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1.
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
2.
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
3.
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
4.
生长素及其信号转导系统对植物的生长发育具有重要的影响。本研究从银腺杨'84K'(Populus alba × P. glandulosa cl. '84K')中分离了生长素受体基因PtrFBL1,利用PMDC32构建了PMDC32-PtrFBL1超量表达载体,并通过遗传转化获得了超量表达植株17个。对温室定植的3个转基因株系和对照植株的根系、生长量和光合指标等性状分析结果显示:转基因株系总根长和总根面积达到显著或极显著差异,而根系干质量、平均不定根系长度、平均不定根直径差异不显著;株高、平均节间长、地径和高径比皆高于对照,且大多数转基因株系达到显著差异;除气孔限制值(Ls)低于对照外,气孔导度(Cd)、水分利用效率(WUE)、光能利用效率(LUE)和叶绿素相对含量皆高于对照,且大多数转基因株系达到显著或极显著差异。以上结果表明,可能是FBL1超表达增加了转基因株系根系面积,提高了水分和养分的吸收利用,进而导致转基因株系光能吸收和转化效率提高,引起转基因株系生长加快。 相似文献
5.
《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
6.
7.
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
8.
Akira Kikuchi Xiang Yu Takayoshi Shimazaki Akiyoshi Kawaoka Hiroyasu Ebinuma Kazuo N. Watanabe 《Journal of Wood Science》2009,55(2):149-153
Allelopathy tests were conducted on salt-tolerant transgenic eucalyptus trees conferring bacterial codA gene in the designated net-house conditions under Type II use (contained use) of the Japanese law on environmental biosafety
aiming for Type I (field use) application. Three transgenic and corresponding nontransgenic genotypes were employed for four
different tests: (1) sandwich bioassay; (2) soil germination method; (3) gas chromatography (GC) for volatile substances from
the plants; and (4) high-performance liquid chromatography (HPLC) on phenolic compounds from fresh leaves, which are the primary
allelopathic substances on the species. The simple approaches, the bioassays, indicated no significant difference between
the transgenic and nongenetically modified genotypes. There was no qualitative difference between the transgenic and nontransgenic
lines by GC or HPLC. Absence of any quantitative difference was suggested by repetitive examination and subsequent analysis
of variance assessments with the chromatographic methods and bioassays. Moreover, it was also indicated that bioassays should
be the primary assessment method for allelopathy in considering the simplicity, speed, low cost, and reproducibility of these
methods. Overall, substantial equivalence was considered on the three transgenic genotypes with codA gene when compared with the nontransgenic Eucalyptus camaldulensis lines. The experiments supported the application to isolated field testing of the transgenic Eucalyptus camaldulensis genotypes as the first case and experience in Japanese regulatory approval processes Type I Use for the deliberate release
to the environment. 相似文献
9.
Toru Taniguchi Ken-ichi Konagaya Manabu Kurita Naoki Takata Katsuaki Ishii Teiji Kondo Fumiaki Funahashi Seiichi Ohta Tomomi Kaku Kei’ichi Baba Rumi Kaida Takahisa Hayashi 《Journal of Wood Science》2012,58(6):550-556
Xyloglucan is thought to be a key hemicellulose cross-linking adjacent cellulose microfibrils in plant cell walls. The growth traits of transgenic poplars (Populus alba) with decreased xyloglucan from overexpression of Aspergillus aculeatus xyloglucanase were characterized during a 4-year field trial. The field-trial site consisted of two blocks, a fertile soil block and a non-fertile soil block, determined by soil analysis. In the fertile block, the growth of aboveground biomass of the transgenic poplars was reduced to 24?C44?% compared to that of wild-type poplars, in contrast to the growth seen in chamber and greenhouse conditions. In the non-fertile block, the aboveground biomass of transgenic poplars was also smaller than that of the wild-type poplars. Because poplars reproduce asexually by root suckers, we also compared the formation of root suckers from transgenic and wild-type poplars. Root suckers formed less frequently from transgenic poplars than from wild-type poplars. The growth rates of root suckers from transgenic poplars were also slower than those from wild-type poplars. The results showed that constitutive degradation of xyloglucan impairs poplar growth and vegetative reproduction ability. 相似文献
10.
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
11.
Amines and their derivatives are known to influence insect behavior involved in feeding and reproduction. In order to examine
the feasibility of improving the resistance of poplar to insect pests by the introduction of a plant-derived amine-generating
transgene, explants from the hybrid poplar clone ‘INRA 717 1B4’ (P. tremula ×P. albo) were transformed with a Camptotheca acuminata tryptophan decarboxylase (TDC, EC 4.1.1.28) cDNA driven by the CaMV 35S promoter. The enzyme TDC catalyzes the decarboxylation
of tryptophan to tryptamine, which, in addition to being a bioactive amine itself, is known to act as a precursor of various
other indole derivatives. Putative transgenic lines were confirmed by PCR for the TDC1 gene sequence and by the expression analysis of the transgene mRNA and encoded protein. No visible phenotypic changes were
associated with ectopic TDC1 expression. Chemical and radiotracer analyses of the transgenic plants revealed tryptamine accumulation as high as 4 mM in
leaf tissue, and suggested that the tryptamine produced by ectopically expressed TDC was not further metabolized. Insect bioassays with the TDC transgenic plants showed that the tryptamine accumulation was consistently associated with adverse effects on feeding potential
and physiology of Malacosoma disstria (forest tent caterpillar). 相似文献
12.
[目的]通过不同群落毛红椿天然林林地土壤及其真菌对种子萌发和幼苗存活影响的研究,探讨影响毛红椿天然更新的障碍因素。[方法]在江西九连山国家级自然保护区内的3个毛红椿天然优势群落中,分别取距离毛红椿母树3个不同距离(2.5、5.0、7.5 m)处的根区土壤和远离毛红椿母树(25 m以外)的非根区土壤,以非林地土为对照开展室内模拟播种和以根区土壤真菌的水或根系分泌物悬液对毛红椿幼苗进行灌根接种,观察种子萌发、幼苗存活和幼苗感病情况。[结果]各群落土壤中种子发芽均呈先上升后下降规律,且基本在第8 10天达到高峰;林地土与非林地土种子发芽率差异不显著,但林地根区土幼苗的存活率极显著低于非林地土和林地非根区土;距母树3个不同距离根区土的幼苗存活率差异不显著。灌根接种不同处理间幼苗存活率差异极显著,具体表现为接种RS2、RS5根区真菌的幼苗感病率显著高于根系分泌物及无菌水空白对照,且RS2真菌的根系分泌物悬浮液幼苗感病率显著高于其无菌水悬浮液。[结论]毛红椿种子萌发不受土壤环境的影响,但幼苗建成受根区范围内土壤的影响,根系分泌物和致病菌的互作显著降低幼苗保存率。由此推断,毛红椿根区土壤内存在幼苗的潜在致病菌及根系分泌物可增强其致病性。 相似文献
13.
[目的]模式植物在木本植物中鉴定的许多重要调控因子家族在木本植物中出现了基因家族成员扩张,但ARRs家族作为细胞分裂素响应调节因子在杨树基因组中成员数量反而减少,其在木本植物中如何行使功能需要进一步研究。[方法]本研究通过生物信息学构建PtRRI启动子与GUS融合表达载体,检测植物激素处理后PtRRI表达量和检测PPtRRI::GUS转基因植株在生根过程中GUS信号等方法,对杨树PtRRI基因的组织特异性表达模式进行分析。[结果]表明:PtRRI在杨树根部、形成层、木质部表达量相对较高,PtRRI转录受6-BA激素诱导,与其在不定根发育过程中激素调控下的表达相一致。[结论]PtRRI基因可能参与杨树的次级生长。 相似文献
14.
[目的]探讨南亚热带西南桦和尾巨桉人工纯林的凋落叶分解动态及其与土壤化学性质之间的相关关系.[方法]采用原位分解袋法研究凋落叶的分解过程.[结果]表明:西南桦、尾巨桉人工林凋落叶分解系数分别为0.96 a-1和0.88 a-1.在为期12个月的分解试验中,2种凋落叶有机C含量在整个分解过程中呈逐渐下降趋势;全K含量和C/N比在分解前期迅速下降,之后趋于平缓;全N含量和全P含量在整个分解过程中呈逐渐上升趋势;2种凋落叶N/P比则呈先升高后下降的趋势.无论是分解前期还是分解后期,凋落叶质量损失与N含量均呈显著正相关(前期R=0.877;后期R=0.855),与C/N均呈显著负相关(前期R=-0.735;后期R=-0.697).与尾巨桉林地土壤性质相比,西南桦凋落叶分解提高了林地0~10、10~20 cm土壤的有机C、全N、全P、全K、N/P,对2030 cm土壤有机C、全K、pH值、C/N、N/P则未产生显著影响.相关分析表明:凋落叶初始有机C含量与土壤有机C、全N、全P、全K、N/P显著相关;凋落叶初始全N含量与土壤全N、pH值、C/N显著相关.[结论]凋落叶的养分含量与土壤养分的关系紧密;与尾巨桉相比,西南桦凋落叶的养分含量明显较高,分解速率更快,释放到土壤中的养分也更多. 相似文献
15.
A 3 125 bp cellulose synthase gene, PtoCesA1, which has a 98% identity to PtrCesA1 from Populus tremuloides, was cloned from cDNA prepared from secondary xylem of P. tomentosa. Four anti-expression vectors with different fragments of PtoCesA1, named as pBIPF, pBICC1, pBIPR and pBIBR, were constructed. Some traits of transformed tobacco of pBICC1, pBIPR and pBIBR
differed from wild types, such as small leaves, “dwarf” phenotype and thinner xylem and fiber cell walls than wild plants
consistent with a loss of cellulose. It indicated that the growth of transgenic tobacco was restrained by the expression of
anti-PtoCesA1. Transgenic tobacco was obtained and the contents of cellulose and lignin were analyzed as well as the width and length of
fiber cells, and xylem thickness for both transgenic and control plants. Transformed tobacco showed a different phenotype
from control plants and it implied that PtoCesA1 was essential for the cellulose biosynthesis in poplar stems.
[Supported by the Hi-Tech Research and Development Program of China (863) (2001AA244060 and 2003AA244020) and National Basic
Research Program of China (973) (J1999016003)] 相似文献
16.
糖基化磷脂酰肌醇锚定蛋白(GPIAP)因其结构和功能的多样性,决定了它在各种生物学过程中都发挥着重要作用。采用同源克隆的方法从绿竹(Bambusa oldhamii)中获得一个GPIAP同源基因,命名为BoGPIAP,cDNA全长1 772 bp,其中包括1 356 bp的开放阅读框,编码一个451 aa的的蛋白。蛋白结构分析表明,该蛋白包含1个典型的GPIAP家族保守区域(47-211)和1个CCVS结构域,在N-端和C-端分别具有1个跨膜信号肽和1个GPI锚定信号肽,属于GPIAP家族。构建BoGPIAP与GFP融合的表达载体,在洋葱表皮细胞中瞬时表达,结果显示BoCOBL::GFP融合蛋白定位于细胞膜上,证明BoGPIAP基因编码的蛋白为膜蛋白。分别构建BoGPIAP的正义、反义表达载体并转化烟草(Nicotiana tabacum)。PCR检测结果表明,BoGPIAP已转入烟草。与野生型相比,转反义基因植株细弱,纤维细胞壁明显变薄;而转正义基因植株粗壮,纤维细胞壁明显变厚。表明BoGPIAP可能对竹子纤维细胞壁的发育具有调控作用。 相似文献
17.
Zhang Qian Lin Shanzhi Lin Yuanzhen Zhang Zhiyi Liu Haijun Zou Yening Wang Zeliang College of Biological Sciences Biotechnology Beijing Forestry University Beijing P. R. China College of Resources Environment Beijing Forestry University Beijing P. R. China 《中国林学(英文版)》2004,6(3)
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
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.
PtLFY, a LEAFY (LFY) gene, was cloned from Populus tomentosa (LM50) by PCR. Sequencing analysis indicated that PtLFY was 2 629 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.
[Supported by the National Natural Science Foundation of China (Grant No. 30371175) and Postdoctoral Foundation of China (Grant
No. 2002032041)] 相似文献
20.
[目的]本研究旨在探索与挖掘胡杨基因组大片段的潜在功能,发掘具有潜在育种价值的胡杨基因簇。[方法]利用已构建的胡杨基因组BIBAC文库,采用花序浸染法,将胡杨基因组大片段78A2D10导入模式植物拟南芥基因组中。采用抗性筛选、分子检测及表型观察等方法鉴定、分析转化型植株。[结果]共获得15株特异表型的转化植株。与野生型相比,转化型植株主侧茎生长受到抑制,莲座叶面积增大近3倍,叶片数量增多,叶边缘皱缩,抽薹推迟约13周,株高增加近32.0 cm,侧茎发育成次生莲座,植株寿命延长约7周。[结论]胡杨基因组片段78A2D10可延长植株营养生长期及植株寿命,据此推测该基因片段可能与营养生长有关。 相似文献