共查询到20条相似文献，搜索用时 22 毫秒
1.
Vipasha Verma Kamlesh Kanwar Mahak Tufchi Monika Kashyap 《Journal of Crop Science and Biotechnology》2014,17(1):1-10
Three different regeneration systems, viz. regeneration through callus cultures using embryonic explant, direct regeneration using shoot bud/nodal segments as explant and regeneration through cell suspension culture using cotyledonary explant (for the induction of transgenic callus for suspension culture) were evaluated to see their effect on transfer of Cry1A(b) gene to Punica granatum L. cv. Kandhari Kabuli through Agrobacterium mediated transformation. Pre-conditioning and co-cultivation durations had a marked effect on transformation frequency of different explants. Out of different explants used (embryo, shoot bud, and cotyledon) for different regeneration systems cotyledonary explant showed highest putative transformation frequency (13.54%) inducing callus on selective medium for carrying out cell suspension culture to regenerate transgenic shoots. Despite of the highest transformation frequency obtained from the cotyledon explant, the plating efficiency of the transgenic cells generated through the transgenic callus (callus formed from the cotyledonary explant) during cell suspension culture was found to be very low (0.7%). Thus the plating efficiency has also played worth mentioning role in the regeneration of transformants following cell suspension culture. Among the three regeneration systems, regeneration through callus cultures using embryonic explant was found to be best for regeneration of transformants. The highest per cent regeneration of 23.33 was obtained from the putative transgenic embrogenic calli. Successful genetic transformation in the transformed plantlets was confirmed by PCR analysis. The transformation system thus developed is valuable and may be used to produce insect resistant trees. 相似文献
2.
为建立以玉米幼苗为外植体的高效再生体系, 克服幼胚取材的限制, 本研究选取萌发3 d的玉米幼苗作为供体材料, 研究其不同部位的再生能力。分别研究了种子萌发时的光照条件, N6培养基中Ca2+浓度, 及6个不同基因型对芽尖和根尖愈伤组织诱导率和分化率的影响。结果显示, 黑暗条件下萌发的幼苗, 其芽尖和根尖更适合作为外植体, 诱导培养基中添加终浓度为5 mmol L-1的CaCl2 时的愈伤组织诱导率最高, 不同基因型的玉米愈伤组织的诱导率和分化率存在差异, 但影响趋势相同, 且同一基因型的不同外植体形成的愈伤组织形态相似, 分化率也相似。最后结论是黑暗条件下萌发3 d的齐319和鲁原92的幼苗, 可以代替玉米幼胚作为外植体用于组织培养研究。 相似文献
3.
辣椒子叶离体培养再生植株 总被引：26，自引：1，他引：26
随着收集到的８个商品辣椒品种的子叶外植体在ＰＤ培养基上形成芽丛，转移到ＰＥ培养基上伸长为具２－３个节的茎芽，茎芽在ＰＲ培养基上生根，得到了再生植株。芽分化率达１００％，移栽成活率１００％。多植体切除芽后回到ＰＤ培养基上的反复操作可重复诱导芽的分化，提高繁殖系数。 相似文献
4.
Aneesha Singh 《Journal of Crop Science and Biotechnology》2018,21(1):89-94
Although several studies have been made on the micropropagation of Jatropha curcas using agar base mediums, none of them have been by using liquid medium systems. The effects of explant type and temporary immersion system (test tube, jar with filter paper boat, and growtek bioreactor) on the micropropagation of J. curcas were studied. The explant type influenced shoot quality, multiplication coefficient (MC), and rooting. Leaf explant produced more and longer shoots than nodal explant. Use of filter paper (FB) boat prevented hyperhydricity and allowed proliferation of nodal explants cultured in liquid MS (Murashige and Skoog) medium supplemented 6-benzylaminopurine (BAP) and Kinetin (KN). The best shoot bud induction (92.1±3.1%) was achieved in liquid MS medium supplemented with 2.0 mg/L KN. Leaf regeneration efficiency was compared in growtek bioreactor and in jar containing liquid MS medium supplemented with 0.5 mg/L Thidiazuron (TDZ). The best shoot bud regeneration (78.7±2.1%) was obtained in growtek bioreactor. Shoot buds achieved from nodal segment and leaf were subcultured on filter paper boats in jar and bioreactor containing liquid MS medium supplemented with BAP, Indole butyric acid (IBA), Indole-3-acetic acid (IAA), and KN. Best shoot proliferation and elongation was obtained in filter paper boats containing liquid MS medium supplemented with 1.5 mg/L BAP, 0.5 mg/L IAA, and 0.2 mg/L KN. The number of multiple shoot buds was higher in leaf explants as compared to nodal explants and the highest number of multiple shoot buds was recorded from leaf explants. Up to 76.4% rooting efficiency was obtained when the shoots were ex vitro rooted. The generated plants well established in the nursery and grew normally in outdoor conditions. The protocol has good potential for application in large-scale propagation of J. curcas using liquid medium. 相似文献
5.
用根癌农杆菌介导法转化大豆萌动子叶节细胞 总被引：2，自引：0，他引：2
利用根癌农杆菌转化萌动大豆成熟子叶节获得了高频率的转基因大豆。从萌发12～16 h的大豆种子切取半片种子作为目标组织,对其子叶节组织进行伤处理后,接种于含有pGB载体的LBA4404农杆菌溶液。pGB载体含有除草剂（phosphinothricin, PPT）抗性基因bar和绿色荧光蛋白基因sgfp。将接种的外植体分别在含有3或5 mg/L PPT的芽诱导培养基、3 mg/L PPT的芽伸长培养基及2～4 mg/L PPT的根诱导培养基上进行了筛选。利用萌发5 d的外植体进行PPT浓度筛选的结果表明,芽诱导、芽伸长及根诱导阶段的PPT浓度分别为5、3及3 mg/L时,转化效率达到最大值,为5.3%。PCR和Southern杂交结果证实了外源基因稳定地整合在大豆基因组中。Northern杂交和GFP分析结果表明被整合的外源基因在大豆细胞中得到了稳定的表达。成熟植株对体外喷洒100 mg/L PPT溶液产生抗性。转化效率达8.9%。 相似文献
6.
Satyajit Saurabh Dinesh Prasad Ambarish S. Vidyarthi 《Journal of Crop Science and Biotechnology》2017,20(2):81-87
The pointed gourd (Trichosanthes dioica Roxb.) is an important cucurbit reported for its medicinal value, therapeutic potential, and as a popular delicacy (especially in Indian cuisine). Being nutritive and desirous, it has potential to feed the nations and addresses their nutritional security and economic prosperity. The plant is usually vegetatively propagated and cultivated for fruits during summer and rainy seasons. The limited supply of planting material, limits cultivation and production. The present study was in anticipation for direct organogenesis, callus induction, and somatic embryo formation from leaf and node explants of T. dioica Roxb. In this study, the MS medium supplemented with 0.5 mg/L BA and 0.5 mg/L 2,4-D was found to be most efficient for callus induction, followed by 0.5 mg/L Kn and 0.5 mg/L 2,4-D. The embryogenic callus was developed by sub-culturing of node callus in the same media. The scanning electron microscopic (SEM) analysis revealed the presence of embryogenic cell clusters having globular embryos, which were found irresponsive to develop further. Through direct organogenesis, the node explants have responded to produce true-to-type plants for propagation. It was observed that MS supplemented with 1.0 mg/L BA was efficient for shoot proliferation, and 0.5 mg/L IAA was found more efficient for root development. Notably, the plant remains unexplored in its potential for improvement involving molecular breeding and tissue culture. These results may be effective to produce genetically stable plants on a large scale and aid the genetic improvement of pointed gourds. 相似文献
7.
Zeynab Shahabzadeh Bahram Heidari Reza Faramarzi Hafez 《Journal of Crop Science and Biotechnology》2013,16(4):263-268
An important feature of A. rhizogenes-induced hairy roots is their unique ability for investigation of gene function and production of secondary metabolites such as diosgenin in fenugreek. In order to evaluate the transformation frequency and the efficiency of transgenic hairy root induction, leaf and stem explants from two fenugreek ecotypes, Karaj and Bushehr, were infected with three concentrations of OD600= 0.8, 1.2, and 1.6 of A. rhizogenes strain K599 harboring a GFP gene. Regardless of ecotype, the ability of stem explants for the induction of hairy roots (8.09) and the transformation frequency (81.3%) was higher compared with leaf explants with the values of 5.97 and 71.88%, respectively. The number of transgenic GFP-positive hairy roots ranged from 4.2 to 13.5 in the Karaj ecotype and 3.8 to 9.9 in Bushehr. The highest transgenic hairy root (8.76), the transformation frequency (79.76%), and the growth rate of transgenic roots (0.77 d?1) were obtained from infection with K599 at OD600= 1.2, while the lowest belonged to the bacterial concentration of OD600=1.6. Although the ecotype Bushehr had lower total roots (7.53) and transgenic hairy roots (6.08), it showed higher transformation frequency (79.56%) than Karaj (73.63%). Therefore, the results indicate the importance of genotype, type of explant and bacterial concentration in breeding for induction of transgenic hairy roots and consequently, production of secondary metabolites in fenugreek. 相似文献
8.
菜豆组织培养及遗传转化研究进展 总被引：3，自引：1，他引：2
笔者概述了近年来国内外菜豆组织培养及菜豆遗传转化研究进展。重点阐述了菜豆组织培养过程中外植体选择、分生组织培养、愈伤组织的诱导、不定芽的诱导、生根培养及移栽等方面的研究成果。同时总结了农杆菌介导法和基因枪法2种遗传转化方法在菜豆转基因方面的应用情况。指出了基因型的差异是菜豆组织培养及遗传转化的主要限制因素,因此如何建立一套不存在基因型差异的菜豆再生体系,是今后菜豆组织培养和遗传转化的研究重点。 相似文献
9.
Data presented herein provides a rapid and efficient method for Agrobacterium rhizogenes-mediated genetic transformation of Arnebia hispidissima for hairy root cultures as well as for enhancing Shikonin production. Etiolated explants viz. shoot tip, nodal, leaf and
internodal segments were co-cultivated with Agrobacterium rhizogenes for induction of hairy root. Among the various explants employed, leaf explant showed maximum 70.7% response followed by
shoot tip 48.3%, nodal segment 38.7% and internodal segment 9.3%. Integration of Ri plasmid rolB gene in the transformed hairy root cultures was confirmed by PCR analysis using forward (FrolB) and reverse (RrolB) primers of rolB gene resulting in the amplification of 0 ∼ 0.8 kb fragments. Medium compositions have been optimized for in vitro induction
of Shikonin in hairy root cultures of Arnebia hispidissima. Hairy roots on hormonefree MS medium showed red spots in the older part of the tissues which turned white after a second
subculture. Whereas hairy roots cultured on RC medium showed faster growth and produced large amount of Shikonin. The Shikonin
content in transformed hairy root culture was estimated by recording absorbance at 620 nm and quantified against authentic
sample of Shikonin. Shikonin content was estimated to be 0.85 mg g−1 fresh weight of tissue at the end of the 50 days of culture. The results presented herein will help to design strategies
for bridging the gap between ever increasing demand and supply of raw products necessary for obtaining Shikonin for cosmetic,
dyeing, food, medicinal, and pharmaceutical industries. 相似文献
10.
11.
Two different protocols for in vitro regeneration of cassava using zygotic embryos and nodal axillary meristems have been developed. In both cases, buds were regenerated directly from excised explants without an intervening callus phase after a two-step culture procedure. In cotyledonary explants derived from zygotic embryos, prolific shoot formation occurred within 2—3 weeks on MS medium supplemented with 0.5—5 mg/1 BAP alone or in combination with 0.1 mg/1 NAA. Nodal explants with axillary meristems derived from aseptically grown seedlings or stem cuttings were used to initiate a round compact bulb-like structure on MS medium containing 10 mg/1 BAP. These latter structures, when cultured on MS medium supplemented with 0.1 mg/1 NAA, 1 mg/1 BAP and 0.1 mg/1 GA3, produced multiple shoots. Somatic embryos isolated at the globular/torpedo stage from zygotic embryo explants were also capable of multiple shoot production on medium with 1 mg/1 BAP. Rooting of regenerated shoots exceeded 95 % in phytohormone-free MS medium. No change in their ploidy levels was observed. Therefore, the protocols developed should be of use in the particle gun and Agrobacterium-mediated genetic transformation of cassava. 相似文献
12.
13.
Sumita Acharjee Pratap Jyoti Handique Bidyut Kumar Sarmah 《Journal of Crop Science and Biotechnology》2012,15(4):311-318
Regeneration has been achieved in blackgram (Vigna mungo) using thidiazuron (TDZ) in the culture medium. The explanted cotyledon with wounded embryonic axes produced the highest number (9.75–10.45) of healthy, elongated shoots when cultured on shoot bud regeneration medium (SRI) composed of 2 μM BAP, 2 μM KIN, 2 μM TDZ, and 0.5 μM NAA followed by multiple shoot regeneration (SRII) medium containing 2 μM BAP, 2 μM KIN, and multiple shoot elongation (SE) medium (0.5 μM of BAP + 0.5 μM of KIN). The presence of TDZ in combination with BAP and NAA in the SRI medium for one sub-culture cycle (10–14 days) significantly increases formation of multiple shoot buds per explant. Independent, healthy shoots obtained were selected for both in vitro rooting and grafting. Establishment of plantlets in the soil was highest (80–100%) in the case of in vitro rooted compared to grafted shoots (40%). The protocol appears to be competent to Agrobacterium-meditated transformation with ‘gus’ as a reporter gene. PCR analysis of the T0 and T1 progenies showed the presence and transmission of the transgene. We document here the regeneration and transformation of blackgram using cotyledons with wounded embryonic axes and the protocol appears to be suitable for genetic transformation of blackgram. 相似文献
14.
The present study describes the procedure for micropropagation of Dracocephalum kotschyi L. using shoot tips from in vitro-germinated plants. The best response was observed for shoot tips on MS medium containing 5 mg 6-benzylaminopurine L?1 and 0.2 mg 1-naphthaleneacetic L?1 acid. Regeneration for other types of the explant hypocotyls and cotyledons did not show satisfactory results so that the explants did not develop into normal shoots and in turn developed into the calli after 12 days of culture. Histochemical analysis showed that only the shoot tip revealed a direct induction of more teratological protuberances that arise around the cut end of the explants. Elongation of shoot buds was obtained on MS medium containing 1 mg BAP L?1 + 0.5 mg IBA L?1. Regenerated shoots rooted best on the same medium of elongation. After hardening, the rooted plants were transferred to the greenhouse where they grew, matured, and flowered normally with a survival rate of 95%. We concluded that the present protocol can be efficiently used for mass propagation of Dracocephalum kotschyi. 相似文献
15.
沙田柚遗传转化再生体系的建立 总被引：4，自引：1，他引：3
以沙田柚的实生苗不同外植体为对象,进行了沙田柚遗传转化再生体系建立研究。研究结果表明：以实生苗上胚轴作为外植体,以MS无机盐＋100 mg/L肌醇＋0.2 mg/L维生素B1+1 mg/L维生素B6+1 mg/L烟酸+3％蔗糖＋0.8％琼脂（pH5.8）＋5 mg/L 6-BA为不定芽诱导培养基,暗培养7 d后,转移至光/暗周期16/8 h的条件下培养,不定芽的分化率高达91.1%;不定芽在1/2MS基本培养基＋3％蔗糖＋0.7％琼脂＋1.5 mg/L IBA＋0.2%活性炭的生根培养基,生根率高达80％;以卡那霉素作选择剂时,选择浓度为50mg/L。 相似文献
16.
Nitish Kumar K. G. Vijay Anand Muppala P. Reddy 《Journal of Crop Science and Biotechnology》2010,13(3):189-194
Jatropha curcas, the energy plant has attained great attention in recent years because of its biodiesel production potential; however, oil and deoiled cakes are toxic. A non-toxic variety of J. curcas is reported from Mexico. A simple and efficient protocol has been developed for plant regeneration using cotyledonary petiole explants of non-toxic variety of J. curcas. The percentage of induction of shoot buds (59.11%), and the number of shoot buds (5.01) per explant was achieved on Murashige and Skoog’s (MS) medium supplemented with 2.27 μM thidiazuron (TDZ). These induced shoot buds multiplied when subcultured on MS medium supplemented with 10 μM kinetin (Kn), 4.5 μM 6-benzyl aminopurine (BAP), and 5.5 μM α-naphthaleneacetic acid (NAA) for 4 weeks and subsequent elongation achieved on MS medium supplemented with 2.25 μM BAP and 8.5 μM indole-3-acetic acid (IAA). Shoots more than 2 cm long were harvested and cultured on MS medium containing different concentrations and combinations of IBA, IAA, NAA, and 0.25 mg L?1 activated charcoal, and 19.91% rooting was achieved in 15 μM IBA, 5.7 μM IAA, and 16.5 μM NAA after 4 weeks with more than 90% survival rate. 相似文献
17.
Sudipta Shekhar Das Bhowmik Adreeja Basu Lingaraj Sahoo 《Journal of Crop Science and Biotechnology》2016,19(2):157-165
A simple and efficient protocol for direct in vitro shoot multiplication and plant regeneration was established for an important aromatic medicinal plant, Alpinia calcarata. Preinduction of rhizome segments in medium containing 8.8 μM 6-benzylamino purine (BAP) rescued the buds from dormancy in 60% of the cultures. An average of 6.2 shoots were produced from rhizomatous bud explants on Murashige and Skoog (MS) medium supplemented with 5 μM BAP, 10 μM kinetin, and 2.5 μM α-Naphthalene acetic acid (NAA). The mother cultures retained their morphogenetic potential upto four subcultures and a maximum of 1.77-fold increase in shoot multiplication was recorded after the 3rd subculture. Rooting was simultaneously induced during subculture on shoot multiplication medium eliminating an additional step for rooting induction. Rooted plantlets were successfully acclimatized in pots in the greenhouse and subsequently established in the experimental garden without any visible symptoms of wilting and necrosis. The genetic fidelity of regenerated plants was evaluated by adapting to two PCR-based DNA marker techniques, Random Amplified Polymorphic DNA (RAPD) and Inter Simple Sequence Repeats (ISSR) which detected no variability in the in vitro multiplied plantlets of A. calcarata. This efficient method of clonal multiplication may be useful for commercial scale multiplication, and in situ and ex situ conservation of elite germplasm of A. calcarata. 相似文献
18.
不同基因型对花生胚小叶植株再生的影响 总被引：1，自引：1，他引：0
以不同类型18个花生品种成熟种子的胚小叶为外植体,对不定芽诱导及植株再生进行了研究,旨在为花生遗传转化和离体诱变等提供培养方法。将胚小叶外植体培养在添加1 mg/L NAA和6 mg/L BAP的诱导培养基上,4周后转移到添加4 mg/L BAP的培养基上进行培养。结果表明,所有的供试品种芽点诱导率均高于60%,但5大类型间及品种间(62.2%~95.5%)存在显著差异,龙生型平均诱导率最高(87.2%),其次是珍珠豆型(81.5%),多粒型最低(63.1%)。将形成芽点的外植体转移到添加4 mg/L BAP的培养基上后,部分外植体从芽点上分化出不定芽,继续培养不定芽伸长并再生植株。植株再生率也存在显著性差异,最高的是珍珠豆型(83.5%~97.1%),最低是多粒型(14.8%~22.0%)。 相似文献
19.
以棉花茎尖分生组织为受体进行基因枪轰击转化的研究 总被引：3，自引：1，他引：3
以棉花茎尖分生组织为受体进行基因枪遗传转化,在不受宿主基因型范围和操作时间的限制等方面优于农杆菌介导法和花粉管通道技术。影响棉花茎尖分生组织基因枪遗传转化率的因素较为复杂,轰击前,外植体制备需考虑茎尖分生组织的取材时间、下胚轴保留的长度、外源生长调节剂及活性炭的使用量,以保证其正常生长;轰击后,抗生素筛选的浓度、筛选培养基的转换时间和抗性苗的壮苗也会影响转化效率;此外,DNA和钨粉或金粉的质量、基因枪参数及操作等对转化效果都有一定的作用。除对影响茎尖分生组织基因枪转化率的因素分析以外,还对近年来在棉花茎尖分生组织遗传转化中的最新动态进行了综述,并对目前存在的问题加以讨论。 相似文献
20.
几个温敏不育水稻品种组织培养特性的研究 总被引：3，自引：2，他引：1
以我国特有的温敏不育系籼稻品种安农S-1,及其转育的不育系安湘S和香125S为实验材料,对愈伤组织的诱导、继代和再生体系的建立进行了研究.结果表明:不同水稻品种和不同的外植体种类愈伤组织的诱导存在比较明显的差别:对成熟种子来说,安农S-1的愈伤组织诱导率最高;而对幼胚而言,香125S的诱导率最高.不同的水稻品种所适用的培养基不同,形成的愈伤组织的状态也存在差别,香125S所适用的培养基种类比较广泛,形成的胚性愈伤组织状态最好,也最容易形成.在愈伤组织的分化阶段,添加3 mg/L 6-BA 1 mg/L NAA分化成苗率最高;随继代时间的延长,愈伤组织的再生率下降,形成的幼苗的状态也存在明显的差别. 相似文献