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.

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.

Efficient Disinfection of Explants in the Tissue Culture of Chinese Orchids

Explants,namely the seeds (Cymbidium faberi) and buds (Cymbidium goeringii) of local Hunan orchids were disinfected with 75% ethanol, 0.1% mercuric chloride,and 10% sodium hypochlorite in six different disinfection treatments. Results showed that The treatment with 75% ethanol (contact time: 45 s) and 0.1% mercuric chloride (contact time: 8 min) proved to be the most effective one in disinfecting the explants of Cymbidium faberi seeds.The combination of 75% ethanol (contact time: 45 s) and 0.1% mercuric chloride (contact time: 5 min) provided the optimal disinfection effects when disinfecting Cymbidium goeringii buds.In general, the disinfection effect of mercuric chloride was found to be superior to that of sodium hypochlorite,but care should be taken with the disinfection time to avoid damage to the explants.

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.

Direct somatic embryogenesis and related gene expression networks in leaf explants of Hippeastrum ‘Bangkok Rose’

Hippeastrum, a highly diverse genus in the Amaryllidaceae family, is a valuable ornamental bulbous flowering plant. Somatic embryogenesis(SE) is an efficient method for mass production of Hippeastrum plantlets. Previous studies have been devoted to the in vitro propagation of Hippeastrum, but the SE and its regulatory networks are rarely reported. In this study, we established a direct SE method of Hippeastrum Bangkok Rose' using leaf bases as explants. MS supplemented with 1.00 mg·L^(-1)NAA +1.00 mg·L^(-1)KT + 0.25 mg·L^(-1)TDZ was the optimal medium for SE. Histological observations showed that the bipolar somatic embryo originated from the epidermal cell layer and underwent initiation,globular, scutellar and coleoptile stages. During SE, endogenous hormones of IAA, CTK, ABA, and SA were highly accumulated. Transcriptomic analysis revealed the genes encoding auxin biosynthesis/metabolic enzymes and efflux carriers were induced, while the auxin receptor of TIR1 and ARF transcriptional repressor of Aux/IAA were down-regulated and up-regulated, respectively, leading to suppression of auxin signaling. In contrast, cytokine signaling was promoted at the early stage of SE, as biosynthesis, transport, and signaling components were up-regulated.Various stress-related genes were up-regulated at the early or late stages of SE. Chromatin remodeling could also be dynamically regulated via distinct expression enzymes that control histone methylation and acetylation during SE. Moreover, key SE regulators, including WOXs and SERKs were highly expressed along with SE. Overall, the present study provides insights into the SE regulatory mechanisms of the Hippeastrum.

Postnatal development of rat retina:a continuous observation and comparison between the organotypic retinal explant model and in vivo development

The organotypic retinal explant culture has been established for more than a decade and offers a range of unique advantages compared with in vivo experiments and cell cultures.However,the lack of systematic and continuous comparison between in vivo retinal development and the organotypic retinal explant culture makes this model controversial in postnatal retinal development studies.Thus,we aimed to verify the feasibility of using this model for postnatal retinal development studies by comparing it with the in vivo retina.In this study,we showed that postnatal retinal explants undergo normal development,and exhibit a consistent structure and timeline with retinas in vivo.Initially,we used SOX2 and PAX6 immunostaining to identify retinal progenitor cells.We then examined cell proliferation and migration by immunostaining with Ki-67 and doublecortin,respectively.Ki-67-and doublecortin-positive cells decreased in both in vivo and explants during postnatal retinogenesis,and exhibited a high degree of similarity in abundance and distribution between groups.Additionally,we used Ceh-10 homeodomain-containing homolog,glutamate-ammonia ligase(glutamine synthetase),neuronal nuclei,and ionized calcium-binding adapter molecule 1 immunostaining to examine the emergence of bipolar cells,Müller glia,mature neurons,and microglia,respectively.The timing and spatial patterns of the emergence of these cell types were remarkably consistent between in vivo and explant retinas.Our study showed that the organotypic retinal explant culture model had a high degree of consistency with the progression of in vivo early postnatal retina development.The findings confirm the accuracy and credibility of this model and support its use for long-term,systematic,and continuous observation.

Differentiation potential of human adipose tissue derived stem cells into photoreceptors through explants culture and enzyme methods

AIM： To investigate the retinal photoreceptor differentiation potential of human orbital adipose tissue-derived stem cells （ADSCs） generated by enzyme （EN） and explant （EX） culture methods.METHODS： We investigated potentials of human orbital ADSCs to differentiate into photoreceptors through EN and EX culture methods. EN and EX orbital ADSCs were obtained from the same donor during rehabilitative orbital decompression, and then were subject to a 3-step induction using Noggin, DKK-1, IGF-1 and b-FGF at different time points for 38d. Stem cell, eye-field and photoreceptor-related gene and protein markers were measured by reverse transcription-polymerase chain reaction （RT-PCR） and immunofluorescent （IMF） staining.RESULTS： Both EX and EN orbital ADSCs expressed CD133, a marker of cell differentiation. Moreover, PAX6 and rhodopsin, markers of the retinal progenitor cells, were detected from EX and EN orbital ADSCs. In EX orbital ADSCs, PAX6 mRNA was detected on the 17th day and then the rhodopsin mRNA was detected on the 24th day. In contrast, the EN orbital ADSCs expressed PAX6 and rhodopsin mRNA on the 31st day. EX orbital ADSCs expressed rhodopsin protein on the 24th day, while EN orbital ADSCs expressed rhodopsin protein on the 31st day. CONCLUSION： Orbital ADSCs isolated by direct explants culture show earlier and stronger expressions of markers towards eye field and retinal photoreceptor differentiation than those generated by conventional EN method.

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.

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.

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.

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.

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.

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.

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.

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.

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.