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1.
Summary A high frequency plant regeneration system via organogenesis and somatic embryogenesis was established with callus cultures derived from mature zygotic embryos of different leek genotypes (Allium ampeloprasum L.). Four different callus types with varying morphogenetic potential were obtained. Relatively high concentrations of the auxin 2,4-dichlorophenoxy-acetic acid reduced callus weight and subsequent shoot regeneration and primordia formation of the callus. Shoot regeneration and primordia formation of the callus decreased after prolonged subculture on media containing 2,4-dichlorophenoxy acetic acid. A callus growth period of six weeks on Murashige and Skoog medium with 0.25–0.5 mg l-1 2,4-dichlorophenoxy acetic acid showed the highest rate of shoot regeneration after transfer of callus to regeneration medium with 1 mg l-1 kinetin.Differences between leek genotypes in callus type, callus weight, shoot regeneration and primordia formation were observed. Histological observations showed that plant regeneration took place, both via the pathway of somatic embryogenesis and organogenesis.Abbreviation 2,4-D
2,4-dichlorophenoxy acetic acid
- MS
Murashige and Skoog (1962) medium 相似文献
2.
Summary Callus was obtained from immature excised embryos of triticale using MS medium supplemented with 3 mg/l 2,4-D and 1 mg/l kinetin. The presence of 2,4-D was essential for continued callus proliferation. Plantlets were induced from the calli by sub-culturing on medium either devoid of auxin or containing 0.1 mg/l 2,4-D. The capacity to produce callus and to form organs and plantlets differed markedly among the genotypes used. Lines also had distinct response to presence and absence of 2,4-D in the regeneration media. The callus of most triticale lines used differentiated into organs more readily on MS medium supplemented with 0.1 mg/l 2,4-D than on medium without growth regulators. Very high frequencies (up to 75%) of plantlet regeneration were observed in several of the triticale lines studied. 相似文献
3.
Summary Immature embryos of seven rye inbred lines were cultured on modified MS medium containing 3 mg/dm–3 2,4-D. According to thein vitro response lines were divided into four groups: (1) those producing non-embryogenic callus (NEC) from above 30% of the embryos and having a high rate of non-responding (NR) embryos, (2) those producing non-embryogenic callus (NEC) from the majority of embryos, (3) those producing NEC by the majority of embryos with a high percentage of calli regenerating roots, (4) those producing embryogenic callus (EC) and regenerating plants by above 50% of the embryos. The inheritance of these response types was analysed in F1, F2, and F3 generations of crosses of some lines. The results obtained indicate that EC production and both plant and root regeneration are determined by recessive genes whereas the reduced ability for NEC production most probably by dominant genes. The lack of response is controlled by at least two interacting genes. 相似文献
4.
Summary Callus induction and green plant regeneration at high frequencies from an interspefic hybrid, Oryza sativa L. x O. rufipogon Griff. has been achieved by simply coordinating the growth regulators in the induction medium. The study was conducted with two different basal media (Potato-2 and N6) and seven different combinations of growth regulators 2,4-D, NAA and kinetin. Synergistic effects of the two auxins in enhanced anther response to callus induction and subsequent green plant regeneration were observed in both media. The highest frequency of callus induction was obtained on Potato-2 medium supplemented with 1 mg/12,4-D, 2 mg/l NAA and 1 mg/l kinetin. The same combination of growth regulators which yielded higher frequencies of callus induction also induced higher mean number of calli per anther. Although the calli formed on N6 medium showed high regenerability, there was a concomitant increase in the number of albinos among the regenerants. The auxins in the induction media had considerable influence on the regeneration capacity of the calli. The regeneration frequencies were higher from calli formed in the presence of both auxins in the induction media. The levels of growth regulator combinations seem to influence the green plant regeneration especially for calli induced on Potato-2 medium. Among the pollen grain derived plants the majority were either haploids or double haploids and very few were chromosomal variants. 相似文献
5.
B. S. Ahloowalia 《Euphytica》1982,31(3):755-759
Summary A procedure for plant regeneration from callus culture of potato, Solanum tuberosum L. is described. Calli were induced from 1–2 mm long shoot apices of potato cultivars Cara and A25/19 on half-strength Murashige and Skoog's medium (half-MS) supplemented with 3.2 mg IAA (indole-3-acetic acid), 1.0 mg kinetin (6-furfurylamino)purine], and 0.5 mg 2,4-D [2,4-dichlorophenoxy)acetic acid]/1. Sixty percent explants produced nodular calli on this medium within 30 days. Calli differentiated into shoot-primordia when subcultured on half-MS medium supplemented with 0.5 mg 2,4-D and 1.0 mg zeatin [6-(4-hydroxy-3-methybut-2 enylamino)amino purine]/1. Differentiated calli on half-MS medium without growth hormones produced complete plantlets which were cloned on the same medium and transferred into soil. 相似文献
6.
B. S. Ahloowalia 《Euphytica》1987,36(2):659-665
Summary Plants were regenerated from callus cultures initiated from immature embryos of barley, Hordeum vulgare L. Immature embryos from seven diverse genotypes were cultured on modified Murashige and Skoog (MS) medium supplemented with 1.5 mg 2,4-D and 6.5 mg IAA/l. Of the 249 embryos cultured, 30% initiated callus within 8 days. Subculture of callus for 80 to 100 days on half-MS medium supplemented with 0.5 mg/l 2,4-D and 1.0 mg/l zeatin resulted in organogenesis. Culture of organogenic calli for 30 days on half-MS medium without growth regulators produced plants which originated mostly via multiple shoot formation. Callusing response of the tested genotypes ranged from zero to 44%; however, only 23% of the calli were regenerative. Regenerated plants included variants for chlorophyll deficiency, plant height, stem thickness, spike shape, pollen fertility, seed set and ploidy. 相似文献
7.
Callus induction and plant regeneration from immature and mature embryos of winter durum wheat genotypes 总被引：9，自引：0，他引：9
Seven genotypes of winter durum wheat (Triticum durum Desf.) were cultured to establish an efficient method of callus formation and plant regeneration from mature embryo culture, and to compare the responses of immature and mature embryo cultures. Immature embryos were aseptically dissected from seeds and placed, with the scutellum upwards, in dishes containing Murashige and Skoog's (MS) mineral salts and 2mg 2,4- dichlorophenoxyacetic acid (2,4-D) per litre. Calli and regenerated plants were maintained on 2,4-D-free medium. Mature embryos were moved slightly on the imbibed seeds. For callus formation, the seeds with moved embryos were placed, furrow downwards, in dishes containing 8 mg 2,4-D per litre. The developed calli and regenerated plants were maintained on the MS medium. Plants regenerated from both embryo cultures were vernalized and grown to maturity in soil. Variability was observed among the wheat genotypes tested for various culture responses in both explant cultures. Callus induction rate and regeneration capacity of callus were independent of each other. Mature embryos have a low frequency of callus induction but a high regeneration capacity. Considering availability, rapidity and reliability, this form of mature embryo culture can be used as an alternative method for immature embryo culture. 相似文献
8.
植物激素配比对大蒜茎盘愈伤组织再生体系的影响 总被引：5，自引：0，他引：5
植物激素对大蒜茎盘愈伤组织再生的各个环节都发挥着重要的作用。试验结果表明：2,4．D对茎盘愈伤组织的诱导具有主要作用，高浓度的2，4-D有利于大蒜愈伤组织的诱导，但并不利于其增殖分化。MS＋NAA 2mg／L＋BA 1mg／L培养基适宜于愈伤组织的继代增殖，MS＋NAA 1mg／L＋BA 5mg／L培养基适宜不定芽的分化，只添加NAA或无激素的MS培养基有利于不定芽生根。 相似文献
9.
Continuous Plant Regeneration from Established Embryogenic Cell Suspension Cultures of Italian Ryegrass and Tall Fescue 总被引：8，自引：0，他引：8
The suitability of different protocols was compared for entire plant regeneration by somatic embryogenesis, of the forage plants Lolium multiflorum Lam. (Italian ryegrass) and Festuca arundinacea Schreb. (tall fescue). In the first protocol, miniature embryos were used as starting material, while mature seeds were retained in the other two. Whichever the considered protocol, undifferentiated calli were produced on Murashige and Skoog MS medium supplemented with 2,4-D. The calli were subcultured in the dark on solid MS agar medium, containing 5 mg/1 2,4-D (protocol 2) or on solid MS medium followed by transfer to a rotated liquid MS medium with 2 mg/1 2,4-D (protocol 1). In these conditions, induction of somatic embryogenesis occurred, and whole plants were regenerated during a limited lapse of time, upon transfer in the light, to MS medium supplemented with BAP but devoid of 2,4-D. The simultaneous elimination of 2,4-D and transfer to light appeared essential for full regeneration of the plants. Using this characteristic, an additional step was added to a new protocol (protocol 3) in which microcalli, cultured on liquid MS medium containing 5 mg/1 2,4-D, were transferred to the same medium with 2 mg/1 2,4-D, in the dark. In these conditions, the suspensions kept their embryogenic potential for months. In all cases, plantlets were successfully transferred into the soil. An evaluation of the somaclonal variation potential of the plants issued from each protocol is now underway. 相似文献
10.
Genotypic and exogenous factors affecting shoot regeneration from anther callus of linseed (Linum usitatissimum L.) 总被引：1，自引：0，他引：1
Summary The objective of this study was to investigate factors affecting the regeneration capacity of linseed anther culture. Four different environmental conditions in a phytotron were tested with regard to their effects on anther donor plants of cv. Hella. Anther response and shoot regeneration from anther callus was maximal when donor plants were grown in a 16 hrs-day at 14°C day/8°C night temperature. Anthers of four linseed genotypes were cultured on different media. Maximum shoot regeneration was achieved when the induced calli were transferred onto a modified N6 medium containing zeatin (1 mg l-1). Most of the calli regenerated shoots in the second subculture on regeneration media. Shoots were rooted on modified B5 or MS media containing NAA (0.1 mg l-1). Cytological examinations of incubated anthers and root tips of regenerated plants indicated that the anther calli were derived from microspores.Abbreviations B5
Gamborg's (1975) medium
- BAP
6-benzylaminopurine
- 2,4D
dichlorophenoxyacetic acid
- N6
Chu's (1978) medium
- NAA
-naphthaleneacetic acid
- MS
Murashige & Skoog's (1962) medium
- ZEA
zeatin 相似文献
11.
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
12.
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
13.
Summary Immature inflorescences of smooth bromegrass were cultured on MS agar media supplemented with varying combinations of 2,4-D and kinetin. Callus was initiated from segments of young inflorescences on each medium. All of the calli were subcultured monthly for 5–6 times and transferred onto hormone-free MS medium for plant regeneration. Addition of kinetin to the basal medium stimulated shoot initiation in the callus cultures. Plantlets were regenerated only from calli grown on media containing 2 and 6 mg I-1 2,4-D with a supplement of 0.2 mg I-1 kinetin. No albino plantlets were produced. Morphological characteristics and dry matter yield of ten somaclones and the parental plant (SBG7) were studied in the greenhouse in a randomized complete block experiment with five replications. There was significant variation (P>0.01) among genotypes for all morphological characteristics studied. Although all somaclone heights and leaf widths were lower than those of the parental plant, the somaclone F9A, F10A, and F10B had larger tiller numbers, and leaf/stem ratio by dry weight than the parental plant. Only somaclone F9B gave higher specific leaf area and leaf area ratio than the parental plant. Almost all somaclones had the same leaf length, total dry weight, and specific leaf weight as the parental plant. The variation found in somaclones should permit selection for desirable agronomic traits. 相似文献
14.
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
15.
This study was conducted to develop an efficient in vitro selection system for scab resistance by using in vitro screening
for tolerance to deoxynivalenol (DON). Immature embryos of two wheat varieties, a scab-resistant variety Sumai 3 and a susceptible
variety Mianyang 11, and their reciprocal F1 hybrids were cultured on MS medium supplemented with 2,4-D 2 mg/l and 0.6 × 10-4 M DON for callus induction. The responses of callus induction and plant regeneration to 0.6 × 10-4 M DON differed significantly between resistant and susceptible varieties, according to observed scab resistance levels at
the plant level in the field. The percentage of callus formation of resistant variety Sumai 3 on induction medium containing
DON was higher than that of susceptible variety Mianyang 11. Regeneration of DON-tolerant calli on DON-containing differentiation
medium differed significantly between Sumai 3 and Mianyang 11. Averaged across the DON-tolerant calli of two varieties and
their reciprocals, regeneration of DON-tolerant calli was decreased 3-fold on DON-containing medium. By an inoculation test
with conidiospores of Fusarium graminearum Schw, we obtained several resistant lines from progenies of regenerated plants
from DON-tolerant calli. These somaclonal lines had lower disease scoring (reaction index, infected spikelets and disease
incidence), shorter plants and better yield components than Sumai 3, a famous Chinese resistant variety.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
16.
Summary Calli were obtained from mesocotyl explants of eight varieties of pearl millet using MS medium supplemented with 2,4-D. Variable frequencies of callus initiation were observed among the varieties. Plantlets were regenerated from the calli by transferring on to the medium devoid of auxins. Incorporation of coconut milk, Kinetin, IAA or NAA did not enhance either the callus initiation or growth rate or plantlet regeneration. The eight genotypes differed among themselves in callus growth rate and frequency of shoot bud production. 相似文献
17.
新疆棉花4个主栽品种的体细胞胚胎发生及植株再生 总被引：15，自引：0，他引：15
以新疆4个主栽棉花品种新陆中20、新陆早24、新陆早33和03298为材料, 通过不同浓度的激素组合成功地诱导获得了体细胞胚并进一步发育成苗。研究发现, 所用的4种激素组合均能有效诱导愈伤组织, 其中又以0.02 mg L-1或0.10 mg L-1 KT和0.1 mg L-1 2,4-D组合的诱导效果最佳; 两个诱导措施有利于胚性愈伤组织的产生, 即沿中柱纵切棉花下胚轴切段, 并以纵切面接触培养基; 愈伤组织诱导培养基中KNO3用量加倍。挑选黄绿色、灰绿色或浅绿色的质地疏松的愈伤组织继代于无激素且KNO3含量加倍的培养基中可产生胚性愈伤组织, 并在高比例KT/2, 4-D(0.05 mg L-1或0.10 mg L-1 KT和0.01 mg L-1 2,4-D)促进下发育成胚。借助在培养基上垫滤纸产生干燥作用, 并间隔使用强透气效果的棉塞对培养三角瓶进行透气处理, 体细胞胚可成熟发育并产生根系发达的正常再生植株。应用此法, 4个实验材料在6~8个月内即可获得大量再生苗。 相似文献
18.
为了建立粳稻恢复系C418转基因再生体系,本研究以不同培养基、不同2,4-D浓度、不同KT浓度及不同6-BA浓度的不同组合培养基来诱导C418成熟胚愈伤组织,以获得最适C418诱导的培养基,然后以获得的愈伤组织利用农杆菌介导法进行转基因再生体系的建立。结果表明:NBD培养基为6种培养基中最适培养基;2,4-D浓度为2.0 mg/L时,诱导率最高;随着6-BA的浓度增加诱导率降低;KT对C418成熟胚愈伤组织的诱导没有影响。本研究成功建立了C418转基因再生体系。 相似文献
19.
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
20.
为探索杜仲胚性愈伤组织诱导的条件,建立杜仲体细胞胚胎发生初步体系,以杜仲幼嫩叶片和未成熟合子胚为外植体、MS为基本培养基,探究外源激素配比、未成熟合子胚发育阶段与基因型对愈伤组织诱导的影响,并从形态学和细胞学对愈伤组织进行胚性的判断。试验结果表明：4种不同激素配比的培养基诱导出的叶片愈伤组织在形态上具有差异,MS+ 2,4-D 2 mg/L+ 6-BA 1 mg/L和MS+ 2,4-D 2 mg/L+6-BA 0.5 mg/L有利于叶片胚性愈伤组织诱导;在培养基MS+ 2,4-D 2 mg/L+ 6-BA 1 mg/L上,以未成熟合子胚为外植体诱导出4种类型愈伤组织,其中圆球形和颗粒型突起的愈伤组织具有胚性;未成熟合子胚采集时间对愈伤组织诱导率具有显著差异,6月14日采集的外植体愈伤诱导率最高,不同基因型差异不显著。 相似文献