排序方式: 共有11条查询结果,搜索用时 15 毫秒
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小叶杨原生质体培养植株再生及其同工酶的变化 总被引:4,自引:2,他引:4
从小叶杨(Populussimonii)胚性悬浮细胞分离得到大量有活力的原生质体,产量高达3.8×107g-1FW。高密度(3×106/mL)液体浅层培养可促进小叶杨原生质体的分裂和生长;以葡萄糖为唯一碳源的Km8p培养基,有利于原生质体的持续分裂;在培养基中添加有机氮,有助于提高原生质体的植板率。原生质体形成细胞团后,逐步降渗和分皿培养,能有效促进细胞团的持续分裂。愈伤组织在不同分化培养基上的分化频率存在着明显的差异,同工酶分析说明,分化能力强的愈伤组织,其TCA循环和磷酸戊糖途径较为活跃,GOT、EST和PEROX的活性比较高。当芽伸长到2—3cm时,从基部切下转移到无激素的1/2MS培养基上诱导生根并形成完整植株。移入盒内,在温室生长良好。 相似文献
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花药特异嵌合启动子的构建及雄性不育转基因拟南芥的获得 总被引:9,自引:0,他引:9
为了克服TA29/Barnase转基因雄性不育植物的温度敏感性,在TA29启动子上游串联了CaMV35 S启动子,同时在CaMV35 S启动子上游串联一个调节序列antherbox,构建成嵌合启动子antherbox-CaMV35 S-pTA29。该启动元件调控的GUS基因瞬间表达实验表明它是花药颈毡层特异的启动子。这个启动元件及其调控下的barnase基因构建植物表达载体转化,拟南芥菜,获得了雄性不育的转基因植株。 相似文献
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紫花苜蓿原生质体培养与植株再生 总被引:20,自引:6,他引:14
由和田苜蓿和单选苜蓿叶肉分离原生质体,采用改良的K8P和MS培养基进行液体浅层培养。原生质体经一次分裂,二次分裂和多次分裂,形成小细胞团,并很快长成小愈伤组织。愈伤组织在MS分化培养基上增殖并分化,产生小植株,经MS无激素培养基生根形成完整的再生植株。 相似文献
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小麦花粉管途径转化及高效筛选体系的建立 总被引:4,自引:0,他引:4
本文报道了一种从大量花粉管途径转化处理后代中快速有效地筛选到转基因植株的方法。转化质粒为含bar筛选标记基因的天麻抗真菌蛋白(GAFP)与兔防御素(NP-1)双价抗病基因植物表达载体(pBI35S-gafp-NP1-bar),受体小麦品种为扬麦158和Alondra。对花粉管途径转化处理获得的种子分3步进行筛选鉴定:首先,转化处理种子密播于塑料盘中,待小苗长到3叶期时,用浓度为120mg/L的Basta除草剂弥雾喷施筛选,约10d后绝大部分小苗变黄枯死,只有约1%的高抗Basta除草剂的T0小苗存活;然后对其Basta抗性植株用gafp基因引物进行PCR检测,其中62.5%的Basta抗性植株为PCR阳性;最后将T0代PCR阳性植株后代种子(T1)分别发芽并按株系混合取样提取DNA,分别用gafp和bar基因引物进行PCR检测验证,结果均为阳性。表明外源基因已整合入小麦基因组并由T0代植株稳定遗传到T1代,同时证明该筛选方法是可靠有效的。 相似文献
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摘 要:小球藻繁殖快,培养简单,用转基因小球藻表达目的基因产物具有重要的应用前景。本研究构建了兔防御素NP-1的植物表达载体,NP-1由Ubiquitin启动子控制,含NPTII基因和硝酸还原酶基因两个筛选标记。以椭圆小球藻(Chlorella ellipsoidea)硝酸还原酶缺失突变体nrm-4为受体,采用原生质体转化的方法将目的基因转入该突变体中。用含G418的硝酸盐培养基筛选出阳性藻落,经PCR和RT-PCR检测,证明获得了转基因藻株。转基因藻株在无抗生素筛选压力的硝酸盐培养基上继代培养后进行抑菌活性检测。研究表明,转基因椭圆小球藻表达的NP-1具有正常的生物活性,并在无抗生素的培养基中稳定遗传。本研究的结果为小球藻表达外源基因提供了新的技术途径,也为NP-1的规模化生产奠定了基础。 相似文献
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Microsatellite or SSR marker is an efficient tool for plant genotype identification, molecular mapping and marker-assisted selection. Objective of this study is to analyze the mutagenized microsatellite variations in soybean genome and reveal nature of these mutations. In the present study, mutations at fifteen microsatellite loci were detected in genomic DNAs of soybean mutant E182 induced by EMS (ethyne metyl sulfate) using PCR amplification of 485 pairs of SSR primers. These fifteen mutagenized microsatellite loci with repeat number variation were Satt005, Sattll7, Satt185, Satt282, Satt290, Satt420, Satt452, Satt483, Satt569, Satt579, Satt600, Satt602, Sat-086, Sat-107 and Sat-135, respectively. Sequencing results of these fifteen loci indicated that microsatellite sequences at Satt282, Satt483, Satt579, Satt600 and Satt602 loci were respectively deleted 1 -, 3 -, 8-, 20 - and 1 - trinucleotide (all [ATT]1-20 except for [CAA]8 [TAA]12 at Satt600 locus) repeats, which made allele sizes at these five loci decrease 3, 9, 24, 60 and 3 bp, respectively. And while microsatellite sequences at the other ten mutated loci, Satt005, Sattll7, Satt185, Satt290, Satt420, Satt452, Satt569 and Sat-086, Sat-107, Sat-135, were respectively inserted 1-, 6-, 6-, 3-, 4-, 3-, 8- trinu-cleotide repeats (ATT)1-8 and 12-, 6-, 16- dinucleotide repeats (AT)6-16, making allele sizes at these ten loci increase 3, 18, 18, 9, 12, 9, 24, 24, 12, 32 bp, respectively. On the other hand, eleven events of base mutations were detected in flanking regions at seven (Sat- 107, Satt185, Satt282, Satt420, Satt569, Satt579 and Satt600) of fifteen mutated microsatellite loci. These base mutations consisted of 6 transitions (4T→C and 2 A→G), 2 transvertions (A→T and T→A), 1 insertion (T) and 2 deletions (A and T). The experimental results proved that EMS mutagenesis could cause different types of mutations at microsatellite multilocus in soybean genome, including repeat number variations in microsatellite regions and random base mutations in flanking regions. We found three mutational biases, which were frequent insertion mutations of repeat units, initiating positions of microsatellite sequences of repeat unit insertions/deletions and both flanking-base T ↓ A of these insertion/deletion positions. In addition, the resolution capacity of high-quality agarose gels was sufficient to distinguish differences of only three base pairs in this experiment. 相似文献