共查询到20条相似文献，搜索用时 15 毫秒
1.
Concentration effects of dicamba on shoot regeneration in wheat 总被引：1，自引：0，他引：1
Shoot formation and regeneration rates of calli derived from immature embryos of three spring wheat varieties were investigated to evaluate the influence on wheat regeneration of different concentrations of dicamba (0.5, 0.1 and 0.02 mg/l) in the regeneration medium. It was found that dicamba concentrations much lower (0.02 and 0.1 mg/l) than the levels reported previously (1 and 0.5 mg/l) induced significantly more shoots per callus in all three varieties, while the enhancement in regeneration rate was variety dependent. Overall, a dicamba concentration of 0.02 mg/l was more favourable for wheat regeneration. 相似文献
2.
Optimization of mature embryo-based high frequency callus induction and plant regeneration from elite wheat cultivars grown in China 总被引：2，自引：0，他引：2
We have developed an efficient procedure for plant regeneration of elite wheat cultivars using mature embryos. Firstly, we established the optimal combination of basal media, inoculation method and pretreatment method using biostatistical methods. The results indicated that the combination of MS medium and longitudinally bisected mature embryos showed the highest culture efficiency, whereas the pretreatment method had no significant effects on callus induction or plant regeneration. A 70% primary callus induction rate was achieved on MS medium containing 2 mg/l 2,4‐d for all tested cultivars. Primary calli were then transferred onto the subculture medium to initiate embryogenic calli. Supplementation of the subculture medium with the appropriate combination of phytohormones (2.0 mg/l 2,4‐d , 0.5 mg/l BA and 0.1 mg/l NAA) significantly enhanced embryogenic callus production. The addition of AgNO3 (10 mg/l) in regeneration medium promoted plant regeneration, whereas CuSO4 stimulated root formation. The use of this protocol achieved successful plant regeneration in eight tested cultivars. The culture efficiency ranged from 15.3% to 36.8%, suggesting this regeneration system may be an effective alternative for wheat genetic transformation. 相似文献
3.
Summary When subjected to micropropagation by tissue culture, the two reputed cultivars of date palm (Phoenix dactylifera L.); Bou-Sthammi noire, resistant to Bayoud disease and Bou-Feggous, of high fruit quality, give rise to three types of calli, called white and root-forming callus, hyperhydric and degenerating callus and friable and embryogenic callus. All explant sources, calli and germinated embryos were analysed by denaturing polyacrylamide gel electrophoresis (SDS-PAGE) for acid soluble protein composition. Phenol-oxidizing enzymes; peroxidase and polyphenoloxidase, were also, evaluated and the isoforms separated by polyacrylamide gel electrophoresis. When compared with the explant and germinated embryos, embryogenic calli of the two date palm cultivars could be identified by a concentrated polypeptide of molecular weight 27 500 and polypeptides of molecular weights 70 000 and 11 500. Hyperhydric and degenerating callus contained the polypeptide exhibiting the molecular weight 32 000. Embryogenic calli showed high levels of soluble, ionically and covalently bound peroxidases. The soluble acidic isoperoxidase of R
f
0.60, revealed in these calli and germinated embryos could be a marker of the two tissues. White and root-forming calli of Bou-Feggous cultivar were typified by soluble acidic isoperoxidases with high mobility (R
f
0.75) and anodic ionically wall-bound polyphenoloxidases similar to those of the explant sources. Polyphemoloxidase activities detected in calli and embryos were very low when compared with those of explants. Used as an early test to screen embryogenic calli of date palm, acid soluble proteins, peroxidase and polyphenoloxidase data could lead to introduce lightening and economy in the tissue culture technique. 相似文献
4.
Sathish Sundararajan Balaji Sivaraman Venkatesh Rajendran Sathishkumar Ramalingam 《Journal of Crop Science and Biotechnology》2017,20(3):175-183
This research was undertaken to find an efficient tissue culture system and Agrobacterium-mediated genetic transformation method for recalcitrant indica rice cultivars. For this, mature seeds of commercially important indica rice varieties, ASD16, ADT43, IR 64, and Pusa Basmati were cultured on MS and N6 medium supplemented with 2 mg l-1 2, 4-D + 30 g l-1 sucrose. The calli grown in N6 medium showed better friability and embryogenic response. Out of the four varieties tested, ASD16 and IR64 showed better callusing and embryogenic capacity as compared to ADT43 and Pusa Basmati. For genetic transformation studies, embryogenic calli of all the cultivars were co-cultivated with the Agrobacterium tumefaciens strain LBA 4404 harboring the binary vector pCambia 1305.1 with GUS gene. GUS assay was performed for the putative transformed calli and its activity was found to be qualitatively higher in ASD16 and IR64 than the other two varieties. The best responsive ASD16 transformed calli was regenerated and the putative transgenic lines were regenerated. ASD16 transformed calli were confirmed by GUS assay. PCR analysis confirmed the presence of both GUS and HPT genes in ASD16 transgenic lines. 相似文献
5.
玉米成熟胚胚性愈伤组织的诱导、高频再生及转化的研究 总被引：4，自引：0，他引：4
以玉米自交系CML295、CML304和18-599R的成熟胚为外植体, 结合幼胚离体培养方法, 探讨并优化了成熟胚来源的胚性愈伤组织诱导及继代培养方法。对其愈伤组织的形态和组织切片的研究结果显示, 继代过程可产生良好的Ⅱ型胚性愈伤组织。在分化培养中分别获得68.6%、75.4%和84.8%的高频再生率, 每愈伤组织块成苗数分别为2.45、2.43和2.75。利用基因枪法转化pCAMBIA1301质粒后的GUS瞬时表达效率分别为57.9%、62.5%和73.1%, 转化pCAMBIA1303质粒后检测GFP的瞬时表达效率分别为23.3%、40%和45.5%。以上3种基因型成熟胚来源的愈伤组织转化率与其对应的幼胚来源的胚性愈伤组织转化率相似。这一技术体系为玉米的遗传改良和功能基因组研究提供了重要的技术平台。 相似文献
6.
Daniel S. Moura Francisco J. Zapata-Arias Akihiko Ando Augusto Tulmann Neto 《Euphytica》1997,94(1):01-05
A plant regeneration system from rice protoplasts using calli derived from mature embryos was established for the two Brazilian
modern rice cultivars IAC-201 and IAC-165. After 30 to 40 days of in vitro culture it was possible to obtain on average 6
million protoplasts per gram of callus. Microscopic selection of embryogenic calli was a key step for protoplast isolation.
The production of embryogenic calli increased when L-proline and casein hydrolysate were used in the callus induction medium.
The Oc or IR52 nurse cell lines were essential for protoplast division. Different regeneration media were studied and 139
plants were regenerated which set seed. Some of the regenerated plants showed morphological variation such as the presence
of awns in spite of the short time of the in vitro culture.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
7.
新陆早42号体细胞胚发生和植株再生 总被引：3，自引：2，他引：1
以新陆早33号为对照研究新疆主栽品种新陆早42号的体细胞胚胎发生。研究表明,所用4种激素组合均能有效诱导愈伤组织,但二者仅在经过IBA 1.0 mg·L-1+KT 0.5 mg·L-1和2,4-D 0.1 mg·L-1+KT 0.1 mg·L-1两种激素组合诱导初生愈伤后,才能胚胎发生。新陆早42号在IBA 1.0 mg·L-1+KT 0.5 mg·L-1培养基上46 d就开始胚胎发生;新陆早33号则不能直接胚胎发生,需继代到MSBP培养基上培养48 d才有胚胎发生。经2,4-D 0.1 mg·L-1+KT 0.1 mg·L-1组合培养的愈伤均需要继代到MSBP培养基上培养才能胚胎发生,新陆早42号和新陆早33号胚胎发生的最早时间分别是71 d和81 d。胚性愈伤在铺有滤纸的MSBF培养基上分化成胚并发育成再生植株。新陆早42号在140 d有根系发育良好的能嫁接植株（株高 7～8 cm）,而新陆早33需180 d。即成功建立了新陆早42号的再生体系,且其胚胎发生能力和再生能力均优于对照。 相似文献
8.
C.E. van Schaik C. van der Toorn M.J. De Jeu C.J.J.M. Raemakers R.G.F. Visser 《Euphytica》2000,115(1):17-26
The successful application of plant biotechnology to Alstroemeria improvement will largely depend on the availability of an efficient regeneration/transformation system. Regeneration in Alstroemeria is accomplished from nodular embryogenic callus initiated from zygotic embryos. Histological studies of embryogenic callus
initiation from 4-weeks old cultured ovules revealed that the outermost layers of the protoderm of the embryogenic nodules
divided to form either a new nodule or aproembryo. Transient gene expression after particle bombardment of nodular embryogenic
callus was optimized using DNA of pAHC25. The highest β-glucuronidase expression was found when the GUS gene was under control of the maize ubiquitin promoter, the target tissue was placed 5 cm below the microcarrier launch assembly
and when the rupture disc-breakage point was between 650–900 psi. Kanamycin blocked regeneration of somatic embryos, however,
did not block growth of nodular embryogenic callus. With phosphinothricin both callus growth and regeneration were blocked.
Bombardment of nodular embryogenic callus with DNA of pAHC25 combined with selection on medium containing phosphinothricin
resulted in putative transgenic chimeric. Friable calli were selected from nodular embryogenic callus and used to initiate
suspensions. These cell suspensions were subjected to transformation by particle bombardment using DNA of pAHC25 and resulted
in a stable transformed friable callus line after selection based on luciferase activity. Even after 2 years of maintenance
this callus line was luciferase positive and the Polymerase Chain Reaction analysis demonstrated the presence of the introduced
gene in this friable callus line.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
9.
为建立一个稳定的谷子遗传转化体系,对216份谷子种质资源进行筛选,确定材料178成熟胚为受体材料,从植物激素浓度、防褐化剂、农杆菌菌液浓度、侵染时间、乙酰丁香酮（AS）浓度等方面对谷子再生及转化体系进行优化。结果表明,在MS培养基中加入生长素2,4-D 9μmol/L、细胞分裂素Kinetin 4μmol/L、硝酸银8mg/L,愈伤组织诱导率最高且能有效防止褐化。在侵染液和共培养基中加入100μmol/L AS,gus表达效果最好。农杆菌溶液OD600为0.3~0.5,侵染15min,共培养3d时,抗性愈伤组织率最高。获得的转基因植株,经PCR检测及gus染色分析,确定外源bar基因已成功整合到谷子基因组中。初步建立了一套谷子遗传转化体系,为今后谷子遗传转化提供一些参考。 相似文献
10.
大蕉未成熟雄花接种到胚性愈伤组织诱导培养基中,4～5个月后可诱导出胚性愈伤组织,并可在继代培养基上长期增殖。这些继代培养了6年多的松软易碎的胚性愈伤组织转移到含有0.2 mg/L 6-BA的分化培养基中,可诱导出芽,得到了53株再生植株,这些再生植株可进一步扩大繁殖。组织学切片证明长期继代培养的愈伤组织维持了胚性的状态。 相似文献
11.
Influence of genotype and culture medium on callus formation and plant regeneration from immature embryos of Triticum turgidum Desf. cultivars 总被引：2，自引：0，他引：2
Twelve durum wheat cultivars were evaluated for their response to in vitro tissue culture. Zygotic immature embryos were used to induce callus formation using four different Murashige and Skoog‐based media. Each contained 9.05 μM 2,4‐dichlorophenoxy acetic acid but differed in their carbon source (sucrose or maltose) and the presence of NaCl (0 mM or 40 mM). The influence of both genotype and medium on the type and percentage of callus produced was observed. Calli were either compact and frequently embryogenic, or soft and watery. Percentages ranged from 54 to 100%, depending upon genotype and induction medium. All calli were then plated on a regeneration medium containing 20 g/l sucrose, 2.68 μM 1‐naphthaleneacetic acid and 2.22 μ 6‐benzylaminopurine. The regeneration of plantlets was higher from compact than from soft calli, with a strong dependence on genotype and type of induction medium used. MSm induction medium (30 g/l maltose) and MS40s (30 g/l sucrose plus 40 mM NaCl) were best for inducing compact calli, and gave the highest proportion of regenerated plants. The in vitro response (number of total shoots from a compact callus/number of embryos plated) was higher for immature embryos of ‘Baztan’, ‘Bradano’ and ‘Don Pedro’. These cultivars are a good starting material for experiments involving transformation of calli from zygotic immature embryos. 相似文献
12.
Summary This article reports the culture and plant regeneration of Tripsacum dactyloides. Mature embryos of Tripsacum dactyloides dactyloides were used to obtain embryogenic callus cultures. Currently, 180 normal plants have been regenerated from these cultures. Callus was initiated on MS medium supplemented with dicamba (10 mol or 20 mol) and sucrose (3% or 6%), and plants were regenerated on hormone free MS medium containing 2% sucrose. No significant differences were found in callus initiation frequency or in embryogenic response of cultures on the four combinations of sucrose and dicamba tested. The embryogenic cultures have been maintained for 9 months (12 subcultures) and have retained regeneration capacity. Plants regenerated from tissue culture of maize-by-Tripsacum hybrids could be useful in maize improvement. 相似文献
13.
大麦原生质体培养再生胚性愈伤组织和白化苗 总被引：2，自引：0，他引：2
从来自春大麦品种“如车”成熟胚的愈伤组织中,挑选出适于悬浮培养的松脆型胚性愈伤组织,在短期内建立胚性细胞悬浮系。此系酶解后分离出的原生质体在修改的MS培养基上能够持续分裂形成愈伤组织。将其直接转至分化培养基上获得结构紧密的胚性愈伤组织并再生白化苗. 相似文献
14.
D. Sihachakr R. Haïcour J.M. Cavalcante Alves I. Umboh D. Nzoghé A. Servaes G. Ducreux 《Euphytica》1997,96(1):143-152
The application of new techniques for improvement of sweet potato crops, particularly including the exploitation of somaclonal
variation, gene transfer by genetic transformation and somatic hybridization, requires the control of plant regeneration from
tissue cultures. Shoots can easily be regenerated from explants of stems, petioles, leaves and roots, while callus cultures
do not produce any shoots. The potential of somatic embryogenesis and plant regeneration via embryogenesis was evaluated for
10 cultivars of sweet potato. Protocols for plant regeneration from cultured protoplasts have also been developed. Since mesophyll
was resistant to enzyme digestion, fragments of stems and petioles, callus and cell suspensions were used as source of protoplasts
of sweet potato. Series of transfers of protoplast-derived calluses, particularly those which had been obtained from in vitro
plants, to media containing a high level of zeatin resulted in successful formation of shoots in only two sweet potato cultivars.
In addition, the embryogenic potential was irreversibly lost through protoplast culture, since protoplasts isolated from embryogenic
cell suspensions developed into non-embryogenic callus. Consequently, an alternative protocol is being successfully developed
to improve plant regeneration from cultured protoplasts of sweet potato, involving first root formation from which shoots
can then be regenerated. Preliminary evaluation in field conditions in Gabon revealed that plants regenerated from cultured
protoplasts exhibited a great genetic variability in their growth and tuber formation in particular.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
15.
Summary Experiments upon in vitro culture of immature durum wheat embryos, harvested at different growth stages, were made in two consecutive years. Callus formation and plantlet regeneration were obtained. The ability to form callus and the degree of morphogenetic processes varied with the different hormonal treatments used and with the age of the embryos. In the first year the best response for callus growth was observed with 2,4-D 2 mg l-1 plus adenine 50 mg l-1 or 2,4-D 5 mg l-1 alone in the more mature embryos (15 and 20 days after anthesis). On the contrary, NAA 5 mg l-1 had a greater shoot regeneration effect. In the next year, at all 2,4-D concentrations and for the two different ages of the embryos tested, all embryos formed callus. Regeneration of plantlets was obtained in higher percentage in calli originated from the more developed embryos. The effect of changed media upon plantlet regeneration was studied after callus transplant.Investigation by cytophotometry and chromosome counts on different calli showed, practically in all cells, a diploid condition. A histological analysis demonstrated embryogenic somatic characteristics in many samples of callus. The pattern of organogenesis seemed to be via adventitious bud formation but structures resembling embryoids were also observed in the callus. 相似文献
16.
To establish a plant regeneration system from embryogenic callus derived from mature rice embryos, the addition of aminoacids
and the effect of two macronutrient solutions MSD and N6D to the basal callus induction medium was tested in three Spanish
varieties, Senia, Tebre and Bahia. Aminoacids enhanced the production of embryogenic callus in Tebre and Senia whereas in
the case of Bahia, embryogenic callus, which gave rise to a high rate of differentiated shoots, was induced without aminoacids.
The macronutrient solution had also to be adjusted for each variety. Pre-regeneration treatment with ABA significantly improved
the regeneration rate in all media tested, independently of the media in which the embryogenic callus were induced. In a comparison
of growth regulators, BA yielded more shoots than Kin in all varieties whereas the effect of the auxins NAA or IAA was dependent
on the variety. Transgenic plants from the three varieties were obtained via an Agrobacterium tumefaciens transformation system, using the optimised culture media.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
17.
Production of embryogenic tissues and regeneration of transgenic plants in cassava (Manihot esculenta Crantz) 总被引：3，自引：0，他引：3
Nigel J. Taylor Munyaradzi V. Masona Rosa Carcamo Thao Ho Christian Schöpke Claude M. Fauquet 《Euphytica》2001,120(1):25-34
Disorganised embryogenic tissues have been utilised as target tissues for transgene insertion and transgenic plant regeneration
in cassava (Manihot esculenta). The production of friable embryogenic callus in fourteen geographically diverse cassava cultivars, from which eleven were
established as embryogenic suspension cultures, is reported. Embryogenic tissues were similar in nature in all cultivars tested
although there was variation in the time required to generate friable callus and the growth rates of suspension cultures.
Regeneration of plants has been achieved from eight cultivars but varied significantly in efficiency, with cv. TMS 60444and
Line 2 from Zimbabwe being the most responsive. Tissues from the remaining eight cultivars became arrested at globular and
torpedo stages of regeneration indicating that they most likely process an inherent ability to produce plants but require
further research to allow this to be realised. Significant numbers of transgenic plants containing transgenes for putative
resistance to important viral diseases of cassava in addition to visual marker genes have been regenerated. Transgenic plants
from three the cultivars TMS 60444, Bonoua Rouge and M.Col 1505 were recovered after particle bombardment of embryogenic suspension
cultures. Correlation's have been made between abnormal leaf morphology and plant vigour with the use of embryogenic suspension
cultures for transgene insertion. As an result friable embryogenic callus is now being successfully utilsed as the target
tissue for genetic transformation and plant regeneration at ILTAB.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
18.
Summary Use of 2,4-D was superior to NAA or IAA for embryogenic callus initiation or maintenance in barley cultivar Bruce. A concentration of at least 2.0 mg/l 2,4-D was desirable for culture initiation. The developmental size of the embryo was more important than embryo age for obtaining embryogenic calli. Even brief exposures (20–40 days) of calli to concentrations of higher than 5.0 mg/l 2,4-D or 10.0 mg/l NAA resulted in inhibition of subsequent plant regeneration and therefore, concentrations above these could not be used for maintenance cultures. In the long-term maintenance cultures, the best production of embryogenic calli was with 0.1 mg/l and 1.0 mg/l 2,4-D. 相似文献
19.
A Yi-Xia-Mu-Gu-Li-·Si-Ma-Yi-Li QU Yan-Ying SE Na-Wa-Er-·Mai-Mai-Ti-Ming MENG Ling-Zhen YANG Ting WANG Li-Ping CHEN Quan-Jia- 《棉花学报》2013,25(4):352-358
The aim of this study was to induce embryogenic callus from various cultivars of cotton in tissue culture, so that a stable and efficient regeneration system could be developed to produce new cotton varieties for cultivation in Xinjiang. The explant materials were hypocotyls of the main cotton cultivars grown in Xinjiang, i. e. Xinhai 25, Xinhai 16, Xinluzao 39, and Xinluzao 42. We tested the effects of different combinations of two hormones (kinetin, KT; 2,4-dichlorophenoxyacetic acid, 2,4-D) on induction of callus from these explants. Calli were produced by the explants under four different combinations of hormones in the media. The optimal hormone combination to induce callus from Gossypium hirsutum explants was 0.1 mg·L-1 KT + 0.05 mg·L-1 2,4-D, while that to induce callus from Gossypium barbadense explants was 0.1 mg·L-1 KT + 0.1 mg·L-1 2,4-D. Hormone-free medium and medium containing double to the normal concentration of KNO3 promoted the emergence of embryogenic callus. Filter paper placed under the medium promoted somatic embryo growth and regeneration of the root system. The differentiation and embryogenesis processes occurred more rapidly in G. hirsutum explants than those in G. barbadense explants. Using this protocol, normal plantlets of these cotton cultivars with strong roots were produced within 10 to 12 months. These methods could be used to increase the number of cotton genotypes that can be regenerated in tissue culture. 相似文献
20.
绿色棉新彩棉7号体细胞胚胎发生及其植株再生 总被引：1，自引：1，他引：0
以绿色棉新彩棉7号的子叶、下胚轴为外植体,MSB(MS培养基附加B5维生素)基本培养基附加不同激素组合,诱导愈伤组织及调控分化,通过体细胞胚胎发生方式获得再生植株.结果表明:0.1 mg· L-1 KT(Kinetin,激动素)+ 0.1 mg·L-12,4-D(2,4-dichlorophenoxyacetic,2,4-二氯苯氧乙酸)为诱导愈伤组织的最适植物激素组合,不同外植体处理出愈率均达到100％,但下胚轴纵切面背向培养基放置培养更有利于诱导愈伤组织形成;分化调控阶段的最佳植物激素组合为0.15 mg·L-1 KT+ 0.3 mg·L-1 IBA(Indole-3-butyric acid,吲哚丁酸),胚性愈伤分化率可达23.33％; MSB中去除NH4NO3同时KNO3加倍,附加0.5 g·L-1Asn(Asparagine,天冬酰胺)和lg· L-1 Gln(Glutamine,谷氨酰胺),胚性愈伤可进一步分化获得体细胞胚,将成熟的子叶胚接种于1/2MS获得完整的再生植株. 本研究通过体细胞胚发生途径获得了新彩棉7号的再生植株,为天然彩色棉基因工程研究奠定了一定基础. 相似文献