Establishment of an in vitro Tissue Culture and Rapid Propagation System using Lonicera hypoglauca and Content Assessment of Total Flavonoids and Chlorogenic Acid

Lonicera hypoglauca is a traditional Chinese medicinal plant.In this study,the tender young leaves of L.hypoglauca were used for the first time as the explants to establish a rapid in vitro propagation and regeneration system.The results revealed that the optimal time for disinfection of the explants was 8 min and the optimal medium for callus induction was MS+2,4-D 4.0 mg·L^(-1)+sucrose 30 g·L^(-1),with an average callus induction rate of 86.67%.The optimal medium to induce differentiation of callus to bud was MS+6-BA 1.0 mg·L^(-1)+NAA 0.10 mg·L^(-1)+sucrose 30 g·L^(-1),with an average germination rate of 83.33%.The optimal medium to induce multiplication was MS+6-BA 1.5 mg·L^(-1)+NAA 0.05 mg·L^(-1)+sucrose 30 g·L^(-1),with a multiplication coefficient of 5.42.The optimal medium for root induction was 1/2 MS+NAA 0.15 mg·L^(-1)+activated carbon 0.3 g·L^(-1)+sucrose 15 g·L^(-1),with an average rooting rate of 91.11%.The survival rate of tissue-cultured seedlings in nutrient soil cultivation medium was as high as 100%.The total flavonoid content and chlorogenic acid content in the explant,callus tissue and regenerated plant were 1.83%,2.27%,1.33%and 2.77%,1.83%,1.74%respectively.This study provides novel insights into the rapid propagation and mass production of L.hypoglauca seedlings at an industrial scale and that it exhibits important application value and future prospects.

Targeting Cancers: Uncovering the Potential Roles of Potato Tissue Culture as Anti-Cancer Agents - A Secondary Publication

Plant tissue culture is a technique that enhances the quality and quantity of potatoes. Potatoes are a significant crop and are primarily used in the world. It is a staple food in many countries, where millions of tonnes are produced annually. It is an essential source of many nutrients, such as proteins, carbohydrates, vitamins, and beta-carotene. In addition, potatoes are being used as therapeutic agents against cancer and other human diseases as well. Potatoes are on the third list after wheat and rice. To overcome food shortages and malnutrition, there are two methods used for producing potatoes: the first is sexual, which is seed propagation, and the second is asexual, which is plant tissue culture propagation. Conventional potato breeding is a uniform method, but it is unsafe because there is a risk of pathogen attack. In a laboratory setting, the tissue culture of potatoes produced millions of plants with nutrient-rich medium under controlled environmental conditions that prevent pest attacks. Some environmental stresses, such as salinity and water scarcity, affect potato yield and production;however, applying nanoparticles like organic, inorganic, and silicon dioxide enhances potato quality and combats stress. Biotechnology has proven to be helpful in addressing all these issues. This review discusses the significance of potatoes, their production through the tissue culture technique, and the application of nanoparticles to improve the growth, and impact of potatoes on human health.

Tissue Culture of Calla Lily (Zantedeschia spreng.): An Updated Review on the Present Scenario and Future Prospects

The calla lily(Zantedeschia spreng.)is a bulbousflower native to the tropical regions of Africa.Calla lily has gained significant popularity in the international market owing to its intricate morphology and prolonged flowering duration.Despite such advantages,for two sub-groups of calla lily,known as group Zantedeschia and group Aestivae,there are challenges in terms of hybrid production due to the‘plastome-genome incompatibility’there-between.Tissue culture is a fundamental biotechnological tool used in gene editing research,with a focus on disease resistance andflower color in calla lily breeding programs.The present review provides a brief background on the history and development of the calla lily,as well as a comprehensive and critical summary of calla lily tissue culture research.The regeneration pathways for both group Zantedeschia and group Aestivae can be divided into de novo organogenesis and somatic embryogenesis.Both groups are capable of obtaining replants through such means.However,only some species in group Aestivae have been reported to be successful in the somatic embryogenesis pathway.In the present review,special attention was paid to the influence of explant types,plant growth regulators,and culture conditions on both de novo organogenesis and somatic embryogenesis in calla lily tissue culture.Ultimately,future research prospects were determined based on integrated analysis of recent progress in calla lily tissue culture research.

Evaluation of Different Substrates Compositions for Acclimatization of Tissue Culture Taro Plantlets in a Propagator

Taro is cultivated in most Regions of Cameroon and it is affected by taro leaf blight disease since 2010 which has decreased its production. Lack of disease-free planting materials has been a main problem to farmers. This study was carried out at International Institute of Tropical Agriculture (IITA) Yaounde and Institute of Agricultural Research for Development (IRAD) Bambui to assess different substrates for acclimatization of tissue culture taro plantlets in apropagator. No information is available on acclimatization of Cameroonian taro plantlets in different substrates. Taro plantlets from tissue culture were acclimatised in a propagator for six weeks under different substrates, the first substrate consisted of sterile three parts of soil and one part of river sand mixed together (3:1), the second substrate consisted of sterile two parts of soil and two parts of river sand mixed together (2:2), the third substrate consisted of sterile two parts of soil, one part of rice husk and one part of river sand mixed together (2:1:1) and the fourth substrate consisted of sterile one part of soil and three parts of river sand mixed together (1:3). After acclimatisation of the different taroplantlets (Dark green petiole with small leaves (L1), Red petiole with small leaves (L2), Light green petiole with large leaves (L3) and Light green petiole with small leaves (L4) in these four substrates, it was observed that the best growth rate of plant was recorded on substrate sand + soil (1:3). The other substrates showed moderate growth of plants. Substrate sand + soil (1:3) can be recommended for acclimatization of Cameroonian taro plantlets.

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 Explants of Lilium lancifolium Have Different Potential to Differentiate and Regenerate in Tissue Culture

[Objective] The aim was to investigate differences in differentiation and regeneration of the explants from different parts of Lilium lancifolium（Yixing Lily） in tissue culture.[Method] The different parts of scale,leaf and root of Yixing Lily were cultured as explants on MS basic medium supplemented with different concentrations of plant growth regulators,so as to compare their capacity to differentiate and regenerate.[Result] The explants had different potential to differentiate（scale root leaf）.The capacity of different scale parts to differentiate was the lower part middle partupper part;the capacity of different leaf parts to differentiate was the leaf base middle part leaf tip;the capacity of different root parts to differentiate was the root base root tip middle part.[Conclusion] Tissue culture could be well applied in propagation of Yixing Lily.

Tissue Culture of Mini-Papaya

[Objective] The aim was to study the tissue culture of mini-Papaya.[Method] The pretreatment seeds of mini-Papaya were cultured in the MS medium containing 6-BA and IBA of different densities for rapid propagation.[Result] In the condition of aseptic strain,the surface of mini-Papaya peel was uniformly wiped by 75% alcohol,then seeds were removed and washed by aseptic water for 3 times,which was the best sterilization method,and the pollution rate of seed was only 2.52%.After seeds which had been soaked by the equi-volume mix-solution of 1 000 mg/L GA3 and 1 mg/L 6-BA for 18 h were further purified in MS medium,the germination rate of seed,the length of embryo bud and radicle and the height of seedling were 68.42%,2.25,0.80 and 1.52 cm respectively,furthermore the total situation of seedling growth was better.When subculture multiplication medium was MS＋0.5 mg/L 6-BA＋0.1 mg/L IBA medium,proliferation coefficient of subculture multiplication reached the highest （7.92）,and the seedlings grew better.The ratio of vitrified shoots decreasing with the increase of light intensity could reach the lowest level （3.21%） under the light intensity of 3 000 lx.[Conclusion] The research provides reference for studies on tissue culture and rapid propagation of mini-Papaya.

Effects of Different Culture Conditions on Vitrification of Lycium barbarum L. Plantlets in Tissue Culture

The tender stems from new Lycium barbarum L. cultivar ＂Ningqi 3＂ released by Ningxia Academy of Agricultural and Forestry Sciences were regarded as explants to investigate the vitrification of Lycium barbarum plantlets in tissue culture under different concentrations of 6-BA, sucrose, agrose, culture temperature, and illumination duration with MS as basic medium. The results show that the conditions for maximal proliferation coefficient and min- imal vitrification are as following： the basic medium with 0.2 mg/L 6-BA, 3% sucrose and 0.65% agarose; culture at 25℃; 12 h/d（ daylight lamp, 2 000 lx） illumination.

Research on Tissue Culture and Rapid Propagation Technology of Superior Individuals of Lonicera edulis Turcz

[Objective] This paper aimed to study the tissue culture and rapid propagation technology of superior individuals of Lonicera edulis Turcz. [Method] Several superior individuals of Lonicera edulis Turcz were used as materials for selecting the primary medium, subculture medium, rooting medium and acclimatization substrate during the tissue culture and rapid propagation. [Result] 6-BA was the optimal cytokinin for tissue culture of Lonicera edulis Turcz, compared with ZT; modified MS＋1.0 mg/L of 6-BA ＋ 0.2 mg/L of IBA was the optimal medium as primary and subculture medium, modified MS＋ 1.5 mg/L of IBA was the optimal medium for rooting of Lonicera edulis Turcz, the rooting rate had achieved 100% after cultured for 30 d. The optimal substrate for transplanting plantlets of Lonicera edulis Turcz was composed of humus and perlite （1∶ 1, V/V）, survival rate was as high as 95% after 30 d. [Conclusion] This study provided basis for the rapid propagation of superior seedlings of Lonicera edulis Turcz, as well as the establishment of industrialized breeding technical system and the implementation of scale production.

Shoot tissue culture of Robinia pseudoacacia f. decaisneana

Robinia pseudoacacia f. decaisneana is a transfiguration of Robinia pseudoacacia. For enhancing propagation coefficient of the species, the experiment of shoot tissue culture of Robinia pseudoacacia f. decaisneana was conducted in Forestry College of Shenyang Agricultural University from July 1999 to July 2001. The experiment included medium selection of explant induction survival, initial culture, subculture as well as rooting culture, and forming seedling with callus. The results showed that shoot segment in vitro survive rate is larger in spring than in autumn, and green dense callus could form plantlet. The best medium for initial culture was SH+0.5mg/L BA+0.05 mg/L NAA, with a propagation coefficient of 4.1 (per micro-cutting in a month), and for subculture it was B5+0.5 mg/L BA+0.05 mg/L NAA+ 10 mg/L Glu., with a propagation coefficient of 4.7. The best rooting medium was 1/2MS+0.5 mg/L NAA+10 mg/L Glu., with a rooting rate of 84.4%. These results provide reference data for reproduction of superior individuals of Robinia pseudoacacia f. decaisneana.

Reproduction of the Three-line Genic Male Sterile Line Parent Mian 7MB-1 （Brasscia Napus L.） and Seed Production of F1 Based on Somatic Tissue Culture

[Objective] The aim was to study the reproduction of the three-line genic male sterile （GMS） lineparent Mian7MB-1 （B. NapusL.） and the seed production of F1 through somatic tissue culture. [Methed] Through hybridization, a new breeding material Mian 7MB-1 in three-line genic temporary maintainer line propagated by tissue culture was used to improve the sterile plant rate of rapeseed in dual-purpose recessive GMS line, such as Mian 7AB type, S45AB type, and etc. And then the variety comparative test was performed. [Result] In order to avoid some fertility restoration phenomena occurring during the process of self-reproduction, Mian 7AB was propagated in bulk with somatic tissue culture of temporary maintainer line plant stem. The propagated temporary maintainer line seedlings were applied to the breeding and seed production of net room male sterile line parent, promoting the sterile plant rate of the male sterile line parent to 91.7% -93.5%. The male sterile line parents per hectare were enough for the seed production of hybrid F1 in 7 500 -15 000 hm^2. [ Conclusion ] Compared with the original dual-purpose GMS line, the seed production ultilizing male sterile line with high sterile plant rate greatly reduced the labor, significantly improved the seed yield, ensuring the seed quality and forming a perfect breeding and seed production system.

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.

Tissue Culture of Lespedeza cyrtobotrya

[Objective] The aim was to carry out study on tissue culture of Lespedeza cyrtobotrya. [Method] The seeds of L. cyrtobotrya were used as materials to study on its tissue culture. [Result] The best sterilization time to L. cyrtobotrya seeds was 8 min with 2.1% NaClO,in which shooting percent reached 37.8% and no polluted situations occurred. In the primary culture with the MS as basal medium,the concentration of 6-BA showed a significant effect on the index of buds differentiation,the optimum differentiation culture medium was MS＋BA 1.0 mg/L＋NAA 0.1 mg/L＋2,4-D 0.01 mg/L,on which the index of generation could reach 6.69. The optimum subculture medium was MS＋6-BA 1.0 mg/L＋2,4-D 0.05 mg/L. The plants can generate the highest roots and rooting percent with IBA 0.50 mg/L. [Conclusion] This study had provided theoretical basis for genetic improvement of L. cyrtobotrya.

A Preliminary Study on the Tissue Culture of Sagittaria trifolia L.

[Objective] This study was to optimize the experimental conditions for large scale propagation of Sagittaria trifolia L via tissue culture.[Method] The dominant S.trifolia cultivar Baoyingziyuan introduced from Jiangsu Province was used as experimental material to study the impacts of various culture conditions on tissue culture of its stem tips and induction of stolons.[Result] Hormone combination 0.10 mg/L 6-BA ＋0.05 mg/L NAA performed best in plantlet regeneration and 2.0 mg/L 6-BA ＋0.5 mg/L NAA best in induction of stolon.Various sucrose concentrations did not show significant difference in the impact on sprouted stolons.[Conclusion] Various culture conditions could to some extent impact plantlet regeneration and stolon induction,and our results reveal the optimal hormone combinations for regeneration and stolon induction of S.trifolia.

Tissue Culture of Toona ciliata

In this study, a screening experiment was carried out among MS, WPM, DCR, MT, B5 and N6. The results showed that for proliferation culture of Toona ciliate, the optimum basic medium was WPM, and the optimum medium was WPM ＋ 6-BA 0.5 mg/L ＋ IBA 0.05 mg/L. The proliferation coefficient reached 4.29. The optimum rooting culture medium was 1/2 MS ＋ IBA 0.5 mg/L ＋ NAA 0.5 mg/L with rooting rate of 100%. In the optimum rooting ＇culture medium, the roots of T. ciliate developed neatly, and the seedlings grew well.

Effect of Bacteriostat(Qianxing No.1) on Open Tissue Culture of Sugarcane

[Objective] The effect of bacteriostat （Qianxing No.l） on open tissue cul- ture of sugarca6e was discussed in this paper. [Method] The axillary bud explants of Qiantang No.5 were used as experimental material. Different concentrations of Qianxing No.1 were added in the media to determine the optimal bacteriostatic con- centrations of Qianxing No.1 for induction and proliferation culture of sugarcane in the open tissue culture. [Result] The results showed Qianxing No.1 with the con- centration of 0.5% could effectively inhibit the medium contamination. Under that bacteriostatic concentration, the surviving rate of explants was up to 80%, and the propagation coefficient was 3.1. [Conclusion] This research would provide a theoretic basis for the simplification of sugarcane tissue culture procedure and the reduction of production costs.

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.

Advances in Tissue Culture Technology of Bletilla striata

The reports about tissue culture technology of Bletilla striata were summa- rized in this paper, and explants for tissue culture, induction differentiation of callus, rooting culture and transplanting, effect of growth regulator and influence of external environment were expounded. This study provides support for further research.

Study on Tissue Culture and Radiation Mutation of Astragalus membranaceus Bge. var. mongholicus(Bge.) Hsiao

[Objective] The research aimed to study the tissue culture technology and callus induction by radiation mutation of A. membranaceus Bge. [ Method ] With the different parts of Astragalus membranaceus Bge. var. mongholicus （ Bge. ） Hsiao aseptic seedling as explants （ leaves, cotyledons, hypocotyls） induced callus, and cotyledon and hypocotyls taken by the method of radiation mutation were studied. [ Result]The results showed that the three explants had relatively high callus induced rate in the medium which respectively made up of MS ＋6-BA 2.0 mg/L ＋ NAA2.0 mg/L, LS ＋6-BA2.0 mg/L ＋NAA0.1 mg/L, MS ＋ 6-BA2.0 rng/L ＋ NAA2.0 rag/L; the optimum mutation time of hypocotyls and cotyledons was 15 minutes; the growth of the callus induced from hypocotyls would be better as the mutation time increased, but when it reached a certain time the growth would be weaken, the induction rate also would be reduced. [ Conclusion] This study will provide the scientific reference in tissue culture and mutation breeding of A. membranaceus Bge.

Optimization of the Method to Obtain Effective Sterile Explants in Tissue Culture of Ardisia mamillata Hance

In the present study, we optimized the procedures to pretreat the parental material, to collect, sterilize and inoculate the explants in the tissue culture of Ar- disia mamillata Hance, so that more effective sterile explants can be obtained. The results revealed the optimal procedures. In detail, the parental materials were trans- ferred and pretreated in laboratory in February. The stem tips of new branches were collected in middle March, cleaned with eradicator and then rinsed with tap water for 1-2 h. After that, the explants were surface sterilized with 70% alcohol for 30 s, rinsed with sterile water 2-3 times, sterilized with 0.1% mercuric chloride for 4 min, rinsed with sterile water three times and sterilized with 0.1% mercuric chlo- ride for 4 min again. Finally, they were inoculated into medium after being rinsed with sterile water 5-8 times. By this method, the pollution rate of explants can be greatly reduced while their survival rate can be significantly improved.