Plant Regeneration from In Vitro Cultured Hypocotyl Explants of Euonymus japonicus Cu zhi

Adventitious shoots were successfully regenerated from hypocotyl explants of in vitro cultures of Euonymus japonicus Cu zhi. Hypocotyl slices were cultured on Murashige and Skoog （MS） and B5 basal medium supplemented with varied concentration of different plant growth-regulators, e.g., α-naphthaleneacetic acid （NAA） and indole-3-butyric acid （IBA） in combination with 6-benzylaminopurine （6-BA） and kinetin. The study showed that shoots could be directly regenerated from hypocotyl explants without the intervening callus phase; MS medium was more suitable for adventitious shoots regeneration. The ability of hypocotyls segments to produce shoots varied depending upon their position on the seedlings. The highest regeneration rate was obtained with hypocotyl segments near to the cotyledon cultured on MS basal medium supplemented with 1.5 mg L^-1 6-BA and 0.05 mg L^-1 NAA （63.64%）. The regenerated shoots were readily elongated on the same medium as used for multiplication and rooted on half-strength MS basal medium supplemented with 1.0 mg L^-1 IBA and 100 mg L^-1 activated carbon. After being transferred to greenhouse conditions, 96% of the plantlets were successfully acclimatized. This regeneration system is applied for genetic transformation now.     

百蕊草无性系建立与瓶外生根研究(英文)

[Objective] The aim of this study is to establish the rapid micro-propagation system in Thesium chinense Turcz.[Method]With stem fragments of wild Thesium chinense Turcz as explants,different culture media were designed to conduct induction culture,strengthening plantlet culture and in vitro rooting.[Result]The optimum medium for inducing clustered shoots was determined to be MS medium appended with 1.5 mg/L 6-BA,0.01 mg/L NAA and 0.3 mg/L 2,4-D;in addition,60 mg/kg ABT was suitable for rooting,by which the percentage of rooted plantlets reached 76.6%.[Conclusion]This study simplified the procedures of tissue culture in Thesium chinense Turcz and enhanced the proliferation rate,providing basis for artificial cultivation and resource protection of Thesium chinense Turcz.     

辣椒胞质雄性不育系(CMS)子叶培养植株再生(英文)

An in vitro shoot regeneration procedure was developed in pepper (Capsicum annuum L.) cytoplasmic male sterility (CMS) lines 9704A and 8214A using cotyledon as explant. The callus and bud cluster derived from cotyledon tissue explants were proliferated on Murashige and Skoog (MS) medium supplemented with different combinations of 6-benzladenine (6-BA), indole-3-acetic acid (IAA), gibberellic acid (GA3) and silver nitrate (AgNO3). From the formula of MS appended with 5.0 mg/L 6-BA, 1.0 mg/L IAA and 5.0 mg/L AgNO3, for the explants callus and bud cluster, the maximum differentiation rates (respectively 100.0% and 58.3%) and average number of adventitious bud from each explant (respectively 18.8 and 13.2) were obtained. The optimum medium combination for the elongation of adventitious bud was determined to be: MS+ 3.0 mg/L 6-BA + 1.0 mg/L IAA+ 5.0 mg/L AgNO3+ 2.0 mg/L GA3, from which the elongation rates of buds from callus and bud cluster were both 100%, and the average number of per explant adventitious bud number reached 6.3 and 5.8, respectively. And all the elongated shoots were successfully rooted on half-strength MS medium supplemented with 0.3-0.5 mg/L IAA.     

泥胡菜的组织培养及高效无性系建立(英文)

[Objective] The research aimed to study the rapid propagation technology and establish effective clone of Hemistepta lyrata Bunge. [Method] With tender stem of Hemistepta lyrata Bunge as material, the conditions needed in calluses induction and differentiation, adventitious bud differentiation and radication, test tube seedling cutting and transplantation were studied. [Result] The results showed that the optimum medium for granulated calluses induction from tender stem was MS+BA 0.3 mg/L+2,4-D 1-1.5 mg/L, for granulated calluses and adventitious bud differentiation was MS+AgNO3 1.5 mg/L +BA 0.4 mg/L +NAA 0.1 mg/L. 1/2 MS+IAA 0.6 mg/L was suitable for test tube seedling rooting and regeneration, and cinder was used as transplantation and cutting substrate. [Conclusion] This study will provide the scientific reference for choosing the feasible medium in tissue culture of Hemistepta lyrata Bunge.     

Somatic Embryogenesis and Plant Regeneration from Leaflets of Fritillaria ussuriensis M.

A method for the production of somatic embryos and subsequent plant regeneration for fritillaria ussuriensis M.is described.Whole leaflet explants,derived from plantlets grown in vitro,formed light yellowith embryogenic calli within one month of culture in the dark.Somatic embryogenesis was obtained after a 28d incubation on MS induction medium supplemented with 2mg/L 2,4-D 0.5mg/L BA,0.5mg/L KT and 500mg/L CH followed by transfer to a second N medium containing 0.5mg/L KT and 100mg/L CH.Somatic embryos were transferred to MS medium with 0.1mg/L NAA placed in the light for regeneration ,After two weeks,mature somatic embryo developed into whole phantlet.     

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 minimal 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.     

不同培养条件对枸杞组培苗玻璃化的影响(英文)

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 minimal 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.     

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.     

Optimization of Agrobacterium tumefaciens-Mediated Immature Embryo Transformation System and Transformation of Glyphosate-Resistant Gene 2mG2-EPSPS in Maize（Zea mays L.）

Since maize is one of the most important cereal crops in the world,establishment of an efficient genetic transformation system is critical for its improvement.In the current study,several elite corn lines were tested for suitability of Agrobacterium tumefaciens-mediated transformation by using immature embryos as explants.Infection ability and efficiency of transformation of A.tumefaciens sp.strains EHA105 and LBA4404,different heat treatment times of immature embryos before infection,influence of L-cysteine addition in co-cultivation medium after transformation,and how different ways of selection and cultivation influence the efficiency of transformation were compared.Glyphosate-resistant gene 2mG2-EPSPS was transformed into several typical maize genotypes including 78599,Zong 31 and BA,under the optimum conditions.Results showed that the hypervirulent Agrobacterium tumefaciens sp.strain EHA105 was more infectious than LBA4404.Inclusion of L-cysteine（100 mg L-1） in co-cultivation medium,and heating of the immature embryos for 3 min prior to infection led to a significant increase in the transformation efficiency.Growth in resting medium for 4-10 d and delaying selection was beneficial to the survival of resistant calli.During induction of germination,adding a high concentration of 6-BA（5 mg L-1） and a low concentration of 2,4-D（0.2 mg L-1） to regeneration medium significantly enhanced germination percentage.Using the optimized transformation procedure,more than 800 transgenic plants were obtained from 78599,Zong 31 and BA.By spraying herbicide glyphosate on leaves of transgenic lines,we identified 66 primary glyphosate-resistant plants.The transformation efficiency was 8.2%.PCR and Southern-blot analyses confirmed the integration of the transgenes in the maize genome.     

Effects of Different Natural Organic Additives on in Vitro Shoot Regeneration of Raphanus sativus L. Var,Beeralu

Abstract： Raphunus sativus L. commonly known as ＂radish＂ is a popular vegetable crop used by people all over the world for its culinary and medicinal properties. Enhancement of in vitro regeneration system for radish variety beeralu is needed to further tissue culture studied. Therefore, the present study was carried out to find out the effects of different organic additives on in vitro shoot regeneration of Radish （Raphanus sativus L.） Var. Beeralu. Hypocotyl explants of aseptic plantlets were cultured on MS basal medium supplemented with 2.5 mg/L BAP （benzyl adenine） and 0.1 mg/L NAA （1-nphthaleneacetic acid） with different natural additives; 20% coconut water, 20% coconut milk, 10% grind spinach leaves, 10% grind potato tubers, 10% grind carrot, 5% rice flour, 10% green gram, 10% grind pumpkin, 10% banana fruit, 10% orange and control （without any additives）. Complete randomized design （CRD） with five replicates was used. After one month the numbers of regenerated shoots were counted and statistical analysis was carried out using the Student Newman-Kuells Means Separation Test of SAS program （9.1.3）. The highest mean number of shoots （12 shoots/explant） from Radish （Raphanus sativus L.） Var. Beeralu observed in MS basal medium with 2.5 mg/L BAP and 0.1 mg/L NAA with 10% orange juice whereas the 2nd highest shoots were obtained with 20% coconut water. The lowest number of shoots （0 shoot/explant） was observed from medium with carrot juice and pumpkin juice, but they induced callus formation. Media with grind spinach leaves, rice flour, green gram, grind potato tubers and banana inhibit the shoot regeneration.     

野百合试管鳞茎诱导与增殖的研究

利用现代生物技术组织培养方法对野百合进行种质保存和快速繁殖研究,对发展野百合的大规模生产及百合育种工作,保护野生资源和生态环境有重要意义。     

野百合试管鳞茎诱导与增殖的研究

以贵州野百合鳞片不同部位为外植体,用3%次氯酸钠进行消毒,以MS为基本培养基,附加不同浓度的激素进行培养。结果表明,用3%次氯酸钠对野百合鳞茎进行外植体进行消毒完全可行,且对操作人员,实验材料和环境都不存在不良影响,价格低廉;最佳的诱导培养基为MS+6-BA1.5mg/L+NAA0.3mg/L;最适合的外植体为百合鳞片基部。用相同的培养基进行增殖培养,25d后,可获得繁殖系数高、生长势好、并有新根生成的试管鳞茎;将直径为1~2cm的试管鳞茎移栽培养,成活率高于90%。该研究得出的方法可在短时间内提供大量的贵州野百合种苗。     

龙牙百合的组织培养和快速繁殖研究

分别以龙牙百合的鳞片、顶芽、茎段和叶片为外植体进行组织培养,结果表明:基本培养基以MS为最适培养基,最易分化出小鳞茎的外植体为秋、冬季的鳞片,不同外植体的最适分化培养基配方不同,鳞片在MS+0.03mg/LNAA上分化效果最好,顶芽和叶片以MS+2mg/L6-BA+0.2mg/LNAA为宜,茎段以MS+0.2mg/LIAA较好。继代增殖培养基MS+0.1mg/LKT+0.15mg/LNAA的效果较好。无根子球和小苗在1/2MS+0.2mg/L6-BA+1mg/LIBA上易生根。     

宜兴百合组培快繁技术的研究

[目的]采用组织培养的方法对宜兴百合(Lilium lancifoium Thumb)进行扩大繁殖研究。[方法]以宜兴百合鳞茎为外植体,在其不同的组织培养阶段采用不同的激素配比,观察培养效果,以建立宜兴百合的组培快繁体系。[结果]宜兴百合鳞茎不定芽诱导的最佳培养基为MS+3.0 mg/L 6-BA+0.5 mg/L NAA,继代培养的最佳培养基为MS+3.0 mg/L 6-BA+0.1 mg/L NAA,生根培养的最佳培养基为1/2MS+0.5 mg/L IBA。[结论]为提高宜兴百合的繁殖系数和加快其产业化生产提供技术支撑。     

卷丹的原球茎诱导及植株再生

以卷丹(Lilium lancifolium)的鳞茎、叶片和珠芽为外植体,建立了卷丹快速繁殖的植株再生体系,探讨了不同外植体、培养基对卷丹原球茎诱导、分化的影响。结果表明,鳞茎是最适合诱导原球茎的外植体,诱导愈伤组织的最佳培养基为MS+1.00 mg/L 6-BA+1.00 mg/L NAA;诱导原球茎的最佳培养基为MS+0.50 mg/L 6-BA+0.10 mg/L NAA+1.00 mg/L 2,4-D;较适合原球茎出芽的培养基为MS+1.00 mg/L 6-BA+0.10 mg/L NAA,较适合不定芽生根的培养基为MS+0.50 mg/L NAA。     

万载百合组织培养快速繁殖的研究

以万载龙牙百合当年成熟的鳞片为外植体 ,接种于不同激素配比的MS培养基上诱导小鳞茎 ,然后 ,转入较高激素配比的继代培养基上诱导不定芽 ,并进行快速增殖。试验结果表明 :在小鳞茎诱导过程中 ,暗处理有利于小鳞茎的生长 ,最佳诱导培养基为 :MS +NAA 0 .5mg/L +CH 30 0mg/L ;快速增殖以培养基MS +NAA 0 .2mg/L +BA 2 .0mg/L +CH5 0 0mg/L的效果最好。百合试管苗容易生根 ,在附加较低浓度IBA或NAA的 1 / 2MS培养基上均能 1 0 0 %生根 ,但以附加IBA 0 .2mg/L及 0 .1 %活性碳的培养基效果最好     

中国柽柳组培快繁技术研究

为了更好的对中国柽柳种质进行保存,以中国柽柳茎段为外植体进行组培快繁研究。结果表明：采取75%酒精浸泡30秒、2%次氯酸钠浸泡3-5分钟和升汞浸泡5分钟为最适宜的灭菌方案。MS+蔗糖20g/L+琼脂6.5g/L为中国柽柳最适初代培养基,MS+蔗糖30g/L+琼脂7g/L+6-BA0.5mg/L+NAA0.1mg/L为其最适增殖培养基,1/2MS培养基为最适生根培养基。     

龙选蕉组培快繁技术

【目的】探讨不同灭菌剂对外植体的灭菌效果及不同激素浓度和组合对不定芽诱导和增殖的影响,建立龙选蕉的高频再生体系。【方法】以龙选蕉吸芽为外植体,以升汞和0.2%次氯酸钠对外植体灭菌;采用0～6.0 mg/L 6-BA、0.1～0.2 mg/L NAA激素组合对不定芽进行诱导或增殖培养。【结果】相对于升汞,次氯酸钠对龙选蕉外植体灭菌的效果更理想,灭菌率达到90.47%,且外植体生长良好,无中毒现象。在不定芽诱导培养基中,随着6-BA浓度的升高,不定芽萌发率呈先增加后降低趋势,其中以添加6-BA 3.0 mg/L的诱导效果最理想,萌发率为66.67%;随着6-BA和NAA浓度的提高,不定芽的增殖系数也随之增加,其中以4.0 mg/L 6-BA+0.2 mg/L NAA为最佳组合,不定芽增殖效果最理想,增殖系数为4.05,芽苗粗壮,叶片颜色正常。【结论】次氯酸钠对龙选蕉吸芽的消毒效果优于升汞,操作简便,易于获得无菌外植体并利于不定芽生长;不定芽诱导最佳培养基为MS+3.0 mg/L 6-BA;不定芽增殖最佳培养基为MS+ 4.0 mg/L 6-BA+0.2 mg/L NAA。     

南川百合资源现状及组培快繁技术研究

研究发现,近年南川百合资源呈明显下降趋势,评估为易危(VU)等级.以鳞茎为外植体进行芽的诱导、增殖培养,筛选出培养配方:MS+6-BA 1.4 mg/L+NAA 0.2 mg/L+白糖30 g/L+琼脂7.0 g/L, pH=5.8;壮苗、生根培养基:MS+NAA 0.3 mg/L+香蕉泥15 g/L+活性炭0.1%+白糖40 g/L, pH=5.8;可以实现南川百合快繁和资源保护.     

香水百合组织培养的试验研究

通过筛选获得最佳的香水百合组培外植体 ,接种于 1 / 2 MS+ 2 ,4- D3 mg/L(单位下同 ) + 6 - BA1+ GA0 .1的诱导培养基上 ,诱导形成不定芽 ,且不定芽迅速形成小鳞茎 ,分化率达 93 .3 3 % .将不定芽接种到 1 / 2 MS+ 6 - BA0 .5的增殖培养基上 ,1个月左右小鳞茎增殖率可达到 1∶ 6 .80 .切取单个小鳞茎 ,考查不同基本培养基、蔗糖、生长素和活性碳浓度对小鳞茎复壮的影响 ,结果在培养基 3 / 4MS+ 6 - BA0 .6+ NAA0 .1+ GA0 .2 中生长的小鳞茎复壮效果最好 ,2 5d左右即可转入1 / 2 MS+ KT0 .3 + NAA0 .5的生根培养基中进行生根 .形成完整的百合小苗移栽至苗圃中 ,可打破休眠 ,使开花期提前