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1.
Temperature-stress Pretreatment in Barley Anther Culture 总被引:7,自引:0,他引:7
Methods of pretreating anthers at different temperatures priorto culture have been tested, with respect to pollen-callus productionand plant regeneration, in Hordeum vulgare cv. Sabarlis. For callus production, pretreatment of excised spikes (in sealedPetri dishes) was more effective than pretreatment of excisedtillers (in water or in polythene) at both 4 and 25 °C.Pretreatment of individual anthers at these temperatures wasdeleterious. Greater callus yields resulted from pretreatmentat 4 than at 25 °C, both for spikes and tillers, 35weeks being required for maximal yields at 4 °C and 35days at 25 °C. At 4 °C, a shorter pretreatment was requiredfor spikes than for tillers. Pretreatment of spikes was alsomore effective at 4 than at 7, 14 or 20 °C. Pretreatmentof individual spikelets at 4 °C was as effective as thatof whole spikes. For plant regeneration, calluses derived from pretreatment ofspikes were more effective than those derived from pretreatmentof tillers. More plants resulted from pretreatment at 4 thanat 25 °C, both for spikes and tillers. Maximal pretreatmenttimes for plant regeneration generally exceeded those for callusproduction. Following spike pretreatment at 4 °C the maximumfor plant regeneration exceeded that for callus production byabout 2 weeks. With this optimal pretreatment approximately60 per cent of the calluses gave rise to plantlets. Among this60 per cent, for every three calluses giving albinos, two gavegreen plantlets, equivalent to five green plantlets on averagefor every 100 anthers (= two spikes) cultured. The ratio ofgreen to albino plantlets was lower for all other pretreatments. Hordeum vulgare L., barley, anther culture, pollen callus, pollen plant-production, temperature stress 相似文献
2.
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
3.
Nuclear growth, microspore cell growth and cell cycle stage were examined in microspores of anthers of Hordeum vulgare L. cv. Klages taken from florets of the middle of the spike as per anther staging methods. Although there was wide variation in nuclear size at all stages of the cell cycle, mean nuclear size appeared to be a good indicator of cell cycle stage for microspores within anthers. Microspore cell size increased considerably during Gl of the cell cycle. Anthers bearing microspores cytologically characterized as in the mid-uninucleate stage, which have proven to yield high levels of callus production, were determined to be in G1 of the cell cycle and were regularly found in spikes taken from tillers in which the base of the flag leaf had emerged 0 to 3 cm above the penultimate leaf. 相似文献
4.
The effect of various experimental parameters upon the frequency of callus formation from cultured anthers of Oryza sativa has been investigated. Although certain medium components were found to be critical to callus formation, the concentration of these components had little effect upon the frequency of callus formation. The degree to which the callus formation frequency was influenced by cold pretreatment of the flowers was variable. Even though plants were grown under uniform conditions and flowers containing pollen in the microspore stage of development were selected for dissection, the frequency of callus formation varied nonrandomly between flowers. In experiments with populations of flowers in a physiological and developmental state favorable to callus formation 35% of plated anthers produced callus and at least 60% of these calluses gave rise to green plants. 相似文献
5.
Embryoid Formation in Pollen Grains of Nicotiana tabacum 总被引:3,自引:0,他引:3
Anthers of Nicotiana tabacum (n = 24) were cultured on nutrientagar and examined at intervals for pollen embryoids. Embryoidswere formed in anthers of varying developmental stage, the youngestof which coincided with the liberation of free microspores fromtetrads, and the oldest with the formation of bicellular grains.This period in the development of the anther occupied 45days. Older anthers within this range were more successful thanyounger anthers. The first mitosis of the pollen was typicallyasymmetric and resulted in the formation of unequal generativeand vegetative cells. Some of the grains then went into a lagphase for at least 56 days, after which the mitotic conditionwas restored. Embryoids were formed by repeated division ofthe vegetative cell. If the generative cell divided, it didso only once or twice. Occasionally the first mitosis was symmetricand gave rise to equal cells, and in these instances both cellsprobably participated in embryoid formation. The youngest anthersexamined were probably less successful because fewer grainssurvived to enter mitosis. The number of embryoids produced varied considerably from oneanther to another both within the same bud and between differentbuds: values ranging from less than 400 to 10 000 per antherwere encountered. Most of these degenerated after the firstfew divisions, partly because they burst prematurely from thepollen grain wall. Embryoids which continued to develop formedplantlets and/or callus. The largest number of plantlets obtainedfrom one anther was 32. Haploid plantlets were also regeneratedfrom callus by transferring it to a low-sugar medium withoutauxin. The behaviour of grains not forming embryoids was also noted. 相似文献
6.
Culture of isolated microspores and of anthers on media containing IAA directed free microspore development to an embryogenic pathway in C. olitorius. The first division of microspores on transfer to culture media was symmetrical in contrast to the asymmetrical division seen in normal development in vivo. Initially, 10-30% microspores divided symmetrically, but only 0.2-1% of the dividing microspores continued dividing and produced multicellular microcalli. About 30% of these microcalli produced callus but only on medium with 2.0 mg/L zeatin and 0.1 mg/L IAA. Incubation in the dark at temperatures of 35 degrees C for 1 day and then 25 degrees C was found effective for induction of first embryonic division in Corchorus. The frequency of microspore callus formation was higher on medium containing either 3% or 5% sucrose. Addition of colchicine and addition of activated charcoal to the above medium did not enhance microspore division in Corchorus olitorius. On transfer to different media most calli produced roots but regeneration of shoots and embryos was not induced. 相似文献
7.
Comparison of Anther and Microspore Culture in the Embryogenesis and Regeneration of Rye (Secale cereale) 总被引:3,自引:1,他引:2
Anther culture in solid and liquid medium and isolated microspore culture were compared in rye genotypes with potential agronomic characteristics. Some important factors influencing androgenic capacity were optimised. Three weeks cold pre-treatment of spikes and two days mannitol pre-treatment of anthers maximized callus and green plant yield in both culture methods. Intensity order of the culture methods in callus and green plant production was: isolated microspore culture, anther culture in liquid medium and anther culture in solid medium. Genotype ability of embryogenesis followed the same pattern in both cultivation methods. Kinetin (BA) with genotype dependent concentrations created the most effective regeneration conditions. 相似文献
8.
Dun well, J. M. and Thurling, N. 1985. Role of sucrose in microsporeembryo production in Brassica napus ssp. oleifera.J.exp. Bot. 36: 14781491. One cultivar of winter oil seed rape (Brassica napus ssp. oleifera)and three cultivars of spring rape were used in a study of theeffects of sucrose on microspore survival and embryo inductionin cultured anthers. A preliminary study on the winter cultivar(Fiona) revealed that the osmotic pressure of the supernatantof anther homogenates was equivalent to a solution of 17% sucrose.A study of microspore survival and embryo induction in thiscultivar on media containing either 8 %, 12%, 16% or 20% sucroserevealed the highest survival (after 16 d) and the greatestnumber of anthers with induced embryos (after 42 d) occurredon the highest sucrose concentration. A subsequent study on three spring cultivars (Willi, Duplo andTower) examined microspore survival at 8 d and embryo induction(42 d) on media containing either 8 % or 16 % sucrose and againrevealed much higher survival and induction at the higher concentration.The variation in response between the cultivars was also reducedby culture at the higher sucrose concentration. The beneficialeffect of the 16% level occurred regardless of the growth environmentof the donor plants and of the stage of pollen development atthe start of culture. However, macroscopic embryos emerged onlyfrom anthers on the 8 % sucrose concentration, suggesting thattransfer of anthers from a high to a normal sucrose concentrationduring culture would ensure that full advantage was taken ofthe much higher initial survival on the higher concentration Key words: Brassica napus, sucrose, microspore embryo production 相似文献
9.
Microspore development in cultured maize anthers 总被引:1,自引:0,他引:1
The present study follows in vivo and in vitro microspore development utilizing an anther culture-responsive maize genotype (Pa91×FR16) and a DNA-specific fluorescent dye (mithramycin). Cultured anthers were sampled at various times and scored for abnormal microspore divisions, multicellular masses, and embryo-like structures. The frequency of abnormal microspore divisions reached a peak during the first 7 days in culture and then declined. The vegetative nucleus was mitotically active in culture with over 50% of the induced microspores exhibiting this type of division. Multicellular masses and embryo-like structures first appeared in the 14 and 25 day samples, respectively. Most of the microspores did not reach the multicellular stage and an even greater mortality occurred during the formation of embryo-like structures. 相似文献
10.
María A. Marassi Adriana Scocchi Ana María Gonzalez 《In vitro cellular & developmental biology. Plant》2006,42(1):31-36
Summary Anther-derived rice (Oryza sativa L. ssp. japonica variety Yerua P.A.) plants were obtained after cryopreservation by an encapsulation/dehydration technique.
Immature anthers, excised from spikelets pretreated at 8°C for 8d, were encapsulated in calcium alginate beads. The beads
were cultured on N6 medium with 11.5 μM naphthalenaecetic acid (NAA) and 2.3 μM 6-furfurylaminopurine (KIN). Fifteen percent of the encapsulated anthers formed calluses when pretreated with sucrose for
3 d in liquid medium, desiccated on silica gel, slowly cooled to −30°C, immersed in liquid nitrogen (LN), thawed, and recultured.
The cryopreserved encapsulated anthers produced 1.67 shoots/callus, in contrast to the control (non-cooled encapsulated anthers),
which produced 6 shoots/callus. Eighty percent of the plantlets developed into normal plants after being transferred to greenhouse
conditions. Histological observations showed that the origin of the plants was not modified by the cryopreservation process. 相似文献
11.
The obtaining of calluses and plantlets from cultured wheat anthersat the stages from pollen mother cell to trinucleate microspore has been reported previously. Haploids as well as diploids existed among the regenerated plantlets derivedfrom anthers at these stages. Present paper reports the study on androgenesis patter-ns of cultured anthers at meiosis, tetrad, early mid- and late uninucleate and trinucleate stage. Cytological evidence of pollen-origin of calluses produced by anthers atthese stages was given. Observation showed that meiosis of wheat anthers was able tocomplete under culture conditions, resulting in releasing microspores, from which multinucleate and multicellular pollen grains formed. In meiosis anthers, abnormal cells,including syncytium and two kinds of binueleate calls were sometimes observed. Theymight be products of abnormal meiosis and abnormal development of tapetum cells. Itwas noted that failure and/or uncomplction of forming callus wall and/or pollen wallin in vitro anthers at meiosis, tetrad and early uninucleate stage occured often. Itmight lead to the low frequency of callus induction. Mature wheat anthers (trinucleate stage) contained both normal and abnormal pollen grains (pollen dimorphism); onlythe abnormal pollen grains developed into embryoids while all the normai trinucleatepollen grains degenerated rapidly. However, the date of the frequency of equal divisionof microspores suggested that abnormal pollen (N pollen, small pollen) could not be theonly source of androgenic pollens in cultured anthers at late uninucleate and other earlier stages. 相似文献
12.
The various pathways of pollen development were investigatedin cultured anthers of Lolium temulentum, Festuca pratensisand the L. multiflorum x F. pratensis hybrid Elmet.In all three, development from the vegetative cell was the predominantpathway of pollen callus development. However, there were characteristicdifferences in the behaviour of the generative cell. In L. temulentumit remained attached to the pollen wall and degenerated, whereasin F. pratensis it divided. In Elmet it detachedfrom the pollen wall and remained undivided. Both polarizedand unpolarized partitioned calluses were observed. Developmentof the fusion product of the vegetative and generative nucleiwere also observed in anthers of L. temulentum. Anomalous grainswere not found to be major source of pollen calluses. Sections of anthers of L. temulentum were used to investigatethe origin of S pollen grains, the small pale-staining grainswhich denote pollen dimorphism. Such grains form out of contactwith the tapetum and are therefore determined before or duringmeiosis (i.e. before harvest of anthers for culture). Sectionswere also used to demonstrate the influence of the durationof pretreatment on the development of the middle layer of theanther wall. Festuca pratensis, Lolium temulentum, Lolium x Festuca, anther culture, haploid, microspore, pollen 相似文献
13.
Bin Huang 《Plant Cell, Tissue and Organ Culture》1987,9(1):45-48
Anthers of wheat cultivars Orofen and Pitic 62 were incubated for 8 days at 15, 20, 25, 30, 35 and 40°C before transfer to 25°C. Compared with anthers cultured at 25°C constantly, anthers treated at 30°C produced 40% more microspore callus and green plants in both cultivars whereas those treated at 35°C produced 2–3 fold more green plants. Treatment at 40°C was deleterious. Possible modes of action of high temperature on callus production and albinism were discussed. 相似文献
14.
大麦不同基因型游离小孢子的直接培养再生及培养体系的优化 总被引:3,自引:0,他引:3
以微搅拌法建立了小孢子直接游离的预处理和培养程序。在大田生长的4个对培养反应不同的大麦基因型上,以新鲜幼穗游离小孢子进行直接培养,均成功地诱导了胚状体并获得再生绿色植株。小孢子的发育进程说明,直接游离的小泡子在预处理过程中的发育要慢于在花药中预处理的小孢子,而且其培养效率也较低。直接游离小孢子的培养密度以0.8~1.0×105/ml较理想,至少应不低于6×104/ml.8%-10%的糖浓度可明显提高小孢子分裂频率和胚状体诱导频率。实验结果也表明两种培养基FHG和MN6无明显差异,均适宜于直接游离的小孢子培养,并对游离小孢子直接培养在理论和应用上的意义进行了讨论 相似文献
15.
Chi-Chang Chen 《In vitro cellular & developmental biology. Plant》1977,13(8):484-489
Summary Rice (Oryza sativa L., 2n=24) anthers containing microspores in the early-uninucleate to first-mitosis stages were induced successfully to develop
into plants in vitro through an intermediary step of callus formation. Callus initiation occurred with highest frequency in
anthers containing mid-uninucleate microspores. The callus derived from different stages of microspore development differed
in the potential to differentiate into plants. The plants regenerated from pollen callus were predominantly haploid or diploid;
polyploid and aneuploid plants were relatively infrequent. The first division of the uninucleate microspores was asymmetrical,
resulting in the formation of large vegetative and small generative nuclei. The vegetative nucleus divided repeatedly and
assumed the major role in the formation of callus, whereas the generative nucleus degenerated rapidly. Simultaneous division
of the two nuclei was observed in a few pollen grains. Nuclear fusion during the very initial stages of pollen development
was postulated to account for the occurrence of the diploid and polyploid plants.
This work was supported by the National Science Council, Republic of China. 相似文献
16.
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
17.
Mingsheng Peng Asma Ziauddin David J. Wolyn 《In vitro cellular & developmental biology. Plant》1997,33(4):263-268
Summary Development of asparagus microspores in cold-treated buds of varying sizes and shed microspores from these buds in in vitro culture were observed cytologically for the G459 genotype. Before cold pretreatment, more than 75% of the microspores in
flower buds of the 1.4–1.6, 1.7–1.9, 2.0–2.2, 2.3–2.5, and 2.6–2.8 mm size classes were at the early-, mid-, late-uninucleate,
early-, and late-binucleate stages, respectively. After 7 d in cold treatment, percentages of microspores at different stages
changed in all flower buds. Most notable was the appearance of binucleate microspores resulting from symmetric rather than
asymmetric division. For flower buds of 1.7–1.9, 2.0–2.2, and 2.3–2.5 mm size classes, 4.9%, 27.2%, and 11.4% of the microspores
had divided symmetrically, respectively. When microspores from buds of each size category were cultured in androgenesis induction
medium, only microspores completing symmetric pollen mitosis I during cold treatment were observed to divide further, and
calluses were only obtained from microspores of flower bud size classes where symmetric divisions were observed after several
days of cold treatment. Significant correlations existed among microspore callus yield, the percentage of late-uninucleate
microspores in vivo before cold treatment, and the frequency of symmetric pollen mitosis I after 7 d of cold treatment. Consequently, asparagus
microspore androgenesis may occur through one developmental pathway, where a symmetric first mitotic division is a prerequisite
for continued development. 相似文献
18.
Shed microspore embryogenesis and fertile plantlet regeneration were observed in a salt susceptible × salt tolerant indica rice F1 hybrid involving IR 24 and CRM 30. The in vitro culture response and regeneration of green plantlets in the hybrid were superior to those of the parents. Direct embryogenesis
and plantlet regeneration with multiple tillers were observed in shed microspore embryos. In intact anther culture, plantlet
development from microspore involved a callus phase. The number of multiple tillers developed through secondary embryogenesis
was almost equal in both the cases. However, the results indicate that regeneration of green plantlets was higher in case
of shed microspore culture in liquid medium containing the synthetic polymer Ficoll 400 than from intact anthers cultured
on a semi-solid system. Shed microspore culture produced a number of double haploids, which may result in far reaching consequences
in genetic improvement of rice.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
19.
20.
The role of ethylene in anther culture of durum wheat ( Triticum durum Desf. cv. Ardente) was analyzed by testing the effects of 2-chloroethylphosphonic acid (ethrel) silver thiosulfate (Ag+ ), a -aminooxyacetic acid (AOA) and 1-aminocyclopropane-l-carboxylic acid (ACC) on microspore division observed after 21 days of culture and on development of calli estimated at day 45. The use of ethrel and Ag+ indicated a positive effect of ethylene on microspore division, whereas the use of AOA, and to a lesser extent ACC, snowed a negative effect. In contrast, the addition of ethrel or Ag+ indicated that ethylene inhibits the development of microspore-derived calli. AOA gave contradictory results. Ethylene production by anthers was about 7 pl anther−1 h−1 and decreased during culture. ACC content in the anthers was maximal at day 9, whereas malonyl ACC (MACC) increased sharply from day 0 to day 3 and then decreased. The addition of AOA or ACC to the culture medium decreased or increased, respectively, ethylene production of anthers and the ACC and/or MACC content, but at concentrations higher than those that modified the formation of calli. This formation seems to occur in two successive phases: induction and initiation of microspore division, which was promoted by ethylene, followed by callus development, which was inhibited by ethylene. 相似文献