 0.6% agar, with a nitrogen ratio of 1:2. The propagated S. tamariscina was then acclimatized in a controlled environment to improve survival in an external environment. These results demonstrate the effective conditions for in vitro mass propagation of S. tamariscina, finding that methods utilizing sporophytes were more efficient than conventional propagation methods and yielded numerous plants in a short period.

Co-expression of hydrolase genes improves seed germination of Sinopodophyllum hexandrum

Sinopodophyllum hexandrum is a perennial rhizomatous herb and its rhizome produces anti-cancer compound podophyllotoxin (PPT). As a result of overexploitation of wild resources and lack of large-scale cultivation, it is currently an endangered species. An effective and conventional propagation method is by seed, but seed germination is erratic attributed to the barriers from seed coat and thick-walled endosperm. While several proteins and genes that are involved in cell wall loosening and endosperm weakening have been identified, the mechanism responsible for regulating germination has not been systematically mapped. Here, transcriptomics profiles of S. hexandrum during seed germination were performed, genes expression was validated as well as metabolites and hormones were measured. Over one thousand genes were observed to be differentially co-expressed at the five germination stages. Functional annotation classified 67 genes associated with cell wall loosening and endosperm weakening (25), primary metabolism and energy (16), hormone biosynthesis and bio-signaling (15), radicle growth (5), and transcription factor (6). The expression levels of genes involved in hydrolases (i.e. XTHs, GLCs, MANs, EXPAs, AGALs, PECSs and PGs), metabolites (i.e. starch, oligosaccharides, proteins, lipids and fatty acids) and hormones (i.e. GA3, ABA and IAA) were almost consistent with changes of seed water uptake, contents of metabolites and hormones, and cell wall modifications. This combinational analysis of transcriptomics with targeted metabolites and hormones provides new insights into seed germination of S. hexandrum.