Establishment of Tissue Culture Regeneration Systems of Stem Segments of Euphorbia tirucalli

[Objective] The aim was to study the conditions of tissue culture regeneration seedling by using the stem segments of Euphorbia tirucalli and determine the optimum culture condition of each culture stage,so as to provide references for the factory production and relative study of tissue culture seedling of E. tirucalli. [Method] Taking the stem segments of E. tirucalli as explants,the effects of various mediums on germination rate,multiplication coefficient and rooting rate were studied. [Result] The optimum induction medium of germination culture was 1/2MS＋NAA 0.02 mg/L＋6-BA 1.0 mg/L,with differentiation rate of 89.7%; the best subculture medium was 1/2MS＋NAA 0.02 mg/L＋6-BA 0.60 mg/L＋AD 3.0 mg/L,with multiplication coefficient of 5.70; the optimum rooting culture medium was 1/2MS＋NAA 0.40 mg/L＋IBA 0.4 mg/L,with rooting rate of 100% and transplanting survival rate of 80%. [Conclusion] The tissue culture conditions of stem segments of E. tirucalli were determined primarily.

Primary Establishment of the Tissue Culture Technique and Regeneration System for Ornamental Lupinns polyphyllus

The purpose of this paper is to develop a system for tissue culture and rapid propagation of two ornamental lupins, Minaretie and Russell Prize. In view of screening out the better explant regeneration and suitable culture medium, through adding hormone 6-BA, NAA and 2, 4-D into MS and B5 basic culture medium, a series of experiments were carried out with the shoot tips, leaves, leaf petioles and stems from the asepsis seedling. The results showed that the shoot tips had favorableness on the rapidly propagation; MS＋6-BA 0.5 mg. L-1 for first generation, the induction rate of Minaretie and Russell Prize was 90.5% and 95.86% respectivdy; Minaretie had the highest propagation index （6.35） on MS＋6-BA 0.5 mg.L^-1＋NAA 0 mg-L^-1＋GA 30.8 mg. L^1＋AC 2 g. L^-1, but Russell Prize had the highest propagation index （7.24） on MS＋6-BA 0.5 mg.L^-1＋NAA 0.15 mg.L^-1＋GA3 1.0 mg.L^-1＋AC 0.5 g.L^-1; 1/2 MS＋NAA 0.25 mg.L^-1 was the best rooting medium. The ratios of getting roots of Minaretie and Russell Prize were 94.78% and 96.32%, respectively.

Establishment of Tissue Culture Regeneration System in Bama Hemp (Cannabis sativa L.)

[Objectives]This study was conducted to establish a tissue culture regeneration system in Bama hemp(Cannabis sativa L.).[Methods]Using hemp seeds as explants,a regeneration system was established through explant sterilization,callus induction,callus differentiation,and rooting culture.[Results]The results showed that the best sterilization effect was achieved when sterilizing with 75%ethanol for 30 s,followed by 0.1%HgCl 2 solution for 9 min,with a contamination rate as low as 11.4%.In presence of 3 mg/L 2,4-D and 0.1 mg/L6-BA,the callus induction effect from hemp seeds was better.The formula for better differentiation of callus was MS+2.0 mg/L 6-BA+0.2 mg/L NAA.IBA had a promoting effect on the rooting of hemp aseptic plantlets.The highest rooting rate reached 80%when MS+0.3 mg/L IBA were used.[Conclusions]This study established a hemp seed regeneration system to provide technical support for the conservation and breeding of hemp germplasm resources.

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.

Optimization of plant regeneration system in vitro culture in wheat

Studies were carried out to establish an efficient regeneration system of three bread wheat cultivars. Results showed induction medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) had a higher plantlet regeneration frequency than Piclorm, with an average frequency of 54% in all treatments. Optimal condition for different genotypic rice was as following: induction medium (MSS 3AA/2) with 0.5 mg L-1 2,4-D, regeneration medium (R) with 0.01 mg L-1 2,4-D and 3 mg L-1 KT. The average regeneration frequency reached 83.3% under the condition. Correlation analysis showed that root differentiation, in different level, correlated with green spot regeneration, and with the number of regenerated plants per callus. No correlation was found between green spots regenerated and the numbers of plants regenerated per callus.

Comparative Study on Antioxidative System in Normal and Vitrified Shoots of Populus suaveolens in Tissue Culture

To explore the physiological and biochemical mechanism of the occurrence of vitrified shoots of Populus suaveolens in tissue culture, the changes in water, chlorphyll, lignin, H2O2, phenylalanine ammonialyase (PAL), malonaldehyde (MDA), protective enzymatic systems, and some key enzymes involved in the ascorbate- glutathione cycle were comparatively studied in both normal and vitrified shoots of P. suaveolens. The results show that the lower activities of peroxidase (POD), catalase (CAT), ascorbate peroxidase (APX), dehydroascorbate reductase (DHAR), glutathione reductase (GR) and PAL, and the less contents of chlorphyll, lignin, ascorbate (ASA) and reduced glutathione (GSH) as well as the lower ratios of ASA / DHA and GSH / GSSG are observed in vitrified shoots than in normal ones during the whole culture period. While in comparison with normal shoots, the higher activity of superoxide dismutase (SOD) and the more concentrations of water, H2O2, MDA, dehydroascorbate (DHA) and oxidized glutathione (GSSG) are found in vitrified shoots. Statistical analysis indicates that the enhanced activity of SOD and the decreased activities of CAT and POD as well as some enzymes involved in the ascorbate-glutathione cycle might be closely correlated to the accumulation of H2O2. The less regeneration of ASA and GSH and the lower capacity of the ascorbate-glutathione cycle observed in vitrified shoots might be due to a significant decrease in APX, MDAR, DHAR and GR activities and a decline in redox status of ASA and GSH. The decreases in chlorphyll content might result in a decline in photosynthesis. The lower activities of POD and PAL could result in the decrease of lignin synthesis and cell wall ligination, which might be the key factor leading to the increase in water content. It is concluded that the deficiency of detoxification capacity caused by the lower capacity of the ascorbate-glutathione pathway and the decreased activity of protective enzymatic system might lead to the large accumulation of H2O2 and the enhancement of membrane lipid peroxidation, which might be the main cause leading to the occurrence of vitrifying shoots of P. suaveolens in tissue culture.

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.

Construction of Rooting System during Camellia oleifera Tissue Culture

An rooting experiment of tissue culture plantlets was carried out with sterile plantlets obtained from the stem segments of a good clone of Camellia oleifera as materials. The results showed that basic medium and illumination condition are factors crucial to rooting of C. oleifera. With 1/4MS as basic medium, the treat- ment with the addition of 0.5 mg/L NAA and soaking in 2 000 mg/L KIBA, containing 30 mg/L sucrose and subjected to dark culture of 20 d was the optimal treat- ment, achieving a rooting rate of 86. 7%.

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).

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 and Rapid Propagation System of Roselle(Hibiscus sabdariffa L.)

[Objectives]This study was conducted to establish a tissue culture and rapid propagation system of roselle(Hibiscus sabdariffa L.)[Methods]A tissue culture and rapid propagation experiment was carried out with roselle plants as materials to study the suitable explants,the best disinfection method,the best growth medium,the best rooting medium and the transplanting and domestication method of roselle.[Results]The lateral buds of roselle were the best explants.Sterilizing with 0.1%mercuric chloride for 7 min showed a contamination rate of 5%and achieved a survival rate of 90%.With MS as the basic medium,adding 1.0 mg/L 6-BA and 0.5 mg/L IBA could obtain the best effect of bud induction.The medium with the highest proliferation rate was MS+6-BA 0.5 mg/L+BA 0.1 mg/L.On the basis of 1/2MS,adding 0.5 mg/L NAA+0.5 mg/L IBA could make adventitious buds root fastest and most,and greatly improve the propagation coefficient.And 1∶1 perlite rock:peat soil was the best transplanting substrate,with which the transplanting survival rate reached 95%.[Conclusions]This study provides technical reference for rapid cultivation and large-scale planting of roselle.

Establishment of a Highly Efficient Regeneration System for the Mature Embryo Culture of Wheat

Establishment of a highly efficient regeneration system for the mature embryo of wheat will provide a convenient tool for wheat tissue culture and transformation, thereby facilitating the transformation of foreign genes into wheat. By using the mature embryos derived from 20 different wheat lines including Shi 4185, Yumai 66, Lunxuan 987, CB037, Yangmai 6, Xinchun 9, Bobwhite, Han 6172, Zheng 9023, Jimai 20, Ningchun 4, and Jing 411, the effects of some factors including inoculation methods, initiating culture media, organic additives, antioxidants, and auxins on the regeneration from the explants were evaluated. The results indicated that the scraping embryo culture was better than the whole embryo culture, the Aa medium was better than the SD2 medium and dicamba was better than 2,4-D in increasing the regeneration frequency. An Adi medium was established in this study by adding silver nitrate, cysteine, ascorbic acid, dicamba, glutamine into the Aa medium at the concentration of 4,40, 100, 2, and 5 mg L^-1, respectively. By using the Adi medium and the scraping technique, the regeneration frequencies of the mature embryos of CB037, Lunxuan 987, Hart 6172, Yangmai 6, Bobwhite, Zheng 9023, Shi 4 185, and Jimai 20 became 85.6, 60,1, 46.0, 42.1,42.0, 34.0, 33.0, and 32.0%, respectively, which were about 5-8 times higher than that obtained from the conventional culture mediums and techniques. This novel regeneration system could be helpful in wheat transformation.

Progress of safflower (Carthamus tinctorius L.) regeneration through tissue culture

Background: Safflower regeneration through tissue culture has long been limited to low frequency and lack of an efficient protocol that suitable for most safflower cultivars. In past decades, researches had been carried out to investigate safflower regeneration through tissue culture and great progress had been made. Objective: To investigate factors that affect safflower regeneration through tissue culture principally. Methods: This article summarized available literatures about advancements in safflower regeneration, especially discussed factors affecting safflower tissue culture in detail. Results: Safflower regeneration was fairly hard than other congeneric plants, such as chrysanthemum. The genotype, seedling age, type of explants, medium components, plant growth regulators and other additives all had specific influences on safflower tissue culture. More deepgoing researches need to be undertaken to establish an effective safflower regeneration system.

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.

Establishment of in Vitro Regeneration System of Triploid Chinese White Poplar

初步建立了三倍体毛白杨 (PopulustomentosaCarr.)组织培养再生系统。组织培养时 ,外植体最好选用在弱光下从休眠芽上萌发的幼芽或嫩茎 ,经HgCl2 溶液表面消毒后 ,接种到大量元素减半、含 0 .2 5mg/L吲哚乙酸和 1.0 0mg/L玉米素的MS培养基上。试管苗的生根可采用含 0 .2 5mg/L吲哚丁酸和 10mg/L维生素B1的MS培养基。试管苗茎段的分化依无性系和外源激素条件的不同而异。无性系B19的茎段较易分化 ,而无性系B30 4和B331分化较难。最适的分化培养基为大量元素减半、含 0 .0 5～ 0 .10mg/L吲哚乙酸和 1.0

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.

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.

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.

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.

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.