Interaction Effect of Auxin and Cytokinin on in Vitro Shoot Regeneration and Rooting of Endangered Medicinal Plant Valeriana jatamansi Jones through Tissue Culture

Micropropagation of Valeriana jatamansi Jones by using small segments of rhizome on full strength MS medium having various concentrations and combinations of auxin Naphthaleneacetic acid (NAA) and cytokinin Benzylaminopurine (BAP) was conducted. The highest mean shoot length (3.71 cm) was achieved when media was fortified with BAP 2 mg/L in combination with NAA 1 mg/L. The highest mean leaf number i.e. 6.00 was observed when BAP was used alone at 2 mg/L. Average root length (0.77 cm) was recorded when BAP 1.5 mg/L along with NAA 0.5 mg/L was used. Maximum mean root numbers 2.57 were obtained when BAP and NAA were used at equal concentrations i.e. 1.5 mg/L. Observed data demonstrated that BAP up to 1 mg/L, 1.5 mg/L and 2 mg/L promotes shoot length, leaf number and leaf growth when used along with NAA at 0 mg/L, 0.5 mg/L and 1 mg/L. However lower quantities of both NAA (0 mg/L, 0.5 mg/L) and BAP (1 mg/L and 1.5 mg/L) produced significantly higher root length of Valeriana jatamansi Jones but the higher concentrations of plant growth hormones BAP (2 mg/L, 3 mg/L) and NAA (1 mg/L, 1.5 mg/L) were found unfavorable for increase in root length but the root number increases at higher concentration of NAA (1 mg/L and 1.5 mg/L).

Coscinium fenestratum: Callus and Suspension Cell Culture of the Endangered Medicinal Plant Using Vermicompost Extract and Coelomic Fluid as Plant Tissue Culture Media

In vitro tissue culture of hard woody, endangered, medicinal plant Coscinium fenestratum is most challenging to plant tissue culturists. In the present study, petiole and leaf explants of Coscinium fenestratum were induced to form callus when cultured on vermicompost extract media along with coelomic fluid. Suspension medium was developed using vermicompost extract and coelomic fluid in 3:1 ratio. Phytochemical analysis of the alkaloid berberine was confirmed from callus, suspension cell culture and suspension medium by Thin Layer Chromatography and High Performance Liquid Chromatography. Vermicompost and its extracts with coelomic fluid have shown maximum (100 per cent) response of callus induction. Callus mass enlarged with increasing concentration of coelomic fluid and callus growth was assessed from the biomass. Incubation of culture tubes in dark supported callus development significantly. The Rf value of 0.36 confirmed the presence of berberine by Thin Layer Chromatography. Qualitative analysis confirmed the presence of alkaloid berberine with the retention time of 2.8 minutes similar to that of standard reference sample from Sigma chemicals, USA. The suspension medium turned deep yellow because of the release of the alkaloid. Vermicompost and its extracts along with coelomic fluid have shown the economical approach for micropropagation of economically and medicinally important plants.

Contributions of Chinese Botanists to Plant Tissue Culture in the 20th entury

This paper looks back to the development of plant tissue culture in China in the last century. Since 1934, tissue culture studies in China has kept up with the international development in the fields. Progress has been made by Chinese in nearly every branches of tissue culture, including in vitro organogenesis, shoot tip culture, anther culture, ovary culture, endosperm culture, protoplast culture as well as mass cell culture. On the basis of reviewing the articles written by Chinese on plant tissue culture, the internationally recognized contributions are specially mentioned. The applications of plant tissue culture to agriculture and industry in China are also introduced.

Plant Tissue Culture: A Recent Progress and Potential Applications

Plant tissue culture systems have enormous potential in fundamental re- search and for commercial applications such as horticultural industry. The process of tissue culture is companied with a series of changes in respect to morphology, physiology, biochemistry, molecule and epigenetics. The changes at molecule levels mainly include genetic variation, DNA sequence variation, chromosomal variation and epigenetic regulation （DNA methylation, histone modification, chromatin remodeling, small RNA regulation）. These changes are believed to facilitate explant adaptation to culture conditions and to help subsequent morphogenesis processes. Nowadays, it has played a crucial part in commercial applications and in basic research into cell biology, genetics and biochemistry, etc. In present review, we shed light on the fun- damental of plant tissue culture, culture medium preparation, explant selection, mechanism of action of various hormones, the three major problems （explant pollu- tion, browning, plantlets vitrification） and the prevention measures in tissue culture, and elaborated on in vitro propagation of plants, virus-free seedling cultivation, cry- opreservation, artificial seeds and molecule levels changes during in vitro culture further.

Advance of New Researches on Tissue Culture of Rare and Endangered Plant Elaeagnus mollis

This review summarized the recent research achievements on the process of tissue culture of the rare and endangered plant Elaeagnus mollis,including sterilizing protocols of different explants derived from adult plants and seeds for tube germination,two types of plants regeneration（organ type and organ genesis type）,methods of rooting and transplanting,factors affecting culture,as well as browning and vitrification phenomena and avoiding measures.And the further biotechnology research fields of E.mollis were prospected.

Tissue Culture and Plant Regeneration of Chicory

[ Objecllve] To establish a high-frequency regeneration system of chicory （ Cichodum intybus L. ） using leaf segments of aseptic seed- lings. E Method] Calluses and adventitious buds of chicory were induced by inoculating explants on MS medium supplemented with 6-BA （6-benayl aminopurine） and NAP, （naphthylacetic acid） at different final concentrations. [ Result] When lower part of leaves derived from 20-day-old seedlings was used as explant and inoculated on MS medium containing 2.0 rng/L 6-BA, 0.5 mg/L NAA and 40 g/L sucrose, the frequency of adventitious bud formation was 90.0%. When the regenerated shoots were cultured in 1/2 MS medium containing 0.1 mg/L NAA, the frequency of root forma- tion was 88.3%. All rooted plants transplanted in pots could survive and grew well without abnormal shape. [ Conclusion] Better differentiation of adventitious buds can be achieved by inoculating the lower part of leaves derived from 20-day-old seedlings on MS medium containing 2.0 mg/L 6- BA, O. 5 mg/L NAP, and 40 g/L sucrose. The 1/2 MS medium containing O. 1 mg/L NAP, is most suitable for rooting.

Pollution and Control Measures in Plant Tissue Culture in Orchid Plant

Contamination is a phenomenon that often occurs in the operation of plant tissue culture,and it is also one of the three major problems in plant tissue culture.Compared with browning and vitrification,contamination is more likely to occur,which brings great harm to scientific research and production practice.Its appearance will greatly affect the normal growth and differentiation of tissue culture materials,and will reduce the yield of cultivated plants to a certain extent.Therefore,we cannot underestimate any step in the tissue culture operation.This study summarized the reasons for its occurrence and how to formulate prevention and control measures based on recent research combined with the actual situation.

Rapid Propagation of Pteris vittata L.via Tissue Culture

[Objective] The study had developed a means of rapid propagation Pteris vittata L.by tissue culture. The species was a perennial fern belonging to the genus Pteris. [Metbed] The leaf bud of P. vittata collected in field conditions as explantsand the 1/2 MS ＋ 3% sucrose ＋ 0.7% agar as the basic medium were used to screen the medium formula of the phytohormone ratio for callus induction and subculture of P. vittata. [Result] The best medium formula for each step was list below： 1/2 MS ＋ 3% sucrose ＋ 0.7% agar ＋ 0.5 g/L PVP ＋ 0.1 mg/L KT ＋ 0.5 mg/L 2, 4-D for in- ducing the callus from explants; 1/2MS ＋ 3% sucrose ＋ 0.7% agar ＋ 0.5 g/L PVP ＋ 1.0 mg/L KT ＋ 0.01 mg/L 2,4-D for inducing the GGB from callus and the seedlings from GGB. In addition, 1/2 MS ＋ 3% sucrose ＋ 0.7% agar ＋ 0.5 g/L PVP ＋ 0.5 mg/L 2,4-D for the subculture could make the continued proliferation of callus. [Cen- clusioa] This study makes an applicable procedure by the direct use of field materi- als, for propagating P. vittata in a simplified and rapid mode.

Experiment on Tissue Culture Technique of Saposhnikovia divaricata(Turcz.) Schischk

The study aimed to optimize the induction and differentiation medium by exploreing different tissue culture of Saposhnikovia divaricata （Turcz.） Schischk. In tissue culture with the root, stem segments, young leaf, cotyledonary node and axillary bud of Saposhnikovia divaricata （Turcz.） Schischk as explants, a lot of plantleles were obtained and the corresponding plant regeneration-system was established. The results showed that when use MS＋1.0 mg·L^-1 6-BA＋0.2 mg·L^-1 NAA as callus induction medium, the cotyledonary node had the highest bourgeon rate, and its callus was better than any others; MS＋2 mg·L^-1 6-BA＋0.4 mg·L^-1 NAA was the best adventitious buds induction medium, and the best adventitious buds induced condition was 3% sucrose as carbon source, illumination for 12-14 h·d^-1 and pH 5.8, The best rootage medium was 1/2 MS＋0.5 mg·L^-1 NAA.

Study on Tissue Culture and Adventitious Bud Induction of Eucheuma striatum

This study aimed to investigate the aseptic treatment and disinfection of E. striatum explants and the effects of three liquid culture medium （ PES, PESI, f/2） and plant hormones on adventitious bud induction from explants. The results showed that the vast majority of explants disinfected by combined treatment exhib- ited good growth and normal color with an average mortality rate of only 9%. E. striatum explants exhibited the best growth in PES liquid medium and the survival rate reached 66.67% at 40 d after culture. Under different concentrations （0.5, 2, 5 mg/L） of 6-BA, the induction rate ofE. striatum explants was significantly higher than that in control group, but no remarkable differences were found in the induction of adventitious buds from E. striatum explants. In PES liquid medium containing 6-BA and 2,4-D, the adventitious bud induction rate was 90% and the average number of induced adventitious buds was 6.63.

Endemic medicinal plant distribution correlated with stable climate,precipitation,and cultural diversity

Medicinal plants provide crucial ecosystem services,especially in developing countries such as China,which harbors diverse endemic medicinal plant species with substantial cultural and economic value.Accordingly,understanding the patterns and drivers of medicinal plant distribution is critical.However,few studies have investigated the patterns and drivers of endemic medicinal plants distribution in China.Here,we linked endemic medicinal plants distribution with possible explanatory variables,i.e.,paleoclimate change,contemporary climate,altitudinal range and ethnic minority human population size at the prefecture city level in China.Our results show that endemic medicinal plants are concentrated in southern China,especially in southwestern China.Notably,both endemic medicinal plant species richness and the ratio of endemic medicinal plant species richness are negatively associated with glacialinterglacial anomaly in temperature,and positively associated with contemporary precipitation and altitudinal range.In addition,we found that endemic medicinal plant species richness is positively associated with ethnic minority population sizes as well as its ratio to the overall population size.These findings suggest that the distribution of endemic medicinal plants is determined by multiple drivers.Furthermore,our findings stress that dramatic future climate changes and massive anthropogenic activities in southern China pose great challenges to the conservation of China's endemic medicinal plants.

Development of an in vitro culture system of Rosa spp.Plants

[Objective] This study was to develop an in vitro tissue culture system of Rosa spp.[Method] Using ten species of Rosa spp.plants as experimental materials,different combinations of hormones were designed to establish their in vitro tissue culture system with the stem segments as explants.[Result] All ten tested varieties germinated when the nodal segment explants were cultured on the sprouting medium MS＋ 0.5 mg/L BA ＋0.01 mg/L NAA and grew vigorous shoots,and the sprouting rate was up to 70%.Of the ten tested rose varieties,each has a respective optimal proliferation medium,and the multiplication rates for all the varieties reached 3.0%.The axillary buds were vigorous and normal in leaf color.The optimal medium for rooting and acclimation was 1/2MS medium containing 0.1 mg/L or 0.2 mg/L NAA,in which the rooting frequency reached 90%-100% and the root system was developed.After acclimation and transplant,the survival rate was as high as 95%.[Conclusion] An in vitro tissue culture system of Rosa spp.has been established in this study,which lays foundation for the molecular breeding of Rosa spp.

Study on Shoot Tip Culture of Illciaceae Ornamental Plants

[Objective] This study aimed to establish a technology system for tissue culture and rapid propagation of Illciaceae ornamental plants. [Method] Effects of medium components and anti-browning agents on the survival and growth of shoot tips were investigated by using apical buds of IItciaceae plant Haierlian as experiment material and MS as basic medium. [Result] The results showed that apical buds at the early germination period in spring were the most suitable explants for tissue culture of IIIciaceae plant Haierlian. Sterilization with 0.1% HgCI2 for 6 min achieved the best effect, while conventional surface-sterilization with ethanol would affect the survival of explants. The optimal medium for primary culture was MS-D （with modifications in major elements and organic components） ＋ anti-browning agents （equa~ volume） ＋ 2.0 mg/L of 6-BA ＋ 0.5 mg/L of NAA. The optimal subculture medi- um was MS-F （with modifications in inorganic and organic components） ＋ anti-brown- ing agents （equal volume） ＋ 2.0 mg/L of 6-BA ＋ 0.1 mg/L of NAA. [Conclusion] This study laid the foundation for establishment of tissue culture and rapid propagation technology system for Haierlian.

Study on Plant Regeneration of Wheat Mature Embryos Under Endosperm-Supported Culture

To reveal the suitability of using mature embryos as an explant source in wheat tissue culture, mature embryos from eight common wheat cultivars （Triticum aestivum L. cv.） were cultured with or without endosperm to test their efficiency of callus induction and plant regeneration. When embryos were cultured together with endosperm （endosperm-supported culture, ES）, the percentage of callus induction was significantly lower than that when embryos were cultured in the absence of endosperm （non-endosperm-supported culture, NES）. This pattern was evident in most genotypes, regardless of whether 2 or 8 mg L^-1 2,4-D was added in the NES culture. However, in ES culture, more induced calli were differentiated into distinct green spots and they further developed into plantlets. Thus, more plants were regenerated in ES culture than in the NES treatment. Most of the eight tested genotypes showed a significant difference in callus induction rate and plantlet regeneration in both ES and NES cultures. In addition, the enzymatic activity of oxalate oxidase in the callus of ES culture condition was obviously higher than that in the callus of NES culture condition, suggesting that the activity of oxalate oxidase may be a parameter for selection of calli with potential for plantlet regeneration. These results indicate that wheat mature embryos are valuable explants for highly efficient callus induction and plant regeneration, if proper treatment and medium are used.

Dicamba and Sugar Effects on Callus Induction and Plant Regeneration from Mature Embryo Culture of Wheat

To establish a highly efficient plant regeneration system for wheat genetic transformation, the effects of three different concentrations of dicamba and two different sugar types on callus induction and plant regeneration from mature embryo cultures were evaluated. Callus induction and plant regeneration were obtained from mature embryos of two commercial cultivars Zhoumai 18 and Yumai 34 （Triticum aestivum L.） cultured on L3 basal medium. The results showed that the efficiency of mature embryo culture was significantly influenced by the genotypes, sugar types and dicamba concentrations. 4 mg L^-1 dicamba proved the best effective for inducing embryogenic callus and also gave the highest proportion of plants regenerated across the two cultivars. Substitution of maltose by sucrose significantly improved the plant regeneration efficiency in both cultivars. There was a significant interaction between genotype-by-sugar types, and sugar types-bydicamba concentrations. Overall, Zhoumai 18 gave the highest frequency of plant regeneration （82.65%） when dicamba concentration was 4.0 mg L^-1 and with sucrose in initial callus induction. These results will facilitate genetic transformation work with elite wheat.

Anticancer Activity of Rice Callus Suspension Cultures from Aromatic Varieties and Metabolites Regulated in Treated Cancer Cell Lines

Tissue culture techniques were used to produce large amounts of bioactive compounds with medicinal potential, overcoming space and time constraints for cancer prevention. Rice callus suspension cultures(RCSC) and seed extracts prepared from aromatic rice varieties were used to evaluate the cytotoxic impact on human colon and lung cancer cell lines, as well as a normal control cell line, using Taxol as a positive control. RCSC and seed extracts from two Indian aromatic rice varieties were applied at different concentrations to treat the cancer cell lines and normal lung fibroblasts over varying time intervals. Apoptosis was assessed in 1:5 dilutions of the A549 and HT-29 cell lines treated with RCSC for 72 h, using propidium iodide staining and flow cytometry. RCSC showed a more potent cytotoxic effect than seed extracts with minimal effect on the normal cell line, in contrast to Taxol. Confocal microscopy and flow cytometry further confirmed the apoptotic effect of RCSC. Gas chromatography-mass spectrometry-based metabolic profiling identified metabolites involved in cytotoxicity and highlighted altered pathways. RCSC is proposed as an alternative source for the development of novel anticancer drugs with reduced side effects.

Different culture conditions applied to in vitro shoot multiplication of two Eucalyptus benthamii explant sources

Eucalyptus adult material requires more successive subcultures in the in vitro multiplication phase for increased vigor and cellular activity. This study evaluated the endophytic manifestation and shoot multiplication of one 13-year-old Eucalyptus benthamii clone under different culture conditions and used canopy branches(CB) and trunk base material as explant sources. The culture media were wood plant medium(WPM), Murashige and Skoog medium(MS) and JADS(Correia and co-authors medium).Based on the results of the initial multiplication experiment, further tests examined sucrose concentrations and p H. Morphophysiology, dry mass production, endophyticmanifestation and histochemical were determined. Explant sources responded differently to MS and JADS media, but the WPM medium promoted homogeneous development.The responses were similar for both explant sources when sucrose concentrations varied. Shoots died in the absence of sucrose, showed high oxidation at 60 g L-1 and optimal development at 30 g L-1. Endophytes were more evident for shoots from the CB origin. Explant sources responded distinctively to treatment due to physiological and intrinsic genetic factors. Therefore, explant sources, different culture media, sucrose concentration and p H may determine micropropagation success and influence the presence and/or intensity of endophytic manifestation.

An Overview of the Use of Medicinal Plants in Regenerative Dentistry

Aim: The oral cavity has the particularity to host multiple hard and soft tissues, in this paper, we will discuss the current therapies that lead to cell differentiation by regenerative therapies and the future alternatives proposed by medicinal plants and all the regenerative potential of these different tissues. Material and Methods: A detailed review of the literature through the various search engines: Scopus, PubMed, google scholar, Cochrane, etc., uses the selected keywords to explore the effect of the regenerative potential of several medicinal plants. Results: Through our research, we have proceeded to sort different medicinal plants, according to their repairing and regenerative potential on the different tissues of the oral cavity. Conclusion: Future studies are conceivable to explore the opportunities and potential provided by medicinal plants in the field of regenerative dentistry.

Production of Active Compounds in Medicinal Plants:From Plant Tissue Culture to Biosynthesis

Over past decades plant tissue culture has emerged as an alternative of whole plant cultivation in the production of valuable secondary metabolites.Adventitious roots culture of Panax ginseng and Echinacea purpure has reached the scale of 1-10 kL.Some molecular biological techniques,such as transgenic technology and genetic stability are increasingly used in the studies on plant tissue cultures.The studies on elicitors have deepened into the induction mechanism,including signal molecules,functional genes,and so on.More and more biological elicitors,such as A.niger and yeast are used to increase the active compounds in plant tissue cultures.We also discussed the application of synthetic biology in the studies on biosynthesis of artemisinin,paclitaxel,and tanshinon.The studies on active ingredients biosynthesis of medicinal plants provide unprecedented possibilities to achieve mass production of active ingredients.Plant tissue cultures can not only produce active ingredients but also as experimental materials for biosynthesis.In order to improve the contents of active compounds in medicinal plants,following aspects could be carried out gene interference or gene silencing,gene overexpression,combination with chemical synthesis,application of elicitors,and site-directed mutagenesis of the key enzymes.

Plant Tissue Culture and Biosynthesis Provide a Fast Way to Produce Active Constituents of Traditional Chinese Medicines

The plant kingdom has provided literally thousands of natural products with widely diverse chemical structures and a vast array of biological activities.Many of them have seen subsequent application in discovery of new druids and the pharmaceutical industry and clinical therapeutic application.