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针叶树体细胞胚胎发生研究进展 总被引：2，自引：0，他引：2
针叶树的体细胞胚胎发生是体细胞在人工无菌培养条件下分化产生体细胞胚胎的过程,体细胞胚胎发生途径是细胞全能性表达的一种方式,重演了合子胚形态发生的过程。自1985年Hakman等首次报道挪威云杉的未成熟合子胚形成体细胞胚并再生植株以来,针叶树体细胞胚的诱导、成熟和植株再生过程的研究取得了巨大进展。综述了针叶树体细胞胚胎发生研究领域的研究进展,总结了影响体细胞胚胎发生的主要因素,展望了这方面的研究在实际生产中的应用前景及发展趋势,并指出了该研究领域中需要解决的主要问题及解决途径。 相似文献
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利用花楸树人工控制授粉后的未成熟合子胚进行体细胞胚的诱导培养,通过石蜡切片、扫描电镜和透射电镜对合子胚和体细胞胚的发生发育过程进行组织、细胞观察。结果表明:1)体细胞胚发生进程与合子胚相似,均经历了球形期、心形期、鱼雷形期,最终发育成成熟胚;2)直接发生和间接发生在愈伤组织表面的体细胞胚均为单细胞起源,间接发生在愈伤组织内部的体细胞胚存在单细胞起源,同时还存在可能的多细胞起源;3)子叶和愈伤组织表面发生的体细胞胚在发育早期均具明显的胚柄,鱼雷形胚期胚柄退化;4)愈伤组织内部发生的体细胞胚没有胚柄;5)与合子不同,胚性细胞的第1次分裂为均等分裂;6)多细胞原胚周围降解的细胞具有细胞程序性死亡的形态特征。 相似文献
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麦芽糖、NAA及ABA对华北落叶松体细胞胚成熟及生根的影响 总被引：11，自引：2，他引：11
研究了麦芽糖、NAA、ABA对华北落叶松体细胞胚发生及体细胞胚根发生和再生完整植株的影响。结果表明 :在胚性愈伤组织继代、增殖过程中 ,以 0 5mg·L- 1 的NAA代替 2 ,4 -D后 ,有利于体细胞胚的成熟和根发生。在ABA 16mg·L- 1 时 ,产生的华北落叶松体细胞胚 ,能够再生良好植株 ,ABA大于 2 0mg·L- 1 时对体细胞胚根的发育不利。在含 3%麦芽糖、4 %PEG4 0 0 0活性炭 1g·L- 1 的成熟培养基上 ,华北落叶松每克愈伤组织子叶胚达 10 6 7个 ,生根率达 5 7 89% ;显著地提高了成熟体细胞胚及其根的质量。 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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Agrobacterium tumefaciens strain LBA 4404 carrying pBI121 plasmid was used to transform mature zygotic embryos of three genotypes (E-Hb, E-Ma, and
E-Mc) of loblolly pine. The results demonstrated that the expression frequency of β-glucuronidase reporter gene (GUS) varied
among genotypes after mature zygotic embryos were infected withAgrobacterium tumefaciens cultures. The highest frequency (27.8%) of GUS expressing embryos was obtained from genotype E-Mc with mean number of 21.9
blue GUS spots per embryo. Expression of β-glucuronidase reporter gene was observed on cotyledons, hypocotyls, and radicles
of transformed mature zygotic embryos, as well as on organogenic callus and regenerated shoots derived from co-cultivated
mature zygotic embryos. Nineteen regenerated transgenic plants were obtained from GUS expression and kanamycin resistant calli.
The presence and integration of the GUS gene was confirmed by polymerase chain reaction (PCR) and Southern blot analysis.
These results suggested that an efficientAgrobacterium tumefaciens-mediated transformation protocol for stable integration of foreign genes into loblolly pine has been developed and that this transformation
system could be useful for the future studies on transferring economically important genes to loblolly pine. 相似文献
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Genetic transformation of loblolly pine using mature zygotic embryo explants by Agrobacterium tumefaciens 总被引：1，自引：0，他引：1
Introduction1Genetictransformationinconifershasthepotentialtoallowtheselectiveimprovementofindividualtraitsinelitecloneswhilestillmaintainingtheexistingcombinationofgenesresponsibleforthesuperiorphenotype(Charestetal.1991;Jamesetal.1996;Walteretal.1999).Atpresent,althoughconsiderableresearchefforthasbeendevotedtothegeneticengineeringofconiferspecies(Sederoffetal.1986;Bekkauoietal.1988,1990;Robertsonetal.1992;Bomminenietal.1993;Shinetal.1994;Klimaszewskaetal.1997),ithaslaggedbehindadvancesma… 相似文献
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表达耐盐基因的转基因火炬松的再生 总被引：2，自引：0，他引：2
盐害是限制作物和树木分布和生产的重要因素。盐分过多导致细胞内水分缺失并影响许多重要的细胞代谢活动。本文利用火炬松作为模式植物建立了一套提高植物耐盐性的新技术。这一技术以火炬松合子胚为材料,利用农杆菌介导的转化方法将山犁醇脱氢酶和甘露醇脱氢酶基因转入火炬松。然后再生转化的愈伤组织和转基因植株。经DNA杂交证实的转基因植株被用于耐盐性试验,结果表明这些转基因的植株的耐盐性有明显的提高。这一技术对针叶树的遗传工程育种有重要的参考价值。图3表2参26。 相似文献
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Ana M. Vieitez Elena Corredoira M. Teresa Martínez M. Carmen San-José Conchi Sánchez Silvia Valladares Nieves Vidal Antonio Ballester 《European Journal of Forest Research》2012,131(3):519-539
The genus Quercus, which belongs to the family Fagaceae, is native to the northern hemisphere and includes deciduous and evergreen species. The trees of the different species are
very important from both economic and ecological perspectives. Application of new technological approaches (which span the
fields of plant developmental biology, genetic transformation, conservation of elite germplasm and discovery of genes associated
with complex multigenic traits) to these long-rotation hardwoods may be of interest for accelerating tree improvement programs.
This review provides a summary of the advances made in the application of biotechnological tools to specific oak species.
Significant progress has been made in the area of clonal propagation via organogenesis and somatic embryogenesis (SE). Standardized
procedures have been developed for micropropagating the most important European (Q. robur, Q. petarea, Q. suber) and American (Q. alba, Q. bicolor, Q. rubra) oaks by axillary shoot growth. Although regenerated plantlets are grown in experimental trials, large-scale propagation
of oak species has not been carried out. The induction of SE in oaks from juvenile explants is generally not problematic,
although the use of explants other than zygotic embryos is much less efficient. During the last decade, enormous advances
have been made in inducing SE from selected adult trees, mainly specimens of pedunculate oak (Q. robur) and cork oak (Q. suber). Advances in the understanding of the maturation and germination steps are required for better use of embryogenic process
in clonal forestry. Quercus species are late-maturing and late-flowering, exhibit irregular seed set, and produce seeds that are recalcitrant to storage
by conventional procedures. Vitrification-based cryopreservation techniques were used successfully in somatic embryos of pedunculate
oak and cork oak, and an applied genbank of cork oak selected genotypes is now under development. The feasibility of genetic
transformation of pedunculate oak and cork oak somatic embryos by means of co-culture techniques with several strains of Agrobacterium tumefaciens has also been demonstrated. To date, most research on the genomics of Quercus species has concerned population genetics. Approaches using functional genomics to examine the molecular and cellular mechanisms
that control organogenesis and or somatic embryogenesis are still scarce, and efforts on the isolation and characterization
of genes related to other specific traits should be intensified in the near future, as this would help improve the practical
application of clonal forestry in recalcitrant species such as oaks. 相似文献
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Robinia pseudoacacia 'Idaho' is one of several multi-purpose trees used in ornamental, soil and water conservation, fodder and nectar sources. Plant abiotic stress tolerance transformed by genes could meet the requirements for reclamation of arid or alkalid lands and vegetation restoration. For this paper, we studied the effects of auxin and cytokine on Idaho locust in vitro regeneration and the establishment of gene transformation systems for plants mediated by Agrobacterium tumefaciens. Results showed that the ratios of cytokinin and auxin were the major factors affecting adventitious bud differentiation on a MS medium; the concentration of 0.5inhibit rooting. The most effective antitoxin for screening transgenic Idaho locust shoots was G418 and the most sensitive concentration of it was 8 mg·L-1. 相似文献
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Production of transgenic adult plants from clementine mandarin by enhancing cell competence for transformation and regeneration 总被引：3，自引：0，他引：3
Genetic transformation of mature trees is difficult because adult tissues are recalcitrant to Agrobacterium tumefaciens infection and transformation and because transgenic mature events are less competent for regeneration. We have shown that reinvigoration allows manipulation of the vegetative phase to increase the potential for transformation and regeneration without loss of competence for flowering and fruiting. To produce transgenic plants from clementine mandarin (Citrus clementina hort. ex Tanaka), we optimized the conditions of the source material both ex vitro and in vitro. Grafting of mature buds on juvenile rootstocks in the spring and preventing multiple bud sprouting by removing all but one bud permitted selection of vigorous first flushes for in vitro culture. Use of additional virulence genes from A. tumefaciens to increase transformation frequency and optimization of culture media and conditions to enhance explant cell competence for T-DNA integration and organogenesis resulted in efficient and reliable transgenic plant production. Transformed regenerants from explants, cultured in media without antibiotics, were identified by a screenable marker (either beta-glucuronidase or green fluorescent protein (GFP)), creating the possibility of generating transgenic clementine plants without antibiotic resistance marker genes. Stable integration of foreign genes was demonstrated by Southern blot analysis, and expression of these foreign genes was confirmed by detection of GFP fluorescence in leaves, floral organs and fruits of the transgenic plants. 相似文献
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Zhang Xiao-ying Wang Hua-fang Yin Wei-lun Zhu Zhen College of Biological Sciences Biotechnology Beijing Forestry University Beijing P. R. China Beijing Langshan Nursery Beijing P. R. China Institute of Genetics Chinese Academy of Sciences Beijing P. R. China 《中国林学(英文版)》2006,8(4):6-9
Agrobacterium-mediated genetic transformation of Sophora japonica was standardized using the Agrobacterium tumefaciens strain LBA4404 that harbored the binary vector pBI121 containing genes forβ-glucuronidase (GUS) and neomycin phos-photransterase (nptⅡ). S. japonica transformants were selected by the ability of the leaf explants to produce kanamycin-resistant calli that regenerated into kanamycin-resistant plantlets. Successful transformation was confirmed by histochemical assay for GUS activity, PCR analysis and Southern blot. The period of nearly two months was required for the regeneration of transgenic plantlets from the explants. The transformed plants resembled their parents in morphology. 相似文献
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Gartland JS McHugh AT Brasier CM Irvine RJ Fenning TM Gartland KM 《Tree physiology》2000,20(13):901-907
A transformation system was developed for English elm (Ulmus procera Salisbury) using Agrobacterium tumefaciens C58 pMP90 p35SGUS/INTRON, allowing for the transfer of foreign genes and regeneration of phenotypically normal elm plantlets. The PCR analysis indicated that both nptII and uidA genes were stably inserted in the plant genome. beta-Glucuronidase histochemical and fluorimetric assays revealed expression of the uidA gene in the shoots, leaves, stems and roots of regenerated transgenic plants. The DNA-DNA hybridizations confirmed the presence of the uidA gene in regenerant plants. Factors influencing successful transformation and regeneration of elms included: identifying gene-transfer-proficient Agrobacterium strains for use with elms; developing an infection protocol allowing T-DNA transfer while retaining the ability to remove inciting bacteria; and identifying selection conditions to eliminate non-transformed material and choice of regeneration medium to allow shoot production. The potential utility of an effective elm transformation and regeneration system in the control of Dutch elm disease is discussed. 相似文献
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Sun Hai-jun Li Min Chen Shou-yi He Si-jie Wang Hua-fang College of Biological Sciences Biotechnology Beijing Forestry University Beijing P. R. China Institute of Genetics Developmental Biology Chinese Academy of Sciences Beijing P. R. China 《中国林学(英文版)》2006,8(4)
Based on the plant regeneration system, a GUS gene transformation system to Idaho locust (Robinia pseudoacacia 'Idaho') mediated by Agrobacterium tumefaciens was established. The successful transformation was confirmed by regenerating the shoots from the infected leaves in the presence of hygromysin; by histochemical X-gluc assays ofβ-glucuronidase (GUS) and by PCR and PCR-Southern blotting analysis. The ratio of positive transgenic plants is 5.8% (5 out of 86 plants). With this system, the target gene DREB was introduced into the leaves of Idaho locust. The transgenic plants regenerated, which was verified by PCR-Southern blotting. It is suggested that the transformation system could be a new, simple, reliable and practical route to gene transformation of R. pseudoacacia 'Idaho' mediated with A. tumefaciens. 相似文献
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This review on current biotechnological methods in forestry for in vitro tissue cultures to define the effect of stress conditions on trees,concentrates on somatic embryogenesis.Callus tissue,the key product of somatic embryogenesis,grows over a tree wound under ex vitro conditions.Callus tissue can be used in research in areas such as pathogenic susceptibility at the embryonic level,effect of heavy metals,influence of low temperatures(cryopreservation),production of secondary metabolites and transformation of plants.Callus of arborescent plants can be induced in vitro by fungal elicitors to produce secondary metabolites for pharmaceutical and cosmetic industries and are strongly repellant to herbivores and can thus act to protect forests.Analyses of dual cultures demonstrated that callus tissue exposed to a pathogenic fungus responds by synthesizing low-molecular-mass proteins belonging to an immune protein class.Cryopreservation of embryonic callus tissue also has broad applications,e.g.,for valuable plant genotypes in gene banks.Without strategies to protect forests against stressfactors,forest ecosystems will degrade to the detriment of all life,including humans.In vitro biotechnological research using callus tissue contributes to progress in forestry and the disciplines of ecology,physiology,phytopathology,culture and selection of plants. 相似文献