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1.
《科学通报(英文版)》1999,44(22):2051-2051
The synthesized Bacillus thuringiensis insecticidal protein gene crylA(b&c) and the synthesized gene GNA, (the mannose specific lectin from snowdrop ( Galanthus nivalis)), tumefaciens have been inserted into plant expression vector pGW4BAI. Leave stripes of Nico-tiana tabacum var. K326 have been transformed with Agrobacterium tumefaciens strain LBA4404 harboring the plant expression vector. 28 kanamycin resistant tobacco plants have been obtained. PCR and Southern blot analyses show that the foreign crylA and GNA genes have been inserted into the genome of transformed tobacco plants. Haemagglutination assays show that GNA has a functional activity. Leaf disc bioassays against cotton bollworm ( H. armigera) show that the transgenic tobacco plants have a high insecticidal activity. The inhibition of aphid population in leaf disc bioassays against Myzus persicae shows that the fecundity of aphid on transgenic plants is lower than that on untransformed plants; the aphid population on the transgenic tobacco plants is 25%-70% that on untransformed tobacco plants. ELISA analysis of CrylA protein in tobcco leaves provides similar data to bioassay results. Through the two bioassays against H. armigera and M. persicae, several transgenic tobacco plants showing high insect-resistant activities to both pests have been obtained. 相似文献
2.
The resistance ratio ofHelicoverpa armigera to Cry1 Ac insecticidal protein fromBacillus thuringiensis (Bt) is 13.1- and 3.02-fold after 18 generations of selection by transgenic tobacco expressing Bt or two (Bt and CpTI) insecticidal
protein genes, in which the average corrected mortality for each selection treatments is about 60%. The mortality of selected
population by transgenic Bt gene tobacco is significantly lower than the control strain when fed on transgenic tobacco plants.
The mortaltty of the selected population by transgenic two genes tobacco was not significantly different from the control
strain. This is the first experiment under laboratory condition which has proved that transgenic two genes tobacco could significantly
delay resistance development ofH. armigera compared with one gene. 相似文献
3.
SHI Hong ZHANG Lifu HUA Baozhen MI Xiangcheng WEI Wei ZHANG Yongjun MA Keping 《科学通报(英文版)》2006,51(8):946-951
Since the first commercial release of transgenic crop expressing genes from Bacillus thuringiensis (Bt), there have been concerns about its potential impact on the environment. Research has focused on the ecological effects from second exposure to Bt prot… 相似文献
4.
GUO Hongnian QIN Hongmin CHEN Xiaoying LI Changqing LU Rui TIAN Yingchuan 《自然科学进展(英文版)》2002,12(5):347-352
A new plant expression vector (pBS29K-BA) containing two insect resistant genes, a synthetic chimeric gene BtS29K encoding the activated insecticidal protein Cry1Ac and a gene API-BA encoding the arrowhead (Sagittaria sagittifolia L.) proteinase inhibitor (API) A and B, is constructed. Transgenic tobacco plants expressing these two genes are obtained through Agrobacterium-mediated transformation of tobacco leaf discs. The average expression levels of Cry1Ac and API-BA proteins in transgenic plants are of 3.2 μg and 4.9 μg per gram fresh leaf respectively. The results of insecticidal assay of transgenic plants indicate that the pBS29K-BA transformed plants are more resistant to insect damage than the plants expressing the Cry1Ac gene or API-BA gene alone. 相似文献
5.
The DNA 4 coding region of banana bunchy top virus from a Chinese Zhangzhou isolate (BBTV-ZZ) is cloned by PCR. The sequencing analysis shows that it is 351 nucleotides long and it putatively encodes a protein of 116 amino acids. On the basis of a plant binary vector pBin438, the plant expression vector pBBTV-4B harboring the BBTV-ZZ DNA 4 coding region has been constructed and then transferred to tobacco (Nicotiana tobacum cv. Xanthi nc) by a Agrobacterium-mediated procedure. Under insect-free condition, movement-defective mutant of CMV-Fny strain (CMV-Fny-△MP) is mechanically inoculated on the lower leaves of transgenic plants. Systemic symptoms with different degrees of severity are developed in the upper uninoculated leaves of transgenic plants at 12 days postinoculation (dpi), while no symptoms can be seen in the uninoculated leaves of untransformed plants at any time. Accumulation of CMV-Fny is detected on the upper uninoculated leaves of transgenic plants, but is not on that of untransformed plants by indirect double antibody sandwich enzyme-link immunosorbent assay (DAS-ELISA). The results reveal that transgenic plants have acquired the property of cell-to-cell movement and systemic spread of CMV-Fny-△MP. This suggests that the protein encoded by BBTV-ZZ DNA 4 might have function of viral movement protein. 相似文献
6.
Transgenic tobacco plants expressing synthetic Cry1Ac and Cry1Ie genes are more toxic to cotton bollworm than those containing one gene 总被引:2,自引:0,他引:2
Yun Lian ZhiWei Jia KangLai He YunJun Liu FuPing Song BaoMin Wang GuoYing Wang 《科学通报(英文版)》2008,53(9):1381-1387
Transgenic tobacco plants carrying CrylAc, Crylle or both genes were obtained. In the leaves of transgenic plants carrying both genes, the contents of CrylAc and Crylle proteins were 0.173% and 0.131% of the total proteins, respectively. CrylAc protein content was 0.182 % and Cry1 le protein content was 0.124% of the total proteins in the leaves of transgenic plants containing only one Bt gene. Fresh leaves of transgenic tobacco and wild-type plants were used for the insect bioassay against wild-type and Cry1Ac-resistant cotton bollworm (Helicoverpa armigera). The bioassay results showed that transgenic plants carrying both genes were significantly more toxic to wild-type and CrylAc-resistant cotton bollworm than those carrying CrylAc or Crylle alone. This study indicates that the higher toxicity of transgenic tobacco plants carrying both genes is caused by the cooperative function of both Bt proteins, thus providing a potential way to delay the development of insect resistance to transgenic crops. 相似文献
7.
Inheritance of resistance toHelicoverpa armigera of 3 kinds of transgenicBt strains available in upland cotton in China 总被引:1,自引:0,他引:1
Canming Tang Jing Sun Xiefei Zhu Wangzhen Guo Tianzhen Zhang Jinliang Shen Congfen Gao Weijun Zhou Zhixian Chen Sandui Guo 《科学通报(英文版)》2000,45(4):363-367
There are 3 kinds of transgenicBt strains, Shanxi 94-24, Zhongxin 94, and R19, in upland cotton in China. Their transgenicBt insect-resistance cultivars or hybrids have been developed and grown by farmers. Genetic studies indicate that the resistance
of the 3 transgenicBt cotton strains toHelicoverpa armigera is controlled by one pair of non-allelic dominant genes. Linkage relationship between the resistant genes of R19 and Shanxi
94-24 transgenicBt strains shows that they may be inserted in the same chromosome. F1 hybrids crossed among the 3 strains show that high levels of protection from feeding damage are the same as that of their
parents. Therefore, there is no co-suppression phenomenon in many transgenic plants. The results presented here afford a fundamental
reliance in developing transgenicBt insect-resistant cultivars and exploiting the heterosis of hybrids in upland cotton. 相似文献
8.
采用农杆菌介导法将含有苏云金芽孢杆菌毒蛋白基因(CryIAc)与半夏凝集素抗虫基因(Pta)的高效植物表达载体pCAMBIA3300转入番茄品系Micro Tom的子叶外植体中。经过共培养、除草剂筛选和分化再生,获得了24个具有除草剂抗性的株系。再将转化后的番茄植株经过PCR检测和Southern Blot检测,确定检测后呈阳性反应的株系为8个。通过小菜蛾幼虫初步抗性试验证明,转基因株系表现出较强的抗虫性。实验结果为进一步研究番茄抗虫性和培育抗虫番茄新品种奠定了重要基础。 相似文献
9.
Shi Chunlin Zhu zhen Chen Lei Xu Honglin Xiao Guifang Feng Pingzhang Illimar Altosaar 《高技术通讯(英文版)》2000,6(1)
To increase the expression level of CryIA(c) gene in transgenic plants, a plant expression vector pBinMoBc carrying the CryIA(c) gene under control of chimeric OM promoter and Ω factor was constructed. As a control, pBinoBc carrying the CryIA(c) gene with the CaMV 35S promoter was also constructed. The vectors were transferred into tobacco plants respectively via Agrobacterium-mediated transformation. ELISA assay showed that the expression level of the CryIA(c) gene in pBinMoBc transgenic tobacco plants was 2.44-times that in pBinoBc transgenic tobacco plants, and it could be up to 0.255% of total soluble proteins. Bioassay showed that pBinMoBc transgenic tobacco plants had more notable insecticidal effect than pBinoBc transgenic tobacco plants. The above results showed that the chimeric OM promoter was a stronger promoter than CaMV 35S promoter that was widely used in plant genetic engineering, and this is very useful in pest-resistant plant genetic engineering. 相似文献
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2—3 anti-fungal disease genes are coinserted with hygromycin phosphotransferase in the same vector. Two insecticidal genes and PPT acetyl transferase genes are placed in another one. The vectors are co-delivered to rice embryonic cellus tissue at a molar ratio of 1︰1 using the particle gun method. 55 independent regenerated lines have been obtained through screening for hygromycin resistance. Of these, 70% transgenic plants harbor 6—7 foreign genes. The genes on the same vectors are always co-delivered to rice plant. Northern blot analysis has indicated that the multiple foreign genes give stable expression. In the 6 transgenic plants carrying 6—7 foreign genes, multiple foreign genes tend to integrate in 1 or 2 genetic loci. Progeny segregation is consistent with Mendel’s 3︰1 segregation law. 8 homozygous R1 transgenic plants harboring 2—3 anti-fungal and 2 insecticidal genes are selected from large number of transgenic progeny screening for hygromycin and Basta resistance. 相似文献
12.
为筛选出烟蚜茧蜂(Aphidius gifuensis Ashmead)规模扩繁中的最佳繁蚜冬寄主,比较了自然条件下7种不同冬寄主上烟蚜的种群数量变化,烟蚜茧蜂对各寄主上烟蚜的防控效果及僵蚜羽化率.结果表明:烟蚜在7种寄主上种群数量的自然增长从大到小依次表现为烤烟K326、芥菜、芥菜型油菜、甘蓝型油菜、萝卜、红菜薹、白菜;芥菜上烟蚜茧蜂的寄生率为70.56%,显著高于其他寄主;7种冬寄主上僵蚜羽化率均高于70%,其中芥菜最高,为97.33%,萝卜最低,为72.87%,二者差异具有统计学意义.烟蚜茧蜂均能寄生7种冬寄主植物上所饲养的烟蚜且僵蚜羽化率相对较高,但总体而言芥菜上烟蚜的种群数量增长较快,烟蚜茧蜂的寄生率和僵蚜羽化率均最高,可利用芥菜来替代烟草作为冬季繁蚜寄主,也可在春烟种植前对芥菜田释放烟蚜茧蜂使其在田间建立自然种群. 相似文献
13.
利用番茄U3snRNA基因上游启动区构建植物表达载体及对烟草的转化 总被引:1,自引:0,他引:1
利用番茄U3snRNA基因上游启动区和ACC合成酶反义RNA-核酶嵌合基因DNA片段,构建含U3snRNA基因上游启动区-ACC合成酶的反义RNA-核酶嵌合序列的表达载体,重组于植物双元表达载体pGA643中,得到pGU3R.用三亲融合法导入农杆菌LBA4404中,采用叶盘法转化烟草,诱导再生小植株,获得了卡那霉素的抗性植株.提取抗性植株总DNA,通过PCR、PCRSouthern杂交检测并分别用启动区序列和ACC合成酶的反义RNA-核酶嵌合序列作探针,通过Southern杂交检测,已筛选出整合有外源基因的转化植株.为进一步研究U3snRNA上游启动区增强反义RNA-核酶基因的表达奠定了基础. 相似文献
14.
CHEN Yan GUO Hongnian JIANG Ying LI Changqing ZHAO Wenming TIAN Yingchuan 《自然科学进展(英文版)》2003,13(5):362-366
An antimicrobial peptide gene from Amaranthus hypochondriacus, Ah-AMP, was amplified by PCR and cloned. Sequence analysis results revealed that this gene is 261 bp in length encoding a precursor polypeptide of 87 amino acid residues. Ah-AMP gene was inserted in the binary vector pBin438 to construct a plant expression vector pBinAH916. Leave explants of Nicotiana tabacum var. SR1 were transformed with Agrobacterium tumefaciens LBA4404 harboring the above expression vector. Results from PCR, Southern and Northern blot analyses confirmed that the Ah-AMP gene had been integrated into the tobacco genome and was transcribed at mRNA level. Two bacterial-resistant transgenic plants were selected by inoculating the plants with Pseudomonas solanacearum and statistic analysis of two T1 lines showed that the resistance increased by 2.24 and 1.62 grade and the disease index decreased by 49.6% and 37.3% respectively when compared with the non-transformed control plants SR1. The results from challenging the plants with inoculums of Phytophthora parasitica showed that the symptom development was delayed and disease index was significantly reduced. These results suggest that Ah-AMP gene may be a potentially valuable gene for genetic engineering of plant for disease-resistance. 相似文献
15.
Function of resveratrol derived from transgenic plant expressing resveratrol synthase gene 总被引:1,自引:0,他引:1
Two genes from grapevine coding for resveratrol synthase, named RS1 and RS2, were cloned by RT-PCR. AnEscherichia coli expression vector was constructed by insertion of RS1 into pBV221. A specific protein with the same molecular weight (42
ku) as the resveratrol synthase was expressed and used to prepare the rabbit antiserum. A plant expression vector was constructed
by inserting the RS1 gene into pBin438 downstream of the doubled CaMV 35S promoter and TMV-Ω fragment. PCR-positive transgenic
tobacco plants were obtained after transformation withAgrobacterium tumefaciens LBA4404 harboring the plant expression vector. Southern blot analysis demonstrated that the foreign gene was integrated into
the tobacco genome. The results of RT-PCR and Western blot indicated that the RS1 gene was transcribed and expressed. Formation
of resveratrol in transgenic tobacco was further determined by thin-layer chromatography of silica gel and HPLC. Increased
accumulation of human breast adenocarcinoma cells in G0 and G1 phases of cell cycle was observed in cells treated with resveratrol purified from transgenic tobacco as compared to the untreated
cells. 相似文献
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Ubi1 intron-mediated enhancement of the expression of Bt cry1Ah gene in transgenic maize (Zea mays L.) 总被引:3,自引:0,他引:3
WANG YueBing ;LANG Zhihong ;ZHANG Jie ;HE KangLai ;SONG FuPing ;HUANG DaFang 《科学通报(英文版)》2008,53(20):3185-3190
The cry1Ah gene was one of novel insecticidal genes cloned from Bacillus thuringiensis isolate BT8. Two plant expression vectors containing cry1Ah gene were constructed. The first intron of maize ubiqutinl gene was inserted between the maize Ubiquitin promoter and cry1Ah gene in one of the plant expressing vectors (pUUOAH). The two vectors were introduced into maize immature embryonic calli by microprojectile bombardment, and the reproductively plants were acquired. PCR and Southern blot analysis showed that foreign genes had been integrated into maize genome and inherited to the next generation stably. The ELISA assay to T1 and T2 generation plants showed that the expression of CrylAh protein in the construct containing the ubil intron (pUUOAH) was 20% higher than that of the intronless construct (pUOAH). Bioassay results showed that the transgenic maize harboring cry1Ah gene had high resistance to the Asian corn borers and the insecticidal activity of the transgenic maize containing the ubil intron was higher than that of the intronless construct. These results indicated that the maize ubil intron can enhance the expression of the Bt cry1Ah gene in transgenic maize efficiently 相似文献
19.
The plant expression vectors pBCT2 and pBT2 were constructed with the cDNA sequence (tin2) and genomic DNA sequence (tin2i) of tomato proteinase inhibitor II gene respectively. Then the two expression vectors were transferred into tobacco via the Agrobacterium tumefaciens strain LBA4404, and transgenic tobacco plants were generated. Molecular analysis and trypsin activity assay showed that both cDNA and genomic DNA were expressed properly in the transgenic plants. Insecticidal activities in these transgenic plants indicated that transgenic tobacco plants carrying tin2i sequence were more resistant to 2-instar larvae of Heliothis armigera Hubner than those carrying tin2 sequence. Therefore the intron of tin2i sequence might be a contributor to insecticidal activity of the transgenic tobacco. 相似文献
20.
转蔗糖: 蔗糖-1-果糖基转移酶基因提高烟草的耐旱性 总被引:4,自引:0,他引:4
蔗糖: 蔗糖-1-果糖基转移酶(sucrose: sucrose 1-fructosyltransferase, 1-SST)以蔗糖为底物催化生成蔗果三糖等低聚合度的果聚糖.将从莴苣中克隆的1-SST基因重组到pCAMBIA1300-als中,构建了在CaMV 35S启动子调控下的植物表达载体,利用农杆菌介导的叶盘转化法将1-SST基因导入烟草中,PCR和Southern杂交检测表明获得了转基因植株,RT-PCR结果表明该基因在烟草中正常表达. 对T0代转基因烟草进行的耐旱性分析结果表明,干旱胁迫6d的转基因植株丙二醛含量和电解质渗漏率显著低于未转基因对照,叶片相对含水量下降速度也明显比对照慢. 对转基因植株叶片糖分分析表明,转基因烟草植株积累果聚糖,并在干旱胁迫后含量明显增加,而未转基因对照植株不积累果聚糖. 在14%PEG溶液中未转基因烟草种子的萌发率仅为转基因烟草种子的一半;在附加200mmol/L甘露醇的培养基中未转基因烟草种子根的生长明显受到抑制,而转基因烟草根的生长发育正常. 以上研究结果表明,转1-SST基因烟草植株耐旱性的提高可能与该基因的表达有关. 相似文献