共查询到19条相似文献，搜索用时 156 毫秒
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提高大麦游离小孢子培养反应的研究 总被引：1，自引：0，他引：1
提取液中添加10mg／L秋水仙碱可提高大麦小孢子游离后的存活率；以含10mg／L秋水仙碱的预处理液预处理小孢子48h可以降低培养72h后小孢子的死亡率；以小花共培养可以提高正常型多细胞结构形成率，因而明显提高胚状体的形成。 相似文献
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甘蓝型油菜小孢子培养中几项技术改进 总被引：8，自引：0，他引：8
以大田种植的2个半冬性甘蓝型油菜品种为供体材料,对游离小孢子培养的合适时期、加倍方法、再生苗的继代与移栽技术进行了研究.结果表明:小孢子培养的最佳时期为初花期前后2周左右,游离小孢子经过含有50mg/L或75mg/L秋水仙碱的NLN-16培养基直接处理48h效果最好,其产胚率和加倍效率均显著高于未加秋水仙碱的对照.在固体B5培养基中加入4.5mg/L多效唑并补充液体B5-3培养基,再生苗可不需继代.在固体B5培养基或者补充养分的液体B5-3培养基中加入低浓度的0.5mg/L NAA 0.1mg/L 6-BA,直接移栽到大田的再生苗成活率可以达到90%以上. 相似文献
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《中国油料作物学报》2016,(2)
为探索适用于长江流域主产区生态条件下的甘蓝型油菜小孢子培养技术中最佳秋水仙碱浓度,以2个适合进行小孢子培养的油菜材料为对象,研究了4种不同浓度(0、100、200和400mg/L)的秋水仙碱处理对小孢子产胚率和二倍体率的影响。结果表明,200mg/L的秋水仙碱处理24h后,小孢子的产胚率和二倍体率都达到最大,与对照有显著的差异。用这一浓度,对40份不同基因型油菜的小孢子进行诱导培养,结果表明不同基因型油菜小孢子产胚率和二倍体率存在较大差异,小孢子产胚率为0.12~10.39胚/蕾,再生植株二倍体率为26.7%~90.0%。按照种皮颜色分类后发现,黄籽油菜与黑籽油菜的小孢子产胚率有显著差异,而二倍体率因材料基因型而异,推测油菜种皮颜色与小孢子的产胚率有一定相关性,而与再生植株二倍体率的相关性不大。 相似文献
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甘蓝型油菜小孢子单倍体二倍化技术的研究 总被引：19，自引：3，他引：19
采用6种方法系统研究甘蓝型油菜（Brassica napus)品系（种）及杂种的小孢子再生苗单倍体染色体加倍技术。结果表明，单倍体苗接种含70－80mg/L秋水仙碱的培养基处理4－5d，植株染色体加倍率50％以上；单倍体小苗移栽前用1－2g/L秋水仙碱液浸泡处理3－6h，加倍率50％以上，由这些方法产生的加倍植株大多是可育花和不育花共生的嵌合体，每株产生的种子很少。分离的小孢子在含10－50mg/L秋水仙三的NLN培养基中处理48h后，接种无秋水仙碱NLN培养基诱导胚状体，再生植株加倍率80％以上，全株的花均能自交结籽，嵌合植株很少。 相似文献
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《热带作物学报》2017,(10)
为优化白菜游离小孢子培养高频胚诱导技术体系,以20个不同基因型白菜栽培品种为供试材料,对影响白菜游离小孢子胚诱导和植株再生因素进行研究。结果表明:有14个基因型产生了小孢子胚,不同基因型之间胚诱导率差异显著,表明基因型是影响小孢子胚发生的主要因素;通过对白菜游离小孢子培养高频胚诱导技术体系的优化,胚诱导率明显提高,最优组合为盛花期取蕾;花瓣长与花药长比为0.5~0.75;4℃低温预处理1 d;150 g/L蔗糖的NLN培养基33℃高温热激2 d,重新离心更换130 g/L蔗糖的NLN培养;激素最优组合为6-BA0.1 mg/L、NAA 0.5 mg/L;在培养基中添加活性炭有利于胚的形成,最佳浓度为0.5 g/L。 相似文献
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不同基因型菜心游离小孢子培养和植株再生 总被引：1，自引：0，他引：1
以11个不同基因型的菜心栽培品种为试材,研究不同培养条件对菜心游离小孢子胚诱导和植株再生的影响。结果有7个基因型材料获得小孢子胚,从不同基因型诱导形成胚的频率存在显著差异,表明基因型是影响菜心小孢子胚发生的主要因素;第1天热激培养时用170 g/L高浓度蔗糖培养之后转换成含130 g/L蔗糖培养基能显著提高小孢子胚诱导率;0.05 mg/L 6-BA+0.2 mg/L NAA能促进菜心小孢子诱导成胚;添加0.4 mg/L GA3可显著提高菜心小孢子胚芽诱导率和平均每胚出芽数。7个基因型材料均诱导出再生植株,植株诱导率为100%。 相似文献
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应用低含油量甘蓝型油菜品系1064与高含油量品系H105杂种F1花蕾进行小孢子培养,研究了两个种植地点的油菜小孢子产胚率的影响。对长度在0.4cm～0.6cm,0.6cm～0.8cm以及0.8cm以上的胚分别培养,并比较了秋水仙碱悬浮处理离体小孢子、浸根和滴芽3种染色体加倍方法的加倍效率。结果表明,种植在西宁的材料小孢子产胚率比南京高,为8.6枚/皿;0.6cm～0.8cm大的胚转接到B5固体培养基,其成苗率高达48.53%;3种加倍处理方法中以50mg/L秋水仙碱悬浮小孢子48h染色体加倍效率最高,达到46.23%。本研究构建了含有123个株系的DH群体,可用于油菜含油量性状的QTL分析。 相似文献
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茄子游离小孢子培养初探 总被引：1，自引：0，他引：1
以10份本地主栽长茄品种为试材,对二倍体茄子游离小孢子培养进行研究.结果表明:7份材料可诱导出愈伤组织,‘黑金刚’长茄愈伤组织诱导率可达8.3个/蕾;单核期小孢子是诱导愈伤组织最佳时期;4～6d蔗糖饥饿结合35℃高温预处理是小孢子脱分化的必要条件;愈伤组织诱导与培养分3段进行,先以1.5 NLN培养基附加甘露醇90g/L、2,4-D 0.5 mg/L、NAA 0.2 mg/L、6-BA 0.2 mg/L 35℃暗培养4～6d,接着添加等体积60g/L蔗糖培养基25℃暗培养3d,再均分成2皿,每皿添加等量30g/L蔗糖的培养基,25℃暗培养5～10d肉眼可见愈伤组织. 相似文献
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单倍体籼稻无性系微芽的离体调控 总被引：5，自引：0，他引：5
以籼稻单倍体无性系微芽为材料,研究了激素及秋水仙碱等处理对籼稻单倍体微芽的诱导、分化、扩增及加倍的影响。2 mg/L的2,4-D可提高籼稻单倍体微芽的培养力。NAA对籼稻单倍体微芽的扩增有明显影响,0.5 mg/L的NAA可成倍扩增微芽,培养35 d后,其芽数比原始芽数增加了46.21倍,比不加NAA的对照增加了2.8倍,且单芽重仅0.079 mg。籼稻单倍体微芽扩增的适宜培养基为：N6附加MET 2.5 mg/L、NAA 0.5 mg/L、6-BA 2 mg/L。500 mg/L秋水仙碱溶液处理籼稻单倍体微芽48 h,其二倍体得率较高,绿苗率及二倍体率分别为42.9%和60.0%;秋水仙碱处理愈伤组织的效果不佳,虽然提高秋水仙碱处理浓度可提高二倍体率,最高可达100%,但由于绿苗分化率下降,使总的二倍体得苗率比不处理的对照低。 相似文献
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以橡胶树品种海垦2号为材料,对橡胶树游离小孢子培养体系进行初步研究,为进一步建立其成熟培养体系提供基础。小孢子的提取采用直接研磨法和间接研磨法进行,结果发现以间接研磨法为佳,虽然工作量大,但污染率低、杂质少、培养效果好。对小孢子分别进行高温热击、Starvation Medium B溶液、对照预处理试验,发现用Starvation Medium B溶液预处理小孢子2 d有利于小孢子分化。不同的碳源比较发现麦芽糖培养效果较蔗糖好。对诱导培养基的组分、pH研究发现,以改良N6为基本培养基添加外源激素2,4-D 0.5 gm/L、KT 0.5 mg/L,pH 6.6时诱导效果好,小孢子分化率达8.33%。小孢子的分化以B途径为主,初步获得小孢子分化的细胞团和微愈伤。同时以橡胶树小孢子为受体进行电激转化试验,瞬时表达结果证明外源基因已导入部分小孢子基因组中。 相似文献
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采用浸根、1g/L秋水仙碱滴生长点、含100mg/L的秋水仙碱培养基处理芸薹属三种单倍体无性系幼芽,诱导染色体加倍。根据处理后材料的田间表型、花粉育性和细胞学检测结果发现,各处理的染色体加倍效率存在显著性差异,其中培养基处理幼芽的加倍效率最高,达到50.75%;浸根和滴生长点的处理加倍效率分别为44.07%和17.54%。 相似文献
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《中国油料作物学报(英文)》2020,5(2):52-57
To facilate breeding process of Brassica napus, a microspore culture and molecular marker-assisted screening combined system were proposed in this research. At early flowering stage, F1 offspring of hybridized combination HY15A × HF06 was used as donor for microspore culture to analyze effects of colchicine concentration on embryogenic and diploid rates of microspore. Treatment with 50 mg/L colchicine resulted in embryogenic rate of 3.56 embryos/bud, which was substantially higher than control (0.78 embryos/bud). A total of 1,387 embryos and 862 single plants were obtained after induction culture. Ploidy detection was performed for the regenerated plants by flow cytometry. Diploid rates of microspores treated with 50 mg/L and 70 mg/L colchicine were 17.2% and 21.0% respectively, which was significantly higher than control (10.5%). Totally 108 single plants that doubled successfully were randomly selected and screened using molecular marker BE10. Approximately 54 of 108 plants generated a 305 bp amplification product, whereas the other 54 plants showed a 398 bp band, thereby satisfying 1:1 separation ratio (x20.05 = 0.0093). These coincided with field identification results. Findings of this study indicated that homozygous breeding material could be obtained by microspore culture in a short time, thereby remarkably accelerate breeding. 相似文献
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The potato plantlets singly infected by PVA, PLRV, PVS, PVX and PVY and mix-infected by PVM, PVS and PVY were cultured on MS medium with different concentration of ribavirin. The effects of ribavirin on growth of the plantlets and efficiency of virus elimination were investigated. Results showed that the plant height and fresh weight obviously decreased with increase of ribavirin concentration from 0 mg/L to 150 mg/L, and most of the plantlets could not survive when the concentration reached 200 mg/L. According to the ELISA tests, ribavirin was more efficient for eradicating PVA, PVM, PVS and PVX than PVY and PLRV, and healthy plantlets could be obtained with high frequency (up to 100 %) by culturing with 75?~?150 mg/L ribavirin after 2?~?3 subcultures. Whereas, only 33?~?66 % PVY and PLRV infected plantlets were found to be virus-free after 3 subcultures with 75?~?150 mg/L ribavirin. The results of quantitative RT-PCR (qPCR) indicated that ribavirin could obviously reduce virus content in the plantlets. Except PLRV was detected positive after 3 subcultures with ribavirin, the healthy seedlings were obtained from infected stocks at the first or end of propagation and no viruses could be detected at the post-eradication stage. No apparent difference of genetic variation resulted from ribavirin treatment was found by SSR analysis between the control and the treated plantlets. All of these results above proved that ribavirin treatment in vitro was an effective method to eliminate viruses in the propagation of potato. 相似文献
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Background: Primary cultures of embryonic neurons have been used to introduce a model of neurons in physiological and pathological conditions. However, age-related cellular events limit this method as an optimal model in adult neurodegenerative diseases studies. Besides, short-interval changing media in previous cultures decreases the effectiveness of this model. As an example of this matter, we can refer to the study on some special neuronal secreted factors or the influence of some experimental materials on neurons. Meanwhile, short-interval changing media could remove the effects of some released factors from the environment. In this study, the method for isolation and culturing adult rat hippocampal neurons with long-intervals medium changing has been described. Methods: The hippocampal neurons of adult male rats were cultured. We used Neurobasal A/B27 culture medium, papain (2 mg/ml), trypsin 0.25% and collagenase (1 mg/ml) for neuronal isolation, OptiPrep density gradient for separation of neurons from other cell types and also debris and FGF2 (10 ng/ml) for increasing neuronal survival and regeneration. Results: The neuronal sprouting and viability were increased by using papain and mild triturating (P<0.05). Adult neuronal culturing and their regeneration were impossible without FGF2. It was shown that adding new fresh medium every 4 days and exchanging half of it every 8 days had no detrimental effect on neuronal viability. Conclusion: This investigation shows the possibility of culturing adult neuronal cells and their maintaining in long-interval media. It could be happened because of adult neurons rely significantly on the neighboring cells secreted factors for living and making synaptic connections. This model is very useful in physiological and pathological studies which need stable conditions of neuronal culture in a long period of time. 相似文献
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毛叶枣与冬枣原生质体分离体系的建立 总被引：1，自引：0，他引：1
研究了毛叶枣及冬枣的原生质体分离条件,为以后应用体细胞融合技术创造优异的毛叶枣体细胞杂种材料提供技术支持。结果表明:枣叶片分离出的原生质体活性显著高于悬浮培养物,但是产量低于悬浮培养物分离出的原生质体。毛叶枣酶解所需酶液最佳浓度为纤维素酶10g/L 离析酶4g/L 甘露醇0.7mol/L;冬枣酶解所需酶液最佳浓度为纤维素酶15g/L 离析酶4g/L 甘露醇0.7mol/L,酶解时间均为14￣16h。 相似文献
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[目的]筛选适宜君子兰叶片离体的条件。[方法]选取君子（Clivia miniata Regel）品种“油匠”的叶片为外植体进行离体培养,在MS培养基中附加不同浓度的2,4～D、BA、NAA和KT,对外植体采用不用时间的低温预处理,并对接种后的外植体分别进行不同时长的暗培养,研究不同培养条件对外植体愈伤组织诱导和分化的影响。[结果]叶片诱导分化的最佳培养基为MS＋BA2．0mg／L＋NAA3．0mg／L＋KT1．0mg／L＋蔗糖3．0％,诱导与分化率分别为59．2％和55．2％;暗培养10d后外植体诱导与分化率较高,为l3．8％和25．0％。[结论]培养基中加入活性炭对叶片诱导与分化不利;接种前低温预处理1d,叶片诱导与分化率较高,分别为3313％和25．0％、 相似文献