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.

Optimization of Rooting Medium for Tissue Culture Seedlings of Dendrobium officinale by Response Surface Methodology

[Objectives] To increase the survival rate of tissue culture seedlings of Dendrobium officinale,and optimize the conditions of rooting medium by the response surface methodology( RSM). [Methods]The effects of 6-BA concentration,NAA concentration,potato amount and the amount of mashed banana on the growth of seedlings were determined by single factor experiment and were analyzed by Box-Behnken design and response surface methodology. [Results]The optimal culture conditions: rooting medium 1/2 MS + 6-BA 0. 24 mg/L + mashed banana 87. 63 g/L + potato 89. 30 g/L + NAA 0. 52 mg/L + sucrose 20. 0 g/L + activated carbon 4. 0 g/L + agar 7. 0 g/L,p H 5. 8,and light intensity 2 000 Lx. [Conclusions]The model established by response surface methodology has a good predictability and could be used to optimize the conditions of tissue culture and rooting medium of D. officinale.

STUDY ON TISSUE CULTURE FOR GELIDIUM SEEDLING

As seedling culture is a crcial factor for successful cultivation of Gelidium, the authors researched tis-sue culture technology for producing seedlings. The merphogeny and experirmental ecology were observedand studied fully in 2-5 mm isoIated tissue fragments, appearaare of branching creepersand attaching structure and new erect seedlings production and development were studied. Fragments weresown on bamboo slice and vinyon rope. The seedlings were cultured 20-30 days indoor, then cultured inthe sea, where the density of erect seedlings was 3- 19 seedlings/cm^2, growth rate was 3.84%/day. The frondarising from seedlings directly was up to 10 cm per year. The ecological conditions for regenerated seedlingsare similar to the natural ones. The regenerated seedlings are sintable for raft culture in various sea areas.

Outdoor Domestication Cultivation and Survival Mechanism for Tissue Culture Seedlings of Paeonia suffruticosa

In order to reveal the differences between different peony varieties and facilities,thousands of tissue culture seedlings of 21 Itoh peony varieties imported from abroad were used as test materials,and the peony domestication cultivation method used in the past was used as a control. The technical measures and mechanisms for the outdoor domestication of seedlings and the survival of field transplantation were studied. The results indicate that the main factors and measures for effectively protecting the outdoor transplanting of tissue culture seedlings include cultivating sound and strong seedlings,transplanting in proper period of spring,and providing good ventilating places and facilities,removing leaves,keeping buds and releasing dormancy before transplantation,planting depth should meet the requirement of exposing to the root neck,changing the pots according to the size of seedlings,the substrate should be loose and permeable,seriously disinfected and sterilized,scientific management,and prevention and control of diseases and insect pests. In addition,the survival mechanism was also analyzed,and the reasons and countermeasures for the successful application of peony tissue culture in China were found. Finally,the differences in survival rates between different varieties and facilities were summarized.

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.

Inhibitory Effect of Tissue Transglutaminase (tTG) Antisense Oligodeoxynucleotides on tTG Expression in Cultured Bovine Trabecular Meshwork Cells

To study the effect of tTG fully phosphorothioated antisense oligodeoxynucleotides （tTG-ASDON） on tTG expression in cultured bovine trabecular meshwork cells （BTMCs） in vitro and explore a new treatment alternative for primary open angle glaucoma （POAG）, the ASDON1 and ASDON2 complementary to the protein codogram region of tTG were designed, synthesized and phosphorothioated according to the secondary structure of tTG. The ASDON1 and ASDON2 were embedded in Lipofectamine and transfected into BTMCs. The untreated group served as negative controls. The expression of tTG in the mRNA and protein level were measured by semi-quantitative RT-PCR and immunohistochemical technique-Supervision method respectively. Our results showed that both the mRNA and the protein of tTG with tTG-ASDON1 and tTCr-ASDON2 were significantly decreased as compared with that of the controls （P〈0.05）. On the other hand, no significant difference was found between the ASDON1 group and the ASDON2 group. It is concluded that the expression of tTG mRNA and protein in cultured BTMC are down-regulated by tTG- ASDON. As a result, tTG-ASDON may be used for the treatment of POAG through the inhibitory effect on the expression of tTG.

Effects of tissue-cultured mountain ginseng （Panax ginseng CA Meyer） extract on male patients with erectile dysfunction

Korean ginseng and mountain ginseng （Panax ginseng CA Meyer） are important traditional herbal plants whose ginsenosides are generally accepted as serving to improve sexual functions, such as penile erection. We investigated the effects of tissue-cultured mountain ginseng extract （TMGE） on male patients with erectile dysfunction （ED）. A double-blind, placebo-controlled study was conducted with 143 patients experiencing ED. Over the course of 8 weeks, one group took 1 000 mg of TMGE twice a day, and the other group took 1 000 mg of placebo twice a day. The effects of the TMGE and the placebo were analyzed using the Korean version of the International Index of Erectile Function （IIEF） questionnaire. A total of 86 patients completed 8 weeks of treatment. The scores on the five domains of the IIEF after medication were significantly higher than the baseline scores in the group treated with TMGE （P 〈 0.05）, whereas no significant improvement was observed in the placebo group （P 〉 0.05）. Erectile function and overall satisfaction scores after medication were significantly higher in the TMGE group than in the placebo group （P 〈 0.05）. Erectile function of patients in the TMGE-treated group significantly improved, suggesting that TMGE could be utilized for improving erectile function in male patients.

Expression of Tissue Transglutaminase in Cultured Bovine Trabecluar Meshwork Cells

Summary: To study whether cultured bovine trabecluar meshwork cells (BTMC) are capable of expressing tTG in protein and at mRNA level, BTMC were cultured in vitro and passaged three times, then the cells were transferred onto or cultured on sterile cover or submitted to isolation of RNA with Trizol, and the expression of tTG was detected by immunohistochemical technique and reverse transcription polymerase chain reaction (RT-PCR) respectively. Our results showed that tTG immunostaining was positive in the cytoplasm and rarely in the nucleus of cultured BTMC. No immunostaining was seen in the negative control. Moreover, a single RT-PCR amplified product whose sequence and size were in accordance with our known parameters was obtained. The expression of tTG in cultured BTMC was confirmed in protein and at mRNA level. BMTC is available more readily for the investigation of the relationship between tTG and primary open-angle glaucoma.

Mechanical Stimulus Inhibits the Growth of a Bone Tissue Model Cultured In Vitro

Objectives To construct the cancellous bone explant model and a method of culturing these bone tissues in vitro,and to investigate the effect of mechanical load on growth of cancellous bone tissue in vitro.Methods Cancellous bone were extracted from rabbit femoral head and cut into 1-mm-thick and8-mm-diameter slices under sterile conditions.HE staining and scanning electron microscopy were employed to identify the histomorphology of the model after being cultured with a new dynamic load and circulating perfusion bioreactor system for 0,3,5,and 7 days,respectively.We built a three-dimensional model using microCT and analyzed the loading effects using finite element analysis.The model was subjected to mechanical load of 1000,2000,3000,and 4000μεrespectively for 30 minutes per day.After 5 days of continuous stimuli,the activities of alkaline phosphatase(AKP)and tartrate-resistant acid phosphatase(TRAP)were detected.Apoptosis was analyzed by DNA ladder detection and caspase-3/8/9 activity detection.Results After being cultured for 3,5,and 7 days,the bone explant model grew well.HE staining showed the apparent nucleus in cells at the each indicated time,and electron microscope revealed the living cells in the bone tissue.The activities of AKP and TRAP in the bone explant model under mechanical load of 3000 and 4000μεwere significantly lower than those in the unstressed bone tissues(all P<0.05).DNA ladders were seen in the bone tissue under 3000 and 4000μεmechanical load.Moreover,there was significant enhancement in the activities of caspase-3/8/9 in the mechanical stress group of 3000 and 4000με(all P<0.05).Conclusions The cancellous bone explant model extracted from the rabbit femoral head could be alive at least for 7 days in the dynamic load and circulating perfusion bioreactor system,however,pathological mechanical load could affect the bone tissue growth by apoptosis in vitro.The differentiation of osteoblasts and osteoclasts might be inhibited after the model is stimulated by mechanical load of 3000 and 4000με.

The Comparison in Tissue Culture Ability of Mature Embryo in Different Cultivars of Rice

In order to study the regeneration technology of mature embryos in different rice varieties,nine japonica,nine indica and eleven hybrid rice varieties of two line or three line or superiority combinations were selected as explants to study the callus induction,differentiation and regeneration rates on different media.The higher callus induction （61.7-89.2%） was observed in japonica rice,when cytokinin was added at lower concentration （0.3 mg L-1 6-BA） in M8 basal medium,supplemented with 30 g L-1 sucrose,8 g L-1 agar and 2 mg L-1 2,4-D.Further,the addition of two cytokinins （2 mg L-1 6-BA,0.5 mg L-1 KT） and 1 mg L-1 NAA in the M8 basal supplemented medium resulted in 9.1-100% of the callus induction in indica rice.The percent callus induction in hybrid rice varieties was 40-86.3% when addition of 1 mg L-1 6-BA and 1 mg L-1 KT was added,and the cytokinins was required by the japonica and indica rice varieties in the M8 basal supplemented medium.It was observed that when the 0.5 mg L-1 2,4-D and 1 mg L-1 6-BA were added in japonica rice,and 0.2 mg L-1 2,4-D and 0.5 mg L-1 6-BA were added in indica and hybrid rice in the MS different media,the regeneration rates were 9.2-59.5%,3.6-87.5% and 17.2-43.2% for japonica,indica and hybrid rice,respectively.Thus,the regeneration technology with higher output is established in the mature embryos of similar rice varieties.

Effects of different culture media on the growth of Indian sandalwood(Santalum album L.) seedlings in Zhanjiang,Guangdong,southern China

We studied the effects of different culture media on the growth of India sandalwood （Santalum album L.） seedlings in Zhanjiang, Guangdong Province in southern China. Five different growth substrates, lateritic subsoil, burnt soil, agricultural soil, peaty soil and coconut dust, were used as the basic culture materials and seven different treatments of composition were used as potting media. Kuhnia rosmarinifolia Vent. was used as a primary host plant for all treatments. Statistically significant differences were found between treatments in respects of survival rate （p 〈 0.001）, height （p 〈 0.001）, ground diameter （p 〈 0.001） and biomass （p = 0.002）, as well as for quality index （p 〈 0.001） of S. album seedlings after 6-month growth in containers with different culture media. Among all treatments, the treatment combining burnt soil, peat and coconut dust in a weight ratio of 1：1：1 plus 2% calcium super-phosphate as basal manure achieved the best performance for most of the seedling growth parameters, including survival rate （98%）, height （35.81 cm）, ground diameter （0.56 cm）, biomass （4.46 g） and quality index （0.65）, followed by the treatment using only burnt soil plus 2% calcium super-phosphate as the culture medium （survival rate 86%, height 29.23 cm, ground diameter 0.48 cm, biomass 3.36 g and quality index 0.52）, while the treatment using only lateritic subsoil plus the basal manure as the medium obtained the poorest results in survival rate （38%）, height （12.04 cm）, ground diameter （0.19 cm）, biomass （0.26 g） and quality index （0.043）. Increasing the proportion of lateritic subsoil in the medium when mixed with peat and coconut dust did not show statistically significant differences in survival, height, ground diameter, biomass nor in quality index. In consideration of cost, using burnt soil plus 2% calcium super-phosphate as basal manure may be the optimum culture medium for large-scale production of Indian sandal- wood seedlings in Guangdong, southern China.

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.

Effects of Toxic Levels of Aluminium on Seedling Parameters of Rice under Hydroponic Culture

The presence of AI in the rhizosphere of rice in acid soil restricts root growth and significantly reduces crop productivity. In this study, the effects of AI （30, 60 and 90 μg/mL） on seedling root growth, number of primary roots per seedling, seedling shoot length, number of leaves per seedling, seedling fresh weight, and seedling dry weight were studied. Rice genotypes were classified into three different classes, namely, tolerant, moderately tolerant, and susceptible, based on root tolerance index. The method of hydroponic culture was modified, and elaborated in the text. Toxic levels of AI in nutrient solution significantly decreased seedling root growth, number of primary roots, seedling shoot length, number of leaves per seedling, seedling fresh weight, and seedling dry weight. Few genotypes showed longer root length at 30 pg/mL AI in nutrient solutions compared with the control. High levels of AI in nutrient solutions were highly toxic for rice seedlings. Based on root tolerance index, Radhunipagal, Gobindobhog, Badshabhog, Kalobhog, UBKVR-11, UBKVR-16, UBKVR-18, Khasha and IVT4007-B were classified as tolerant genotypes, and these genotypes may be used as donors for breeding of Altoxicity tolerance.

Contamination and browning in tissue culture of Platanus occidentalis L.

Twigs of 2-3-year-old Platanus occidentalis L. were used as experimental material to find the causes for the contamination and browning in the initial stages of tissue cultures. To compare the degree of browning of explants picked off from different growing seasons, the experimental material was excised from trees on each of the first ten days in January, March, May and July, 2006. The results indicated that the contamination and browning rates of the material cut off in January （14.2% and 30.6%, respectively） and March were somewhat lower than those in July. The pretreatment of soaking the explants in different anti-oxidants and absorbents at the same time could diminish some side effects. The pretreatment of using 10 g·L^-1 vitamin C reduced the contamination and brown- ing rate effectively. An orthogonal experiment showed that the optimal factor and level arrangement is 0.5 mg·L^-1BA, 2.0 g·L^-1 active carbon and 1.5 g·L^-1 PVP which resulted in a browning rate of only 16.5%. In general, sampling period, physical properties and pretreatment of explants are the main factors responsible for the contamination and browning of material in the initial stages of P. occidentalis tissue cultures.

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 and Rapid Propagation of Spathiphyllum kochii Engl. et Krause

[Objectives] This study was conducted on tissue-culture rapid propagation techniques of Spathiphyllum kochii Engl. et Krause. [Methods] With lateral buds of S. kochii as explants, the effects of such four basic media as MS, B5, Nitsch and Wpm and the ratio of two hormones(1, 2, 3 mg/L 6-BA and 0.1, 0.3, 0.5 mg/L NAA) on bud proliferation of S. kochii were studied by the complete test method, and the effect of the ratio of the two hormones(0.25, 0.50, 1.00 mg/L NAA and 1.0, 1.5 mg/L IBA) on rooting of S. kochii as well as the effect of substrate ratio(perlite and peat soil at 1∶9, 2∶8, 3∶7, 4∶6 and 5∶5) on its transplanting survival rate were also studied. [Results] The best basic medium for the rapid propagation of S. kochii was MS. The best hormone ratio suitable for bud proliferation was 2 mg/L6-BA+0.1 mg/L NAA, and the average number of buds proliferated at 45 d was 3.04. The average bud height was 2.05 cm; the most suitable medium for rooting was 1/2 MS+1 mg/L IBA+0.25 mg/L NAA, and its rooting rate was 100%; and the best transplanting substrate was 5∶5 perlite and peat. The soil ratio had a survival rate of 94%. [Conclusions] This study provides a theoretical basis for the improvement of the transplanting survival rate of test-tube S. kochii plantlets.

Effect of Coconut Milk and Rare Earth on Proliferation, Rootage and Isozymes of Dendrobium in Tissue Culture

In tissue culture, effects of coconut milk, rare earth and medium on proliferation, rootage and isozymes of Dendrobium were studied. The result indicated that coconut milk and RE could promote proliferation and rootage, with their combined treatment the most effective. In proliferation culture, coconut milk enhanced peroxidase (POD) and esterase(EST) activity and types, RE decreased POD, but increased EST activity and types and combined treatment did not show the same influence on POD and EST too. In rootage culture, adding coconut milk decreased POD activity, but improved EST activity in 1/2MS medium and declined EST activity, expression type in WPM medium. when using RE, POD and EST activity both reduced, types of the former decreased, types of the latter changed slightly with the basic medium. And combined treatment would decreased POD activity and types.