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1.
The putative transgenic hybrid triploid poplars [(P. tomentosa × P. bolleana) × P. tomentosa] with CpTI gene have been outplanted in test field for 2 years. Although the authors’ previous studies have proved that they are highly resistant to 3 species of poplar-threatening insect pests and contain high content of CpTI protein in foliage, incorporation status of foreign CpTI gene in poplar genome is uncertain. In this present study, the incorporation of foreign CpTI gene in genome of 5 transgenic poplars was confirmed by PCR and Southern blotting analysis. DNA amplification showed that there were clear DNA bands of about 450bp specific to CpTI gene in transgenic lanes, while no corresponding band in non-transgenic lane was observed. Correspondingly, clear DNA hybridization signals and no signal were exhibited on film for DNA Southern blotting analysis in transgenic lanes and non-transgenic lane, respectively, which further confirmed the stable integration of foreign CpTI gene in genome of 2-year-old transgenic poplar. 相似文献
2.
对转豇豆胰蛋白酶抑制剂基因的三倍体毛白杨杂种 [(毛新杨×毛白杨 )×毛白杨 ]的可溶性总蛋白和胰蛋白酶抑制剂蛋白的含量进行测定 .结果发现 ,与未转基因植株相比 ,所有供试转基因株系叶片中的总可溶性蛋白含量明显增加 ,但老叶的含量高于嫩叶 ,这表明转基因株系可溶性总蛋白含量增加可能是CpTI基因表达的结果或由于外源基因导入后引起杨树基因组中某些自身基因的表达所致 .转基因株系叶片中有较高含量CpTI,而对照叶片则检测不到CpTI,这进一步证实了CpTI基因已在转基因株系中得到稳定表达 .进一步比较发现 ,在供试的 5个无性系中 ,TG0 7、TG0 4和TG71在总蛋白含量和CpTI含量增加较为明显 .另外 ,聚丙烯酰胺凝胶电泳胶分析发现 ,与未转基因植株相比 ,在所有供试转基因株系叶片中均出现一条分子量为 11.3kD的清晰蛋白带 相似文献
3.
表达豇豆胰蛋白酶抑制剂转基因杨树的虫试试验 总被引：2，自引：0，他引：2
将转豇豆胰蛋白酶抑制剂基因的三倍体毛白杨回交杂种 [(毛新杨×毛白杨 )×毛白杨 ]]于春季萌发前截干 .用萌条上的树叶对 3种主要杨树害虫 :天幕毛虫 (Malacosomadisstria)、舞毒蛾 (Lymant riadisparL .)和杨雪毒蛾 (StilpnotiacandidaStaudinger)进行离体虫试试验 ,年底测量各转基因株系萌条树高和地径 ,以衡量其生长性状 .结果发现 ,与对照相比 ,转基因株系的树高和地径没有显著降低 ,说明外源基因的导入没有影响转基因杨树的速生特性 ,相反 ,TG0 4的树高和地径较对照有明显的提高 ,这可能是外源导入后促进转基因杨树生长的结果 ;转基因植株均能显著提高害虫的死亡率 ,降低幼虫的排粪量、蛹重、体重增加量和成虫的产卵量 ;其中 ,TG0 7、TG0 8和TG713个转基因株系的抗虫特性较为突出 ,表明CpTI基因在它们的树体内表达较为活跃和稳定 相似文献
4.
To have a preliminary insight into biosafety of genetically transformed hybrid triploid poplars (Populus tomentosa × P. bolleana) × P. tomentosa with the cowpea trypsin inhibitor (CpTI) gene, two layers of rhizospheric soil (from 0 to 20 cm deep and from 20 to 40 cm deep, respectively) were collected for
microorganism culture, counting assay and PCR analysis to assess the potential impact of transgenic poplars on non-target
microorganism population and transgene dispersal. When the same soil layer of suspension stock solution was diluted at both
1:1 000 and 1:10 000 rates, there were no significant differences in bacterium colony numbers between the inoculation plates
of both transgenic and non-transgenic poplars. The uniform results were revealed for both soil layer suspension solutions
of identical poplars at both dilution rates except for non-transgenic poplars at 1:10000 dilution rates from the same type
of soil. No significant variation in morphology of both Gram-positive and Gram-negative bacteria was observed under the microscope.
The potential transgene dispersal from root exudates or fallen leaves to non-target microbes was repudiated by PCR analysis,
in which no CpTI gene specific DNA band was amplified for 15 sites of transgenic rhizospheric soil samples. It can be concluded that transgenic
poplar with the CpTI gene has no severe impact on rhizospheric microorganisms and is tentatively safe to surrounding soil micro-ecosystem.
[Supported by the National Project in Transgenic Plant and Application (Grant No. J2002-2003)] 相似文献
5.
To have a preliminary insight into biosafety of genetically transformed hybrid triploid poplars (Populus tomentosa × P. bolleana) × P. tomentosa with the cowpea trypsin inhibitor (CpTI) gene, two layers of rhizospheric soil (from 0 to 20 cm deep and from 20 to 40 cm deep, respectively) were collected for microorganism culture, counting assay and PCR analysis to assess the poten-tial impact of transgenic poplars on non-target microorganism population and transgene dispersal. When the same soil layer of suspension stock solution was diluted at both 1:1 000 and 1:10 000 rates, there were no significant differences in bacterium colony numbers between the inoculation plates of both transgenic and non-transgenic poplars. The uniform results were revealed for both soil layer suspension solutions of identical poplars at both dilution rates except for non-transgenic poplars at 1:10 000 dilution rates from the same type of soil. No significant variation in morphology of both Gram-positive and Gram-negative bacteria was observed under the microscope. The potential transgene dispersal from root exudates or fallen leaves to non-target microbes was repudiated by PCR analysis, in which no CpTI gene specific DNA band was amplified for 15 sites of transgenic rhizospheric soil samples. It can be concluded that transgenic poplar with the CpTI gene has no severe impact on rhizospheric microorganisms and is tentatively safe to surrounding soil micro-ecosystem. 相似文献
6.
转基因杨树对杨小舟蛾幼虫解毒酶活性的影响 总被引：3，自引：2，他引：3
用转Bt单基因和转Bt CpTI双价基因杨树叶片饲喂杨小舟蛾,幼虫中肠酯酶的活力在饲喂初期显著升高,但在饲喂一定时间后受到抑制:转Bt单基因杨树在饲喂24 h开始受到抑制,48 h受到明显抑制,较对照下降了15.8%;而转双价基因杨树在饲喂12 h受到明显抑制,较对照下降了38.1%.转双价基因杨树对中肠酯酶的抑制作用大于转Bt单基因杨树.转Bt单基因杨树对幼虫中肠羧酸酯酶的抑制能力不强,而转双价基因杨树对中肠羧酸酯酶的抑制作用大于转Bt单基因杨树,饲喂12 h后活力开始受到抑制,饲喂24、36、48 h的活力分别较对照下降了33.4%、22.5%、29.6%,与对照均有显著差异.转基因杨树主要通过抑制幼虫中肠酯酶和羧酸酯酶这2种解毒酶活性而干扰昆虫正常的生理代谢,从而起到毒杀害虫的作用. 相似文献
7.
Li Min Wang Hua-fang Yin Wei-lun He Si-jie Chen Shou-yi College of Biological Sciences Biotechnology Beijing Forestry University Beijing P. R. China Institute of Genetics Developmental Biology Chinese Academy of Sciences Beijing P. R. China 《中国林学(英文版)》2006,8(4):20-24
Transgenic lines were achieved by transforming the E. coli 1-phosphate mannitol dehydrogenase gene (mtl-D) into the Populus tomentosa Carr. genome. An Agrobacterium tumefaciens strain (AGL1), constructed by cloning mtl-D into the disarmed plasmid pBin438, was used to infect leaves of the clone YW2. The infected leaf discs were cultured on a medium containing 30 mg·L-1 kanamycin and 500 mg·L-1 cefotaxime. Transgenic plantlets regenerated from the infected leaves, rooted on the medium containing 30 mg·L-1 kanamycin. PCR and a Southern blotting test verified that the exogenous mtl-D gene had integrated into the transformation plants of the P. tomentosa genome. The mannitol content in control plant was 69μg·g-1 FW, and the mannitol contents of the transgenic lines T1 to T5 ranged between 103.7 and 289.5μg·g-1 FW. Of the shoots of the control plants 20% survived; on the medium containing 0.6% NaCl, 60% and 70% of two transgenic shoots survived on a medium containing 0.8% NaCl. 相似文献
8.
PtLFY, a LEAFY （LFY） gene, was cloned from Populus tomentosa （LM50） by PCR. Sequencing analysis indicated that PtLFY was 2629 bp long, composed of three exons and two introns and encoded 378 amino acids. The splice donor sites and the splice acceptor sites were in identical positions to the LFY and its homologues. The amino acid sequence inferred was 68%-99% homologous to those of LFY and its homologues by blast analysis in GenBank. The Southern blot analysis indicated that there was a single copy of the PtLFY gene in genomic DNA of male and female P. tomentosa （LM50 and 5082）. The pBI121-Ptalfy （reverse）-intron-Ptlfy-GUS-nos was constructed using RNA interference （RNAi） technique and verified by PCR and digestion identification and transformed into tobacco. Some transgenic tobacco plants were obtained by PCR and PCR-Southern identification. The growth was generally repressed in transgenic tobacco plants compared with wild-type ones and some phenotypic differences were observed. 相似文献
9.
转基因杨树对美国白蛾幼虫中肠保护酶系统的影响(英文) 总被引：10，自引：0，他引：10
以转Bt基因欧洲黑杨(P.nigra L.)和转CpTI基因毛白杨(Populus tomentosa)叶片饲喂4-5龄美国白蛾（Hyphantria cunea Drury）幼虫，对其体内保护酶系统活性进行测定。结果表明，饲喂两种转基因杨树叶片的幼虫中肠保护酶表现出相似的变化规律，SOD、CAT和POD三种酶的活性在饲喂后数小时内逐渐增加，某一时刻达到最高值，此后突然下降。饲喂转Bt基因杨树叶片的幼虫，其中肠SOD、CAT活性峰值出现在饲喂后的24小时，POD活性峰值出现在饲喂后的12小时；饲喂转CpTI基因杨树叶片的幼虫，其中肠三种保护酶活性高峰出现时间均较前者滞后12小时。本文还比较了饲喂两种转基因叶片不同中毒程度的美国白蛾幼虫体内保护酶活性，发现不论饲喂那种转基因叶片，中毒较轻者其体内保护酶活性显著高于中毒较重者，这种差异在饲喂CpTI叶片的处理株表现尤为明显。 相似文献
10.
ZHANG Xiao-fen CHEN Min ZHANG Bin-yu LUO You-qing HOU Ying-jie SU Xiao-hua 《中国林学(英文版)》2009,11(3):164-167
Laboratory feeding experiments with the poplar aphid, Chaitophorus populeti (Panzer), feeding on transgenic poplar (P. alba × P. glandulosa) varieties C13-5 and C013-5, were carried out to study the effect of transgenic poplar on the ladybird Harmonia axyridis (Pallas). The mortality and development time of the immature stages, the eclosion rate and body mass of H. axyridis were measured. The results indicated that C. populeti feeding on different varieties of transgenic plants had no statistically significant effect on the mortality of H. axyridis larvae. The development time of larval and pupal stages were not significantly different between the two transgenic poplars
and a non-transgenic poplar. Furthermore, the body mass and eclosion rate did not show any difference between the H. axyridis feeding on aphids reared on transgenic plants and those from non-transgenic plants. It is suggested that transgenic plants
have no deleterious effect on the predatory ladybird. 相似文献
11.
利用ＲＴ－ＰＣＲ技术克隆了番茄ＡＣＣ氧化酶基因ＬＥＥＴＨＹＢＲ编码区０．９ｋｂ的ｃＤＮＡ片段,经酶切图谱分析和序列分析鉴定后,反向插入到植物表达载体ｐＢｉｎ４３８中,构建了表达ＡＣＣ氧化酶反义ＲＮＡ的二元载体。用农杆菌侵染美洲黑杨叶片,在含卡那霉素的ＭＳ培养基上选择转化子和植株再生,通过ＰＣＲ检测筛选到１６株转基因杨树植株,Ｓｏｕｔｈｅｒｎｂｌｏｔ分析初步确证了外源基因是以单拷贝插入到杨树基因组中;对杨树幼苗乙烯释放量的测定结果表明转基因杨树的乙烯释放量为对照植株的２８％。 相似文献
12.
《Forest Ecology and Management》2005,217(1):95-102
Ecological risk analysis for transgenic conifers is more complex compared to transgenic food crops. With added age and size, greater quantities of seed and pollen travel even farther via long-distance dispersal. Perennial reproduction in conifers means that the same plant disperses annually for one or more decades prior to harvest. Although complex, gene flow in transgenic trees will have adverse consequences only if the following conditions are met: (1) the inserted gene or DNA construct itself inflicts harm or (2) the DNA insertion event causes unintended or secondary effects on fitness traits. Here, transgene seed escape into less managed ecosystems was modeled across 5 and 50 generations using Pinus taeda, an indigenous US timber commodity species. The inserted DNA construct itself was assumed to exert no direct effects on fitness but rather caused secondary or unintended effects on one or more fitness traits. The deterministic population genetics model, based on one or two fitness components, predicted the transgene allele frequency for a single colony of escaped transgenic seedlings through time. The first scenario modeled commercial transgenic plantations. A transgene allele escaping from a 100-ha plantation spread rapidly through time even if it exerted slight, unintended secondary effects on viability and fecundity. The second scenario modeled the case of a 10-ha field trial surrounded by non-transgenic or wild-type forests. Here, transgene spread was slowed by dilution from non-transgenic sources. 相似文献
13.
With the widespread distribution of transgenic cotton plants, concerns over their potential effects on beneficial insects
and other non-target pests have been raised. In the present study, the field and laboratory effects of Cry1Ac/CpTI transgenic
cotton pollen on the pollinating beetle Haptoncus luteolus were investigated. Abundance of adult beetles was studied in cotton plots at two sites in Wuhan in 2009, and the average
number of adults per flower was recorded. Life-history parameters including developmental duration, hatch rate, pupation rate,
emergence rate and egg number per female, as well as the survivorship curves of H. luteolus were studied in laboratory experiments. The results showed that the adult abundance in the field did not differ significantly
between non-transgenic and transgenic cotton, the exposure to Cry1Ac/CpTI pollen in laboratory had no effect on the developmental
time, hatching rate, pupation rates and emergence rates of H. luteolus, no significant differences were found in the mating rates and the fecundity between treatments, and the survival curves
of both treatments also resulted in no significant differences. It indicated there were no significant direct adverse effects
of transgenic cotton pollen on H. luteolus both in the field and in the laboratory. Our study bridged the gap of environmental risk assessment of transgenic cotton
for pollinators other than pollinating bees. 相似文献
14.
以毛白杨为转基因受体材料,利用根癌农杆菌介导法转化来源于益母草的阳离子抗菌肽基因LJAMP2。经卡那霉素筛选,共获得50株抗性植株。GUS组织染色和PCR检测显示有30株抗性植株呈阳性,初步证明外源目的基因已整合到毛白杨基因组中。RT-PCR证实抗菌肽基因LJAMP2在转基因植株中能大量表达。离体抗病性试验表明:转基因毛白杨细胞粗提液的抑菌能力明显强于非转基因植株。进一步将溃疡病菌接种在转基因和野生型毛白杨茎段上培养30天,转基因植株的病级指数均低于非转化植株。上述抗性试验结果表明:在毛白杨中超量表达益母草抗菌肽基因LJAMP2能显著提高其溃疡病抗病性。 相似文献
15.
We have extracted total DNA from different fractions of fresh wood as well as from cold-stored and air-dried wood harvested from transgenic aspen grown in the field. The highest amounts of DNA were obtained from bark/cambium tissue; the DNA quality, however, was poor. Best results in PCR and Southern blot analyses were obtained from DNA extracted from early wood. Using appropriate primer pairs, amplification products were obtained from both the foreign gene (transgene) and aspen genomic sequences. In Southern blot analyses transgene-specific hybridisation signals were obtained. This is the first report on the detection of foreign genes in wood sampled from genetically modified trees. 相似文献
16.
Ferritin, a universal intracellular protein, can store large amounts of iron and improve plant resistance to abiotic and biotic stress. In this study, a ferritin gene(TaFer) from Tamarix androssowii Litv. was transferred into Populus tomentosa Carr. cv 'BJR01' via Agrobacterium. Six independent transgenic lines were obtained with a tolerance to kanamycin and three were randomly selected for further analysis. The PCR and RT-PCR results indicate that the TaFer gene had been integrated into the poplar genome. The effect of the gene on abiotic stress tolerance was tested, and the results show that transgenic plants improve growth, had higher chlorophyll and lower MDA contents, and higher relative electrical conductivity,fewer changes of SOD and POD activities, higher iron content, higher root ferric reductase activity and lower levels of ROS accumulation and cell death in response to drought, Fe-insufficient or Fe-excess tolerance. These results indicate that the TaFer gene can improve abiotic stress tolerance in transgenic Populus tomentosa. 相似文献
17.
以转Bt基因南林895杨株系B1、B4、B17、B21扦插苗为试验材料,分析其在室内和野外自然条件下对靶标害虫杨小舟蛾幼虫的抗虫性。结果表明:转Bt基因杨树4个株系均有一定的杀虫活性,其中株系B21对杨小舟蛾1龄幼虫12 d校正死亡率高达95.3%;虫试表明转Bt基因杨树各株系扦植苗在野外自然条件下的12 d幼虫校正死亡率8月份为35.0%88.8%,9月份为40.5%95.3%。用转Bt基因杨树叶片饲养杨小舟蛾,对照植株杨小舟蛾幼虫化蛹率为83.3%96.0%,而转Bt基因株系幼虫化蛹率为8.0%76.7%,二者有显著差异。转Bt基因杨树对杨小舟蛾幼虫的生长发育有抑制作用,饲喂8 d后害虫取食量、体质量增长速率显著低于对照。此外,为了解转Bt基因杨树对土壤微生物的影响,对转Bt基因杨树株系及对照进行了根际土壤微生物可培养类群的分析,初步结果显示:转Bt基因杨树根际土壤微生物中细菌、真菌、放线菌的数量与对照比较无显著差异。 相似文献
18.
Expression of the two coleopteran-resistant proteins (Bt-Cry3A and oryzacystatin I) was detected in the leaves of field-grown transgenic poplar (BOGA-5) in two or three subsequent years. The BOGA-5 contained ～10 μg g(-1) of Cry3A over the individual years with no detection in the control, and protein extracts from BOGA-5 displayed a higher reduction in papain activity (～42%) compared with ～21% in the control. Laboratory feeding experiments showed that the total mortality of the target pest Plagiodera versicolora (Coleoptera, Chrysomelida) larvae fed with BOGA-5 leaves was 76.7%, significantly higher than that of the control (P相似文献
19.
以转PEPC基因南林895杨和对照(南林895杨)为材料,分析不同转基因植株的光合生理参数。结果显示:转PEPC基因杨树与C4光合途径相关的酶活性比对照增高,PEPC活性增加较为明显,最高达38.6%。转PEPC基因杨树表现较强的对强光利用能力,其光饱和点比对照提高约10%～20%,饱和点时的光合速率比对照高17.7%～58.1%。转PEPC基因杨树光补偿点低于对照,利用弱光的能力增强;CO2饱和点低,比对照低22.2%～44.4%;CO2补偿点也较低,比对照降低58.1%～76.5%,表明对CO2的利用效率较高。此外,转基因杨树羧化效率增高,比对照增加最高达62.3%。 相似文献