共查询到19条相似文献，搜索用时 593 毫秒
1.
2.
RNA干涉下调RACK1基因表达增强水稻抗旱能力 总被引：1，自引：0，他引：1
RACK1是一种多功能支架蛋白,广泛参与植物生长发育过程的调节。利用RNA干涉技术抑制水稻RACK1基因的表达,分析了RACK1基因在响应干旱胁迫中的功能。实时定量PCR对获得的转基因植株的RACK1基因表达分析结果表明,转基因水稻RACK1基因表达受抑制程度达50%左右。与非转基因水稻（对照）相比,转基因水稻耐干旱能力显著强于对照,其膜的过氧化酶和丙二醛的产生显著低于对照,而超氧化物歧化酶活性极显著高于对照。表明RACK1蛋白质调节水稻对干旱胁迫的耐性,并且这种调节在很大程度上与植株体内的氧化还原系统有关。 相似文献
3.
OsBTF3过量表达和RNAi转基因水稻抗盐和抗低温胁迫鉴定 总被引：2，自引：0，他引：2
为评估水稻OsBTF3基因在水稻对非生物逆境胁迫应答中的功能,利用RT Q PCR技术,对经高盐(200 mmol/L NaCl溶液)和低温(4℃)胁迫处理后水稻的OsBTF3基因表达动态进行了测定,并对前期获得的OsBTF3过量表达和RNAi转基因水稻株系进行了抗高盐和抗低温胁迫的鉴定。结果表明,高盐胁迫处理显著抑制了OsBTF3基因的表达,低温胁迫对其表达影响有一定的变化;转基因T2代过量表达株系对高盐和低温胁迫抗性显著增强,而RNAi转基因株系抗性减弱,说明OsBTF3可能在水稻对高盐和低温胁迫反应中起着重要的调控作用。 相似文献
4.
活化C激酶1受体(RACK1)属于含有色氨酸-天冬氨酸结构域(WD)重复结构的蛋白质亚家族.为研究GmRACK1在干旱胁迫中的调控作用,本研究构建GmRACK1基因过表达与沉默两种双元表达载体并转化大豆,对转基因和对照大豆株系进行干旱和复水处理,测定株系的干重、存活率、叶绿素含量、过氧化氢含量和抗氧化酶活性等指标;分析GmRACK1基因对大豆抗旱性的调控作用.结果 显示:在干旱胁迫下,大豆GmRACK1基因沉默株系(RNAi)的存活率和干重明显高于野生型(WT)和过表达(OE)株系.干旱10 d后RNAi株系的叶绿素含量显著高于WT和OE株系,OE株系的DAB和NBT染色颜色比WT和RNAi株系深.土壤干旱胁迫10 d后复水4d,RNAi株系恢复好于WT和OE株系.在15％ PEG 4000处理下,RNAi株系抗氧化物酶SOD、POD和CAT的活性和ABA含量显著高于WT和OE株系,而MDA和电导率显著低于WT和OE株系.结果 说明RNAi株系能清除过多活性氧,导致植物的氧化伤害减弱.GmRACK1基因对大豆耐旱性存在调控作用. 相似文献
5.
同源异型域(Homeobox,HB)转录因子对于植物的生长发育具有重要的调控作用,主要涉及细胞分化、形态建成、内外环境信号应答等多个方面。为了研究该转录因子家族成员在水稻中的功能,构建了该家族成员OsHox9基因的RNAi表达载体,利用反向遗传方法分析该基因的功能。与野生型对照植株相比,RNAi转基因植株株高变矮,分蘖数减少。实时PCR表达分析表明OsHox9基因在有表型转基因植株中的表达下调,但在没有表型转基因植株中OsHox9表达量与野生型植株差异不显著,说明RNAi载体起到了干涉作用,转基因植株的表型是由RNAi造成的。组织特异性表达分析表明OsHox9在水稻的根、茎、叶、茎顶端分生组织及不同时期的幼穗中均有表达,但在茎顶端分生组织及幼穗中表达量较高。为了查明OsHox9的亚细胞定位,构建了OsHox9与绿色荧光蛋白GFP的融合载体并导入烟草叶片中进行瞬时表达,结果表明OsHox9定位于细胞膜上,在细胞膜上起作用。 相似文献
6.
外源木聚糖酶基因atx在水稻中的表达 总被引：1，自引：0，他引：1
为通过在转基因植株中表达木聚糖酶来提高木聚糖酶的生产效率,将具有较高热稳定性和催化活性的杂合木聚糖酶基因atx连接到双元表达载体pCAMBIA1301上,成功构建了木聚糖酶植物表达载体atx Ru3ep 1301。然后以水稻成熟胚的愈伤组织作为转化受体,采用农杆菌介导法将木聚糖酶基因导入水稻(中花11)中。经过潮霉素抗性检测和PCR鉴定,证实目的基因已经整合到转基因水稻基因组中。RT PCR分析结果显示,外源木聚糖酶基因能够在CaMV 35S启动子的引导下在转基因水稻中正常转录。转基因水稻能够正常生长和繁殖。木聚糖酶活性分析表明,转基因植株最高木聚糖酶活性约为4.37 U/g（鲜叶片）。因此,利用转基因水稻生产木聚糖酶将会是一种经济、有效的方法。 相似文献
7.
为进一步研究RNAi介导的Δ12-脂肪酸脱饱和酶（FAD2）干扰基因对油菜内源FAD2基因表达的影响,以油菜肌动蛋白(β-Actin)基因为内参照基因,提取转基因油菜幼嫩种子总RNA,通过半定量RT-PCR检测内源FAD2基因的相对表达量。结果表明,T1,T3代转基因种子中FAD2基因的相对表达量与对照相比明显降低。对T3代种子的油酸含量的分析表明：转基因油菜种子中油酸的含量比野生型增加了13.90到32.20个百分点,直接说明RNAi干扰体下调了FAD2基因的表达。因此,种子内源表达的FAD2基因被RNAi干扰体有效沉默,并且产生了能够稳定遗传两代的表型变化。 相似文献
8.
《中国油料作物学报》2017,(3)
大豆Kunitz型胰蛋白酶抑制剂(trypsin inhibitor,Kti)是大豆品质的主要营养抑制因子之一,培育低Kti含量的大豆品种是提升大豆品质的关键。本研究采用RT-PCR技术克隆得到大豆Kti基因的核心保守序列,并构建成种子特异性表达的、具有Kti基因反向重复结构的RNAi载体,利用农杆菌介导法侵染大豆品种吉林30子叶节,通过潮霉素筛选和PCR检测,获得4株转基因阳性植株。RT-PCR检测表明,构建的种子特异性RNAi载体可以在转基因植株的种子中特异表达,且Kti基因表达量均有降低;通过胰蛋白酶抑制剂活性检测发现,转基因大豆籽粒中胰蛋白酶抑制剂活性平均降低了77.5%。为进一步评价RNAi技术对大豆Kti基因的表达调控效果奠定良好基础。 相似文献
9.
《杂交水稻》2015,(3)
结合表达谱数据分析,参考公共数据库中的序列信息设计引物,利用RT-PCR技术,从东乡野生稻中获得了1个在低温诱导条件下高表达的螺旋—环—螺旋结构域蛋白基因BGIOSGA013293-DX的全长c DNA,并构建了其过表达载体。序列比对分析表明,该基因位点在93-11中编码308个氨基酸,而在东乡野生稻中编码335个氨基酸,比93-11多27个氨基酸。且这27个氨基酸连续分布在一起,另外还有1个氨基酸的差异。利用农杆菌介导法将BGIOSGA013293-DX转入水稻受体品种93-11,获得了9株过表达转基因水稻植株,经潮霉素抗性标记基因PCR阳性检测和GUS检测,获得的转基因植株为携带BGIOSGA013293-DX的阳性植株。 相似文献
10.
《中国油料作物学报》2015,(5)
为幼龄动物提供优质的大豆饲料,同时有效抑制α'亚基和β亚基基因表达量,根据dsRNAi原理,以α'亚基基因RNAi重组植物表达载体p CAMBIA3301-PFNZ-BADH为基础,构建以BADH安全标记为筛选标记的α'亚基和β亚基基因双价RNAi植物表达载体。用根癌农杆菌介导法转化大豆子叶节,并对再生植株进行分子生物学检测。结果表明:成功构建β-伴大豆球蛋白α'亚基和β亚基基因双价RNAi表达载体,并获得大豆转基因植株。T1转基因大豆植株经PCR检测得到11株阳性转化株,经Southern杂交检测证明构建的双价植物表达载体已经以1~2个拷贝整合到大豆基因组中,实时荧光定量PCR检测表明β-伴大豆球蛋白α'亚基和β亚基基因的表达被明显抑制。 相似文献
11.
Down-Regulated Expression of RACK1 Gene by RNA Interference Enhances Drought Tolerance in Rice 总被引：2，自引：0，他引：2
The receptor for activated C-kinase 1 (RACK1) is a highly conserved scaffold protein with versatile functions, and plays important roles in the regulation of plant growth and development. Transgenic rice plants, in which the expression of RACK1 gene was inhibited by RNA interference (RNAi), were studied to elucidate the possible functions of RACK1 in responses to drought stress in rice. Real-time PCR analysis showed that the expression of RACK1 in transgenic rice plants was inhibited by more than 50%. The tolerance to drought stress of the transgenic rice plants was higher as compared with the non-transgenic rice plants. The peroxidation of membrane and the production of malondialdehyde were significantly lower, and the superoxide dismutase activity in transgenic rice plants was significantly higher than those in non-trangenic rice plants. It is suggested that RACK1 negatively regulated the redox system-related tolerance to drought stress of rice plants. 相似文献
12.
13.
The gene Pi-d2, conferring gene-for-gene resistance to the Chinese blast strain ZB15, was isolated from a rice variety (Digu) by the map-based cloning strategy. Here, we constructed a control plasmid pZH01-pi-d2tp309 (pZH01-tp309) and three different expression constructs, pCB-Pi-d25.3kb (pCB5.3kb), pCB-Pi-d26.3kb (pCB6.3kb) and pZH01-Pi-d22.72kb (pZH01-2.72kb) of Pi-d2, driven by Pi-d2 gene's own promoter or CaMV35S promoter. These constructs were separately introduced into japonica rice varieties Lijiangxintuanhegu, Taipei 309, Nipponbare and Zhonghua 9 through Agrobacterium- mediated transformation. A total of 150 transgenic rice plants were obtained from the regenerated calli selected on hygromycin. PCR, RT-PCR and Southern-blotting assay showed that the gene of interest had been integrated into rice genome and stably inherited. Thirty-five transgenic lines independently derived from T1 progeny were inoculated with the rice blast strain ZB15. Transformants exhibited resistance to rice blast at various levels. The lesions on the transgenic plant leaves were less severe than those on the controls and the resistance level of transgenic plants harboring the gene of interest from three vectors had no difference. The own promoter of Pi-d2, about 2.2 kb or 3.2 kb, had the similar promoter function as CaMV35S. Field evaluation for three successive years supported the results of artificial trial, and some lines with high resistance to rice leaf blast and neck blast were obtained. 相似文献
14.
CHEN Zhuanzhuan # LI Xianfeng # ZHONG Min GE Jiaqi FAN Xiaolei ZHANG Changquan LIU Qiaoquan 《中国水稻科学》2019,33(6):513-522
【Objective】Gelatinization temperature (GT) is one of the key physicochemical properties in rice quality, which is mainly regulated by ALK (SSII-3) gene. In general, there are two ALK alleles among indica cultivars. To detect their functional differentiation in indica rice,【Method】Zhenshan 97B (a high GT variety carrying ALKc allele) and Longtefu B (a low GT variety carrying ALKb allele), were used as receptors for the generation of transgenic rice with down-regulation of ALK expression by RNA interference (RNAi).【Result】Down-regulation of ALK gene significantly decreased the GT of the transgenic lines. Due to the difference of original GT between the two receptors, the GT of transgenic rice lines derived from Zhenshan 97B (a high GT variety) decreased significantly, but it is slightly decreased in transgenic plants derived from Longtefu B (a low GT variety). The differential scanning calorimetry (DSC) results showed that the initial temperature of RNAi transgenic rice was significantly lower than the corresponding control and the transgenic lines were gelatinized in advance. The peak value of GT(Tp) in RNAi rice grains was significantly lower than that of the control under Zhenshan 97B background. However, Tp of RNAi rice grains under Longtefu B background was significantly lower than the control to a lesser extent. Also, down-regulation of ALK expression had a significant effect on rice physical-chemical characteristics. An increase of apparent amylose content in RNAi transgenic plants was detected due to the decreased expression of ALK gene. Besides, the pasting properties showed that down-regulation of ALK gene had obvious effects on peak viscosity and breakdown value, improving the taste of the transgenic rice. The gel consistency was significantly different among Zhenshan 97B RNAi lines and their parents, but no difference was found in Longtefu-derived transgenic lines.【Conclusion】RNA interference to ALK allele expression had a significant effect on rice quality, especially the gelatinization characters. Down-regulated expression level of ALKc allele would cause larger variation of physical-chemical characteristics between transgenic rice and their parent than that of ALKb allele. 相似文献
15.
【目的】稻米糊化温度是影响稻米品质的重要指标，该性状受主效基因ALK/SSII-3调控，ALK基因具有多个复等位基因，本研究旨在通过RNAi技术明确籼稻亚种中两个不同ALK等位基因的效应。【方法】以分别含有ALKc和ALKb等位基因的高糊化温度品种珍汕97B和低糊化温度品种龙特甫B为试验材料，使用RNAi技术构建ALK表达下调的转基因株系，通过对其稻米理化品质的测定来明确不同等位基因表达下调对稻米品质的影响。【结果】对不同转基因水稻目的基因的表达分析显示本研究中转基因株系的ALK基因受到了不同程度的干扰。重点分析了不同RNAi株系稻米的糊化温度，结果表明珍汕97B的RNAi转基因稻米的糊化温度极显著降低，而在低糊化温度品种龙特甫B背景中下调表达ALK基因后对糊化温度的影响较小；转基因株系与未转化亲本相比，米粉的起始糊化温度都显著降低，表现为提前糊化；在珍汕97B背景下干扰系的峰值温度与未转化亲本相比极显著降低，而在龙特甫背景下米粉的峰值温度与未转化对照相比显著降低。对不同转基因系的理化品质分析表明，ALK下调表达植株稻米的表观直链淀粉含量显著增加，下调表达ALK后会引起米粉峰值黏度和崩解值的改变。高糊化温度品种珍汕97B干扰系与未转化对照相比胶稠度呈现极显著性差异，而低糊化温度品种龙特甫干扰系的胶稠度与未转化对照相比没有差异。【结论】下调表达ALK等位基因对稻米理化品质产生显著影响，并且干扰不同等位基因的效应存在明显差异，即籼稻中的两个ALK等位基因的效应存在显著差异。 相似文献
16.
1-磷酸甘露醇脱氢酶基因转化水稻的研究 总被引：14，自引：0，他引：14
摘要：PCR和Southern blotting检测表明,来自大肠杆菌的[i]mtlD[/i]基因已通过农杆菌介导整合进水稻基因组。[i]mtlD[/i]基因在T1代出现分离, T2代出现纯系。在0.75% NaCl胁迫下,7个转基因T3代株系都能检测到mtlD酶活性,与对照相比细胞膜的相对电导率和大分子渗漏值明显降低。部分转基因株系能在 1.0% NaCl浓度下正常生长,而对照在0.5% NaCl浓度下已不能存活。通过有性杂交途径实现了[i]mtlD[/i]和[i]gutD[/i]两个基因的聚合,部分杂交后代株系能在1.25%NaCl胁迫下正常生长结实。 相似文献
17.
18.
19.
水稻精细胞优势表达基因RSG6启动子的克隆及表达 总被引：1，自引：0，他引：1
利用已知的[i]RSG6[/i]基因序列和GenBank中提供的[i]RSG6[/i]前导序列设计引物,通过基因组DNA的PCR扩增得到长度为1 408 bp和1 173 bp的两个[i]RSG6[/i]启动子片段PB、PS,测定序列分析发现,PB、PS分别位于水稻第10和第4染色体上,PB、PS序列之间具有较高的同源性,且都含有大量的启动子元件。通过设计带有酶切位点的引物扩增出PB和PS的各两条缺失片段PB1、PB2、PS1、PS2,与质粒pBI221连接后得到中间载体pBI221 PB1、pBI221 PB2、pBI221 PS1、pBI221 PS2,再与质粒pBIN19连接构建出双元表达载体pBIN GUSB1、pBIN GUSB2、pBIN GUSS1、pBIN GUSS2,转化农杆菌EHA105,感染烟草叶片和烟草花粉,瞬时表达显示缺失片段PB1、PB2、PS1、PS2均具有启动子功能,且PB1、PS1的启动效率较PB2、PS2高。 相似文献