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1.
The influence of culture medium on induction of androgenic calli was examined with five different basal media. MS medium was the most responsive in inducing callus. Differences in induction of calli among ten genotypes revealed that the most responsive genotype was a local cultivar, Mangira, with 48.6% anthers initiating callus formation. The influence of temperature pre-treatment (5°±1°C) for varying periods (0 to 15 days) on immature capitula prior to inoculation of anthers on the medium revealed that the percentage of anthers inducing callus increased till 3–5 days of pre-treatment. The effect of physiological conditions of anther donor plants grown in the field and in green houses on induction and re-differentiation have shown that the field grown anther donor plants exhibited optimum response. Shoot regeneration was observed on MS supplemented with BAP (2.0 mg/l) and NAA (0.5 mg/l) and rhizogenesis on MS (half-strength) medium, supplemented with NAA (0.1 mg/l) and 1% sucrose. Cytological studies of anther derived plants showed two ploidy levels, where the haploids were predominant (64%). 相似文献
2.
In
vitro culture ofBrassica
alba anthers on a growth medium containing inorganics of KB5 and organics, iron, sucrose and hormones of B5 resulted in a very
high response of anthers (93.75%) towards callus induction. All the calli transferred to regeneration media responded favourably
even after six months of callus induction. Numerous torpedo-shaped embryoids developed in clusters at many sites from each
callus mass. Secondary embryogenesis and multiple shoot formation was also observed in many cases. The number of embryoids
and plantlets produced by one embryogenic anther were as high as 169.8 and 17 respectively. 87% of the regenerated plants
were haploids. 相似文献
3.
This study concerns anther culture and the production of microspore-derived calluses and plants of the opium poppy (Papaver somniferum L.). It was confirmed that growth regulators were necessary for microspore callus production. Cold treatment (7 d at 7°C) of the buds prior to culture lead to a twofold increase in the frequency of responsive anthers and in the number of calluses per 100 anthers plated. Callus was produced from cultured anthers of several genotypes, covering a wide genetic background. Step by step removal of growth regulators from the culture medium promoted organogenesis and plant regeneration. Most regenerated plants were diploid. The overall process of microspore embryogenesis closely resembled that described in previous reports on somatic callus production and plant regeneration from poppy hypocotyls in vitro. 相似文献
4.
Direct microspore-derived embryo formation in anther cultures of two cultivars of Brassica juncea was obtained. Preliminary culture of anthers at 35°C for 1–5 days prior to maintenance at 25°C stimulated embryogenesis. Embryogenesis was also stimulated by an initial culture at 5°C for 3 days. Analysis of squashed anthers revealed that approximately 10% of the microspores began dividing, but less than 1% developed into macroscopic embryos. All embryos transferred to embryo culture medium survived, but only 30% of these developed directly into normal plantlets. The androgenic plants were haploid (2n=18). 相似文献
5.
The development of haploid callus, embryos and plantlets from cultured anthers and the various factors affecting androgenesis in Peltophorum pterocarpum (Copper pod), a tropical legume tree is reported. A pretreatment of flower buds at moderate temperature of 14°C for 8 days was most effective for callus production. The colour of the anther was found to be a reliable and efficient indicator for identification of suitable stage of anther for culture. The frequency of anthers which produced callus and shoots was highest when anthers were cultured at mid or late-uninucleate stage. A high sucrose concentration of 10% is a specific media requirement for androgenesis. The haploid nature of the embryos, callus and regenerated plants (n=14) were confirmed by chromosome count.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- KN
kinetin
- NAA
naphthaleneacetic acid
- BAP
bezylaminopurine 相似文献
6.
Edyta Skrzypek Ilona Czyczyło-Mysza Izabela Marcińska Maria Wędzony 《Plant Cell, Tissue and Organ Culture》2008,94(2):131-137
Anthers cultures of six Polish cultivars of pasture lupin (Lupinus L.) were examined for their androgenic response. Anthers with microspores at the uninucleate stage were isolated from flower
buds and cultured in liquid media. Better viability of androgenetic structures was obtained when donor plants had grown under
field as opposed to greenhouse conditions. A density of five anthers per 0.5 ml medium was more conducive to androgenetic
induction than 25 anthers per 0.5 ml medium. Addition of 5% maltose to the induction medium and culture at 25°C without pre-treatment
of flowers, buds or anthers promoted microspore release and division. The greatest frequency of androgenic callus, ~70% was
developed from cvs. Katon, Wat (white lupin), in contrast to cvs. Legat, Juno (yellow lupin), Polonez and Sonet (narrow-leafed
lupin) with callus induction ~30–40%. Despite various combinations of media tested, plant regeneration was not obtained from
anther derived callus. 相似文献
7.
L. A. Shtereva N. A. Zagorska B. D. Dimitrov M. M. Kruleva H. K. Oanh 《Plant cell reports》1998,18(3-4):312-317
The influence on androgenesis of donor plant growth conditions, anther size and developmental stage of the microspore, medium
composition and different anther treatments prior to culture was investigated in L. esculentum Mill. cv Roma and its hybrids. Growth conditions of donor plants affected the induction of tomato androgenesis. Anthers isolated
from plants grown in the greenhouse during winter at high humidity and in short days possessed higher androgenetic ability
than those grown in the field. The physiological state and age of the donor plants also influenced the processes investigated.
Regarding the developmental stage of microspores, the period from prophase to telophase II is optimal for tomato anther implantation.
More then 20 culture media were tested. Two, based on Murashige and Skoog medium were selected as most favourable for callus
induction, organogenesis and regeneration. The effect on callus induction of 2ip in combination with indole-3-acetic acid
(IAA) was greater than that of zeatin and IAA. Zeatin promoted entire plant regeneration. A highly significant interaction
between genotype and medium was observed. Temperature and gamma ray treatments of anthers enhanced callus production, shoot
formation and plant regeneration. Treatments at 4 °C (48 h) and 10 °C (9 days) stimulated these processes. Combined treatment
of anthers with 4 Gy and 10 °C for 9 days was the most efficient.
Received: 5 September 1997 / Revision recieved: 5 June 1998 / Accepted: 15 June 1998 相似文献
8.
The present study involves in vitro androgenesis of Zea mays L. using anther culture. We tested combinations of single factors and their influence on microspore induction. Embryogenic induction of microspores within anthers in in vitro conditions was the best when combination of cold treatment, TIBA (0.1 mg l–1) in media and colchicine (0.02% during first 3 days of culture) was applied, but colchicine alone can be factor, which can stimulate or initiate embryogenesis in anther culture of maize. 相似文献
9.
Taina Tiainen 《Plant Cell, Tissue and Organ Culture》1992,30(3):211-219
The effect of media manipulatioss, temperature pretreatment, carbohydrate source, and seasonal variation on tetraploid potato anther cultures was investigated. The anther culture responses of three commercial Nordic potato varieties from Scandinavia and two from Germany were compared on different media manipulations. With most of the varieties, solid MS media gave better yields than other published media manipulations. Pretreatments at +6°C and at +30°C were studied on Pito and Danva varieties. The +6°C pretreatment and no pretreatment had the same effect on the anther culture response of cv. Pito, while with cv. Danva pretreatment at +6°C promoted embryogenesis. The +30°C pretreatment had no positive effect on anther culture response on either cultivar. The effect of maltose, melibiose and mannitol individually and in combination with sucrose were compared to normal sucrose medium in cv. Pito anther cultures. Anthers incubated on normal sucrose medium gave the highest embryoid and plant yields; the second highest plant yields were obtained on pure maltose medium. Strong seasonal variation was observed throughout the year in cv. Pito anther cultures. The percentage of anthers producing embryoids ranged from 15–20% during September and October to just 1–3% from February through May. The annual average embryoid production rate was 6.18%. 相似文献
10.
Summary The effect of daylenght and temperature for the donor plants (Nicotiana tabacum var. Badischer Burley) on the formation of pollen competent for embryogenesis (P-pollen) by the three possible routes (during normal flower developmentin situ (pollen dimorphism), during cold-treatment of excised flower buds, in cultured anthers) was studied. In all three routes, P-pollen frequency (premitotic pollen, before 1. sporophytic division, PPF) was affected in essentially the same way. At 24 °C and long days, PPF was low and short days had only a slightly increasing effect. At 18 °C and long days, PPF was higher and short days further increased it. Correlated with PPF under the different growth regimes was the percentage of units with more than one vegetative-type nucleus (normal embryos + abortive embryos + multinucleate pollen) in 3 weeks old anther cultures. Under greenhouse conditions, PPF was generally higher than at 24° in growth rooms and showed a maximum in the winter months. Plant age did not affect PPF. These results give further evidence that pollen embryogenesis is predetermined before excision and culture of the pollen or anthers. 相似文献
11.
Efficient callus induction and plant regeneration from anther of Chinese narcissus (Narcissus tazetta L. var. chinensis Roem) 总被引:2,自引:0,他引:2
Callus culture has, to date, been reported only in a few species of Narcissus. We used anthers of Chinese narcissus (Narcissus tazetta L. var. chinensis Roem) as explants for callus induction and plant regeneration. A high percentage of anthers at the early- to mid-uninucleate microspore stage were responsive on the basal MS medium supplemented with 0.5–1 mg l–1 2,4-dichlorophenoxyacetic acid and 0.5–2 mg l–1 6-benzyladenine under dark conditions. Calli were initiated from anther connective tissue or anther wall tissue, and no division of microspores occurred during callus formation, as determined by histological observation. Using 20 random amplified polymorphic DNA primers, we verified the genetic integrity of the anther-derived plants of Chinese narcissus with respect to the donor plants. These results suggest that anther culture in vitro can provide an efficient new micropropagation technique for Chinese narcissus as well as a new strategy for in vitro mass propagation of other daffodils. 相似文献
12.
Z. M. Wei 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1982,63(1):71-73
Summary Pollen shed between 4–8 d from anthers of Triticum aestivum cultured in liquid medium gave rise to calluses. Tillers were harvested at the mid-to late-unicellular pollen stages and chilled for 8 d at 4–5 °C before the anthers were dissected out. Pollen cultures gave about 6 times as many calluses on a per anther basis as anthers cultured on solid medium. With the most productive of 5 cultivars tested, pollen culture results in roughly one callus for each anther used, though the calluses formed by pollen culture were less productive for the regeneration of shoots than calluses derived from anthers cultured on solid medium. The ratio of green to albino shoots is roughly 1 1 for anther cultures but considerably less for pollen cultures. 相似文献
13.
Hui Song Qun-Feng Lou Xiang-Dong Luo Joseph N. Wolukau Wei-Ping Diao Chun-Tao Qian Jin-Feng Chen 《Plant Cell, Tissue and Organ Culture》2007,90(3):245-254
Six experiments (including pretreatment, embryonic callus induction media, preculture conditions, embryo induction media,
embryo germination media, and genotypic effects) were conducted to develop an efficient cucumber (Cucumis sativus L., 2n = 2x = 14) anther culture protocol. Pretreatment and embryo induction were key factors for successful anther culture. Suitable
temperature stress depended on the ecotype, i.e., cucumbers from cold areas responded well to cold shock whereas those from
temperate areas responded well to heat treatment. The best medium for embryonic callus induction was MS medium supplemented
with 4.44 μM BA, 2.26 μM 2, 4-D, 4.64 μM KIN, 3% sucrose and 0.8% agar. For embryo induction, MS medium supplemented with
0.54 μM NAA, 13.32 μM BA, 3% sucrose and 0.8% agar was optimal, and for embryo germination MS medium containing 2.22 μM BA,
6% sucrose and 1.2% agar was best. Using this protocol, we produced callus from 16 genotypes and regenerated plants from three
of 20 evaluated. Three embryos per anther and 42 DH per 45 anthers (93% success) were obtained for cv. Ningjia No. 1, which
was an improved result over a previous report. The origin of regenerants from microspores was determined by cytological, morphological
and AFLP analyses. 相似文献
14.
XiaoPeng Fu ShuHua Yang ManZhu Bao 《In vitro cellular & developmental biology. Plant》2008,44(3):194-202
In this study, we aimed to maximize the rates of somatic embryogenesis achievable in anther cultures of Chinese pink (Dianthus chinensis L.) (2n = 2x = 30). The genotype of the donor plant was found to be a major factor in determining the success rate. Conditions
imposed during anther culture (notably medium composition and light conditions) and pretreatments (namely, cold, heat, and
mannitol incubations) were also found to influence somatic embryo induction. For example, the highest levels of embryogenic
callus induction were achieved when the donor buds had been cold pretreated and the subsequent anther culture was maintained
in darkness. Furthermore, there appeared to be an interaction of genotype with culture conditions. Thus, in cultures of the
cultivar (cv.) ‘Carpet’, the highest rates of embryogenesis were obtained when the anthers had received a 5-d heat-shock,
but such a thermal treatment did not generally produce a significant effect. Likewise, a 3-d mannitol pretreatment was optimal
only for the cross-hybrid line ‘HC’. Assessment of the ploidy of the plants regenerated from the anther cultures revealed
both diploid and tetraploid plants. Histological and cytological observations showed that all of these (both from n-pollen
and 2n-pollen lines) derived from anther wall cells. Spontaneous chromosome doubling was inferred to have occurred during
the embryogenic callus culture period. 相似文献
15.
Iva Smykalová Ji?í Horá?ek Michaela Kubo?iová Prokop ?mirous Jr Ale? Soukup Nikol Gasmanová Miroslav Griga 《In vitro cellular & developmental biology. Plant》2012,48(1):30-39
Plant regeneration was obtained from cultured anthers and hypocotyl segments of caraway (Carum carvi L.). Microspore- and somatic tissue-derived embryos were compared by observation of the regeneration process under identical
induction conditions. Fluorescent microscopy with DAPI staining showed initiation of cell divisions and formation of embryogenic
callus and somatic embryos from anther sacs, with production of embryos of both microspore and somatic origin. Induction of
somatic embryos from hypocotyl-derived callus was also demonstrated. Isozyme native polyacrylamide gel electrophoresis was
used to identify haploids and doubled haploids, and to determine the frequency of spontaneous diploidization of regenerated
plants of microspore origin. Donor plants (2n = 20) and their anther-derived derivative plants (n = 10, 2n = 20, 4n = 40) in callus stage or leafy rosette stage were compared. The esterase (EST) band patterns of regenerated plants differed
from the heterozygous parental material, suggesting that the regenerated plants were microspore-derived haploid/doubled haploid
plants. The similar profile of EST bands between the diploid anther-derived plants and a sample of the donor plants corresponded
to a somatic regeneration pathway. Although the selected induction conditions revealed no preference for induction of microspore
embryogenesis, the anther culture protocol established for caraway utilizing isozyme segregating EST loci markers is suitable
for DH production. 相似文献
16.
J'Aiti F. Benlhabib O. Sharma H. C. El Jaafari S. El Hadrami I. 《Plant Cell, Tissue and Organ Culture》1999,59(1):71-76
The response of anthers to in vitro culture and the effect of coculture of ovaries on anther culturability have been studied in responsive and recalcitrant cultivars
of durum wheat (Triticum turgidum ssp. durum) from Morocco and ICARDA. A large genotypic-dependence of anther culture has been shown in 18 cultivars. Their response in
term of callus and embryo induction varied from 0 to 13%. Coculture of ovaries with anthers enhanced the response of the most
responsive genotype (cv. Sarif) and removed the recalcitrance in Cocorit and Isly cultivars. However, there was no effect
of anther-ovary coculture on green plant regeneration. The implication of the genome and the media conditioning by the ovaries
on anther response is discussed.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
17.
Antimitotic and hormone effects on green double haploid plant production through anther culture of Mediterranean japonica rice 总被引:1,自引:0,他引:1
Isidre Hooghvorst Eduardo Ramos-Fuentes Camilo López-Cristofannini Mirari Ortega Raimon Vidal Xavier Serrat Salvador Nogués 《Plant Cell, Tissue and Organ Culture》2018,134(2):205-215
Rice double haploid (DH) plants are produced mainly through anther culture. In order to improve the anther culture protocol, microspores of two japonica rice genotypes (NRVC980385 and H28) were subjected to three growth regulator combinations and four colchicine treatments on induction medium. In addition, a post anther culture procedure using colchicine or oryzalin was tested to induce double haploid plantlets from haploid plantlets. A cold pre-treatment of microspores for 9 days at 10 °C increased callus induction 50-fold in the NRCV980385 genotype. For both genotypes, 2 mg L?1 2,4-D and 1 mg L?1 kinetin on colchicine-free induction medium gave the best culture responses. The culturability of both genotypes changed on colchicine-supplemented induction media. A high genotype dependency was recorded for callus induction, callus regenerating green plantlets and regeneration of green double haploid plantlets. Colchicine at 300 mg L?1 for 48 h enhanced callus induction 100-fold in H28. Colchicine-supplemented media clearly improved green double haploid plantlet regeneration. We showed that the post-anther culture treatment of haploid plantlets at 500 mg L?1 of colchicine permitted fertile double haploid plantlets to be generated. Finally, an enhanced medium-throughput flow cytometry protocol for rice was tested to analyse all the plantlets from anther and post anther culture. 相似文献
18.
C. Pugliesi P. Megale F. Cecconi S. Baroncelli 《Plant Cell, Tissue and Organ Culture》1993,33(2):187-193
A method of plant regeneration from cotyledons ofHelianthus tuberosus, Helianthus annuus ×Helianthus tuberosus and for the backcross of the interspecific hybrids onH. annuus was developed. Induction of somatic embryogenesis and plantlet regeneration from anther culture of the interspecific hybridsH. annuus ×H. tuberosus is reported.Cotyledons were cultured on Murashige and Skoog basal medium (MS) supplemented with indole-3-acetic acid (IAA) and 6-furfurylaminopurine (kinetin) or N6-benzylaminopurine (BAP). Shoot regeneration occurred on most of the media tested, but the best results were obtained on media with a high concentration of cytokinins (BAP or kinetin: 4 mg l–1) and lower concentration of auxin (IAA: 0.5–1 mg l–1).Embryogenic callus and adventitious buds were initiated from only two anthers of the hybridH. annuus ×H. tuberosus cultured on the MS medium containing BAP (0.2 mg l–1) and 1-naphtalenacetic acid (NAA: 0.1 mg l–1). Prolonged culture of these embryogenic calli and buds on the original medium with successive subculture on MS basal medium without growth regulators resulted in embryo formation and shoot differentiation. The plantlets, after rooting, were established in soil. 相似文献
19.
M. Cristina Pedroso Salomé Pais 《In vitro cellular & developmental biology. Plant》1993,29(4):155-159
Summary Embryogenesis and callus formation inCamellia japonica L., cv. Elegans and cv. Ville de Nantes (>50 yr old), were higher when anthers at tetrad stage or early uninucleate microspore
stage were used. Direct embryo formation was obtained, both in anthers and anther-petaloids. Embryogenesis only occurred under
light on modified Murashige and Skoog medium supplemented with 6-benzylaminopurine (MS10). Embryo production was higher from
petaloids (2.2 to 3.5 embryos/petaloid) than from anther locules (1.2 to 1.5 embryos/anther). However, petaloid-derived embryos
aborted by Week 10 of culture. The embryogenic efficiency of anthers was 5.3% for cv. Elegans and 4.1% for cv. Ville de Nantes.
Scanning electron microscopy images showed that anther-derived globular embryos were covered by a layer of material of smooth
texture. Shoot regeneration efficiency by organogenesis was 6.8%. Regeneration by embryogenesis was 60% for cv. Ville de Nantes
and 97.5% for cv. Elegans. 相似文献
20.
Zhuang Jiajun Jia Xu Chen Guoqing Sun Shancheng 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1985,70(3):294-299
Summary Experimental results showed that the use of potato extract as a basic component of culture medium had a promoting effect on producing calli in anther culture of the intergeneric hybrids of Triticum aestivum × Triticum-Agropyron (intermediate type). The induction frequencies of pollen callus on the Potato-II medium containing potato extract as the main component was much higher than that found on N6 and W5 media. The induction frequencies of pollen callus and green plantlets in four intergeneric hybrid material inoculated at the late-uninucleate pollen stage were all higher than those inoculated at the mid-uninucleate stage. Appropriate increases in culture temperature significantly increased pollen callus induction frequencies of the intergeneric hybrids. The genotype and physiological state of anther donor plants also influenced pollen callus and green plantlet induction frequencies. 相似文献