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农业创新技术成果转化是与时俱进、不断创新过程，必须坚持辩证唯物论的认识论，正确对待技术创新所涉及到的传统观念转变、技术新课题的产生及技术的优点与存在问题，关键在于看准、坚持、不断开拓创新。 相似文献
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Genetic transformation is a useful technique to complement conventional breeding in crop improvement. Although carrot has been a model organism for in vitro embryogenesis study, genetic transformation of carrot is still lengthy and labor intensive. An efficient transformation and detection system is desirable. Direct infection of Agrobacterium to carrot calli has provided an easy way for carrot genetic transformation. To improve the efficiency of antibiotic selection in this method, we report the combined use of an improved green-fluorescent protein, referred to as smGFP, to establish a versatile selection method for carrot callus transformation system. By combining antibiotic selection with the bright fluorescence observed in the callus tissue, we were able to easily identify stable transformants in early stage of the transformation process. In addition to the GFP expression of the callus cells, the transgenic nature of callus cells was confirmed with Southern and Western analysis. We found we can link the simplicity of carrot-callus-cell transformation, early detection of stable transformants with antibiotic selection, visualization of GFP fluorescence, and molecular analysis （Southern and Western） of callus tissue （non-photosynthetic tissue） to provide a more efficient way in identifying stable transformants at early stage of carrot transformation. 相似文献
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The regeneration ability of four alfalfa （Medicago sativa L.） cultivars, Xinjiang Daye, Longdong, Gannong 1 and Gannong 3, was studied, and the effects of various cultivars, explant sources and medium recipes on regeneration were compared. The better callus forming frequency obtained from hypocotyls of Xinjiang Daye is 88.5% and regeneration frequency is 9.8% in our initial experiments. To further optimize regeneration system for genetic transformation, we therefore changed concentrations of plant growth regulators and supplemented with glutamine into callus-induction and shoot-regeneration media. Callus forming frequency and shoot differentiation frequency were increased to 100%. The time taken to generate transgenic plants （16 weeks） was shorter than that for previouse procedure （25 weeks） and regeneration frequency was promoted to 15.1%. The results show that addition of glutamine is particularly important for shortening period of regeneration and promoting regeneration frequency. For study of genetic transformation of alfalfa, Agrobacterium tumefaciens-mediated transformation of Xinjiang Daye was developed based on this optimized regeneration system. The plant expression vector carrying two glutamine synthetases （GS 1 and GS2） and △1-pyrroline-5-carboxylate synthetase （P5CS） gene was used for alfalfa in vitro transformation. Six transgenic alfalfa plantlets with resistance to PPT were obtained. The introduction of foreign genes into plants was assessed in the transformants by PCR analysis and Southern hybridizations. 相似文献
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Effects of Environmental Temperature on the Regeneration Frequency of the Immature Embryos of Wheat （Trificum aestivum L.） 
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下载免费PDF全文 WANG Xin-min REN Xian YIN Gui-xiang WANG Ke LI Jia-rui DU Li-pu XU Hui-jun YE Xing-guo 《农业科学学报》2014,13(4):722-732
The immature embryos （IEs） of wheat are the most widely used tissues for in vitro culture and genetic transformation due to its high regeneration competency. However, this explant can only be maintained in 4℃ daily cooler for a short period time for its use in plant tissue culture or transformation experiments. This study aimed to investigate the effects of environmental temperature, cryopreservation storage temperature, and heat shock culture （HSC） temperature on the regeneration frequency of wheat IEs. Results indicated that environmental temperature significantly affected the induction of embryonic calli. The optimum total accumulated temperature （TAT） during the time of anthesis and sampling for regeneration of these tissues was around 280℃ for spring wheat type cv. CB037 and approximately 300℃ for winter wheat type cv. Kenong 199. Regeneration ability obviously declined when the highest environmental temperature was over 35℃ for 1 d or a high temperature between 30 and 33℃ lasted for 5 d during anthesis and sampling. This finding was verified by culturing the freshly isolated IEs under different temperatures from 29 to 37℃ in different controlled growth incubators for 5 d; the IEs almost completely lost regeneration ability when the temperature rose to 37℃. Cryopreservation of-20℃ caused the wheat samples lost ability of producing callus or embryonic callus in a few days, and cryopreservation of-10℃ more than 10 d made the regeneration potential of the tissues dramatically declined. Comparatively, the temperature that best maintained high regeneration ability was -5℃, at which the materials can be maintained for around 1 mon. In addition, the preservation of the immature samples at -5 or -10℃ inhibited the direct germination of the IEs, avoiding the embryo axis removing process. Our results are useful for ensuring that field collection and cryopreservation of the wheat IEs are done correctly to enable tissue culture and genetic transformation. 相似文献
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Meropenem as an Alternative Antibiotic Agent for Suppression of Agrobacterium in Genetic Transformation of Orchid 总被引：1，自引：0，他引：1
CAO Ying Niimi Yoshiyuki HU Shang-lian 《中国农业科学(英文版)》2006,5(11):839-846
A case of Meropenem as a novel antibacterial agent to suppress and eliminate Agrobacterium tumefaciens in the Agrobacterium-mediated transformation of orchid protocorm-like bodies （PLBs） has been reported in this article. The in vitro activities of meropenem and four comparator antibacterial agents against three Agrobacterium tumefaciens strains, LBA4404, EHA101, and GV3101, were assessed. In addition, the effect of meropenem on the growth of Dendrobium phalaenopsis PLBs was determined. Compared with other commonly used antibiotics （including ampicillin, carbenicillin, cefotaxime, and cefoperazone）, meropenem showed the highest activity in suppressing all tested A. tumefaciens strains （minimum inhibitory concentration [MIC] 〈 0.5 mg L^-1, which is equal to minimum bactericidal concentration [MBC]）. Meropenem, at all tested concentrations, except for 10 mg L^-1 concentration, had little negative effect on the growth of orchid tissues. The A. tumefaciens strain EHA101 in genetic transformation with vector plG121Hm in infected PLBs of the orchid was visually undetectable after a two-month subculture in 1/2 MS medium with 50 mg L^-1 meropenem and 25 mg L^-1 hygromacin. The expression and incorporation of the transgenes were confirmed by GUS histochemical assay and PCR analysis. Meropenem may be an alternative antibiotic for the effective suppression of A. tumefaciens in genetic transformation. 相似文献
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甘蓝下胚轴的高效再生和农杆菌介导B.t.基因转化甘蓝 总被引：31，自引：3，他引：28
分别对取材时间和培养基组成进行研究,建立了甘蓝下胚轴外植体的高效再生系统,当选取６－７ｄ苗龄的下胚轴,将其置于ＭＳＢ＋ＢＡ２．５ｍｇ／Ｌ＋ＺＴ１ｍｇ／Ｌ＋ＩＢＡ０．１ｍｇ／Ｌ的芽分化培养其中,并于培养基中附加ＡｇＮＯ３７．５ｍｇ／Ｌ＋ＺＴ１ｍｇ／Ｌ＋ 相似文献
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As a result of interspecific hybridization of lentil Lens culinaris with wild species L. orientalis, breeding lines recombinant on the basis of flower color, seed coat, and cotyledon have been created. The availability of recombinations in progeny of hybrids is proven by RAPD-analysis. According to morphological characteristics, the best breeding lines are close to cultural varieties and may be used as initial material for breeding. 相似文献
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Li S Bramley P M Smith J Cannon P F 《浙江大学学报(农业与生命科学版)》2004,30(4):455-455
A series of highly taxonomically diverse Trichoderma strains were investigated using proteomic approaches, to investigate the utility of protein profiles as taxonomic markers and to identify proteins of potential economic importance. Initial studies have focused on a comparison of single strains of T. aureoviride, T. saturnisporum, T. polysporum, T. longbrachiatum and T. spirale, along with two strains of T. harzianum. All seven strains were grown in synthetic medium supplemented wi… 相似文献
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采用适于研究石灰性土壤中锌形态的分组方法,利用盆栽试验研究锌在土壤中的动态及有效性。结果表明,水溶性锌施入土壤后,大部分(77%以上)迅速转化为碳酸盐结合态和三二氧化物结合态,前者又向残渣态转化。种植玉米可促进碳酸盐结合态和三二氧化物结合态向残渣态转化;土壤各组分锌之间存在着相互转化、相互依赖的平衡关系。本文同时对各种形态锌的有效性提出了某些看法。 相似文献
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Transformation of Upland Cotton （Gossypium hirsutum L.） with gfp Gene as a Visual Marker 总被引：1，自引：0，他引：1
The green-fluorescent protein (gfp) gene was evaluated as a screening marker during cotton (Gossypium hirsutum L.) transforming and plant regeneration.High expression of GFP (green-fluorescent protein)... 相似文献
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YU Gui-rong LIU Yan DU Wen-ping SONG Jun LIN Min XU Li-yuan XIAO Fang-ming LIU Yong-sheng 《中国农业科学(英文版)》2013,(12):2134-2142
Since maize is one of the most important cereal crops in the world,establishment of an efficient genetic transformation system is critical for its improvement.In the current study,several elite corn lines were tested for suitability of Agrobacterium tumefaciens-mediated transformation by using immature embryos as explants.Infection ability and efficiency of transformation of A.tumefaciens sp.strains EHA105 and LBA4404,different heat treatment times of immature embryos before infection,influence of L-cysteine addition in co-cultivation medium after transformation,and how different ways of selection and cultivation influence the efficiency of transformation were compared.Glyphosate-resistant gene 2mG2-EPSPS was transformed into several typical maize genotypes including 78599,Zong 31 and BA,under the optimum conditions.Results showed that the hypervirulent Agrobacterium tumefaciens sp.strain EHA105 was more infectious than LBA4404.Inclusion of L-cysteine（100 mg L-1） in co-cultivation medium,and heating of the immature embryos for 3 min prior to infection led to a significant increase in the transformation efficiency.Growth in resting medium for 4-10 d and delaying selection was beneficial to the survival of resistant calli.During induction of germination,adding a high concentration of 6-BA（5 mg L-1） and a low concentration of 2,4-D（0.2 mg L-1） to regeneration medium significantly enhanced germination percentage.Using the optimized transformation procedure,more than 800 transgenic plants were obtained from 78599,Zong 31 and BA.By spraying herbicide glyphosate on leaves of transgenic lines,we identified 66 primary glyphosate-resistant plants.The transformation efficiency was 8.2%.PCR and Southern-blot analyses confirmed the integration of the transgenes in the maize genome. 相似文献
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本试验采用土壤恒温恒湿培养和土壤中锌的分组方法,研究碳酸钙对土壤中加入锌的形态转化以及 pH 值的影响,进而探讨碳酸钙对有效锌的影响。试验结果证明:碳酸钙可引起土壤pH 值上升,交换态锌下降以及碳酸盐结合态锌增加,从而使有效锌降低。在 CaCO_2含量1%左右时,各种形态锌分布及 pH 值对 CaCO_2增加较为敏感;在1%～7%时,其变化比较缓慢。 相似文献
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An investigation on the proteome of drone egg development of native Italian bee （Apis mellifera ligustica Spinola,1806） was carried out in order to prove up the characteristics in protein expression and regulation at egg stage and open out the molecular mechanism of the development. The experiment was carried out by two-dimensional gel electrophoresis. The results showed that there were 200, 242 and 233 proteins in a wide rang of molecular weight （12.42-169.60 kDa） and in a relatively narrow scope of pI （4.50-9.00） detected on day 1, day 2 and day 3, respectively, during the developmental process of the drone egg. Meanwhile, 164 protein spots were resolved at all the images （i.e., the protein was consistently expressed） along with the egg development, among which 7 were significantly up-expressed （P 〈 0.05） and 4 were significantly down-expressed （P 〈 0.05） while 79 had no significant differences （P 〉 0.05）. In addition, the specific proteins expressing proteins on day 1, day 2 and day 3 were 11, 18 and 18, respectively. Besides, 17 proteins expressed both on day 1 and day 2 but silenced on day 3, and 43 proteins expressed both on day 2 and day 3 but silenced on day 1, while only 8 proteins expressed both on day 1 and day 3 but silenced on day 2. The results indicate that 2-d-old eggs are at the most active expressional stage in the development of drone egg. The protein expressing at all images suggests that it should be indispensable for drone egg development, but their expression pattern is different. The proteins expressing at a specific age of egg suggest that specific proteins are needed in different developmental stages to regulate. And there are more house-keeping proteins in the developmental process of the drone egg than that of worker egg, and it will provide more targets for gene improvement. 相似文献
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1 《中国农学通报》是中国农学会主办,由两院院士、著名农业科学家石元春先生任主编,国内外公开发行的国家级农业学术期刊(每月5日发行)。也是国家科技部"中国科技核心期刊"、中国科协优秀学术期刊和全国优秀农业期刊。2 本刊特点:以中青年学科带头人和博士、硕士为主要作者群,以省部级以上科研课题或基金项目论文为刊文重点,出版周期短,载文容量大。 相似文献
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Mervis J 《Science (New York, N.Y.)》2004,304(5675):1280