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根癌农杆菌介导转化获得耐逆性增强的高羊茅转基因植株 总被引：13，自引：2，他引：13
为改良草坪型高羊茅（Festuca arundinacea Schreb.）的耐逆性，以成熟种子来源的胚性愈伤组织为受体材料，通过农杆菌介导法将拟南芥（Arabidopsis thaliana）耐逆相关CBF1基因导入4个供试品种的基因组，经GUS染色、PCR检测和Southern杂交分析验证，获得了112株转基因植株，转化频率为0.92% ~2.87%，不同品种间存在差异。试验表明，在高盐与高渗胁迫下，转基因植株具有显著生长优势，存活率极显著高于非转化对照植株，经低温、高温、干旱和高盐等逆境胁迫处理后的叶片相对电导率平均较对照植株低25% ~ 30%，证明转基因植株的耐逆性有所增强。考察发现，非胁迫条件下CBF1基因的组成型超表达使转基因植株的生长受到抑制。 相似文献
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CBF/DREB基因是一类植物特异性转录因子,通过调控下游抗逆相关基因的表达,在植物逆境胁迫应答过程中发挥重要作用。为了研究新型的禾本科模式植物二穗短柄草DREB基因的功能,克隆BdDREB1G的cDNA序列,分析该基因在各种非生物胁迫条件下的表达,构建过表达载体,获得转基因拟南芥。结果表明:高温、低温和水杨酸胁迫处理显著诱导BdDREB1G的表达,暗示该基因响应高温和低温胁迫。初步观察结果说明,该与野生型拟南芥相比,转基因植株生长发育迟缓。这些为研究该基因响应非生物胁迫的功能奠定基础。 相似文献
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DREB/CBF转录因子响应植物干旱、高盐和低温等非生物胁迫应答是通过特异结合DRE/CRT顺式作用元件。利用RT-PCR技术从毛果杨克隆PtrDREB28转录因子基因,用基因枪法对其进行亚细胞定位分析,结果表明其定位在细胞核中;利用农杆菌介导法对野生型大青杨进行遗传转化,并对转基因株系进行抗逆性分析,结果显示经高盐胁迫处理的转基因株系的耐受性明显高于野生型植株;通过测定和比较经盐胁迫处理后转基因株系和野生型植株的丙二醛质量摩尔浓度和相对电导率,发现转基因株系的丙二醛质量摩尔浓度和相对电导率均显著低于野生型植株,表明PtrDREB28基因的过表达可以显著提高杨树耐盐性,由此推测PtrDREB28转录因子可能参与杨树逆境胁迫应答过程。 相似文献
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Improvement of organic phosphate acquisition in transgenic tobacco plants by overexpression of a soybean phytase gene Sphy1 总被引：1，自引：1，他引：0
Li Guo Yuxin Zhao Shuhua Zhang Haina Zhang Kai Xiao 《Frontiers of Agriculture in China》2009,3(3):259-265
Due to the huge amount in the soil, phytate is an important potential source for providing the plants with available phosphorus
(Pi) by the involved catalytic reaction of phytase. In this study, a construct fusing the open reading frame (ORF) of Sphy1 into corresponding positions in the fragment of binary expression vector pBI121 was created and used to transform tobacco.
Molecular identification by PCR and RT-PCR indicated the target gene Sphy1 in the transgenic tobacco plants was transcribed under the regulation of an upstream promoter. Compared with the control
plants, the phytase activities in all the transgenic plants were increased, with the increased range consistent with the expression
levels in the transgenic plants. Under the growth conditions with phytate as the sole phosphorus source, the transgenic line
1 plants displayed a high expression level of Sphy1 and shows notable improved growth performance, such as higher fresh weight and dry weight, as well as higher total P content
and more accumulative P amount per plant than CK. This clearly indicated that overexpression of Sphy1 could improve the phosphorus acquisition by the extruded Sphy1 phytase in the rhizosphere, where this enzyme could catalyze the degradation of the phytate and release the available Pi
for plants. The Sphy1 gene seemed to have a potential value in the creation of new crop cultivars with high phosphorus use efficiency. 相似文献
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双价抗寒基因CBF3/COR15A转化大白菜的初步研究 总被引：1，自引：0，他引：1
为探索拟南芥的冷诱导转录因子CBF3和抗寒基因COR15A在"黔白"系列大白菜的转化体系,以"黔白"系列大白菜3个优良自交系作为研究对象,分别用大白菜4d苗龄的带柄子叶、半子叶、下胚轴作为外植体,利用农杆菌介导的方法将同时含有CBF3和COR15A双价抗寒基因的表达载体转入大白菜,就影响大白菜再生和遗传转化的因素进行研究。通过抗生素筛选得到大白菜T0代转基因抗性植株36株,经PCR检测后获得转基因阳性植株6株。经抗寒性初步鉴定,转基因大白菜植株抗寒性超过对照。 相似文献
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采用盆栽试验,研究0.3%NaCl胁迫下全红苋菜(低Cd累积品种,简称全红)和柳叶苋菜(高Cd累积品种,简称柳叶)的有机酸代谢差异及其与Cd累积和耐盐性的关系。结果表明:相比对照,盐胁迫后两个品种根部、地上部和根际土壤溶液中的Cd含量均呈增加趋势,且表现为全红显著低于柳叶(P0.05),同时全红和柳叶的Cd转运系数分别提高2.84、5.58倍;盐胁迫促进了苋菜对Na~+的积累且明显干扰K~+的吸收,K~+/Na~+均显著降低(P0.05),且全红根部K~+/Na~+显著高于柳叶(P0.05)。盐胁迫后,两个品种根部、地上部及根际土壤溶液中均检测到多种有机酸,其中根际土壤溶液中苹果酸和草酸含量均显著升高(P0.05),且与根际土壤溶液中的Cd含量显著正相关;柳叶地上部草酸及根部乙酸的含量均显著升高(P0.05),且分别与地上部及根部Cd含量显著正相关。研究表明,盐分胁迫下,全红具有较低的Cd累积性且表现出更强的耐盐性,不同品种的有机酸积累和分泌的差异是导致这一结果的重要原因之一。 相似文献
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Overexpression of a Cytosolic Ascorbate Peroxidase Gene,OsAPX2, Increases Salt Tolerance in Transgenic Alfalfa 
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下载免费PDF全文 Alfalfa (Medicago sativa L.) is an important forage crop in the world and it is of great significance for the improvement of its salt tolerance. To improve salt tolerance in alfalfa, a rice ascorbate peroxidase gene (OsAPX2) was introduced into alfalfa using Agrobacterium tumefaciens-mediated transformation with marker gene bar. The different T-DNA insertions in T1 transgenic alfalfa were identified by Southern hybridization. Three independent T2 transgenic lines were selected for stress analysis and the results showed that all of them were salt tolerant compared with wild-type plants. The transgenic plants had low levels of H2O2, malondialdehyde and relative electrical conductivity under salt and drought stresses. Moreover, the contents of chlorophyll and proline, and APX activity were high in transgenic plants under salt and drought stresses. Taken together, the overexpression of OsAPX2 enhances salt tolerance in alfalfa through scavenging reactive oxygen species. 相似文献
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为探究马铃薯硫氧还蛋白基因(StTrxF)的耐盐生理机制,通过RT-PCR方法从马铃薯品种中薯5号中克隆得到全长549bp硫氧还蛋白基因(StTrxF)。该基因编码182个氨基酸,预测蛋白质分子量为44.17ku,理论等电点(pI)为5.23,具有典型的Thioredoxin结构域。由StTrxF基因推导的氨基酸序列与番茄、拟南芥、芝麻和赤霞珠等TrxF蛋白质氨基酸序列的同源性为96.02%～59.28%。构建植物过表达载体,导入拟南芥中,获得转基因拟南芥纯系植株。通过对转基因拟南芥植株的耐盐离体和盆栽鉴定,表明转基因植株的耐盐性显著提高。同时盐胁迫下,转基因植株的超氧化物歧化酶(SOD)活性和脯氨酸含量显著提高,丙二醛(MDA)含量显著降低。结果表明,表达StTtxF基因通过增加转基因植株脯氨酸含量,提高SOD活性,降低MDA含量,以维持细胞渗透平衡并激活ROS清除系统,具有提高转基因拟南芥植株耐盐性的显著效果。 相似文献
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Ying HUANG Xiao-xia ZHANG Yi-hong LI Jian-zhou DING Han-mei DU Zhuo ZHAO Li-na ZHOU Chan LIU Shi-bin GAO Mo-ju CAO Yan-li LU Su-zhi ZHANG 《农业科学学报》2018,17(12):2612-2623
Maize is one of the most important crops worldwide, but it suffers from salt stress when grown in saline-alkaline soil. There is therefore an urgent need to improve maize salt tolerance and crop yield. In this study, the SsNHX1 gene of Suaeda salsa, which encodes a vacuolar membrane Na+/H+ antiporter, was transformed into the maize inbred line 18-599 by Agrobacterium-mediated transformation. Transgenic maize plants overexpressing the SsNHX1 gene showed less growth retardation when treated with an increasing NaCl gradient of up to 1%, indicating enhanced salt tolerance. The improved salt tolerance of transgenic plants was also demonstrated by a significantly elevated seed germination rate (79%) and a reduction in seminal root length inhibition. Moreover, transgenic plants under salt stress exhibited less physiological damage. SsNHX1-overexpressing transgenic maize accumulated more Na+ and K+ than wild-type (WT) plants particularly in the leaves, resulting in a higher ratio of K+/Na+ in the leaves under salt stress. This result revealed that the improved salt tolerance of SsNHX1-overexpressing transgenic maize plants was likely attributed to SsNHX1-mediated localization of Na+ to vacuoles and subsequent maintenance of the cytosolic ionic balance. In addition, SsNHX1 overexpression also improved the drought tolerance of the transgenic maize plants, as rehydrated transgenic plants were restored to normal growth while WT plants did not grow normally after dehydration treatment. Therefore, based on our engineering approach, SsNHX1 represents a promising candidate gene for improving the salt and drought tolerance of maize and other crops. 相似文献
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为评价 ZmDREB2.7优异等位基因型对玉米耐旱性的作用，选择携带优异等位基因型的玉米自交系CIMBL70、CIMBL92做父本及敏感基因型的玉米自交系SHEN5003做母本，用于构建近等基因系群体。结果表明，NIL- ZmDREB2.7CIMBL70和NIL- ZmDREB2.7CIMBL92恢复率分别为 94.96%和95.11%；苗期存活率分别是81.34%和83.33%，显著高于轮回亲本（SHEN5003，37.3%）。为更加明确 ZmDREB2.7的耐旱效应，构建了 ZmDREB2.7pro: ZmDREB2.7转基因玉米植株。转基因玉米材料 ZmDREB2.7pro: ZmDREB2.7-1和 ZmDREB2.7pro: ZmDREB2.7-10相对表达量更高，苗期存活率分别是68.05%和 70.83%，显著高于野生型（A188，58.33%）。在新疆大田干旱条件下， ZmDREB2.7pro: ZmDREB2.7-1和 ZmDREB2.7pro: ZmDREB2.7-10两个株系均比野生型小区少减产20%以上。因此，研究结果可能为玉米耐旱新品种的培育提供有价值基因资源和可行的实践方案。 相似文献
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通过农杆菌介导的方式将獐茅液泡膜Na~+/H~+逆向转运蛋白基因(AlNHX)表达在马铃薯中,并对影响马铃薯转化的几种因素(抗生素浓度、农杆菌菌液浓度、共培养时间等)进行优化。结果表明:最适农杆菌菌液浓度是OD600=0.6,最适侵染时间是5min,最适共培养时间是2d,马铃薯转化中最适头孢霉素浓度是400mg/L;经PCR检测确认的转基因马铃薯植株在含0.7%NaCl的培养基中可以生长,而野生型马铃薯无法在此培养基中正常生长。在盐胁迫条件下,转基因马铃薯的Na~+、K~+含量及K~+/Na~+的比值均高于野生型马铃薯。研究证实马铃薯植物的的耐盐性可以通过农杆菌转化导入獐茅液泡膜Na~+/H~+逆向转运蛋白基因而得到提高。 相似文献
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You-wen QIU Zhe FENG Ming-ming FU Xiao-han YUAN Chao-chao LUO Yan-bo YU Yan-zhong FENG Qi WEI Feng-lan LI 《农业科学学报》2019,18(2):372-380
Salt stress is one of the major factors affecting plant growth and yield in soybean under saline soil condition. Despite many studies on salinity tolerance of soybean during the past few decades, the detailed signaling pathways and the signaling molecules for salinity tolerance regulation have not been clarified. In this study, a proteomic technology based on two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS) were used to identify proteins responsible for salinity tolerance in soybean plant. Real-time quantitative PCR (qRT-PCR) and Western blotting (WB) were used to verify the results of 2-DE/MS. Based on the results of 2-DE and MS, we selected glucosyltransferase (GsGT4), 4-coumarate, coenzyme A ligase (Gs4CL1), mitogen-activated protein kinase 4 (GsMAPK4), dehydration responsive element binding protein (GsDREB1), and soybean cold-regulated gene (GsSRC1) in the salinity tolerant soybean variety, and GsMAPK4 for subsequent research. We transformed soybean plants with mitogen-activated-protein kinase 4 (GsMAPK4) and screened the resulting transgenics soybean plants using PCR and WB, which confirmed the expression of GsMAPK4 in transgenic soybean. GsMAPK4-overexpressed transgenic plants showed significantly increased tolerance to salt stress, suggesting that GsMAPK4 played a pivotal role in salinity tolerance. Our research will provide new insights for better understanding the salinity tolerance regulation at molecular level. 相似文献
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利用农杆菌介导的叶盘法将类番茄茄(Solanum lycopersicoides)和多毛番茄(Lycopersicon habrochaites)的CBF1基因转入普通烟草(Nicotiana tabacum L.),通过分析转基因烟草在低温胁迫条件下目的基因的表达水平并结合转录组数据分析差异表达的基因,揭示CBF1基因在烟草中发挥抗冷作用的机制与相关性。结果显示:低温胁迫下,转基因烟草的抗低温能力高于非转基因的野生型;SlCBF1和LhCBF1基因在普通烟草中过量表达后,转录组测序分析表明SlCBF1基因转化的烟草植株与野生型烟草植株相比,差异表达基因为53个,其中33个表达上调,20个表达下调;LhCBF1基因转化的烟草和野生型烟草植株相比,差异表达的基因为106个,表达上调的有62个,表达下调的有44个。表明在烟草中过表达SlCBF1和LhCBF1会影响转基因烟草中受其调控的下游基因的表达水平,进而提高烟草的抗冷能力。 相似文献
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紫花苜蓿抗旱耐盐碱转基因抗性苗耐盐性研究 总被引：7，自引：1，他引：7
在紫花苜蓿抗旱、耐盐碱转基因抗性苗和对照苗的组织培养过程中模拟盐胁迫生境,经0%、0.3%、0.5%、0.7%、0.9%、1.1%、1.3%、1.5%、1.7%、1.9%NaCl处理15 d后,苜蓿转化苗和苜蓿对照苗的盐适应性生长性能发生变化.观察统计了转基因抗性苗和无菌苗在盐胁迫下的成活率;测定了这2组苜蓿植株的根长和叶片的电导率.结果表明:转化苗比对照苗耐受盐胁迫的能力要高,转化苗能够耐受1.3%的NaCl盐浓度,在NaCl浓度为1.5%时其生长才开始受到损伤,而对照苗只能在NaCl浓度0.5%以下正常生长,在NaCl浓度为0.7%时其生长即开始受到伤害.同时,在盐胁迫下,转化苗的根生长能力要强于对照苗.在同一盐浓度下转化苗的细胞膜透性比对照苗的细胞膜透性要低,表明苜蓿转基因抗性苗比对照苗耐受盐伤害的能力强. 相似文献