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1.
This study was conducted to evaluate the effects of proline and glutamine on in vitro callus induction and subsequent regeneration and to develop a reproducible and highly efficient plant regeneration protocol in four rice genotypes, viz. Pawana, Jaya, Indrayani and Ambemohar. Considerable variation in response to plant growth regulators and amino acid supplements used was observed in all the four genotypes. Medium supplemented with proline and glutamine was shown to be superior to medium without proline and glutamine. The best callusing from mature embryo was observed on Murashige and Skoog(MS) medium supplemented with 2.0 mg/L 2,4-dichlorophenoxyacetic acid(2,4-D), 500 mg/L proline and 500 mg/L glutamine. Shoot induction was higher in the callus obtained from medium supplemented with 500 mg/L proline and 500 mg/L glutamine. The highest shoot regeneration frequency(83.2%) was observed on MS medium with 2.0 mg/L benzylaminopurine, 0.5 mg/L 1-naphthaleneacetic acid, 500 mg/L proline, and 500 mg/L glutamine in the callus obtained from MS medium supplemented with 2.0 mg/L 2,4-D, 500 mg/L proline and 500 mg/L glutamine. Among the four genotypes, Pawana has the highest regeneration efficiency(83.2%), whereas the regeneration efficiency of the rest three rice genotypes was in the range of 32.0% to 72.3%. This optimized regeneration protocol can be efficiently used for Agrobacterium mediated genetic transformation in rice.  相似文献   

2.
A productive genotype of Irvingia gabonensis were cultured in vitro for induction embryogenic calli, somatic embryogenesis and regeneration of plantlets. Fragments of young leaves were used as primary explants. Callogenesis was initiated by culture of explants during 30 days on Murashige and Skoog medium half strength (MS/2) supplemented with 1-6 mg L(-1) of 2,4-dichlorophenoxyacetic acid (2,4-D). The highest percentage of explants forming calli is 85.1% at 3 mg L(-1) of 2,4-D. Somatic embryos were obtained after a subculture of embryogenic calli during 60 days on MS/2 supplemented with 1-3 mg L(-1) of BAP. The highest percentage of embryogenic calli which differentiates somatic embryos is 63.8 +/- 2.3% at 1 mg L(-1) of 6-benzylaminopurine (BAP). The highest number of somatic embryos per callus which is 43.6 is obtained with 2 mg L(-1) of this phytohormone. When isolated from calli and sub-cultured during 30 days on MS/2 supplemented with 2 mg L(-1) of BAP, somatic embryos germinate with a highest percentage of 83%. The subculture of germinated somatic embryos on the same Basal Medium (BM) supplemented with 4 mg L(-1) of BAP and 2 mg L(-1) of Naphthalene Acetic Acid (NAA) during 80 days gives rise to the plantlets with 82.7 +/- 4.8% of success. With this combination, each plantlet has average length of 5.6 cm, bears 3.3 leaves and 7.2 roots with 1 or 2 pivoting roots. Plantlets acclimatized on a mixture sterilized soil/vermiculite at equal volume survive at 93%. Results of this study constitute a new way for a production of Irvingia gabonensis seedlings with pivoting root and they permit to arrest the difficulties of natural and horticultural reproduction.  相似文献   

3.
The objective of the present study was to develop an effective protocol for optimum callus induction and complete plant regeneration for four varieties of rice (Oryza sativa L.) i.e., Super Basmati, Basmati-370, Basmati-371 and Fakhre Malakand. Calli were induced from mature seed scutelum. The Murashige and Skoog (MS) and Chu's N6 media containing hormone 2, 4-D (2, 4-Dichlorophenoxy acetic acid) in different concentrations were used for callus induction. Fakhre Malakand produced maximum calli on N6 media containing 3 mg L(-1) 2,4-D. while other three varieties showed maximum callus induction on N6 media containing 2.5 mg L(-1) 2,4-D. N6 media was found better than MS media for callus induction. For complete plant regeneration the calli of two varieties i.e., Basmati-370 and Basmati-371 were plated on N6 media containing different concentrations of NAA (1-Naphthalene acetic acid) and BAP (6-benzyl aminopurine). The maximum regeneration frequency (%) was observed on N6 media containing NAA 1 mg L(-1) and BAP 2.5 mg L(-1). It took 27-30 days for the callus to regenerate into a complete plant. Basmati-370 produced 4-7 plantlets per callus whereas Basmati-371 produced 4-8 plantlets per callus with regeneration frequencies of 61 and 69%, respectively.  相似文献   

4.
尾叶桉U6遗传转化再生体系的建立   总被引：1，自引：0，他引：1  
以桉树品种尾叶桉U6叶盘为外植体,选用MS培养基为基本培养基,附加激素6-BA,IAA,IBA,NAAA,通过研究不同激素浓度组合对外植体愈伤诱导及不定芽分化的影响,建立了较好的遗传转化再生体系.结果表明,尾叶桉U6叶盘最适分化培养基为MS 6-BA 2.0 mg/L 2,4-D 1 mg/L IAA 0.2 mg/L 蔗糖30 g/L 琼脂粉5 g/L;小苗的最佳生根培养基为MS NAA 0.2 mg/L mA 0.5 m/L 蔗糖30 g/L 琼脂粉5 g,L.40mg/L卡那霉素可以抑制叶盘的分化;20 mg/L卡那霉素可以抑制再生植株的生根.2种抑菌抗生素中,头孢霉素对叶片再生影响较大;而250 mg/L的羧苄青霉素能有效地抑制农杆菌菌株EHA105的生长,却对尾叶桉叶盘的芽分化影响不大,为适宜的抑菌抗生素.3种农杆菌LBA4404,EHA105,GV3101的菌液浸染桉树叶盘,统计其愈伤组织GUS染色率,以EHA105对外植体的浸染能力最强,达83.3%.  相似文献   

5.
With the objective to promote in vitro callus induction, leaf segments of Achyranthes aspera were inoculated on basal MS medium supplemented with 3.0% sucrose and 0.8% agar with different concentrations of 2,4-D alone and in combination with NAA, BAP, IAA, IBA and Zeatin. The explants were maintained in growth room at 25 +/- 1 degrees C and 16 h light cycle. The best callus induction was obtained with 2,4-D (1.0 and 2.0 mg L(-l)) in combination with NAA (0.5 mg L(-1)). Callus induction and good texture from leaf explant was also observed at 2,4-D with BAP. On these combinations morphologically, light green, soft, compact and non-embryogenic callus (Type III callus) was observed. While morphology of callus and callogenic response was poor at 2,4-D alone or in combination with other hormones at different concentrations.  相似文献   

6.
用水田七成熟种子为材料进行无菌播种,得到无菌苗再用幼叶、叶柄为材料进行组织培养和植株再生。经试验得出各阶段适宜的培养基分别为：(1)诱导愈伤组织：MS＋2.0 mg/L TDZ;(2)诱导芽分化：MS＋1.5 mg/L 6-BA＋0.5 mg/L NAA;(3)生根培养：1/2MS＋0.5 mg /L IBA＋0.1 mg/L NAA。  相似文献   

7.
In this study, in vitro organogenesis of Gladiolus grandiflorus cultivar pink corm segments were evaluated by culturing corm calli in modified MS medium supplemented with 3% sucrose and 0.7% agar with different concentration of BAP (0, 1, 2 and 4 mg L(-1) medium) and NAA (0, 0.5, 1 and 2 mg L(-1) medium) in factorial experiment of Completely Randomized Design (CRD). In order to obtain Gladiolus calli, corm segments (Aprox. 5 x 5 x 1 mm in size) were kept in modified MS medium (Murashige and Skoog, 1962) that was supplemented with 1 mg L(-1) 2, 4-D, 3% sucrose and 0.7% agar. The results showed that increasing the concentration of BAP from 0 to 2 mg L(-1) medium simulated plantlet regeneration but no significantly effect was obtained on shoot and cormel organogenesis between 2 and 4 mg L(-1) BAP concentration in medium. Increasing of NAA content in media without BAP developed rootlet significantly. Interaction results showed that increasing BAP content against decreasing of NAA concentration stimulates the shoot and cormel proliferation.  相似文献   

8.
Summary Shoots, roots and callus were formed from tuber discs of potato, cultivar Désirée, when grown in vitro on the basal medium of Murashige & Skoog (1962) (MS) supplemented with 2,4-D and/or BAP. Callus was formed in MS medium with 1 mg l−1 BAP plus 0.5 mg l−1 2,4-D, callus and roots were formed in MS with 1 mg l−1 BAP plus more than 0.5 mg l−1 2,4-D and shoots were formed directly on tuber discs cultured on MS medium with 1 mg l−1 BAP without the addition of 2,4-D. Nodules produced at the explant surface after the 4th week increased in size following subculture onto the same medium (MS+BAP alone), and 2 to 6 shoots developed from each nodule. After 9 weeks total time in culture, these shoots were excised and transferred as cuttings to MS medium without growth regulators, after which roots developed and plantlets were formed. A histological study of the explants at the sites of nodule formation indicated that the shoots developed from meristematic zones initiated within small outgrowths of tissue similar to those occuring in adventive organogenesis but the presence of shoot and root meristems associated with the same axis suggests the formation of somatic embryos.  相似文献   

9.
小麦不同外植体的组织培养效果研究   总被引：12，自引：5，他引：12       下载免费PDF全文
良好的组织培养效果是提高植物基因转化效率的基础。以山东省近年来育成并大面积推广的10个优良品种（系）为材料，对这些品种的花药、幼穗、幼胚和成熟胚的组织培养效果进行研究，旨在筛选每一基因型最适合于组织培养的外植体类型，为应用基因工程技术进行小麦遗传改良提供基础材料。结果表明，禽伤诱导率和再生成苗率与基因型和外植体类型（花药、幼穗、幼胚和成熟胚）密切相关。8802在花药和成熟胚培养中表现突出，花药出愈率达119．5％，愈伤分化成苗率19．9％;烟农19的幼穗愈伤直接分化成苗率最高，这43．5％；8802、烟农19和潍麦8号三个基因型的幼胚培养效果差异不显著，其愈伤分化成苗率分别为26．3％、24．5％和24．8％。蔗糖浓度在3％～9％之间，对成熟胚的愈伤组织诱导率影响很小。高浓度蔗糖降低愈伤生长速度。在一定蔗糖浓度下，愈伤诱导率与2，4-D浓度密切相关，高浓度的2，4-D对愈伤组织再生不利。MS＋4mg／L2，4-D对于成熟胚的脱分化相对较好，MS和MS＋0．4mg／L NAA＋0．6mg／L KT作为成熟胚的愈伤组织再生培养基，对不同基因型具有一定的适用性。  相似文献   

10.
The influence of media composition on callus induction and subsequent regeneration of Rauwolfia serpentina L. Benth has been studied. High frequency (96.43%) callus induction was obtained when nodal segments from in vitro raised shoots were cultured on MS medium supplemented with 0.5 mg L(-1) BA and 2.0 mg L(-1) NAA. The callus differentiated into adventitious shoots when it was subcultured on MS medium supplemented with 2.0 mg L(-1) BA with 0.2 mg L(-1) NAA. Regenerated shoots were best rooted on half-strength MS medium with 1.0 mg L(-1) each of IBA and IAA.  相似文献   

11.
以贵港报春苣苔的叶片为外植体,研究不同培养基对其不定芽诱导和增殖、愈伤组织诱导与分化以及生根的影响。结果表明,外植体叶片以纵切为宜,不定芽诱导最适培养基为MS+6-BA 4.0 mg/L+IAA 1.5 mg/L,愈伤组织诱导最适培养基为MS+6-BA3.0～5.0 mg/L+2,4-D0.5～1.0 mg/L,不定芽诱导率以及愈伤组织诱导率均为100.00%;愈伤在MS+KT 1.0 mg/L+NAA 0.2 mg/L+potato 30 g/L+banana 30 g/L+apple 20 g/L+coconut juice 100 mL/L培养基上分化系数达12.64;不定芽在MS+ZT 1.0 mg/L+NAA 0.10 mg/L+potato 30 g/L+banana 30 g/L+apple 20 g/L+coconut juice 100 mL/L培养基的增殖系数为8.55;不定芽在3/4 MS+NAA 0.01～0.05 mg/L+活性炭1.0～3.0 g/L培养基上的生根率为100.00%。综上所述,叶片纵切后能通过不定芽途径以及愈伤组织途径建立贵港报春苣苔的组织培养技术体系,在该体系下不定芽增殖系数高、愈伤分化高,组培苗生根好。  相似文献   

12.
Jatropha curcas, a multipurpose shrub has acquired significant economic potential as biodiesel plant. The seeds or pressed cake is toxic due to the presence of toxic substances and is not useful as food/fodder despite having the best protein composition. A simple, efficient, and reproducible method for plant regeneration through direct organogenesis from petiole explants of non-toxic J. curcas was developed using Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ). The best induction of shoot buds (57.61%), and number of shoot buds (4.98) per explant were obtained when in vitro petiole explants were placed horizontally on MS medium supplemented with 2.27 μM TDZ. The Induced shoot buds were transferred to MS medium containing 10 μM kinetin (Kn), 4.5 μM 6-benzyl aminopurine (BA), and 5.5 μM α-naphthaleneacetic acid (NAA) for shoot proliferation and subsequent elongation was achieved on MS medium supplemented with 2.25 μM BA and 8.5 μM IAA. The elongated shoots could be rooted on half-strength MS medium with 15 μM IBA, 11.4 μM IAA and 5.5 μM NAA with more than 90% survival rate.  相似文献   

13.
Broccoli (Brassica oleracea var. italica) is an important nutritionally rich vegetable cole crop grown in the world. Environmental stress, pests, and diseases cause enormous yield losses because of a limited gene pool. Genetic manipulation is becoming an important method for broccoli improvement. The objective of present study was to evaluate the potency of thidiazuron (TDZ) as a plant growth regulator in evoking morphogenic responses in leaf and petiole explants of broccoli. An efficient, reproducible, and high frequency plant regeneration protocol has been standardized in broccoli cv. Solan green head. Leaf and petiole explants were cultured on Murashige-Skoog (MS) medium, supplemented with a wide range of TDZ concentrations. The following treatments were designed for efficient in vitro shoot regeneration: TDZ alone, TDZ with adenine, TDZ with naphthalene acetic acid (NAA), and TDZ with indole acetic acid (IAA). Among the 36 combinations of growth regulators used, the highest percentage of leaf explants producing shoot (89.25%) was recorded on MS medium containing 1.0 μM TDZ and 0.107 μM NAA. The multiple shoot regeneration response of petiole explant producing shoots (91.55%) was obtained on MS medium containing 2.0 μM TDZ and 0.107 μM NAA. Shoot multiplication and elongation were obtained on the same medium. For root regeneration in in vitro regenerated shoots, different concentrations of NAA were applied. High frequency (100%) root regeneration response with healthy and vigorous roots was observed on MS medium supplemented with 0.54 μM NAA. The regenerated plantlets with well-developed shoots and root system were transferred to pots containing cocopeat and successfully acclimatized. We recommend 1.0 μM TDZ with 0.107 μM NAA and 2.0 μM TDZ and 0.107 μM NAA combinations for adventitious shoot regeneration from leaf and petiole explants in broccoli cv. Solan green head respectively. This is the first report on high frequency organogenesis from leaf and petiole explants of broccoli cv. Solan green head using thidiazuron.  相似文献   

14.
[目的]筛选适宜君子兰叶片离体的条件。[方法]选取君子（Clivia miniata Regel）品种“油匠”的叶片为外植体进行离体培养,在MS培养基中附加不同浓度的2,4～D、BA、NAA和KT,对外植体采用不用时间的低温预处理,并对接种后的外植体分别进行不同时长的暗培养,研究不同培养条件对外植体愈伤组织诱导和分化的影响。[结果]叶片诱导分化的最佳培养基为MS＋BA2．0mg／L＋NAA3．0mg／L＋KT1．0mg／L＋蔗糖3．0％,诱导与分化率分别为59．2％和55．2％;暗培养10d后外植体诱导与分化率较高,为l3．8％和25．0％。[结论]培养基中加入活性炭对叶片诱导与分化不利;接种前低温预处理1d,叶片诱导与分化率较高,分别为3313％和25．0％、  相似文献   

15.
In order to investigate the genetic variation in tissue culture response and to find the cultivars with high regeneration ability for genetic transformation, twenty-five turf-type tall fescue ( Festuca arundinacea Schreb.) cultivars, including many elite ones released recently, were evaluated for their callus induction and plant regeneration responses. Callus induction was initiated from mature seeds on a Murashige and Skoog (MS) medium containing 9·0 mg l–1 2,4-dichlorophenoxyacetic acid (2,4-D). Induced calli were subcultured on the same medium with 2·0 mg l–1 2,4-D and then transferred to a MS medium supplemented with 2·5 mg l–1 6-benzylaminopurine (BAP) for plant regeneration. Significant differences were observed among the twenty-five cultivars in both callus induction and plant regeneration ( P  < 0·001). Callus induction rate of viable seeds varied from 4·4% to 51·9%. Callus regeneration rates ranged from 16·7% to 58·8%. Overall regeneration rates (number of regenerated calli over number of cultured viable seeds) ranged from 1% to 22%. Approximately 94% of the regenerants were green plantlets.  相似文献   

16.
青葙愈伤组织诱导研究   总被引：1，自引：0，他引：1  
目的 筛选诱导青葙愈伤组织的最佳外植体和最佳诱导条件.方法 选取青葙饱满成熟种子培养无菌苗,以其叶片和茎段为外植体,分别接种于添加NAA、6-BA、2,4-D组合的MS培养基中诱导愈伤组织,通过正交试验筛选出诱导愈伤组织的最佳外植体和培养基.结果诱导愈伤组织的最佳外植体为试管苗茎段,MS+0.5mg/L 6-BA+0.5mg/L NAA是愈伤组织诱导的最佳培养基,其愈伤组织诱导率为92.5％.结论该研究建立的青葙的愈伤组织培养方法,为青葙的快速繁育体系及工厂化生产提供了技术参考.  相似文献   

17.
以桂竹香花柄，花瓣，花丝为外植体，研究不同激素对诱导愈伤组织和不定芽的影响，研究结果表明，4种外植体均能形成愈伤组织，但愈伤组织的发生率，发生部位及形态等表现差异，花柄愈伤组织继代MS＋2，4－D＋BA培养基，可分化产生不定芽，出芽率78％－82％，再生小苗在1／2MS＋NAA 0．5mg/L培养基中生根形成再生植株。  相似文献   

18.
对海南粗榧茎段外植体灭菌和愈伤组织诱导实验研究表明,以海南粗榧温室扦插苗的嫩茎作为外植体,70％酒精1min,0．1％氯化汞溶液12min表面消毒处理效果最好;最适愈伤组织诱导培养基为MS＋0．5mg／LNAA＋0．5～1．0mg／LBA＋2．5～4．0mg／L2,4～D。  相似文献   

19.
苗永美  简兴  何华奇  汪娜  唐靖 《热带作物学报》2010,31(11):1975-1980
为建立白头翁再生体系,以白头翁主根的切段和试管苗叶片为外植体,比较2种外植体离体培养差异,探讨不同植物生长调节剂对不定芽和愈伤组织诱导、愈伤增殖和再分化及芽苗生根的影响。结果表明:叶片可以通过愈伤组织再分化和直接产生不定芽2种途径建立再生体系,而根只诱导出了愈伤组织,增殖培养中逐渐褐化死亡;在含6-BA培养基上,叶块死亡,而根只诱导出少量愈伤组织;叶片诱导不定芽的培养基为MS+TDZ 0.3 mg/L+NAA 0.1 mg/L;愈伤组织增殖的培养基为MS+TDZ 0.2 mg/L+2,4-D 0.2 mg/L;再分化时,需要转接到TDZ和NAA组合的培养基上,其中处理组合TDZ 0.3 mg/L+NAA0.1 mg/L的再分化率达100%;生根培养基为1/2MS+NAA 0.2 mg/L+IBA 0.2 mg/L+蔗糖20 g/L。不同外植体离体培养存在差异,叶片较根易培养;TDZ对白头翁叶片培养效果较好,2,4-D对愈伤组织的诱导能力较强,而NAA适合于不定芽分化,NAA与IBA组合使用生根效果较好。  相似文献   

20.
荔枝"妃子笑"品种花药培养及其体胚发生   总被引：2，自引：0，他引：2  
以荔枝(LitchichinesisSonn.)品种“妃子笑”为试材,研究其花药离体培养及植株再生的影响因素。结果表明,荔枝花药在MS 2,4-D2mg/L NAA0.2mg/L以及含50g/L蔗糖的培养基上诱导胚性愈伤组织效果较好,把胚性愈伤组织转移到MS BA1mg/L NAA0.5mg/L,谷氨酰胺500mg/L,蔗糖50g/L分化培养基上培养1个月后,体胚大量萌发,再将成熟体胚转移到附加500mg/L谷氨酰胺的MS无激素培养基上,培养1￣2个月后,能再生成完整植株。  相似文献   

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