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离体培养条件下核桃器官发生和体细胞胚胎发生 总被引：11，自引：0，他引：11
采自幼树、成年树及成年树伐根萌条的嫩梢腋芽,在改良DKW培养基+6-BA1.0mg/L中可伸长生长;在加有6-BA1.0mg/L+IBA 0.01mg/L的继代培养基中,可保持腋芽的生长并形成不定芽。芽增殖率为每月600％左右。芽苗经5.0mg/L IBA处理7天,然后在含活性炭的无激素培养基中培养20天,有54％可生根成苗。5月中旬至6月上旬的幼胚,在改良DKW培养基+6-BA1.0 mg/L中,黑暗培养30天左右可产生体细胞胚,并可连续多代保持胚性分生能力。体胚在无激素的发芽培养基中黑暗培养7天左右可发芽成苗。胚性愈伤组织在悬浮振荡培养中可保持分生能力。 相似文献
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Differentiation of adventitious buds and somatic embryos from mature zygotic embryos of Picea sitchensis (Bong.) Carr. is described. Adventitious buds were formed on embryos pulse-treated with 250 microM benzyladenine for 2 h and cultured on medium lacking growth regulators. Buds were initiated at different frequencies and sites depending on when the BA-pulsed embryos were transferred to fresh culture medium. Embryogenic callus was formed when the zygotic embryos were cultured on medium containing 10 microM 2,4-dichlorophenoxyacetic acid and 5 microM benzyladenine. Although 50% of the embryos gave rise to embryogenic callus, only 20% of the callus continued to proliferate after subculture. Proliferation of new somatic embryos occurred from both the embryonic region and the suspensor region on previously formed somatic embryos. The pattern for development of adventitious buds and somatic embryos in Picea sitchensis is compared to that in Picea abies under similar culture conditions. 相似文献
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Of the various alternatives for cloning elite conifers, somatic embryogenesis (SE) appears to be the best option. In recent years, significant areas of lodgepole pine (Pinus contorta) forest have been devastated by the mountain pine beetle (MPB) in Western Canada. In an attempt to establish an SE propagation system for MPB-resistant lodgepole pine, several families displaying varying levels of resistance were selected for experimentation involving shoot bud and immature seed explants. In bud cultures, eight embryogenic lines were induced from 2 of 15 genotypes following various treatments. Genotype had an important influence on embryogenic culture initiation, and this effect was consistent over time. These lines were identified by microscopic observation and genetic markers. Despite the abundance of early somatic embryos, the cultures have yet to develop into mature embryos. In contrast, immature zygotic embryos (ZEs) cultured from megagametophytes initiated SE at an early dominance stage via nodule-type callus in 1 of 10 genotypes. As part of the study, putative embryogenesis-specific genes, WOX2 (WUSCHELL homeobox 2) and HAP3A, were analyzed in cultures of both shoot bud explants and ZEs. On the basis of these analyses, we postulate that PcHAP3A was expressed mainly in callus and may be involved in cell division, whereas WOX2 was expressed mainly in embryonal mass (EM)-like tissues. The findings from this study, based on molecular assessment, suggest that the cell lines derived from bud cultures were truly EM. Moreover, these experimental observations suggest that PcWOX2 could be used as an early genetic marker to discriminate embryogenic cultures from callus. 相似文献
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Two embryogenic cell lines characterized by different morphology and color, white and red, were obtained from an immature
zygotic embryo of Japanese larch (Larix leptolepis Gord.). Mature somatic embryos with cotyledons and regenerated plants were obtained from both cell lines. However optimal
concentration of abscisic acid (ABA) for maturation varied depending on morphology of ECs. From the white ECs which intermingled
with somatic embryos of very early stage, mature somatic embryos were induced on maturation medium containing 50 μM of ABA.
On the other hand, mature somatic embryos with cotyledons were observed from the red ECs which consisted of somatic embryos
of more developed stage on hormone-free medium or 0.1 μM ABA containing-medium. 相似文献
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Toru Taniguchi Kazuo Tabuchi Kazuho Yamaguchi Yoshitake Fujisawa 《Journal of Forest Research》1996,1(1):51-55
Somatic embryos ofAcanthopanax sciadophylloides Franch. et Sav. were differentiated from both zygotic and somatic embryos and calli, and plants were regenerated from these
somatic embryos. A zygotic embryo, enclosed within a small portion of the endosperm, was incubated on Murashige and Skoog
(MS) media supplemented with various combinations (range 0–10.0 mg/l) of 6-benzylaminopurine (BAP) and 2,4-dichlorophenoxyacetic acid (2,4-D). After 4 months, swelling of the zygotic embryos
and callus formation was observed. When the swollen embryos were transferred to MS medium supplemented with 0.5 mg/l of 2,4-D, somatic embryos were formed in one to two months. After subculture on the same medium, new embryos were differentiated
from various parts of the older somatic embryos. The calli were cultured on medium supplemented with 2.0 mg/l of 2,4-D and BAP for three weeks. Proliferated calli were transferred to medium supplemented with 1.0 mg/l of 2,4-D and BAP. Somatic embryos were differentiated from the calli within one to two months. Somatic embryos were germinated
on half-strength MS medium without plant growth regulators and the plantlets were grown in soil.
A part of this paper was presented at the 106th Annual Meeting of the Japanese Forestry Society (1995) & First Asia-Pacific
Symposium on Forest Tree Genetic Improvement (Beijing). 相似文献
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花曲柳体胚发生和植株再生 总被引：1，自引：0，他引：1
以花曲柳合子胚的单片子叶为外植体成功诱导出体胚并获得再生植株。未成熟合子胚的子叶在添加400mg·L-1水解酪蛋白、0.25mg·L-16-BA、1.5mg·L-1NAA、70g.L-1蔗糖和6g·L-1琼脂的MS1/2培养基上可以成功诱导产生体胚,诱导率达到34.7%,每个外植体上体胚数量为2～9个。成熟合子胚的子叶在添加0.25mg·L-16-BA、2mg·L-1NAA的MS1/2培养基(其他成分同上)上可以成功诱导产生体胚,诱导率为10.0%。体胚在MS1/2培养基上经过成熟培养后可以正常萌发,萌发率87.6%。萌发的体胚植株在MS1/2+0.01mg·L-1NAA培养基上生长较好,具备实生幼苗的外观特征。经炼苗后的体胚苗移植到栽培基质(草炭土:蛭石:珍珠岩体积比为5:4:1)中可以正常生长,成活率为75.0%。 相似文献
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【目的】建立天山云杉体细胞胚成熟及萌发阶段的适宜培养条件,为该树种的体细胞胚大规模繁殖和工厂化生产提供基础。【方法】以天山云杉成熟合子胚为外植体所诱导形成的胚性愈伤组织为材料,通过14个株系、4种成熟培养基、接种量(100、150、200mg的新鲜胚性愈伤组织)、ABA浓度(1.9、3.8、7.6、19、38μmol·L^-1)等体细胞胚成熟培养条件,以及对不同起源(体细胞胚、合子胚和种子)的幼苗形态特征的比较,筛选出最优的天山云杉体细胞胚成熟与萌发条件。采用SPSS18.0等统计软件对体细胞胚进行统计分析。【结果】1)天山云杉14个株系诱导体细胞胚的结果表明,不同株系在同一成熟培养基上诱导体细胞胚的数量存在差异;其中4个株系及其接种量对体细胞胚形成数量影响的结果表明,不同株系及其接种量诱导体细胞胚的数量也存在差异,当接种量为100mg时,T36株系的体细胞胚诱导数量最高,为3470±546个,显著高于其他接种量和株系(P<0.05)。2)在4种不同成熟培养基上,均形成了成熟的体细胞胚,在1/2BLG+30g·L^-1麦芽糖培养基上形成的体细胞胚数量高于其他培养基,分别为2044个(T6株系)和2282个(T36株系),且与其他培养基差异显著(P<0.05)。3)不同ABA浓度对天山云杉体细胞胚诱导数量的影响无显著性差异(P>0.05),在ABA浓度为1.9~38μmol·L^-1时,其体细胞胚平均诱导数量为2850~2913个,其中以7.6μmol·L^-1ABA的浓度处理效果较好,平均获得2913个正常体细胞胚,高于其他处理。4)将不同起源(体细胞胚、合子胚和成熟种子)萌发的小植株进行对比,结果表明,体细胞胚和合子胚幼苗发育阶段基本一致,但合子胚的幼苗比体细胞胚的幼苗约大1倍,但二者间差异不显著(P>0.05);体细胞胚与合子胚子叶的平均数量一致,二者无显著差异(P>0.05),多数幼苗子叶数量为7~8个。【结论】天山云杉体细胞胚成熟阶段的适宜培养条件为胚性愈伤组织接种量100mg,1/2BLG培养基添加7.6μmol·L^-1的ABA及75%PEG4000、30g·L^-1麦芽糖,并添加750mg·L^-1的L-谷氨酰胺或50mg·L^-1L-天冬酰胺。不同株系在诱导体细胞胚的数量上存在差异。体细胞胚再生植株与合子胚植株在形态上相似。研究结果可为天山云杉体细胞胚的大规模繁殖和工厂化生产提供基础。 相似文献
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Somatic embryogenesis in Pinus thunbergii was initiated from megagametophytes containing immature zygotic embryos. Embryogenic cultures were maintained and proliferated in medium supplemented with 2,4-dichlorophenoxyacetic acid and 6-benzylaminopurine. High maturation frequencies of somatic embryos were obtained on maturation media containing maltose, activated charcoal, abscisic acid, and polyethylene glycol as osmotic agent. The best result among the cell lines tested was achieved with the cell line T-205-3. More than 900 somatic embryos per petri dish, on average, were obtained after about 8 weeks of culture on maturation medium. Sixty percent of somatic embryos tested germinated after transfer to plant growth regulator-free medium and then 85% of them converted into plantlets. 相似文献
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云南腰果开花授粉习性 总被引：2，自引：0，他引：2
为了找出腰果产量低的原因,对云南省勐拉乡的腰果开花授粉特性进行观察。结果表明:腰果为典型虫媒花植物;腰果的花期较长,90~149 d不等;两性花比雄花多,两性花开花高峰期比花药开裂高峰期提前1 h,给腰果的同株授粉带来了可能;自然授粉受到很大的限制,人工授粉能提高稔实率。这说明腰果的开花授粉习性与腰果产量有着必然联系,建议引进虫媒媒介,选出开花期集中适合推广的优良品种,通过改进栽培方式来提高腰果授粉质量,从而提高产量。 相似文献
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《Scandinavian Journal of Forest Research》2012,27(4):318-321
Abstract Pinus contorta plants regenerated from adventitious buds induced on zygotic embryos were planted in a field trial where growth, straightness and flowering were followed over 7 years. Seedlings were taller than the plants derived from adventitious buds at the start of the field trial. Adventitious plants did not catch up with seedlings in height after 7 years in the field, but the relative height growth was equal. The higher frequency of plagiotropic growth that was displayed by adventitious plants the first year in the field was not reflected by differences in the frequency of basal sweeps 6 years later. However, the frequency of crooked stems was comparatively high but similar among the plant types. Flowering was more abundant for the seedlings than for adventitious plants. Taken together, the results show that P. contorta can be cloned via adventitious buds from zygotic embryos, and that the resulting plants show satisfactory growth for inclusion in breeding. 相似文献
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The focus of the current project was to establish somatic embryogenesis protocols for the tropical pine species Pinus oocarpa using immature zygotic embryos (ZEs) as explants. Somatic embryogenesis is best supported by mimicking natural seed-embryo developmental conditions, through a tissue culture medium formulation based on the mineral content of the seed nutritive tissue [megagametophyte (MG)]. A novel culture medium (P. oocarpa medium, PO) was tested in combination with different plant growth regulator (PGR) concentrations and compared with standard Pinus taeda media for the initiation of somatic embryogenesis from immature ZEs of P. oocarpa. Immature MGs containing immature ZEs of two mother trees were used with 12 and 8% extrusion rates for mother tree genotypes 3 and 5, respectively. In both mother trees the percentage capture was 2%. Multiplication of two captured cell lines (T5C2S01 and T5C1S12) was improved by lowering the concentrations of PGRs to 2.5 μM each 2,4-dichlorophenoxyacetic acid and abscisic acid (ABA) plus 1.0 μM each 6-benzylaminopurine and kinetin. Mature somatic embryos formed on 40 μM ABA, 6% (w/v) maltose, 12% (w/v) PEG 8000 and 0.6% (w/v) Phytagel. While PO medium appeared suboptimal for somatic embryo induction, it did exhibit potential for enhanced culture proliferation and subsequent improved maturation with cell line T5C2S01, where microscopic analysis revealed better embryo morphology on PO medium than on 1250 medium. However, this enhancement was not observed with cell line T5C1S12. Germination was preceded by partial desiccation for a period of 2-3 weeks before transferring the embryos to germination medium. Germination was observed after 7 days under low light, and apical primordia slowly expanded after transfer to ex vitro conditions. To our knowledge, this is the first report on the production of somatic seedlings in P. oocarpa. 相似文献
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Pine somatic embryogenesis: analyses of seed tissue and medium to improve protocol development 总被引：1，自引：0，他引：1
The shift from vegetative to embryogenic growth requires tissue to enter a radically different program of development and can be studied in vitro through the development of somatic embryos. From an applied perspective somatic embryogenesis (SE) is expected to play an important role in increasing productivity, sustainability, and uniformity of future forests. For commercial use, SE technology must work with a variety of genetically diverse trees. Since the first reports of SE in Picea abies and Larix decidua in 1985, many different coniferous species have shown the ability to produce embryogenic tissue. However, initiation frequency is often low, many desired seed sources are recalcitrant, and culture survival is often poor, raising costs of somatic seedlings produced from successful genotypes. A number of tools are now available to improve embryogenic tissue initiation and somatic embryo development in vitro that have resulted from analytical studies of seed tissues, the seed environment and gene expression in megagametophyte, zygotic embryos and somatic embryos. Benefits have occurred from medium supplementation with hormones, plant growth regulators, hormone inhibitors and polyamines. Somatic embryo growth has been enhanced with medium supplementation of nutritional components including specific sugar types, vitamins, organic acids, and redox potential modifiers. Control of environmental factors including, water potential, pH, adsorption of medium components by activated carbon and liquid versus gelled medium have also led to SE protocol improvements. The use of analytical studies to duplicate the seed environment in vitro is improving protocol development resulting in increased initiation, improved yields and higher-quality somatic embryos. 相似文献
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We examined the somatic embryogenesis from and histological studies of zygotic embryos of seeds in European Grape 'Moldova' (Vitis vinifera L. 'Moldova'). Primary calli were initiated on Nitsch and Nitsch (NN) medium supplemented with 1.0mg'L-1 2,4-D and 0.5 mg'L-1 6-BA. Embryogenic calli were produced upon transfer to a NN medium with 0.5 mg-L-1 6-BA and 2 mg.L-1 NAA and somatic embryos were obtained on a half strength MS medium without plant growth regulators. During the somatic embryo germination, an addition of 1.0 mg.L-1 6-BA in the medium could accelerate somatic embryos to develop into normal plants and increase the conversion rate from 0 to 43.3%. Histological studies of embryogenic calli and somatic embryos demonstrated dy-namic changes of proteins and starch grains. The developmental processes of somatic embryos were similar to those of zygotic em-bryos, including typical epiderma, cotyledon primordium and vascular tissue. 相似文献
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以西黄松成熟胚为外植体诱导不定芽,在GD 9.85～19.70 μmol·L'-1 6-BA 0.0～14.42 μmol·L-1 NAA上不定芽诱导率最高达65.8%,平均增殖率为7,最大增殖率达10;不定芽形成有2种途径,即子叶直接形成不定芽和子叶组织再分化形成不定芽;NAA不利于外植体不定芽的诱导;不定芽的生长和扩繁采用不加生长调节剂的1/2 GD和1/2 SH培养基;培养基中加入适量的活性炭有利于不定芽和根的生长.不定嫩梢在1/2 GD和1/2 SH附加不同浓度NAA和GA,3的培养基上进行生根诱导,试验结果表明:NAA对不定根的形成起主要作用,在1/2 GD 28.84 μmol·L'-1 NAA 4.17 μmol·L'-1 GA,3培养基中不定梢的生根率为16.7%.在离体培养条件下,以西黄松成熟胚为外植体获得了再生植株. 相似文献