共查询到18条相似文献，搜索用时 984 毫秒
1.
转基因水稻中重组植酸酶的表达 总被引：5，自引：0，他引：5
为通过在转基因植株中表达植酸酶降解植酸来提高水稻中无机磷利用率,构建了由玉米泛素基因Ubi启动子控制的植酸酶基因植物表达载体,并以来源于水稻未成熟胚的愈伤组织作为转化受体,经农杆菌介导法将植酸酶基因导入水稻中,共获得15个独立的转基因株系。对转基因水稻总DNA的PCR和Southern 杂交分析证明目的基因已整合入转基因水稻植株基因组中,并能稳定遗传。对部分转基因水稻未成熟种子总RNA进行RT PCR分析,表明导入的植酸酶基因能够在转基因水稻种子中正常表达。无机磷含量分析表明含目的基因的转基因水稻种子及其后代叶片中的无机磷含量较未转化植株均有了明显的提高。 相似文献
2.
甘露糖阳性选择系统在水稻遗传转化中的应用研究进展 总被引：1，自引：0，他引：1
甘露糖阳性选择系统是近几年应用较多的一种植物转基因安全筛选体系,该体系是以大肠杆菌6-磷酸甘露糖异构酶基因pmi为筛选标记基因,D-甘露糖为筛选剂进行筛选。本文简要综述了甘露糖阳性选择系统在水稻转基因研究中的应用情况。 相似文献
3.
《热带作物学报》2017,(12)
为研究磷酸甘露糖变位酶基因(PMM)的功能及其在多糖生物合成中的表达调控机制,以霍山石斛原球茎为材料,利用RT-PCR技术,克隆PMM基因,对其cDNA序列和表达情况进行分析。结果显示:霍山石斛PMM基因cDNA序列长949 bp,包含一个747 bp的开放阅读框,共编码248个氨基酸,GenBank中的登录号为KY912084。生物信息学分析表明:该蛋白属于HAD超家族,可能是一种稳定的、不具有信号肽和跨膜结构的亲水蛋白。系统进化树分析表明,霍山石斛PMM与铁皮石斛、海枣、油棕、芦笋亲缘关系较近,处于同一分支。qPCR结果显示:PMM基因在茎中的相对表达量最高;在4种不同药用石斛比较中,鼓槌石斛的相对表达量最高,霍山石斛的表达量最低。本研究结果表明PMM基因可能不仅参与到石斛多糖的合成,还与其它化合物的生物合成相关。 相似文献
4.
利用共转化和花药培养技术快速获得无选择标记的三价转基因水稻 总被引：7，自引：0，他引：7
采用农杆菌介导的水稻转化体系,将包含选择标记潮霉素磷酸转移酶基因（hpt）和目的基因人乳铁蛋白(hLF)、高赖氨酸(SB401)、高甲硫氨酸(RZ10)基因的双T DNA表达载体p13HSR转化水稻。筛选潮霉素磷酸转移酶基因和目的基因都是阳性的转基因植株,按单株进行花药培养,快速得到无选择标记而目的基因阳性的转基因纯合植株,得率为987％。RT PCR检测结果显示外源基因已整合到转基因水稻基因组中并转录。同时观测到三价表达载体在转化过程中部分目的基因丢失。 相似文献
5.
6.
玉米磷酸烯醇式丙酮酸羧化酶基因高效表达载体构建及其导入小麦的研究 总被引：7，自引：2，他引：5
为获得具有C4光合特征的高光效转基因小麦材料,利用从玉米中克隆的磷酸烯醇式丙酮酸羧化酶(PEPC)基因(pepc,GenBank接受号为FJ415327)构建高效双元表达载体,通过基因枪介导法将其导入小麦品种(系)中,利用Real-time PCR方法分析外源基因在转基因植株中的拷贝数.PCR和双酶切鉴定表明,玉米PEPC cDNA序列已插入双元表达载体pCAMBIA3301中,命名为p3301-pepc;对获得的抗性再生植株进行PCR扩增,其中有342株扩增出目的条带;在选取的15株转基因植株中7株拷贝数为1,3株拷贝数为2,2株拷贝数为3,拷贝数为5、8和17的分别为1株.初步证明玉米pepc基因已整合到小麦基因组中,且外源基因在转基因植株中既有单拷贝插入,也有多拷贝插入,这为进一步研究该基因在小麦基因组中的功能表达提供了试验材料. 相似文献
7.
大豆异黄酮是苯丙氨酸代谢途径中合成的一类次级代谢产物,在苯丙氨酸代谢途径中,苯丙氨酸解氨酶(PAL)是关键酶和限速酶。本课题组前期研究发现PAL基因的相对表达量与大豆异黄酮含量具有明显的协同增减趋势,并且在PAL基因家族成员时空表达模式分析中发现,PAL2-3(XM_003542493)是PAL基因家族中相对表达量较高的主要表达成员之一。本试验首先通过克隆大豆中PAL2-3基因;然后构建pCAMBIA3301-GmPAL2-3植物过表达载体,将构建好的pCAMBIA3301-GmPAL2-3表达载体重组质粒转化到根癌农杆菌EHA105中;采用农杆菌介导的大豆子叶节转化体系获得转化植株,并对T1代转化植株进行PPT检测,外源标记基因Bar检测以及荧光定量PCR检测,对转基因阳性植株进行大豆籽粒异黄酮含量的测定。结果表明T_1代转基因植株中PAL2-3基因的表达量是对照的5.11~11.24倍,总异黄酮含量最高的(2 587.63μg·g~(-1))是对照(1 616.90μg·g~(-1))的1.6倍。因此在大豆中过量表达PAL2-3基因可以提高大豆籽粒中异黄酮含量。 相似文献
8.
9.
【目的】水稻Os08g44770.1基因编码一个铜锌SOD酶,但其在响应亚砷酸盐[As(Ⅲ)]胁迫中的生物学功能未知。本研究旨在深入揭示由该基因调控水稻砷耐性改变的分子机理并为水稻抗逆育种提供理论参考。【方法】以野生型日本晴(WT)和2个Os08g44770.1过表达转基因株系为试材,通过胁迫处理、生理指标测定和基因表达分析等,系统探究了转基因植株对As(Ⅲ)的耐受性表现,并初步揭示了其调控水稻砷耐性的生理和分子机理。【结果】与WT相比,过表达转基因株系对As(Ⅲ)更加敏感;转基因植株在砷胁迫下的根系相对伸长量、生物量(干质量)、叶绿素含量、根系细胞质膜完整性、叶片抗氧化程度等均显著低于WT;Os08g44770.1在砷处理后的WT和转基因植株叶片中的表达模式略有不同,但均表现为处理24 h时被显著诱导表达。【结论】过度表达Os08g44770.1基因可导致水稻的砷耐受性极显著下降。 相似文献
10.
为进一步探讨外源玉米pepc基因改良小麦光合性能的机制,以基因枪介导T4代的转玉米pepc基因小麦为试验材料,研究了抽穗期和灌浆期外源pepc基因对小麦内源光合相关酶基因表达的影响,并分析了转基因小麦的光合生理特性及其产量性状表现。结果表明,小麦抽穗期,pepc基因的表达上调了小麦C4微循环ppdk基因(丙酮酸磷酸双激酶基因)、nadp-me基因(NADP-苹果酸酶基因)、ca基因(碳酸酐酶基因)和C3循环rbcL基因(Rubisco大亚基基因)的表达;小麦灌浆期,pepc基因的表达仍显著上调C4微循环ppdk基因和nadp-me基因的表达,而ca基因和C3循环rbcL基因、rbcS基因(Rubisco小亚基基因)的表达量与对照差异不大。相应的酶活性在转基因植株中比对照有所提高,灌浆期增幅最大。两个测定时期的转基因小麦旗叶净光合速率(Pn)均比对照显著提高,在灌浆期增幅最大,比对照提高10.35%~22.77%。产量性状方面,转基因小麦的单穗粒数和收获指数均有所提高。上述研究结果表明,玉米C4型pepc基因导入小麦后,促进了小麦原有的C4循环,从而提高了小麦的光合效率和籽粒产量。 相似文献
11.
分子标记辅助选择改良C418对红莲型粳稻不育系的恢复力 总被引：2，自引：0，他引：2
【目的】高效选育红莲型(Honglian,HL)粳稻恢复系有助于HL型杂交粳稻育种,对促进三系杂交粳稻的发展具有重要的意义。【方法】Rf6是一个HL型恢复基因,来源于HL型籼稻强恢复系9311。前期研究中,在以9311为供体、日本晴为受体的一套染色体片段代换系中鉴定出携带Rf6的株系R1093。本研究利用R1093与BT型粳稻恢复系C418(携带Rf1)杂交,通过常规回交育种结合分子标记辅助选择技术,将Rf6导入C418中,进行Rf6与Rf1聚合育种;利用BT型、HL型六千辛A进行测交鉴定改良系的恢复力。【结果】共获得12个BC3F4株系和55个BC4F3株系,其中6个改良系的农艺性状已基本接近C418;测交鉴定结果表明聚合Rf6的改良系对HL型粳稻不育系的恢复度达到85%以上,可应用于水稻生产;对BT型粳稻不育系的恢复度提升效果不显著。【结论】聚合Rf6能有效改良BT型粳稻恢复系对HL型粳稻不育系恢复力,是选育HL型粳稻恢复系的一条重要途径。 相似文献
12.
Endosperm-Specific Expression of Serotonin <Emphasis Type="Italic">N</Emphasis>-Hydroxycinnamoyltransferase in Rice 总被引：1，自引：0，他引：1
Lee K Kang K Park M Woo YM Back K 《Plant foods for human nutrition (Dordrecht, Netherlands)》2008,63(2):53-57
Serotonin N-hydroxycinnamoyltransferase (SHT) is a key enzyme in the synthesis of feruloylserotonin (FS) and 4-coumaroylserotonin (CS). These serotonin derivatives show strong antioxidant activity, making them valuable for both nutritional and pharmacological use in humans. Ectopic expression of SHT under the control of the endosperm specific-glutelin and prolamin promoters from rice was produced via Agrobacterium-mediated transformation. SHT expression was confirmed by Southern blot analysis, followed by Northern blotting and SHT enzyme activity analyses using total RNA and protein, respectively, extracted from transgenic seeds. The glutelin A3 (GluA3) promoter produced low SHT mRNA expression in rice seeds, whereas the prolamin promoter expressed high levels of SHT mRNA. In spite of the ectopic expression of SHT in rice seeds, both transgenic genotypes accumulated levels of serotonin derivatives similar to those found in wild-type rice. Furthermore, our data suggest that serotonin, rather than phenylpropanid-CoAs, is the rate-limiting substrate in the biosynthesis of serotonin derivatives in SHT-overexpressing transgenic rice seeds. 相似文献
13.
LIU Qiao-quan LI Qian-feng JIANG Li ZHANG Da-jiang WANG Hong-mei Gu Ming-hong YAO Quan-hong 《水稻科学》2006,13(2):79-84
In many plants, phytic acid (phytate, 1, 2, 3, 4, 5, 6-hexakisphosphate) is one of the main storage forms of phosphate. About 80% of phosphorus (P) in cereal plants, including rice is stored as phytic acid [1-2]. P in phytic acid can’t be utilized by monogastric animals including human, while it was estimated that only 1/3 of the total P in most of the vegetal feedstuff could be efficiently utilized by the livestock. Therefore, for animal feed with P supplementation is expected to meet the d… 相似文献
14.
ZHU Jinling WEI Ruping WANG Xin ZHENG Chaoqun WANG Mengmeng YANG Yicheng YANG Liuyan 《水稻科学》2023,30(3):235-247
Crop yield and quality are often limited by the amount of phosphate fertilizer added to infertile soils, a key limiting factor for sustainable development in modern agriculture. The polyphosphate kinase(ppk) gene-expressing transgenic rice with a single-copy line(ETRS) is constructed to improve phosphate fertilizer utilization efficiency for phosphorus resource conservation. To investigate the potential mechanisms of the increased biomass in ETRS in low phosphate culture, ETRS was cultivated in ... 相似文献
15.
应用水稻种子生物反应器开发口服重组胰岛素原具有重要应用前景。通过分子设计保证重组胰岛素原在人体肠道内的自主加工成熟,根据水稻密码子偏爱性人工合成了霍乱毒素β亚基和人胰岛素原的融合基因(cholera toxin B subunit fused with human proinsulin,CTBIN),并在C末端添加内质网滞留信号KDEL。通过PCR技术从粳稻品种日本晴全基因组中克隆谷蛋白启动子及其信号肽序列pGluB1sig(GluB1 promoter and its signal peptide)用于驱动融合基因CTBIN的表达,插入载体pCAMBIA1302,构建了水稻种子蛋白体靶向表达口服重组胰岛素原的载体pCAMBIA1302-pGluB1sig-CTBIN-Nos。采用农杆菌介导法转化日本晴,获得了46株转基因水稻植株,Western杂交检测到CTB-人胰岛素原融合蛋白在水稻种子中表达。 相似文献
16.
17.
18.
ZHANG Yuping WANG Junke WANG Yaliang CHEN Yanhua ZHU Dengfeng CHEN Huizhe XIANG Jing ZHANG Yikai LIU Xiaojun ZHU Yan CAO Weixing 《中国水稻科学》1986,34(6):525-538
【Objective】To clarify the effect of night temperature changes on rice starch accumulation, and to explore circadian changes of physiological characteristics of rice amylose/amylopectin formation at various night temperatures.【Method】High-quality japonica rice ‘Zhehexiang 2’ was subjected to three night temperature gradients of 31℃/20℃ (LT), 31℃/24℃ (NT), and 31℃/28℃ (HT) at the beginning of grain ripening stage, and the amylose and amylopectin contents, the key enzyme activities involved in amylose and amylopectin formation and the expression of related genes were measured at noon and midnight. 【Result】 The results showed that 1) compared with NT, LT and HT treatments significantly reduced grain weight and starch accumulation, LT and HT both reduced amylopectin content and increased amylose content, meanwhile the effect of HT was greater than LT. LT and HT treatments decreased grain pasting temperature and gel consistency, and had a significant influence on branched chain length. 2) The net photosynthetic rate of leaves under LT and HT presented no significant difference compared to NT, but LT and HT significantly reduced the accumulation of non-structural carbohydrates, meanwhile down-regulated the expression level of sucrose transporter genes OsSUT1, OsSUT2, and OsSUT4 both at noon and midnight. 3) LT and HT treatment reduced sucrose hydrolysis-related enzyme activities, meanwhile promoted starch hydrolase activities, resulting in increased soluble sugar contents, sugar utilization in grain was blocked. 4) Compared with NT, the adenosine diphosphate glucose content showed a downward tendency during the day and a rising trend at night under the treatment of LT and HT, which presented that the accumulation and utilization of adenosine diphosphate glucose were inhibited. And the granules bound starch synthetase activity were significantly reduced under LT and HT compared to NT with the prolongation of the treatment, and the enzyme activity at daytime was significantly influenced by temperature changing at night. 5) Compared with NT, LT and HT reduced the activities of night-time amylopectin synthesis-related enzymes and inhibited the expression of night-time related genes, which retarded formation of amylopectin, but the enzyme activities related to amylopectin synthesis was not significantly influenced by night temperature changing. 【Conclusion】High night temperature had a less effect on starch accumulation than low night temperature. High or low night temperature inhibited sucrose transport and metabolism, resulting in declined starch accumulation. The retarding of amylopectin formation was the main reason for the increase of relative content of amylose at high or low night temperature. Night temperature changes directly affected the metabolic process of starch formation at night, but the daytime activities of amylose-related enzymes were affected by night temperature changes, while the daytime activities of amylopectin-related enzymes were not significantly affected by night temperature changes. 相似文献