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1.
Summary

In vitro culture methods were used to germinate olive embryos prior to maturation. Fruit, seed and embryo development were established with consecutive sampling from 20 to 100 days after bloom. For that same period, embryo development and germination success were determined by in vitro culture trials using one-third strength MS medium with or without the addition of zeatin. For early developmental stages, when isolation of the embryo was difficult, a cut portion of the seed containing the embryo was used for culture. The embryos cultured within the cut seed portions germinated and formed normal plantlets. Histological observations indicated a close similarity between the natural and in vitro immature embryo differentiation pattern, progressing through preglobular, globular, heart-shaped and torpedo-shaped stages. In some cases, however, the in vitro immature embryos developed or germinated abnormally. The presence of zeatin (0.25 mg l–1) in the culture medium and the use of a cut seed-portion containing the immature embryo allowed in vitro germination sooner after bloom than previously obtained. On the contrary, zeatin was a handicap for mature olive embryo in vitro germination, which reached 100% seedling formation when no plant growth regulators were used.  相似文献   

2.
冬枣极早期幼胚培养成苗技术研究   总被引：1，自引：0，他引：1  
梁春莉  赵锦  刘孟军 《园艺学报》2014,41(10):2115-2124
以胚龄15 ~ 20 d的球形期后期‘冬枣’幼胚为试材,对培养基种类、种皮保留与否、接种方式、生长调节剂种类、水解乳蛋白（LH）和活性炭（AC）用量、蔗糖浓度等影响极早期幼胚培养的诸因素进行了系统研究。结果表明,采用MS + 3.0 mg ? L-1 IAA + 0.5 mg ? L-1 6- BA + 1 g ? L-1 AC + 0.8 g ? L-1 LH + 30 g ? L-1蔗糖的培养基,去除种皮后将带有胚乳的幼胚合点端接入培养基,在21 ℃下进行培养,胚萌发率和正常胚比率最高可分别达到93%和84%。萌发后的幼胚在MS + IBA 0.4 mg ? L-1的培养基中继续培养,生根率可达40%。其中,去除种皮带胚乳培养和幼胚合点端朝下接种对极早期幼胚的培养至关重要。  相似文献   

3.
SUMMARY

We investigated the effects of dry-storage of Musa velutina seeds on seed and embryo germination. Seeds were collected immediately after harvest and stored in dry vermiculite at 25°C, in the dark, for 0, 1, 2, or 4 months. The germinability of dry-stored seeds was assessed by incubation in vermiculite at 25°C for 4 months, with 75% [(w/w); –0.01 MPa] moisture content to promote embryo development in viable seeds. Seeds that had not been dry-stored and stored in vermiculite at 75% (w/w) moisture content for 4 months showed 90% germination by 20 d after sowing, but no seeds that had been dry-stored for longer than 1 month germinated. Seeds were also dry-stored at 25°C, in the dark, in Petri dishes for 0, 1, 2, 3, or 4 weeks after harvest and the embryos removed from these seeds were cultured on MS medium. All embryos that had been isolated from seeds at harvest (i.e., 0 weeks in dry-storage) germinated during embryo culture. However, embryos removed from seeds that had been dry-stored for 1 week showed a substantial decrease in both size and germinability. Seeds were also stored at different moisture contents for 4 months after harvest. No seeds that had been stored at 0% moisture content (i.e., dry) germinated, but > 85% of seeds stored at ≥ 25% (w/w) moisture contents germinated. Higher moisture contents increased the uniformity of seed germination, showing that M. velutina seeds rapidly lose their germinability through dehydration, indicating that they are sensitive to desiccation.  相似文献   

4.
Summary

Efficient in vitro procedures for mass propagation of four commercially important Echinacea species have been deveoped. Plants of E. angustifolia, E. pallida, E. paradoxa and E. purpurea were regenerated by three methods, namely axillary bud proliferation, adventitious shoot formation and somatic embyrogenesis. Shoot tips obtained from in vitro germinated seedlings, adventitious shoots or somatic embryo-derived plantlets, when cultured on Murashige and Skoog medium enriched with 1 μM 6-benzylaminopurine, 2 μM kinetin, 0.5 μM indole-3-butyric acid and 4 mg–1 paclobutrazol multiplied three-fold within 3–4 weeks in culture. Incorporation of paclobutrazol in the shoot multiplication medium was necessary to recover healthy and robust shoots suitable for rooting. Direct, high-frequency shoot formation on intact leaves of shoots grown on 6-benzylaminopurine and kinetin-supplemented media, an unusual and novel observation made in this study, occurred in all the species studied. Rooting of in vitro developed shoots was achieved relatively easily with Murashige and Skoog basal medium rather than with auxin-enriched media. Culturing of hypocotyl explants on medium containing 3,6-dichloro-o-anisic acid (commonly known as dicamba), or 2,4-dichlorophenoxyacetic acid, resulted in direct somatic embryogenesis in all the species examined. The presence of cytokinin was required for somatic embryo germination, but further development of germinated somatic embryos into normal plantlets occurred in Murashige and Skoog medium. We conclude that the procedures described here could be used for rapid propagation as well as genetic transformation of commerically cultivated Echinacea species.  相似文献   

5.
An efficient protocol for in vitro maturation of very immature, <10 mm, avocado embryos has been developed. The efficiency of plant recovery as well as the quality of the resulting plants was greatly improved by including a maturation phase prior to induction of germination. The influence of different factors, such as the gelling agent, organic supplements or abscisic acid, on embryo maturation and subsequent germination was tested. Optimum conditions were met when maturation was carried out in B5m medium supplemented with the Jensen's amino acids, an extra 88 mM sucrose and 6 g l−1 agar as gelling agent. At these conditions, embryos which had been collected 68 days after pollination germinated at a 65% rate in solid medium, giving rise to healthy and vigorous plantlets. Anatomical differentiation and storage product accumulation occurring during the in vitro maturation phase were studied by means of histological techniques. Results obtained revealed that, at the end of the in vitro maturation period, embryos resembled the pattern previously established for avocado embryos matured under in vivo conditions: histodifferentiation had been accomplished and starch granules and protein bodies were abundant.  相似文献   

6.
Embryo development followed by seed germination was investigated in seeds of Christmas rose (Helleborus niger) held under different temperature conditions. The seeds had a heart-shaped embryo when dispersed from parent plants. That embryo developed rapidly to the torpedo stage at 25 °C, after which no further development occurred. When seeds with a torpedo-stage embryo were held at 15 °C, the embryos developed to the cotyledon stage, but no further germination occurred. Similar phenomena in the course of embryo development were observed in seeds held in a non-heated polyethylene house, in which the embryos developed to the torpedo stage at 20 °C or more, while further development to the cotyledon stage was attained only at temperatures of 15 °C or less. Seeds with a cotyledon-stage embryo rarely germinated until they were treated at 4 °C for more than 8 weeks. These results suggest that seeds of H. niger have a deep, simple morphophysiological dormancy caused by the combination of rudimentary embryos and a physiological dormancy that can be broken by cycles of warming and chilling.  相似文献   

7.
Summary

The effects of seed coat removal and chilling on the germination of seeds of ten cultivars of ornamental peach (Prunus persica Batsch) were investigated. Seeds were rinsed in running tap water for 48 h in order to facilitate seed coat removal. Only a few non-chilled, intact seeds germinated (e.g., 6% of ‘Hito’ seeds). Seed coat removal and no chilling resulted in some seed germination in eight of the ten cultivars, ranging from 6% to 83%. Chilling intact seeds at 5°C for up to 10 weeks resulted in greater frequencies of germination (average = 85% germination) than seed coat removal before (average = 80%) or after chilling (average = 73%). There was a linear decrease in the germination percentage with an increase in abscisic acid (ABA) content for ‘Yaguchi’,‘Kanpaku’, and ‘Kikumomo’ seeds (R = –0.66; P <0.001). The most consistent decrease in total seed ABA content (average of 64%) occurred during the 48 h rinsing period. In ‘Kanpaku’, dry seeds had an intermediate ABA content (13 ng per seed); but, in the embryonic axes, this increased from 0.03 ng to 1.2 ng per seed with an increase in the duration of the chilling period. The lowest germination percentages were found in this cultivar. These results suggest that ABA synthesis in the embryonic axes during chilling may affect the varietal characteristic of seed dormancy, and that rinsing seeds for >48 h could remove sufficient ABA to allow seed germination with minimum chilling.  相似文献   

8.
Summary

A protocol was developed for induction, maturation and germination of somatic embryos from the tissues of germinating seeds of black pepper (Piper nigrum L.). Explants were cultured on growth regulator – free solid SH medium maintained in the dark. The first somatic embryos developing directly from the explant tissue were noticed after 60 d of culture. Somatic embryos originated from a ring-like tissue on the micropylar region of the seeds. Sucrose concentration of the medium was found to be crucial for the induction of somatic embryos, and 30 g l–1 was found to be the optimum. Maturation and germination of somatic embryos were achieved on the same medium. Suspension culture enhanced the process of maturation and germination. Regenerated plants were established in soil. Histology confirmed the ontogeny and each stage of development. Growth regulators were found to inhibit the induction of somatic embryogenesis. Cytological analysis of the regenerated plants revealed the normal chromosome number of 2n=52.  相似文献   

9.
The application of gibberellic acid (0, 5, 20 and 80 µg) to seedlings of Syringa vulgaris L. about two weeks after germination increased significantly the total length, the length of internodes and the dry weight (d.w.) of shoots and the net assimilation rate. GA3 also had a small but significant positive effect on the number of pairs of leaves, especially at high temperatures; it increased the girth, but this effect was not significant.

GA3 reduced significantly the d.w. of roots and leaves but did not affect the leaf/root ratio. GA3 had no effect on the total plant d.w. or the relative growth rate.

The effect of GA3 on shoot growth was dependent on temperature and on the stage of growth. One and two weeks after its application it had the maximum relative effect at high temperatures (21–24 °C) but at the end of the experiment (8.5 weeks) the maximum effect was reached at 12 °C; it decreased with increasing temperature and was not significant at 24 °C. By this stage there were, however, no statistical interactions between temperature and GA3 for total length and for d.w. of shoots, roots, leaves and of the whole plant.

Increasing temperatures over the range 12–24 °C resulted in increases in the following characteristics: the number of pairs of leaves; length of internodes, diameter and total length of the shoot; the d.w. of shoots, roots, leaves and of the whole plant; the d.w. ratios of leaves/roots and shoots/roots; and the relative growth rate and net assimilation rate. High temperatures reduced the root/whole plant dry weight ratio. The effect of temperature on the number of pairs of leaves was linear, and results at alternating temperatures (24°/18° and 21°/15 °C, 8 hr/16 hr) did not deviate significantly from values expected on the basis of mean daily temperature.  相似文献   

10.
Seedless grape cultivars (Vitis vinifera L.) are widely grown in Europe, America and Asia. Fungal diseases are a great threat of them. Several wild Chinese Vitis species showed high resistance to many fungal diseases. Therefore, an investigation was conducted to assess the potential to incorporate these species in a breeding project for development of the disease-resistant seedless cultivars. Hybridization was conducted using V. vinifera as female parents and the wild Chinese Vitis spp. as male parents. In-ovulo embryo rescue was used to develop hybrid plants from the seedless females. An efficient protocol is reported here for the in vitro embryo rescue and plant development from the cross Emerald Seedless × Beichun. Ovules were excised from immature fruits 7 weeks after pollination (WAP) and cultured in the double-phase ER medium supplemented with 6.0% sucrose and 0.3% activated charcoal (AC). Following 8–12 weeks of culture, embryos were removed from the ovules and transferred onto WPM supplemented with 1.0 μM 6-benzyladenine (6-BA), 2.0% sucrose and 0.2% AC and solidified with 0.6% agar. After 8 weeks of culture, the embryos germinated and subsequently grew into whole plantlets. With the optimized parameters developed in the present study, about 34.0%, 91.2% and 77.4% of embryo formation, embryo germination and plant development were obtained, respectively. When this protocol was applied to 11 other cross combinations, genotype was found to significantly influence embryo formation, embryo germination and plant development, with different frequencies of hybrid plants successfully obtained in all crosses.  相似文献   

11.
In vitro embryo culture enabled satisfactory germination of immature seeds produced in crosses from early ripening sweet cherry varieties (Prunus avium L.). Three varieties —‘Rita’, ‘Bigarreau Burlat’ and ‘Carmen’— were crossed with ‘Early Star’ as male parent. Germination rate was affected by the developmental stage of both fruit and embryo. Fruit ripening stage was a critical factor for culture infection rate that increased with maturity. In-ovule embryo culture on Murashige and Skoog medium without hormones improved the embryo size but did not increase the germination rate due to a further increase in infection rate. Ex-ovule embryo culture on Murashige and Skoog medium supplemented with BA 1 mg L?1, NAA 0.5 mg L?1, 20 g L?1sucrose, 10 g L?1 sorbitol and 6 g L?1agar during the stratification time increased embryo length. Germination was performed on Brooks and Hough medium at the 22?±?1?°C with 16/8 h light/dark photoperiod. The highest germination rate (75?%) was reached in embryos that were 3?4 mm in length, after 30-days stratification at 4?°C. Embryos in fruits at green-yellow stage that were 3?4 mm long were morpho-physiologically developed to produce bipolar seedlings, without combined application of embryo culture and micropropagation.  相似文献   

12.
柿胚胎发生发育的研究   总被引：10，自引：1，他引：10  
钟云  刘勇  蒋侬辉  刘善军 《园艺学报》2004,31(3):353-356
 以甜柿品种‘卡迟莎’和涩柿品种‘高安方柿’为材料，采用常规石蜡切片法对胚胎发生发 育进行了研究。结果表明：柿胚珠倒生，双珠被，薄珠心，单孢原细胞；大孢子四分体为直线型，仅合点端的一个具功能作用；蓼型胚囊，两个助细胞具角状丝状器，两极核在受精前融合为次生核，3个反足细胞呈“品”字形排列；胚胎发育属于茄型，合子约休眠两周后开始分裂；胚乳为核型，花后3 d开始分裂，在球形胚早期开始形成细胞壁，在子叶形胚期胚乳组织基本形成。在胚胎发生发育的过程中有胚乳退化，合子不分裂等败育现象。  相似文献   

13.
Summary

We examined changes in leaf growth and chemical composition, including chlorophyll content, ribulose bisphosphate carboxylase-oxygenase (RuBisCO), and glycine betaine (GB) in relation to photosynthesis and transpiration responses to salt stress in Amaranthus tricolor leaves. To induce salt stress, plants were transferred to a growth medium containing 300 mM NaCl for 7 d followed by 7 d of relief from salinity. A decrease in leaf enlargement began 3 d after salt stress, and leaves subsequently showed the same degree of regrowth as controls after relief in non-salt medium. Chlorophyll content expressed on a leaf-area basis increased under conditions of salinity due to a reduction in leaf tissue water content. The decrease in chlorophyll content continued throughout the 7 d of relief from salinity. The RuBisCO and soluble protein contents when expressed on a leaf dry-weight basis decreased in response to salinity, and then gradually increased during the relief period. GB content increased slightly up to 3 d of salt stress, and showed typical accumulation during salt stress. GB content decreased sharply immediately after plants were transferred to non-stress medium, but remained at a higher level throughout the relief period. A decrease in photosynthetic activity and transpiration rate preceded any changes in leaf area, RuBisCO or GB content. During relief from salinity, photosynthesis and transpiration rates gradually recovered to control levels with restoration of stomatal conductance. The above findings suggest that the increase in GB content is important in adaptation to salt stress in Amaranthus plants, although photosynthesis and transpiration responses occurred immediately after salt-stress.  相似文献   

14.
Summary

Low-temperature sweetening of parsnip (Pastinaca sativa L.) roots, which may occur both pre-harvest and post-harvest, is a well-known phenomenon, but is poorly understood. ‘White King’ parsnip roots were analysed over two seasons (2008 and 2009) during field growth and over six months cold storage at 1ºC for their dry matter content, as well as their starch, sucrose, glucose, fructose, and malto-oligosaccharide (MOS) concentrations and for their changes in amylolytic enzyme activities (2009 only). In 2008, when the air temperature fell below 5ºC for 12 d in the last 8 weeks before harvest, the concentration of starch decreased significantly (P ≤ 0.05) and the concentration of total sugar increased significantly. In 2009, when temperatures did not fall below 5°C during the last 8 weeks before harvest, the concentrations of starch (339 + 17 mg g–1 DW) and total sugars (168 + 12 mg g–1 DW) did not change significantly. However, in both years, during 24 weeks in cold storage at 1ºC, starch levels were almost completely depleted, whereas sucrose concentrations and, to a lesser extent, MOS concentrations increased at least two-fold. Glucose and fructose concentrations also increased significantly during 24 weeks in cold storage, but to a much lower level than sucrose or MOS. Total amylolytic activity and α-amylase activity doubled during the first 8 weeks in cold storage, whereas β-amylase activity did not change significantly. As products of α-amylase activity, changes in MOS concentration paralleled the changes in α-amylase activity. The accumulation of sucrose and MOS may raise the culinary quality of cold-stored parsnip.  相似文献   

15.
In order to improve somatic embryogenesis production in date palm Phoenix dactilyfera L. cv. Deglet Nour (DN), a comparative study between somatic (SE) and zygotic (ZE) embryos developments was carried out. The data showed that ZE maturation occurred from 10 to 19 weeks after pollination (WAP). During this period, the fresh weight (FW) and the dry weight (DW) of ZE increased progressively to reach a maximum level at 19 WAP. SE development occurred in three distinct stages. The DW remained constant during the two first stages, and declined slightly during the third and final stage. Embryo protein analysis revealed significant differences between ZE and SE. The ZE total protein level was initially low and increased to the maximum at mature stage. However, no significant change in total protein was detected during SE development. SDS-PAGE analysis showed a poor protein profile for SE, compared to that of ZE. In the latter, a 22 kDa protein was identified by N-terminal sequencing as a glutelin. This protein was accumulated rapidly during early development and remained at a relatively constant level during ZE development, and then declined progressively 12 days after embryo germination (DG). This protein seems to be absent in SE.  相似文献   

16.
Gentians are alpine plants and two endemic species, Gentiana triflora and G. scabra, are cultivated as ornamentals in Japan. Because most of the Japanese cultivars are F1 hybrids, seed germination rate is an important factor for maintaining these cultivars. However, germination rates frequently differ among cultivars. Poor germination is a severe problem for gentian production, although there is almost no difference in their germination period and growth rate. In this study, we compared seed germination rates among three cultivars of G. triflora. A high rate of seed germination was observed in ‘Majel’ (MJ) and ‘Iwate’ (IW), but a low rate was recorded in ‘Iwate-Yumeminori’ (YM). We conducted a targeted metabolome analysis using these cultivars to elucidate the potential cause of inhibited seed germination. Multivariate analysis revealed that the aberrant accumulation of specific amino acids and a decrease in energy metabolites were observed in YM seed. Furthermore, protein concentrations and proteolytic activities in YM seed were lower than in MJ seed. These results imply that energy depletion was the main reason for the decreased rate of germination and that this depletion inhibited processes involved in seed germination, including de novo synthesis of the proteins necessary for germination.  相似文献   

17.
Most of the achenes produced by Rosa multibracteata Hemsl. & E. H Wilson are dormant on maturity and require pretreatment to stimulate germination. To investigate the mechanism of dormancy and to develop effective methods of improving germination, roles of the pericarp, testa, and embryo of R. multibracteata in regulating dormancy were studied by investigating the effect of different pretreatments on germination. The effects of temperature and water stress were also tested with achenes treated by warm plus cold stratification. In freshly harvested achenes, pericarps are permeable and the embryo fully developed, which eliminates the possibility of physical, morphological, or morphophysiological dormancy. Germination percentage remained low (<5%) despite softening the pericarp or even removing it fully. However, fully removing the testa improved germination significantly (39%), indicating the possible presence of germination inhibitors in the testa. Dry storage, scarification with sulphuric acid (H2SO4), and warm stratification proved ineffective by themselves but when combined with cold stratification, improved germination and shortened the cold stratification period needed to break dormancy. Dry storage for 68 weeks followed by cold stratification for 16 or 24 weeks resulted in maximum germination (72–79%) among all the treatments. In achenes scarified with H2SO4, germination increased with an increase in the duration of cold stratification. Neither gibberellic acid (GA3) nor ‘smoke water’ (water through which smoke had been bubbled for 2 h) had any positive effect on germination even on seeds that had been mechanically scarified or stratified. Both high temperature and water stress lowered germination in achenes treated with warm plus cold stratification. Our results suggest that R. multibracteata achenes have an intermediate physiological dormancy, and that dry storage for 68 weeks followed by cold stratification for 16 or 24 weeks is the best method for propagating R. multibracteata from seed.  相似文献   

18.
Plant growth regulators were applied to the foliage and immature fruit clusters of the stenospermic grape selection ‘C35-33’ at various periods before bloom to stimulate viable seed development. In the 1987 season five different plant growth regulators were used, but in 1988 the growth retardants Cycocel and XE-1019 were used exclusively. Chemical treatments applied 35 days after bud break increased significantly germination percentage. Experimental results indicate that the use of certain plant growth regulators may aid in increasing the efficiency of seedless grape breeding by providing an alternative to in-ovulo embryo culture.  相似文献   

19.
沙芥花粉萌发特性和柱头可授性的研究   总被引：8，自引：0，他引：8  
 用培养基发芽法研究沙芥花粉萌发特性;用人工异株授粉法和联苯胺—过氧化氢法测定柱头可授期。结果表明：沙芥花粉萌发的最佳培养基是20%蔗糖+0.001%硼酸;花粉能够萌发的温度范围为15～38 ℃,最适萌发温度为25 ℃;从花蕾充分膨大期至开花后16 h花粉活力较强,开花后48 h花粉基本无活力;花粉储藏的适宜条件为低温（4 ℃）,储藏期为7 d。柱头可授期为开花前48 h至开花后96 h,其中开花后24～72 h可授性强。  相似文献   

20.
Summary

Dehydrins are a group of plant proteins which respond to any type of stress that causes dehydration at the cellular level, such as cold and drought stress. Previously, three dehydrins of 65, 60, and 14.kDa were identified as the predominant proteins present in cold acclimated blueberry (Vaccinium corymbosumLinn.) floral buds. Levels were shown to increase with cold acclimation and decrease with deacclimation and resumption of growth. In the present study, to determine if dehydrins are induced in other organs in response to low temperature treatment (48C) and in response to drought, accumulation of dehydrins was examined in leaves, stems, and roots of two cultivars and one wild selection (a V. corymbosum cultivar, a V. ashei Reade cultivar, and a V. darrowi Camp selection) of blueberry by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) followed by immunoblotting. Cold treatment involved placing plants in a cold room maintained at 48C for five weeks; drought stress was imposed by withholding water from potted, greenhouse-grown plants for 34.d. Relative water content (RWC) of shoots was determined periodically throughout the drought treatment. Dehydrins accumulated with both cold and drought stress but their molecular masses varied depending upon blueberry species. Dehydrins accumulated to higher levels in stems and roots than in leaves with cold stress and to higher levels in stems than in either roots or leaves with drought stress. Furthermore, cold treatment combined with dark treatment induced higher levels of dehydrins than cold treatment combined with a 10.h light/14.h dark photoperiod, suggesting that dehydrins may be responsive to changes in photoperiod as well. In the cold-stress experiment, the level of dehydrin accumulation was correlated with expected level of plant cold hardiness in the three genotypes. In the drought stress experiment, dehydrins accumulated prior to significant changes in RWC, and dehydrin levels did not appear to be closely correlated with RWC either among or within genotypes.  相似文献   

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