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1.
植物生长调节剂优化不同品种冬小麦成熟胚再生体系 总被引：1，自引：0，他引：1
《山东农业科学》2019,(5):19-23
小麦愈伤组织再生能力是遗传转化的重要基础。为优化小麦成熟胚再生体系,以科农199、轮选987和济麦22三个冬小麦品种的成熟胚为外植体,在愈伤组织分化阶段添加不同浓度的植物生长调节剂TDZ(N-苯基-N′-1, 2, 3-噻二唑-5-脲,thidiazuron),统计和分析分化率。结果表明,TDZ最适响应浓度存在品种间差异,科农199、轮选987和济麦22成熟胚愈伤组织分化的最适TDZ浓度分别为1.5、1.0、0.5 mg/L,愈伤组织分化率分别为84.61%、71.88%和71.77%,均极显著高于对照。本研究结果可为优化冬小麦成熟胚愈伤组织再生体系提供参考。 相似文献
2.
为探讨AgNO3对不同发育期银杏胚愈伤组织诱导及分化的影响,以银杏3个不同发育程度合子胚为外植体,添加0～5 mg/L AgNO3做处理。结果表明：AgNO3能显著提高胚的愈伤诱导率：早期心形胚鱼雷形胚,早期子叶胚,成熟胚子叶分别在浓度1,0．5,0．1 mg/L时达到最大诱导率,为87％,77％,98％。隔代添加AgNO3促进了早期子叶胚愈伤的分化：1．0 mg/L处理在第90天分化出愈伤化银杏叶和胚类似物,3．0 mg/L处理分化出短小叶片;常规添加AgNO3的处理在0．1 mg/L分化出芽点。遮光处理表明：黑暗培养对愈伤诱导率影响不明显;添加活性炭时,AgNO3促进胚轴愈伤而抑制子叶愈伤,胚轴愈伤组织白色透明。综合分析表明：AgNO3能提高银杏胚的愈伤组织诱导率并促进其分化。 相似文献
3.
为提高粳稻抗性愈伤分化率和遗传转化率,以农杆菌介导遗传转化得到的水稻成熟胚抗性愈伤(Hyg)为材料,研究了影响粳稻成熟胚抗性愈伤组织再生频率的因素。结果表明分化培养基添加0.5～25 mg.L-1的CuSO4.5H2O,增加分化培养基中的琼脂浓度,分化前对抗性愈伤进行高渗处理,采用6-BA和KT两步分化法和干燥除菌法均能提高抗性愈伤组织的分化率。 相似文献
4.
以籼稻、粳稻两种类型的成熟种子为外植体研究了不同浓度的AgNO3在水稻农杆菌转化中的作用。结果表明，在农杆菌转化前添加AgNO3可以提高愈伤组织诱导率，最适浓度为8～10mg／L，但对愈伤组织的分化没有明显促进作用；转化后添加低浓度的AgNO3可以减轻愈伤组织的褐化，提高抗性愈伤率和绿苗分化率，最适浓度为2-8mg／L。同时发现，无论是转化前还是转化后，添加高浓度的AgNO3对水稻愈伤组织的分化均具有毒害作用。 相似文献
5.
褐化是植物组织培养中一种常见的问题,直接影响组织培养能否成功。为了控制植物组织培养中的褐化现象,选取‘丹黄25’、P138、H21 3个玉米自交系幼胚为外植体,着重探讨胚龄,2,4-D、甘露醇、L-脯氨酸和AgNO3浓度对玉米幼胚愈伤组织褐化的影响。结果表明,选取授粉11天的‘丹黄25’幼胚、13天的P138和H21幼胚接种到诱导培养基中,待长出愈伤后转移到添加2 mg/L 2,4-D、10 g/L甘露醇、10 mg/L L-脯氨酸、10 mg/L硝酸银的继代培养基上,愈伤组织褐化率显著降低,表明培养基的种类及添加物对愈伤组织的生长有重要影响。 相似文献
6.
以79个品种的小麦成熟胚为材料,用切碎处理的方式对参试基因型进行愈伤组织诱导和分化,统计出愈率、胚性愈伤率、褐化率、分化率和成苗率并筛选出高频再生基因型。结果表明,出愈率与胚性愈伤率、褐化率、分化率、成苗率均无相关性。胚性愈伤率与分化率、成苗率都呈极显著正相关。成熟胚愈伤组织分化率与成苗率呈极显著正相关。褐化率与分化率、胚性愈伤率呈显著负相关。在供试的79个小麦品种中再生成苗率大于20%的有8个基因型分别是西农2611、XNS150、XN60、XNYS3、XNZK4、XN105、XNZK10和西农1376,参考愈伤组织出愈率和褐化率等指标筛选出XNZK4,XNS150,XN60,XNYS3,XN105,XNZK10和西农1376等小麦品种作为小麦转基因成熟胚受体材料。 相似文献
7.
不同培养基及激素配比对小麦成熟胚离体培养的影响 总被引：1，自引：0，他引：1
[目的]建立适于优良小麦品种遗传转化的高效组织培养再生体系。[方法]以小麦品种豫麦18和豫麦70成熟胚为外植体,探讨了2种培养基和4种激素配比对小麦成熟胚离体培养的影响。[结果]L3培养基的平均愈伤组织诱导率和分化率都明显高于MS培养基,供试2个品种平均出愈率达91.0%以上,平均分化率达46.4%。不同激素及其配比均可诱导小麦成熟胚愈伤组织分化,但不同处理分化率差别很大(10.5%~50.0%);以L3为分化培养基,同时添加2,4-D 0.1mg/L,6-BA 1.5 mg/L的激素配比更有利于愈伤组织的分化,平均分化率达到50.0%。[结论]MS、L3两种基本培养基对小麦成熟胚愈伤组织诱导率均比较高,且差别不大。不同激素配比对小麦成熟胚愈伤组织分化率影响很大。 相似文献
8.
为了建立寒地粳稻成熟胚的高频再生体系,以寒地粳稻空育131和东农425为试验材料进行成熟胚培养,研究CuSO4和烯效唑的单独和组合使用对水稻成熟胚培养的影响。结果表明:在培养基中单独添加15.0μmol/L的Cu2+或添加0.50 mg/L的烯效唑有助于寒地粳稻成熟胚的诱导继代和分化;在诱导和分化培养基中同时添加CuSO4和烯效唑时,空育131在CuSO4浓度为20.0μmol/L且烯效唑浓度为1.00 mg/L时,愈伤组织的诱导率和分化率都达到最高;CuSO4浓度为10.0μmol/L,烯效唑浓度为1.00 mg/L时,东农425愈伤组织的诱导率最高,CuSO4浓度为15.0μmol/L,烯效唑浓度为1.00 mg/L时,东农425愈伤组织的分化率最高。 相似文献
9.
本研究以6种玉米自交系的成熟胚为外植体,通过优化玉米成熟胚愈伤组织诱导、继代及分化等培养环节,建立了高效的苗再生体系。结果表明,使用切除胚芽和胚根的半胚为外植体,以MS(Murashige&Skoog)培养基作为基础培养基诱导形成的愈伤组织最好,初级愈伤组织率高达80%。在继代培养过程中添加0.5 mg/mL 2,4-D和18mg/mL Dicamba有利于提高胚性愈伤组织诱导率,将胚性愈伤组织诱导率提高到88.89%。添加20 g/L甘露醇可以将褐化率降低到19.44%。分化培养过程中加入2 mg/L Zeatin和6 mg/L TDZ可以将愈伤组织的绿苗分化率提高到48.88%。 相似文献
10.
小麦成熟胚愈伤组织的诱导及植株再生体系的研究 总被引：1，自引：0，他引：1
以小麦的成熟胚为外植体,通过胚性愈伤组织诱导进行植株再生.研究结果表明,在MS+2,4-D(3.0 mg/L)+VB1(1.0 mg/L)的培养基中,成熟胚可较高频率的诱导产生愈伤组织,诱导率为72.3%,但愈伤组织质量较差;在低浓度2,4-D和添加了ABA的继代培养基中,愈伤组织质量明显改善,胚性愈伤组织增加.筛选继代的胚性愈伤组织置于分化培养基MS+6-BA(1.0mg/L)+KT(4.0 mg/L)+NAA(0.3 mg/L)中,分化率达73.18%.该研究建立了一套有效的以成熟胚为外植体的小麦组织培养再生体系. 相似文献
11.
小麦幼胚培养特性对外源物质响应的研究 总被引：8，自引：1，他引：8
以豫麦49、豫麦18和兰考906为试验材料,研究了不同外源物质(2,4-D、ABA和AgNO3)对小麦幼胚愈伤组织培养特性的影响。结果表明不同浓度外源物质间小麦幼胚胚性愈伤组织诱导率和愈伤组织绿点率差异均达到0.01显著水平,其中以2.0~3.0 mg/L 2,4~D、2.0 mg/L2,4-D 0.1 mg/L ABA和2.0mg/L 2,4-D 0.1 mg/L ABA 5.0mg/L AgNO3浓度处理适宜小麦幼胚胚性愈伤组织诱导和愈伤组织分化。在适宜浓度2,4-D、ABA和AgNO3培养条件下,3个供试品种间胚性愈伤组织诱导率和愈伤组织绿点率差异达到0.01显著水平,其中以豫麦18胚性愈伤组织诱导率和愈伤组织绿点率最高。 相似文献
12.
ZHANG Wei WANG Xin-min FAN Rong YIN Gui-xiang WANG Ke DU Li-pu XIAO Le-le YE Xing-guo 《农业科学学报》2015,14(1):11-19
The regeneration rate of wheat immature embryo varies among genotypes, howbeit many elite agriculture wheat varieties have low regeneration rates. Optimization of tissue culture conditions and attempts of adding signal molecules are effective ways to increase plant regeneration rate. Inter-culture is one of ways that have not been investigated in plant tissue culture. Moreover, the use of arabinogalactan proteins(AGPs) and hydrogen peroxide(H2O2) have been reported to increase regeneration rate in a few plant species other than wheat. The current research pioneeringly uses inter-culture of immature embryos of different wheat genotypes, and also investigates impacts of AGP and H2O2 on the induction of embryogenic calli and plant regeneration. As a result, high-frequency regeneration wheat cultivars Kenong 199(KN199) and Xinchun 9(XC9), together with low-frequency regeneration wheat line Chinese Spring(CS), presented striking increase in the induction of embryogenic calli and plant regeneration rate of CS through inter-culture strategy, up to 52.19 and 67.98%, respectively. Adding 50 to 200 mg L–1 AGP or 0.005 to 0.01 ‰ H2O2 to the callus induction medium, enhanced growth of embryogenic calli and plant regeneration rate in quite a few wheat genotypes. At 50 mg L–1 AGP application level in callus induction medium plant regeneration rates of 8.49, 409.06 and 283.16% were achieved for Jimai 22(JM22), Jingdong 18(JD18) and Yangmai 18(YM18), respectively; whereas at 100 mg L–1 AGP level, CS(105.44%), Chuannong 16(CN16)(80.60%) and Ningchun 4(NC4)(62.87%) acted the best. Moreover CS(79.05%), JM22(7.55%), CN16(101.87%), YM18(365.56%), Yangmai 20(YM20)(10.48%), and CB301(187.40%) were more responsive to 0.005 ‰ of H2O2, and NC4(35.37%) obtained the highest shoot regeneration rates at 0.01 ‰ of H2O2. Overall, these two methods, inter-culture and AGP(or H2O2) application, can be further applied to wheat transgenic research. 相似文献
13.
In order to establish the system of high frequency plant regeneration for japonica rice mature embryos, the effects of different concentrations of CuSO4 and uniconazole on in vitro culture of mature embryos were studied using three rice cultivars of Kongyu 131, Longjing 24, and Dongnong 425 as test materials. The results showed that callus induction and differentiation of japonica rice mature embryos were apparently improved on the medium with 10-15 μmol·L-1 CuSO4 and 0.50-1.00 mg·L-1 uniconazole. Induction and differentiation rates of different genotype rice mature embryos displayed different sensitivities to CuSO4 and uniconazole. For the callus induction frequency of three varieties, the optimal concentration of CuSO4 was 15.0 mol·L-1. When the concentration of CuSO4 was 15 μmol·L-1, the plantlet differentiation rates of Kongyu 131 and Dongnong 425 got to the highest, while the concentration of CuSO4 was 10 μmol·L-1 for Longjing 24. For the callus induction and plantlet differentiation rates of Kongyu 131 and Dongnong 425, the ideal concentration of uniconazole was 0.50 mg·L-1 and for Longjing 24 was 1.00 mg·L-1. 相似文献
14.
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
15.
小麦大龄幼胚再生性能改进与农杆菌转化 总被引：1，自引：0，他引：1
【目的】小麦幼胚再生率与幼胚大小关系密切,改进小麦大龄幼胚再生性能促进小麦转基因研究。【方法】以18个普通小麦基因型和5个硬粒小麦基因型为材料,对其开花授粉后15—17 d的大龄幼胚进行破碎处理、组织培养和农杆菌转化,对转化后的幼胚组织和获得的抗性再生植株分别进行组织化学染色检测和PCR检测。【结果】大龄幼胚培养2 d后破碎处理的再生率为18.2%,显著高于完整胚对照(.7%),培养2 d后进行破碎处理的再生效果高于4和6 d后破碎处理;不同基因型小麦大龄幼胚破碎处理后再生率为16.9%—46.7%,其中,Bobwhite和中8423大龄幼胚再生率达到了40%以上;大龄幼胚破碎后在弱光条件下培养,再生率进一步提高,较对照(完整胚黑暗培养)提高5.4%—47.4%;农杆菌侵染后GUS瞬时表达率为0—76.7%,其中科农199高达76.7%,其次为Verry(64.4%)和Alondra(47.2%);经PCR检测,农杆菌转化小麦大龄破碎幼胚获得了候选转基因植株。【结论】破碎处理和弱光培养显著提高了小麦大龄幼胚再生率,比对照提高5—10倍;科农199、Verry和Alondra大龄幼胚对农杆菌侵染比较敏感,适宜作为农杆菌转化的受体材料;农杆菌转化小麦大龄幼胚可以获得转基因植株。 相似文献
16.
Using mature embryos (MEs) as the explants, the callus induction, embryogenic callus differentiation, plantlet regeneration
and culture efficiency in diploid and tetraploid wheat of four genotypes were studied. The tested four genotypes wheat included
cultivable emmer wheat (Triticum dicoccum Schuble), durum wheat (Triticum durum Desf.) and the common wheat progenitors Triticum dicoccoides and Triticum aegilopides. Results indicated that there were significant differences in the efficiency of callus induction, callus differentiation
and plant regeneration among the tested genotypes. The efficiency of differentiation and regeneration shows strong genotype
dependence. The rates of callus induction, embryogenic callus differentiation, plantlet regeneration and culture efficiency
respectively were 95.00%, 90.00%, 32.40%, and 27.70% in cultivable emmer wheat, which were significantly higher than other
tested genotypes. Therefore, this study has provided a basis for genetic transformation, gene cloning and molecular plant
breeding in wheat and other related species.
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Translated from Molecular Plant Breeding, 2007, 5(4): 480–484 [译自: 分子植物育种] 相似文献
17.
通过在培养基中加入不同浓度AgNO3,对离体条件下诱导马铃薯四倍体(S.tuberosum.L.)花药产生双单倍体(dihaploid),双单倍体花药产生一单倍体(monohaploid)的效应进行了研究.在基本诱导培养基中加入50～100μmol/LAgNO3可显著促进四倍体花药和双单倍体花药产生胚状体,胚状体出现的频率最高可达16%.同时可提高胚状体的分化率.AgNO3还可延缓花药的褐化,从而延长胚状体出现的时间. 相似文献
18.
以欧洲卫矛生长健壮的叶片愈伤组织为材料,通过不同激素配比筛选出适宜其分化的培养基,将愈伤组织置于不同条件下培养。结果表明,适宜欧洲卫矛愈伤组织分化的培养基为MS+1.0 mg·L-1 6-BA+0.1 mg·L-1 NAA;欧洲卫矛愈伤组织适宜在光照条件下进行培养,生长周期为28 d,28 d后愈伤组织出现不同程度的褐化;可溶性糖含量先升后降;SOD和POD活性均出现2个峰值,可溶性蛋白变化趋势与POD变化趋势基本一致;可溶性蛋白在分化中起重要作用,且光照条件下,可溶性糖和可溶性蛋白含量、SOD和POD活性均高于黑暗条件下,PPO在培养后期活性下降,可能与愈伤组织褐化有一定关系。 相似文献
19.
褐变是导致青钱柳组织培养失败的主要原因之一。研究维生素C(VC)、植酸(PA)和柠檬酸(CA)3种抗氧化剂对青钱柳愈伤组织生长、褐化及其生理生化特性的影响。结果表明,适当浓度的抗氧化剂处理能有效抑制愈伤组织褐化率,促进愈伤组织生长,抑制培养过程中细胞质膜透性的增加。其中VC的处理效果较为明显,PA其次;最适浓度分别为VC 100 mg·L-1、PA 250 mg·L-1和CA 50~100 mg·L-1。VC 100 mg·L-1处理的愈伤组织鲜重增长率比对照增加73.88%,褐化率和细胞膜透性分别比对照低56.52%和45.75%,均达到显著或极显著水平。适当浓度的抗氧化剂使苯丙氨酸解氨酶(PAL)的活性在一定程度上受到抑制,但对多酚氧化酶(PPO)活性的抑制作用总体上不明显。总之,适当浓度的3种抗氧化剂处理能有效降低青钱柳愈伤组织褐化及其产生的伤害,促进生长,对PPO活性的调控可能不是其控制褐化的主要方式,有关机制尚待进一步探讨。 相似文献
20.
[目的]以广西贵港市白玉蔗为材料,探讨影响白玉蔗离体培养的主要因素,建立其快速再生繁殖体系.[方法]以白玉蔗尾梢心叶为外植体,诱导愈伤组织并进行继代培养后,分别进行不同培养基和激素组合对白玉蔗愈伤组织芽分化培养、不定芽增殖和生根壮苗等影响试验.[结果]白玉蔗心叶切片供诱导愈伤组织是方便快捷的有效方法.心叶切片接人培养基(MS+2,4-D 3 mg/L,琼脂5g/L,糖30 g/L)30 d后,外植体迅速膨胀,边缘出现颗粒状、淡黄或黄色、质地均一的胚性愈伤组织或糊状的愈伤组织.诱导白玉蔗芽分化以培养基MS+6-BA 1.0 mg/L+ NAA0.2 mg/L的效果最佳;培养不定芽增殖以MS+6-BA 3.0 mg/L+KT 0.5 mg/L+NAA 0.6 mg/L+糖30.0 g/L培养基的效果最佳.PVP和活性炭对防止组培苗褐变的效果有差异,1%活性炭能有效抑制白玉蔗增殖培养基中丛生芽的褐化.在生根壮苗培养中,最优培养基为MS+ NAA 0.6 mg/L+糖50.0 g/L+活性炭0.1%.[结论]通过正交试验设计,建立了白玉蔗的再生繁殖体系,为加快白玉蔗繁殖速度、提高繁殖系数、节约种茎提供有效途径. 相似文献