Development of the graft union of explanted internode autografting of cucumber ( Cu-cumis sativus Linn.) cultured in vitro is regulated by plant hormones exogenously added to the media. The levels of auxin (indole-3-a...Development of the graft union of explanted internode autografting of cucumber ( Cu-cumis sativus Linn.) cultured in vitro is regulated by plant hormones exogenously added to the media. The levels of auxin (indole-3-acetic acid, IAA) and zeatin plus zeatin riboside (Z+ ZR) in graft unions and in 4 parts of the graft union have been analyzed by ELISA assay.展开更多
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 contin...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.展开更多
The dynamic changes of IAA in graft union of explanted internode autografting of Cucumber (Cucumis sativus Linn.) have been investigated using the immunohistochemical localization technique. It is shown that the effic...The dynamic changes of IAA in graft union of explanted internode autografting of Cucumber (Cucumis sativus Linn.) have been investigated using the immunohistochemical localization technique. It is shown that the efficiency of IAA fixation using lyophilization-gas fixation is higher than that using liquid chemical fixation. In contrast to few silver particles and no significant changes during the development of graft union cultured in hormone-free medium, more silver particles in graft union and significant changes of IAA related to graft union development have been found when graft union was cultured in medium supplemented with appropriate hormones. The fixation procedure of plant hormones and the roles of IAA in graft union are discussed.展开更多
Retinitis pigmentosa is a hereditary retinal disease that affects rod and cone photoreceptors,leading to progressive photoreceptor loss.Previous research supports the beneficial effect of electrical stimulation on pho...Retinitis pigmentosa is a hereditary retinal disease that affects rod and cone photoreceptors,leading to progressive photoreceptor loss.Previous research supports the beneficial effect of electrical stimulation on photoreceptor survival.This study aims to identify the most effective electrical stimulation parameters and functional advantages of transcorneal electrical stimulation(tcES)in mice affected by inherited retinal degeneration.Additionally,the study seeked to analyze the electric field that reaches the retina in both eyes in mice and post-mortem humans.In this study,we recorded waveforms and voltages directed to the retina during transcorneal electrical stimulation in C57BL/6J mice using an intraocular needle probe with rectangular,sine,and ramp waveforms.To investigate the functional effects of electrical stimulation on photoreceptors,we used human retinal explant cultures and rhodopsin knockout(Rho^(-/-))mice,demonstrating progressive photoreceptor degeneration with age.Human retinal explants isolated from the donors’eyes were then subjected to electrical stimulation and cultured for 48 hours to simulate the neurodegenerative environment in vitro.Photoreceptor density was evaluated by rhodopsin immunolabeling.In vivo Rho^(-/-)mice were subjected to two 5-day series of daily transcorneal electrical stimulation using rectangular and ramp waveforms.Retinal function and visual perception of mice were evaluated by electroretinography and optomotor response(OMR),respectively.Immunolabeling was used to assess the morphological and biochemical changes of the photoreceptor and bipolar cells in mouse retinas.Oscilloscope recordings indicated effective delivery of rectangular,sine,and ramp waveforms to the retina by transcorneal electrical stimulation,of which the ramp waveform required the lowest voltage.Evaluation of the total conductive resistance of the post-mortem human compared to the mouse eyes indicated higher cornea-to-retina resistance in human eyes.The temperature recordings during and after electrical stimulation indicated no significant temperature change in vivo and only a subtle temperature increase in vitro(~0.5-1.5°C).Electrical stimulation increased photoreceptor survival in human retinal explant cultures,particularly at the ramp waveform.Transcorneal electrical stimulation(rectangular+ramp)waveforms significantly improved the survival and function of S and M-cones and enhanced visual acuity based on the optomotor response results.Histology and immunolabeling demonstrated increased photoreceptor survival,improved outer nuclear layer thickness,and increased bipolar cell sprouting in Rho^(-/-)mice.These results indicate that transcorneal electrical stimulation effectively delivers the electrical field to the retina,improves photoreceptor survival in both human and mouse retinas,and increases visual function in Rho^(-/-)mice.Combined rectangular and ramp waveform stimulation can promote photoreceptor survival in a minimally invasive fashion.展开更多
To establish an efficient regeneration method for the rare and endangered plant Mussaenda anomala to address problems regarding its reproductive obstacles and scarce populations.In this study,the terminal buds,axillar...To establish an efficient regeneration method for the rare and endangered plant Mussaenda anomala to address problems regarding its reproductive obstacles and scarce populations.In this study,the terminal buds,axillary buds,stem segments with two axillary buds,stem segments with two axillary buds and one terminal bud,and leaves of M.anomala were used as explants.The effects of different explants and disinfection methods,plant growth regulators and substrates on plant regeneration were explored.The following results were obtained:(1)The terminal bud was a suitable explant for M.anomala tissue culture,and the disinfection method utilized was treatment with 0.2%HgCl2 for 8 min.(2)Initiate medium:MS basic medium supplemented with 0.5 mg/L 6-BA and 0.2 mg/L IBA for the high germination rate(100%)and the maximum bud height(1.70 cm)of the terminal bud.(3)Proliferation medium:MS basic medium supplemented with 3.0 mg/L 6-BA and 0.2 mg/L IBA for a high proliferation rate(96%)and proliferation time(6.0)of terminal buds.(4)Proliferation medium supplemented with 0.7 mg/L GA3 significantly increased the bud heights of multiple buds.(5)Rooting medium:MS basic medium supplemented with 0.5 mg/L IBA and 0.5 mg/L IAA for a high rooting rate(88%),root number(12.0)and root length(5.07 cm).(6)The optimal substrate for seedling acclimation and transplanting was perlite:vermiculite(1:1),which resulted in the highest survival rate(97%)and plant height(5.89 cm),as well as better growth potential for seedlings.The surfaces of M.anomala explants are densely covered with trichome,which increased the difficulty of disinfection;the plant growth regulators directly affected the growth and development in the regeneration process of M.anomala,and the substrate significantly affected the survival rate and height growth for seedling acclimation.展开更多
Fucoidan,a sulfate polysaccharide obtained from brown seaweed,has various bioactive properties,including anti-inflammatory,anti-cancer,anti-viral,anti-oxidant,anti-coagulant,anti-thrombotic,anti-angiogenic,and anti-He...Fucoidan,a sulfate polysaccharide obtained from brown seaweed,has various bioactive properties,including anti-inflammatory,anti-cancer,anti-viral,anti-oxidant,anti-coagulant,anti-thrombotic,anti-angiogenic,and anti-Helicobacter pylori properties.However,the effects of low-molecular-weight fucoidan(LMW-F)on melanoma cell lines and three dimensional(3D)cell culture models are not well understood.This study aimed to investigate the effects of LMW-F on A375 human melanoma cells and cryopreserved biospecimens derived from patients with advanced melanoma.Ultrasonic wave was used to fragment fucoidan derived from Fucus vesiculosus into smaller LMW-F.MTT and live/dead assays showed that LMW-F inhibited cell proliferation in both A375 cells and patientderived melanoma explants in a 3D-printed collagen scaffold.The PTEN/AKT pathway was found to be involved in the anti-melanoma effects of fucoidan.Western blot analysis revealed that LMW-F reduced the phosphorylation of Bcl-2 at Thr 56,which was associated with the prevention of anti-apoptotic activity of cancer cells.Our findings suggested that LMW-F could enhance anti-melanoma chemotherapy and improve the outcomes of patients with melanoma resistance.展开更多
[Objectives]The paper was to study the tissue culture nursery technology of Curcuma alismatifolia‘Kimono Rose’.[Methods]By sterilizing and inoculating explants derived from disparate regions of C.alismatifolia,we id...[Objectives]The paper was to study the tissue culture nursery technology of Curcuma alismatifolia‘Kimono Rose’.[Methods]By sterilizing and inoculating explants derived from disparate regions of C.alismatifolia,we identified the most optimal explants and optimized the culture conditions for cluster buds induction and proliferation.This was achieved by incorporating MS medium with varying concentrations of 6-BA and NAA,thereby establishing a foundation for the large-scale production of C.alismatifolia tissue culture seedlings.[Results]The optimal explant for C.alismatifolia was identified as a lateral bud.The most effective cluster buds induction medium was determined to be MS+6-BA 5 mg/L+NAA 0.1 mg/L+sucrose 25 g/L+agar 6.5 g/L.The optimal cluster buds proliferation medium was found to be MS+6-BA 3 mg/L+NAA 0.1 mg/L+sucrose 25 g/L+agar 6.5 g/L.[Conclusions]The findings of this study can provide a foundation for the enhancement of the industrialized breeding system of tissue culture propagation of C.alismatifolia.展开更多
[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,...[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.展开更多
[Objective] The aim was to study the explants sterilization and callus induction of Aquilegia oxysepala.[Method] the seeds of Aquilegia oxysepala were sterilized by different kinds and concentrations of disinfectants,...[Objective] The aim was to study the explants sterilization and callus induction of Aquilegia oxysepala.[Method] the seeds of Aquilegia oxysepala were sterilized by different kinds and concentrations of disinfectants,and the pollution rate and pollution speed were investigated so as to find the best way to build sterile seedling setup.Taking the roots,stem segments and leaves of the sterile seedlings from Aquilegia oxysepala seeds as explants,the optimum explants and medium were screened by adding MS basic medium with different hormone proportions.[Result] The best germicidal treatment was as follows：explants were soaked in 75% alcohol for 30 s firstly,washed by sterile water for 5 times,then soaked in 0.2% mercuric chloride liquid for 2 min,finally washed by sterile water for 5 times again.The sterilization treatment could get the lowest pollution rate,the highest germinating capacity and the best sterile seedling.Roots were the optimum explants for the callus induction of Aquilegia oxysepala,meanwhile the optimal medium was MS＋0.6 mg/L 2,4-D＋0.5 mg/L 6-BA.[Conclusion] The research provides technical support for the large scale production of Aquilegia oxysepala and also makes a contribution to the medicinal and ornamental value of Aquilegia oxysepala.展开更多
The establishment of high frequency regeneration system is a foundation for Agrobacterium mediated genetic transformation. In this work, several important factors influencing the efficiency of regeneration of pla...The establishment of high frequency regeneration system is a foundation for Agrobacterium mediated genetic transformation. In this work, several important factors influencing the efficiency of regeneration of plants, such as concentration of plant growth regulators, leaf explant orientation, leaf growth sequence and leaf segment, were studied. The results indicated that the differentiation rate of adventitious shoots was 90% on basal MS medium only supplemented with 1 5?mg·L -1 BA (6 benzyladenine) and reached the highest level(95%) when 1 0?mg·L -1 BA and 0 3?mg·L -1 NAA (naphthaleneacetic acid) were added to MS medium. 90% of differentiation rate of adventitious roots were obtained when 0 3?mg·L -1 NAA was only added to MS medium. It was also found that more adventitious shoots were regenerated from the lower segment of leaf (with petiole) than the other segments, the number of adventitious shoots decreased from top to base of leaf growth sequence and the percentage of adventitious shoot induction with adaxial side downward was higher than that with adaxial side upward.展开更多
In this experiment, floral development from tissue culture of bread wheat (Triticum aestivum L.) was investigated. Immature embryos of 45 wheat cultivars were cultured, and 11.1% of the genotypes regenerated floral or...In this experiment, floral development from tissue culture of bread wheat (Triticum aestivum L.) was investigated. Immature embryos of 45 wheat cultivars were cultured, and 11.1% of the genotypes regenerated floral organs from the calli near the bases of the green buds or plantlets regenerated. The floral buds were morphologically incomplete with the appearances of unisexual pistillate flowers which were naked, clustered with normal ovaries and exuberant feather-like stigmas, but without stamens, paleas, lemmas and glumes. The histological examination showed that the pistils originated from the meristematic cells near the green buds or plantlets, and the clustered pistils were formed by secondary pistillate regeneration. The development of the feather-like structures was earlier than that of the ovules. Biovule developed from an ovary besides normal uniovule. Statistical analysis by X 2 test for independency demonstrated highly significant difference of flower regeneration among the tested genotypes. Wheat cultivar YA-1 revealed higher percentage (44.4%) than other genotypes, and the response could well be repeated in different years. It was indicated that the floral regeneration of immature embryo explants of YA-1 is relatively stable. The frequency of floral regeneration was mainly regulated by the components in the subculture media, compared with the response of the dedifferentiation media, despite the obviously different components involving basal medium type, inorganic Fe2+ concentration and plant growth regulators. The results suggested the combination of 6-benzylaminopurine, alpha-naphthalene acetic-acid and doubled inorganic Fe2+ might be more beneficial to inducing the floral development than that of 2,4-D and normal inorganic Fe2+ concentration in subculture medium. However, both immature inflorescence and mature embryo, as cultured explants of YA-1, did not regenerate any flower organs. It is believed that the immature embryo culture of YA-1 can be used to establish ideal experimental system for the study of floral developmental mechanism in wheat.展开更多
This paper summarized the technology of haploid production, protoplast culture, organ regeneration culture of pepper and the key factors affecting in vitro regeneration culture of pepper, including explants, seedling ...This paper summarized the technology of haploid production, protoplast culture, organ regeneration culture of pepper and the key factors affecting in vitro regeneration culture of pepper, including explants, seedling age, medium,genotype and plant growth regulator, then pointed out several main problems, in order to provide the reference for building an efficient in vitro regeneration culture system of pepper and its application in breeding.展开更多
[Objective] The aim was to look for an effective way of decreasing explants contamination in the root tissue culture of Hevea brasiliensis.[Method] In order to study root tissue culture of H.brasiliensis,new roots fro...[Objective] The aim was to look for an effective way of decreasing explants contamination in the root tissue culture of Hevea brasiliensis.[Method] In order to study root tissue culture of H.brasiliensis,new roots from forest section after treatment for one month were as explants,and 1 g/L carbendazim,75% ethanol,0.1% mercuric chloride and Yipeilong with different concentrations were used as disinfectants for roots disinfection.[Result] Before conventional disinfection,root explants were treated by 1 g/L carbendazim for 2.5 h,and disinfected by 75% ethanol for 30 s and 0.1% HgCl2 for 6 min,then cultured on the callus induction medium supplemented with 0.1% Yipeilong.The results showed that the contamination rate of explants decreased to 44.59%,and 25.60% explants survived after cultured for 30 d.[Conclusion] The study could provide theoretical foundation for the decrease of explant contamination in the root tissue culture of H.brasiliensis.展开更多
Plant tissue culture systems have enormous potential in fundamental re- search and for commercial applications such as horticultural industry. The process of tissue culture is companied with a series of changes in res...Plant tissue culture systems have enormous potential in fundamental re- search and for commercial applications such as horticultural industry. The process of tissue culture is companied with a series of changes in respect to morphology, physiology, biochemistry, molecule and epigenetics. The changes at molecule levels mainly include genetic variation, DNA sequence variation, chromosomal variation and epigenetic regulation （DNA methylation, histone modification, chromatin remodeling, small RNA regulation）. These changes are believed to facilitate explant adaptation to culture conditions and to help subsequent morphogenesis processes. Nowadays, it has played a crucial part in commercial applications and in basic research into cell biology, genetics and biochemistry, etc. In present review, we shed light on the fun- damental of plant tissue culture, culture medium preparation, explant selection, mechanism of action of various hormones, the three major problems （explant pollu- tion, browning, plantlets vitrification） and the prevention measures in tissue culture, and elaborated on in vitro propagation of plants, virus-free seedling cultivation, cry- opreservation, artificial seeds and molecule levels changes during in vitro culture further.展开更多
[ Objective] This study aimed to investigate the differences in morphological characteristics of Dendrobium officinale plantlets propagated from different explants. [ Method] Randomly 1 000 D. offtcinale plantlets pro...[ Objective] This study aimed to investigate the differences in morphological characteristics of Dendrobium officinale plantlets propagated from different explants. [ Method] Randomly 1 000 D. offtcinale plantlets propagated via different regeneration pathways were selected for morphological investigation and classification. [ Result] D. officinale plantlets propagated from stem segment explants exhibited highly consistent morphological characteristics, while those propagated from seed explants exhibited a variety of morphological characteristics. [ Conclusion] Therefore, using seed explants for regeneration can effectively broaden the germplasms resources of D. officinale.展开更多
Populus species are important resources for industry and in scientific study on biological and agricul- tural systems. Our objective was to enhance the frequency of plant regeneration in Himalayan poplar （Populus cil...Populus species are important resources for industry and in scientific study on biological and agricul- tural systems. Our objective was to enhance the frequency of plant regeneration in Himalayan poplar （Populus ciliata wall. ex Royle）. The effect of TDZ alone and in combi- nation with adenine and NAA was studied on the regen- eration potential of petiole explants. The explants were excised from Himalayan poplar plants grown in glass- houses. After surface sterilization the explants were cul- tured on shoot induction medium. High percentage shoot regeneration （86 %） was recorded on MS medium sup- plemented with 0.004 mg L-1 TDZ and 79.7 mg L-1 adenine. The regenerated shoots for elongation and multi- plication were transferred to MS ＋ 0.5 mg L-1 BAP ＋ 0.2 mg L-1 IAA ＋ 0.3 mg L-1 GA3. Root re- generation from shoots developed in vitro was observed on MS medium supplemented with 0.10 mg L-1 IBA. Hi- malayan poplar plantlets could be produced within 2 months after acclimatization in a sterile mixture of sand and soil. We developed a high efficiency plant regeneration protocol from petiole explants of P. ciliata.展开更多
Neurotrophins play a major role in the regulation of neuronal growth such as neurite sprouting or regeneration in response to nerve injuries. The role of nerve growth factor, neurotrophin-3, and brain-derived neurotro...Neurotrophins play a major role in the regulation of neuronal growth such as neurite sprouting or regeneration in response to nerve injuries. The role of nerve growth factor, neurotrophin-3, and brain-derived neurotrophic factor in maintaining the survival of peripheral neurons remains poorly understood. In regenerative medicine, different modalities have been investigated for the delivery of growth factors to the injured neurons, in search of a suitable system for clinical applications. This study was to investigate the influence of nerve growth factor, neurotrophin-3 and brain-derived neurotrophic factor on the growth of neurites using two in vitro models of dorsal root ganglia explants and dorsal root ganglia-derived primary cell dissociated cultures. Quantitative data showed that the total neurite length and tortuosity were differently influenced by trophic factors. Nerve growth factor and, indirectly, brain-derived neurotrophic factor stimulate the tortuous growth of sensory fibers and the formation of cell clusters. Neurotrophin-3, however, enhances neurite growth in terms of length and linearity allowing for a more organized and directed axonal elongation towards a peripheral target compared to the other growth factors. These findings could be of considerable importance for any clinical application of neurotrophic factors in peripheral nerve regeneration. Ethical approval was obtained from the Regione Piemonte Animal Ethics Committee ASLTO1(file # 864/2016-PR) on September 14, 2016.展开更多
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 investig...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.展开更多
An efficient in vitro regeneration system by direct organogenesis from mature nodal and internodal stem segments ofNewhall navel orange (Citrus sinensis L. Osbeck) was developed. Illuminating conditions together with...An efficient in vitro regeneration system by direct organogenesis from mature nodal and internodal stem segments ofNewhall navel orange (Citrus sinensis L. Osbeck) was developed. Illuminating conditions together with plant growthregulators affected the adventitious bud regeneration frequency and efficiency. The initial 15 d darkness inoculation isbeneficial for the adventitious bud regeneration. The highest regeneration frequency (85.2%) and bud formationefficiency (3.7 per responsive internodal stem segment) were obtained in the media supplemented with 1.0 mg L-1 BAPand 0.5 mg L-1 NAA. ABA at 0.2 mg L-1 positively affected the bud formation efficiency, which amounted to 8.5 buds perinternodal segment in the presence of BAP at 1.0 mg L-1. The adventitious shoots successfully rooted and weretransferred to the soil.展开更多
Objective:To determine the effects of different cytokinins at various concentrations onin vitro shoot multiplication of an important medicinal plant.Methods:Nodal explants(1.5-2.0 cm)of Sophora tonkinensiswere used.Mu...Objective:To determine the effects of different cytokinins at various concentrations onin vitro shoot multiplication of an important medicinal plant.Methods:Nodal explants(1.5-2.0 cm)of Sophora tonkinensiswere used.Multiple shoots were induced from nodal explants cultured onthe Murashige and Skoog(MS)medium supplemented with 0.0,0.5,1.0,2.0,4.0,8.0,or 16.0μmol2-isopentyladenine(2iP),N6 benzyladenine,kinetin or thiadiazuron.Results:Among the fourinvestigated cytokinins,2iP showed the best response for shoot multiplication.Maximum shootinduction(75%)was achieved on the MS medium supplemented with 2.0μmol 2iP,with a meannumber of 5.0 shoots per explant.In comparison to other cytokinins tried,2iP showed the highestshoot elongation with a mean shoot length of 4.8 cm.Root initiation was observed within 15 dwithin the transfer of shoots onto the MS basal medium,and the rooting percentage was 100%with a mean number of 5.4 roots per shoot and root length of 6.2 cm over a period of 4 weeks.Thehealthy plants,hardened and transferred to a greenhouse for proper acclimatization,exhibited100%survival.Conclusions:It can be summarized that 2iP is the optimal plant growth regulatorforSophoramultiplication.展开更多
文摘Development of the graft union of explanted internode autografting of cucumber ( Cu-cumis sativus Linn.) cultured in vitro is regulated by plant hormones exogenously added to the media. The levels of auxin (indole-3-acetic acid, IAA) and zeatin plus zeatin riboside (Z+ ZR) in graft unions and in 4 parts of the graft union have been analyzed by ELISA assay.
基金supported by the National Natural Science Foundation of China,Nos.81901156(to ZZ),82271200(to ZZ),82171308(to XC)the Fundamental Research Funds for the Central Universities,No.xzy012022035(to ZZ)+1 种基金the Natural Science Foundation of Shaanxi Province,Nos.2021JM-261(to QK),2023-YBSF-303(to ZZ)Traditional Chinese Medicine Project of Shaanxi Province,No.2019-ZZ-JC047(to QK)。
文摘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.
文摘The dynamic changes of IAA in graft union of explanted internode autografting of Cucumber (Cucumis sativus Linn.) have been investigated using the immunohistochemical localization technique. It is shown that the efficiency of IAA fixation using lyophilization-gas fixation is higher than that using liquid chemical fixation. In contrast to few silver particles and no significant changes during the development of graft union cultured in hormone-free medium, more silver particles in graft union and significant changes of IAA related to graft union development have been found when graft union was cultured in medium supplemented with appropriate hormones. The fixation procedure of plant hormones and the roles of IAA in graft union are discussed.
基金supported by The Norwegian Research CouncilDepartment of Ophthalmology,Oslo University Hospital,Oslo,Norway(to TPU)+10 种基金Department of Medical Biochemistry,Oslo University Hospital,Oslo,Norway(to TPU)The Norwegian Association for the Blind and Partially Sighted(to TPU)The Ministry of Science and Technology of Taiwan,China MOST 105-2917-I-002-031,MOST 109-2917-I-564-032(to KC)The Scientific and Technological Research Council of Turkiye-TUBITAK(to KG)BrightFocus Foundation(to KSC)the Massachusetts Lions Foundation(to KSC)National Eye Institute Grant EY031696(to DFC)Harvard NeuroDiscovery Center Grant(to DFC)Department of Defense(USA)HT9425-23-1-1045(to DFC and AL)Core Grant for Vision Research from NIH/NEI to the Schepens Eye Research Institute(P30EY003790)South-Eastern Norway Regional Health Authority and the Norwegian Society of the Blind(to TPU).
文摘Retinitis pigmentosa is a hereditary retinal disease that affects rod and cone photoreceptors,leading to progressive photoreceptor loss.Previous research supports the beneficial effect of electrical stimulation on photoreceptor survival.This study aims to identify the most effective electrical stimulation parameters and functional advantages of transcorneal electrical stimulation(tcES)in mice affected by inherited retinal degeneration.Additionally,the study seeked to analyze the electric field that reaches the retina in both eyes in mice and post-mortem humans.In this study,we recorded waveforms and voltages directed to the retina during transcorneal electrical stimulation in C57BL/6J mice using an intraocular needle probe with rectangular,sine,and ramp waveforms.To investigate the functional effects of electrical stimulation on photoreceptors,we used human retinal explant cultures and rhodopsin knockout(Rho^(-/-))mice,demonstrating progressive photoreceptor degeneration with age.Human retinal explants isolated from the donors’eyes were then subjected to electrical stimulation and cultured for 48 hours to simulate the neurodegenerative environment in vitro.Photoreceptor density was evaluated by rhodopsin immunolabeling.In vivo Rho^(-/-)mice were subjected to two 5-day series of daily transcorneal electrical stimulation using rectangular and ramp waveforms.Retinal function and visual perception of mice were evaluated by electroretinography and optomotor response(OMR),respectively.Immunolabeling was used to assess the morphological and biochemical changes of the photoreceptor and bipolar cells in mouse retinas.Oscilloscope recordings indicated effective delivery of rectangular,sine,and ramp waveforms to the retina by transcorneal electrical stimulation,of which the ramp waveform required the lowest voltage.Evaluation of the total conductive resistance of the post-mortem human compared to the mouse eyes indicated higher cornea-to-retina resistance in human eyes.The temperature recordings during and after electrical stimulation indicated no significant temperature change in vivo and only a subtle temperature increase in vitro(~0.5-1.5°C).Electrical stimulation increased photoreceptor survival in human retinal explant cultures,particularly at the ramp waveform.Transcorneal electrical stimulation(rectangular+ramp)waveforms significantly improved the survival and function of S and M-cones and enhanced visual acuity based on the optomotor response results.Histology and immunolabeling demonstrated increased photoreceptor survival,improved outer nuclear layer thickness,and increased bipolar cell sprouting in Rho^(-/-)mice.These results indicate that transcorneal electrical stimulation effectively delivers the electrical field to the retina,improves photoreceptor survival in both human and mouse retinas,and increases visual function in Rho^(-/-)mice.Combined rectangular and ramp waveform stimulation can promote photoreceptor survival in a minimally invasive fashion.
基金funded by the National Natural Science Foundation of China–Guizhou Provincial People’s Government Karst Science Research Center Project(U1812401)the National Natural Science Foundation of China(31760124),the Department of Education of Guizhou Province(grant code qianjiaoji(2022)136)the New Seedling Program of Guizhou Normal University(grant code 2021-B05).
文摘To establish an efficient regeneration method for the rare and endangered plant Mussaenda anomala to address problems regarding its reproductive obstacles and scarce populations.In this study,the terminal buds,axillary buds,stem segments with two axillary buds,stem segments with two axillary buds and one terminal bud,and leaves of M.anomala were used as explants.The effects of different explants and disinfection methods,plant growth regulators and substrates on plant regeneration were explored.The following results were obtained:(1)The terminal bud was a suitable explant for M.anomala tissue culture,and the disinfection method utilized was treatment with 0.2%HgCl2 for 8 min.(2)Initiate medium:MS basic medium supplemented with 0.5 mg/L 6-BA and 0.2 mg/L IBA for the high germination rate(100%)and the maximum bud height(1.70 cm)of the terminal bud.(3)Proliferation medium:MS basic medium supplemented with 3.0 mg/L 6-BA and 0.2 mg/L IBA for a high proliferation rate(96%)and proliferation time(6.0)of terminal buds.(4)Proliferation medium supplemented with 0.7 mg/L GA3 significantly increased the bud heights of multiple buds.(5)Rooting medium:MS basic medium supplemented with 0.5 mg/L IBA and 0.5 mg/L IAA for a high rooting rate(88%),root number(12.0)and root length(5.07 cm).(6)The optimal substrate for seedling acclimation and transplanting was perlite:vermiculite(1:1),which resulted in the highest survival rate(97%)and plant height(5.89 cm),as well as better growth potential for seedlings.The surfaces of M.anomala explants are densely covered with trichome,which increased the difficulty of disinfection;the plant growth regulators directly affected the growth and development in the regeneration process of M.anomala,and the substrate significantly affected the survival rate and height growth for seedling acclimation.
基金supported by the Priority Research Centers Program through the National Research Foundation of Korea(NRF)funded by the Ministry of Education,Science and Technology(Grant 2017R1A6A03015562 and RS-2023-00237386).
文摘Fucoidan,a sulfate polysaccharide obtained from brown seaweed,has various bioactive properties,including anti-inflammatory,anti-cancer,anti-viral,anti-oxidant,anti-coagulant,anti-thrombotic,anti-angiogenic,and anti-Helicobacter pylori properties.However,the effects of low-molecular-weight fucoidan(LMW-F)on melanoma cell lines and three dimensional(3D)cell culture models are not well understood.This study aimed to investigate the effects of LMW-F on A375 human melanoma cells and cryopreserved biospecimens derived from patients with advanced melanoma.Ultrasonic wave was used to fragment fucoidan derived from Fucus vesiculosus into smaller LMW-F.MTT and live/dead assays showed that LMW-F inhibited cell proliferation in both A375 cells and patientderived melanoma explants in a 3D-printed collagen scaffold.The PTEN/AKT pathway was found to be involved in the anti-melanoma effects of fucoidan.Western blot analysis revealed that LMW-F reduced the phosphorylation of Bcl-2 at Thr 56,which was associated with the prevention of anti-apoptotic activity of cancer cells.Our findings suggested that LMW-F could enhance anti-melanoma chemotherapy and improve the outcomes of patients with melanoma resistance.
基金Supported by Special Project of Public-interest Scientific Institutions of Fujian Province(2021R1011003).
文摘[Objectives]The paper was to study the tissue culture nursery technology of Curcuma alismatifolia‘Kimono Rose’.[Methods]By sterilizing and inoculating explants derived from disparate regions of C.alismatifolia,we identified the most optimal explants and optimized the culture conditions for cluster buds induction and proliferation.This was achieved by incorporating MS medium with varying concentrations of 6-BA and NAA,thereby establishing a foundation for the large-scale production of C.alismatifolia tissue culture seedlings.[Results]The optimal explant for C.alismatifolia was identified as a lateral bud.The most effective cluster buds induction medium was determined to be MS+6-BA 5 mg/L+NAA 0.1 mg/L+sucrose 25 g/L+agar 6.5 g/L.The optimal cluster buds proliferation medium was found to be MS+6-BA 3 mg/L+NAA 0.1 mg/L+sucrose 25 g/L+agar 6.5 g/L.[Conclusions]The findings of this study can provide a foundation for the enhancement of the industrialized breeding system of tissue culture propagation of C.alismatifolia.
基金Supported by Fund for Scientific Research in Yangtze University(CDKF2283)Program of Engineering Research Center of Wetland Agriculture in the Middle Reaches of the Yangtze River of Ministry of Education~~
文摘[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.
基金Supported by the Fundamental Research Funds for the Central Universities(DL09BA16)Science Research Foundation for Young Scientists of northeast Forestry University(09047 )Graduate Project Technology Innovation Funds of Northeast Forestry University~~
文摘[Objective] The aim was to study the explants sterilization and callus induction of Aquilegia oxysepala.[Method] the seeds of Aquilegia oxysepala were sterilized by different kinds and concentrations of disinfectants,and the pollution rate and pollution speed were investigated so as to find the best way to build sterile seedling setup.Taking the roots,stem segments and leaves of the sterile seedlings from Aquilegia oxysepala seeds as explants,the optimum explants and medium were screened by adding MS basic medium with different hormone proportions.[Result] The best germicidal treatment was as follows：explants were soaked in 75% alcohol for 30 s firstly,washed by sterile water for 5 times,then soaked in 0.2% mercuric chloride liquid for 2 min,finally washed by sterile water for 5 times again.The sterilization treatment could get the lowest pollution rate,the highest germinating capacity and the best sterile seedling.Roots were the optimum explants for the callus induction of Aquilegia oxysepala,meanwhile the optimal medium was MS＋0.6 mg/L 2,4-D＋0.5 mg/L 6-BA.[Conclusion] The research provides technical support for the large scale production of Aquilegia oxysepala and also makes a contribution to the medicinal and ornamental value of Aquilegia oxysepala.
文摘The establishment of high frequency regeneration system is a foundation for Agrobacterium mediated genetic transformation. In this work, several important factors influencing the efficiency of regeneration of plants, such as concentration of plant growth regulators, leaf explant orientation, leaf growth sequence and leaf segment, were studied. The results indicated that the differentiation rate of adventitious shoots was 90% on basal MS medium only supplemented with 1 5?mg·L -1 BA (6 benzyladenine) and reached the highest level(95%) when 1 0?mg·L -1 BA and 0 3?mg·L -1 NAA (naphthaleneacetic acid) were added to MS medium. 90% of differentiation rate of adventitious roots were obtained when 0 3?mg·L -1 NAA was only added to MS medium. It was also found that more adventitious shoots were regenerated from the lower segment of leaf (with petiole) than the other segments, the number of adventitious shoots decreased from top to base of leaf growth sequence and the percentage of adventitious shoot induction with adaxial side downward was higher than that with adaxial side upward.
文摘In this experiment, floral development from tissue culture of bread wheat (Triticum aestivum L.) was investigated. Immature embryos of 45 wheat cultivars were cultured, and 11.1% of the genotypes regenerated floral organs from the calli near the bases of the green buds or plantlets regenerated. The floral buds were morphologically incomplete with the appearances of unisexual pistillate flowers which were naked, clustered with normal ovaries and exuberant feather-like stigmas, but without stamens, paleas, lemmas and glumes. The histological examination showed that the pistils originated from the meristematic cells near the green buds or plantlets, and the clustered pistils were formed by secondary pistillate regeneration. The development of the feather-like structures was earlier than that of the ovules. Biovule developed from an ovary besides normal uniovule. Statistical analysis by X 2 test for independency demonstrated highly significant difference of flower regeneration among the tested genotypes. Wheat cultivar YA-1 revealed higher percentage (44.4%) than other genotypes, and the response could well be repeated in different years. It was indicated that the floral regeneration of immature embryo explants of YA-1 is relatively stable. The frequency of floral regeneration was mainly regulated by the components in the subculture media, compared with the response of the dedifferentiation media, despite the obviously different components involving basal medium type, inorganic Fe2+ concentration and plant growth regulators. The results suggested the combination of 6-benzylaminopurine, alpha-naphthalene acetic-acid and doubled inorganic Fe2+ might be more beneficial to inducing the floral development than that of 2,4-D and normal inorganic Fe2+ concentration in subculture medium. However, both immature inflorescence and mature embryo, as cultured explants of YA-1, did not regenerate any flower organs. It is believed that the immature embryo culture of YA-1 can be used to establish ideal experimental system for the study of floral developmental mechanism in wheat.
基金Supported by Graduate Research Innovation Fund of Guizhou Academy of Agricultural Sciences(2011004)~~
文摘This paper summarized the technology of haploid production, protoplast culture, organ regeneration culture of pepper and the key factors affecting in vitro regeneration culture of pepper, including explants, seedling age, medium,genotype and plant growth regulator, then pointed out several main problems, in order to provide the reference for building an efficient in vitro regeneration culture system of pepper and its application in breeding.
基金Supported by Natural Science Foundation of Hainan Province(30821)Basic Scientific Research Business Fees of Chinese Academy of Tropical Agricultural Sciences(XJSYWFZX2008-02)~~
文摘[Objective] The aim was to look for an effective way of decreasing explants contamination in the root tissue culture of Hevea brasiliensis.[Method] In order to study root tissue culture of H.brasiliensis,new roots from forest section after treatment for one month were as explants,and 1 g/L carbendazim,75% ethanol,0.1% mercuric chloride and Yipeilong with different concentrations were used as disinfectants for roots disinfection.[Result] Before conventional disinfection,root explants were treated by 1 g/L carbendazim for 2.5 h,and disinfected by 75% ethanol for 30 s and 0.1% HgCl2 for 6 min,then cultured on the callus induction medium supplemented with 0.1% Yipeilong.The results showed that the contamination rate of explants decreased to 44.59%,and 25.60% explants survived after cultured for 30 d.[Conclusion] The study could provide theoretical foundation for the decrease of explant contamination in the root tissue culture of H.brasiliensis.
基金Supported by Natural Science Foundation of Inner Mongolia(2012MS0516)~~
文摘Plant tissue culture systems have enormous potential in fundamental re- search and for commercial applications such as horticultural industry. The process of tissue culture is companied with a series of changes in respect to morphology, physiology, biochemistry, molecule and epigenetics. The changes at molecule levels mainly include genetic variation, DNA sequence variation, chromosomal variation and epigenetic regulation （DNA methylation, histone modification, chromatin remodeling, small RNA regulation）. These changes are believed to facilitate explant adaptation to culture conditions and to help subsequent morphogenesis processes. Nowadays, it has played a crucial part in commercial applications and in basic research into cell biology, genetics and biochemistry, etc. In present review, we shed light on the fun- damental of plant tissue culture, culture medium preparation, explant selection, mechanism of action of various hormones, the three major problems （explant pollu- tion, browning, plantlets vitrification） and the prevention measures in tissue culture, and elaborated on in vitro propagation of plants, virus-free seedling cultivation, cry- opreservation, artificial seeds and molecule levels changes during in vitro culture further.
基金Supported by of Spark Program of Guangdong Department of Science and Technology"Development of Cultivation Techniques of Dendrobium officinale Seedlings"(2013B020503062)Special Innovation Fund for Small and Medium Enterprise in Maoming City"Research and Demonstration of Alpine Organic Efficient Cultivation Technology of Precious Chinese Herb Dendrobium officinale"(2012B01088)
文摘[ Objective] This study aimed to investigate the differences in morphological characteristics of Dendrobium officinale plantlets propagated from different explants. [ Method] Randomly 1 000 D. offtcinale plantlets propagated via different regeneration pathways were selected for morphological investigation and classification. [ Result] D. officinale plantlets propagated from stem segment explants exhibited highly consistent morphological characteristics, while those propagated from seed explants exhibited a variety of morphological characteristics. [ Conclusion] Therefore, using seed explants for regeneration can effectively broaden the germplasms resources of D. officinale.
文摘Populus species are important resources for industry and in scientific study on biological and agricul- tural systems. Our objective was to enhance the frequency of plant regeneration in Himalayan poplar （Populus ciliata wall. ex Royle）. The effect of TDZ alone and in combi- nation with adenine and NAA was studied on the regen- eration potential of petiole explants. The explants were excised from Himalayan poplar plants grown in glass- houses. After surface sterilization the explants were cul- tured on shoot induction medium. High percentage shoot regeneration （86 %） was recorded on MS medium sup- plemented with 0.004 mg L-1 TDZ and 79.7 mg L-1 adenine. The regenerated shoots for elongation and multi- plication were transferred to MS ＋ 0.5 mg L-1 BAP ＋ 0.2 mg L-1 IAA ＋ 0.3 mg L-1 GA3. Root re- generation from shoots developed in vitro was observed on MS medium supplemented with 0.10 mg L-1 IBA. Hi- malayan poplar plantlets could be produced within 2 months after acclimatization in a sterile mixture of sand and soil. We developed a high efficiency plant regeneration protocol from petiole explants of P. ciliata.
基金supported by the research start-up and the MWU’s intramural grant(to MF)the Italian MURST-MIUR foundation(to SG and IP)
文摘Neurotrophins play a major role in the regulation of neuronal growth such as neurite sprouting or regeneration in response to nerve injuries. The role of nerve growth factor, neurotrophin-3, and brain-derived neurotrophic factor in maintaining the survival of peripheral neurons remains poorly understood. In regenerative medicine, different modalities have been investigated for the delivery of growth factors to the injured neurons, in search of a suitable system for clinical applications. This study was to investigate the influence of nerve growth factor, neurotrophin-3 and brain-derived neurotrophic factor on the growth of neurites using two in vitro models of dorsal root ganglia explants and dorsal root ganglia-derived primary cell dissociated cultures. Quantitative data showed that the total neurite length and tortuosity were differently influenced by trophic factors. Nerve growth factor and, indirectly, brain-derived neurotrophic factor stimulate the tortuous growth of sensory fibers and the formation of cell clusters. Neurotrophin-3, however, enhances neurite growth in terms of length and linearity allowing for a more organized and directed axonal elongation towards a peripheral target compared to the other growth factors. These findings could be of considerable importance for any clinical application of neurotrophic factors in peripheral nerve regeneration. Ethical approval was obtained from the Regione Piemonte Animal Ethics Committee ASLTO1(file # 864/2016-PR) on September 14, 2016.
文摘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.
基金The authors acknowledge the financial support by the National Natural Science Foundation of China(002002)
文摘An efficient in vitro regeneration system by direct organogenesis from mature nodal and internodal stem segments ofNewhall navel orange (Citrus sinensis L. Osbeck) was developed. Illuminating conditions together with plant growthregulators affected the adventitious bud regeneration frequency and efficiency. The initial 15 d darkness inoculation isbeneficial for the adventitious bud regeneration. The highest regeneration frequency (85.2%) and bud formationefficiency (3.7 per responsive internodal stem segment) were obtained in the media supplemented with 1.0 mg L-1 BAPand 0.5 mg L-1 NAA. ABA at 0.2 mg L-1 positively affected the bud formation efficiency, which amounted to 8.5 buds perinternodal segment in the presence of BAP at 1.0 mg L-1. The adventitious shoots successfully rooted and weretransferred to the soil.
基金This study was carried out with the support of"On-Site Cooperative Agriculture Research Project(Grant No.006330)",RDA,Republic of Korea
文摘Objective:To determine the effects of different cytokinins at various concentrations onin vitro shoot multiplication of an important medicinal plant.Methods:Nodal explants(1.5-2.0 cm)of Sophora tonkinensiswere used.Multiple shoots were induced from nodal explants cultured onthe Murashige and Skoog(MS)medium supplemented with 0.0,0.5,1.0,2.0,4.0,8.0,or 16.0μmol2-isopentyladenine(2iP),N6 benzyladenine,kinetin or thiadiazuron.Results:Among the fourinvestigated cytokinins,2iP showed the best response for shoot multiplication.Maximum shootinduction(75%)was achieved on the MS medium supplemented with 2.0μmol 2iP,with a meannumber of 5.0 shoots per explant.In comparison to other cytokinins tried,2iP showed the highestshoot elongation with a mean shoot length of 4.8 cm.Root initiation was observed within 15 dwithin the transfer of shoots onto the MS basal medium,and the rooting percentage was 100%with a mean number of 5.4 roots per shoot and root length of 6.2 cm over a period of 4 weeks.Thehealthy plants,hardened and transferred to a greenhouse for proper acclimatization,exhibited100%survival.Conclusions:It can be summarized that 2iP is the optimal plant growth regulatorforSophoramultiplication.
