共查询到20条相似文献，搜索用时 15 毫秒
1.
林业生物技术的研究已经取得了许多重要的进展。遗传转化和转基因树的应用是当代林业研究中的重要领域。尽管微弹和农杆菌介导的转化已经被应用于重要的针叶树种,转基因植株仅仅在少数几个针叶树种上获得成功。大多数转基因针叶树来自微弹介导的转化,例如转基因的挪威云杉、白云杉、黑云杉和辐射松等。利用转基因技术可以改良树的形状、生长率、木材质量、抗虫性、抗除草剂能力和对不良环境的抵抗力。本文对遗传转化技术在这些领域中所取得的进展做了简短的综述。 相似文献
2.
[目的]通过植物转基因技术获得抗病毒大花蕙兰种质资源,优化转化体系和鉴定方法.[方法]本研究克隆了齿兰环斑病毒外壳蛋白基因,并构建了该基因的pBI121表达载体,用根癌农杆菌介导法转化大花蕙兰,尝试以巢式PCR方法检测转基因再生植株.[结果]优化了大花蕙兰遗传转化体系,建立了利用巢式PCR技术检测转基因大花蕙兰植株的方法,获得了32株转基因株(系).[结论]优化了以类原球茎为外植体的农杆菌介导转化大花蕙兰的方法,确定以5%~10%类原球茎存活时的抗生素(卡那霉素)浓度为筛选浓度;获得了转ORSV CP基因大花蕙兰植株;对大花蕙兰转基因植株检测时,巢式PCR较普通PCR更灵敏、准确. 相似文献
3.
The AhDREB1 gene, cloned from Atriplex hortensis L., was transferred into black locust （Robiniapseudoacacia L.） by an Agrobacterium-mediated transformation. The results suggest that stems of black locust sub-cultured in vitro for 20 d are suitable for genetic transformation. The optimum concentrations of kanamycin and cefotaxime were 30 and 150 mg.L-1, respectively. Important factors affecting the transformation efficiency were studied by means of a L9（3^4） orthogonal design. An effective system for genetic transformation in black locust was developed as follows： the stems were pre-cultured for 2 d, immersed in the Agrobacterium solution （OD600 = 0.7） with 10 mg·L^-1 acetosyringone for 21 min and then co-cultured for 2 d. The selection pressures, changing from low to high, could improve transformation efficiency. The transgenic plants were identified by a PCR method. The PCR results indicated that the AhDREB1 gene had been integrated into the genome of black locust and two lines of the transgenic plants were obtained. 相似文献
4.
《Scandinavian Journal of Forest Research》2012,27(2):151-160
A particle inflow gun enabled efficient production of transgenic plantlets of Picea abies from embryogenic suspension cultures. In transient assays, the Zea ubiquitin promoter was 12-16 times as active as the 35S promoter. For stable transformation, the plasmid pAHC25 contained the bar gene and the gusA gene, both driven by the Zea ubiquitin promoter. Cells were maintained from 1 to 3 h before bombardment on proliferation medium supplemented with 0.25 M myoinositol and, from day 8, supplemented with Basta as selective agent. Embryogenic colonies resistant to Basta appeared from two months after bombardment. Of over 100 independent Basta-resistant sublines tested, 65% expressed the co-transformed reporter gene, even when it was not linked to the selectable marker. Over 80% of the sublines retained their embryogenic potential. Of 11 transformants analyzed, 4 contained transgenes in low copy number (1-3), the rest contained transgenes in up to 15-20 copies. Over 200 Basta-resistant sublines from four cell lines have been established, of which 138 are confirmed as transformed. Plantlets have been regenerated and grown on in pots. 相似文献
5.
The binary vector plasmid pIG121-Hm carrying the chimeric neomycin phosphotransferase (npt II), β-glucuronidase (gusA) and hygromycin phosphotransferase (hpt) genes was delivered intoRobinia pseudoacacia L. callus using theAgrobacterium tumefaciens-mediated transformation method and particle gun transformation method. It was determined that adding 10 mg/L of acetosyringone
toAgrobacterium infection medium and callus-Agrobacterium co-culture medium could increaseAgrobacterium-mediated transformation efficiency. In contrast, particle gun transformation could successfully deliver plasmid DNA into
the callus, but its transformation efficiency was lower and gene expression was transient in comparison with theAgrobacterium-mediated method, suggesting that substantially stronger promoters might be required. 相似文献
6.
Agricultural productivity may be raised in a sustainable way by many different technologies such as biological fertilizers,
soil and water conservation, biodiversity conservation, improved pest control, and changes in land ownership and distribution.
Of these measures, biotechnology applications probably hold the most promise in augmenting conventional agricultural productivity,
because biotechnology applications give not only the need to increase production, but also protect the environment and conserving
natural resources for future generations. Biotechnology applications will have the possibilities to increase productivity
and food availability through better agronomic performance of new varieties, including resistance to pests; rapid multiplication
of disease-free plants; ability to obtain natural plant products using tissue culture; diagnosis of diseases of plants and
livestock; manipulation of reproduction methods increasing the efficiency of breeding; and the provision of incentives for
greater participation by the private sector through investments. Insect resistance through the transfer of a gene for resistance
fromBacillus thuringiensis (Bt) is one of the most advanced biotechnology applications already being commercialized in many parts of the world. This paper
reviews the development and the status ofBt technology and application ofBt transgenic plants in current agriculture, and discusses specific issues related to the transfer of the technology to the
future of genetic engineered trees with emphasis on conifers.
Biography: Tang Wei (1964-), male, Ph. Doctor, Research associate, Department of Biology, Howell Science Complex, East Carelina University,
Greenville, NC 27858-4353, USA.
Responsible editor: Chal Ruihai 相似文献
7.
杨树是重要的栽培树种,也是研究林木基因工程的重要模式植物。杨树转基因研究可以打破种属限制,具有高效性和专一性的特点,是对杨树进行遗传改良的重要手段。为了更好的进行该方面的工作,本文介绍了杨树转基因涉及到的抗虫、抗除草剂、木材材性改良、抗逆、抗病、激素调控、开花调控和植物修复等应用领域的研究进展及现状,分析了影响杨树农杆菌转化效率的主要因素,并探讨了杨树转基因研究存在的问题及发展方向,希望能为后期从事林木基因工程研究的科研工作者提供依据。 相似文献
8.
9.
TANGWei LatoyaHarris RonaldJ.Newton 《林业研究》2003,14(3):185-190
Three antibiotics ampicillin, carbenicillin, and cefotaxime were evaluated for their effects on induction, growth, and differentiation of organogenic calli, as well as rooting of regenerated shoots of three Ioblolly pine (Pinus taeda L.) genotypes. Of the antibiotics administered, cefotaxime maximally increased the frequency of callus formation and growth rate of organogeni ccalli, carbenicillin maximally increased the frequency of shoot regeneration and the average number of adventitious shoots per piece of organogenic callus, ampicUlin maximally decreased the rooting frequency of regenerated shoots and mean number of roots per regenerated shoot, in comparison with antibiotic-free media. Compared with the control, ampicillin minimally increased the frequency of callus formation, cefotaxime minimally increased the frequency of shoot regeneration, and carbenicillin minimally decreased the rooting frequency of regenerated shoots in three Ioblolly pine genotypes tested. All three antibiotics increased the frequencies of callus formation and shoot regeneration, and reduced the rooting frequency ot regenerated shoots suggested that the establishment of an efficient Agrobacterium tumefaciens-mediated transformation protocol for stable integration of foreign genes into Ioblolly pine need to select a suitable antibiotic. This investigation could be useful for optimizing genetic transformation of conifers. 相似文献
10.
Katsuaki Ishii 《Journal of Forest Research》2002,7(2):99-104
Hinoki cypress (Chamaecyparis obtusa) is one of the most important timber resource forest trees in Japan. Because seed production from a seed orchard of hinoki
cypress is not constant every year, micropropagation from a limited amount of material is useful. Up to now, the conventional
tissue culture method using solid medium has been used. Here a new method using liquid culture in tubes rotated vertically
is described. Shoot primordium of hinoki cypress was inoculated in Campbell and Durzan’s (CD) liquid medium containing different
cytokinins (6-benzylaminopurine (BAP), Zeatin, thidiazurone (TDZ)), and the container tubes were rotated vertically around
the axis at 2 times / min. Culture room temperature was 25°C and light condition was 16 h photoperiod per day of fluorescent
lamps. Zeatin at 1μM concentration was the best for maintaining the shoot primordium production and TDZ induced callus on the surface of the
shoot primordia. After shoot primordium multiplication in the liquid culture, they were transplanted to agar medium for shoot
elongation. A high concentration of agar (up to 16 g/L) or AVF (anti vitrification factor from Dr. Nairn, 1995) was effective
to prevent vitrification of the shoots. Transformation of shoot primordium was done using particle bombardment with vectors
containingβ-glucuronidase (GUS) gene or herbicide resistance gene (bar). Positive result for transient transformation was observed with the histo-chemical study for transformation with GUS. Integration
of a useful herbicidebar gene into the shoot primordium culture system was also tried and stably transformed plants were obtained. This is the first
report of stable transformation of Japanese conifer using practically useful gene.
The generous supply of AVF-B from Dr. B.J. Nairn, Tasman Forestry, NZ is also appreciated. 相似文献
11.
T. S. Aronen 《Forest Pathology》1997,27(1):55-67
The ability of Agrobacterium species to infect gymnosperms is generally poor, but as a gene transfer vector Agrobacterium would have several advantages. This study characterizes interactions between coniferous defence compounds, a monoterpene α-pinene and a phenolic constituent trans-stilbene, and Agrobacterium tumefaciens. Of the chemical constituents studied, α-pinene in doses equal to the concentrations found in wounded trees prevented the growth of A. tumefaciens. Neither α-pinene nor trans-stilbene inhibited virulence gene induction in A. tumefaciens, but both compounds caused a reduction in the frequency of gall formation in susceptible woody model plants (birch seedlings) when applied after the vir-induction. This effect was smaller with the strain A281(pTVK291) containing extra copies of the virA, virB, virG, and virC genes than with the wild-type strain A28l. The efficiency of Agrobacterium-mediated transformation in a model gymnosperm, Scots pine, could not be enhanced by using the strains containing a constitutive mutant of virG gene or extra copies of virA and virG. The results suggest that the inefficiency of Agrobacterium spp. in infecting many conifers is not the result of insufficient vir-gene induction but rather is the result of interference by chemical defence compounds during the later stages of T-DNA-transformation. 相似文献
12.
[目的]本研究拟对Thb HLH1所识别的顺式作用元件进行鉴定,进一步揭示Thb HLH1调控抗逆基因表达的机理。[方法]利用以转录因子为中心的酵母单杂交系统鉴定Thb HLH1所识别的顺式作用元件;将元件与报告基因融合构建报告载体(p CAM-Cis),通过基因枪法将报告载体与效应载体35S∶Thb HLH1共转化烟草叶片,在盐、干旱胁迫下比较GUS酶活。[结果]鉴定出2段能够与Thb HLH1转录因子结合的DNA序列:分别为CCGAAA(LTRE1)和TGAC(WRKY710S)。[结论]在盐或干旱胁迫下,Thb HLH1通过与LTRE1或WRKY710S元件作用来激活基因表达。 相似文献
13.
本文建立了一个微弹介导的火炬松遗传转化系统。这个系统解决了火炬松遗传转化过程中存在的许多困难。运载抗虫基因的质粒载体经微弹转化法进入火炬松成熟合子胚,然后在添加了卡那霉素的培养基上从转化的成熟胚上诱导出有器官发生潜力的愈伤组织,再从转化的愈伤组织上产生转基因植株。利用这一系统生产的转基因植株已经被随机扩增技术、Southern杂交技术和虫试验所证实,并且转化的植株已在土壤中成活。图3表2参28。 相似文献
14.
表达耐盐基因的转基因火炬松的再生 总被引：2，自引：0，他引：2
盐害是限制作物和树木分布和生产的重要因素。盐分过多导致细胞内水分缺失并影响许多重要的细胞代谢活动。本文利用火炬松作为模式植物建立了一套提高植物耐盐性的新技术。这一技术以火炬松合子胚为材料,利用农杆菌介导的转化方法将山犁醇脱氢酶和甘露醇脱氢酶基因转入火炬松。然后再生转化的愈伤组织和转基因植株。经DNA杂交证实的转基因植株被用于耐盐性试验,结果表明这些转基因的植株的耐盐性有明显的提高。这一技术对针叶树的遗传工程育种有重要的参考价值。图3表2参26。 相似文献
15.
Xu Wen-li Liu Mei-qin Shen Xin Lu Cun-fu 《中国林学(英文版)》2005,7(4):11-15
Antifreeze proteins （AFPs） enable organisms to survive under cold conditions, and have great potential in improving cold tolerance of cold-sensitive plants, In order to determine whether expression of the carrot 36 kD antifreeze protein gene confers improved cold-resistant properties to plant tissues, we tried to obtain transgenic tobacco plants which expressed the antifreeze protein. Cold, salt, and drought induced promoter Prd29A was cloned using PCR from Arabidopsis. Two plant expression vectors based on pBI121 were constructed with CaMV35S：AFP and Prd29A：AFP. Tobacco plantlets were transformed by Agrobacterium-medicated transformation. PCR and Southern blotting demonstrated that the carrot 36 kD afp gene was successfully integrated into the genomes of transformed plantlets. The expression of the afp gene in transgenic plants led to improved tolerance to cold stress. However, the use of the strong constitutive 35S cauliflower mosaic virus （CaMV） promoter to drive expression of afp also resulted in growth retardation under normal growing conditions. In contrast, the expression of afp driven by the stress-inducible Prd29A promoter from Arabidopsis gave rise to minimal effects on plant growth while providing an increased tolerance to cold stress condition （2℃）. The results demonstrated the prospect of using Prd29A-AFP transgenic plants in cold-stressed conditions that will in turn benefit agriculture. 相似文献
16.
We used in vitro callus and shoot cultures as target material for genetic transformation of silver birch (Betula pendula Roth) by particle bombardment. Cultivation of in vitro shoot cultures before particle bombardment and a long selection period, combined with a high concentration of selective agent after bombardment, led to the production of transformed plantlets that were stable, and no escapes were found among the tree lines produced. Clonal variation in transformation efficiency was found in transient expression of the beta-glucuronidase gene in callus cultures and in plantlets transformed by stable integration of the ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (RbcS) and neomycin phosphotransferase (npt2) genes. 相似文献
17.
18.
Kasim Bajrovic Kemal Kazan Zeliha İpekçi Nermin Gözükırmızı 《Journal of Forest Research》1999,4(2):161-166
Poplar (Populus tremula) was transformed with a construct carrying an antisense caffeic acidO-methyltransferase (COMT) cDNA (pOMT8) from a tropical pasture legume,Stylosanthes humilis. pOMT8 shows 83% overall homology to the corresponding COMT gene (pPCLA) of poplar. Of the 200 putatively-transformed plants regenerated on selective media after co-cultivation of poplar stem explants
withAgrobacterium tumefaciens harbouring a CaMV 35S-antisensepOMT8 construct, a subset of 20 plants were randomly chosen for further analysis. PCR and Southern blot analysis demonstrated the
stable integration of T-DNA into the genome of these plants. Antisense expression ofpOMT8 resulted in reductions in total COMT activity in the majority of the transgenic plants with the lowest total COMT activities
(61–70% of untransformed control plants) being observed in four transgenic plants. The composition of lignin in transgenic
plants was also changed, as detected by reductions in the content of syringyl units using infrared spectroscopy. However,
no changes were found in the amount of insoluble lignin in transgenic plants as compared to untransformed control plants.
These results indicate the potential of thepOMT8 gene to partially suppress COMT activity and modify the composition of lignin in transgenic poplar.
This work was partly supported by General Management of Turkish Pulp and Paper Mills. 相似文献
19.
[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
20.
A transformation system was established for loblolly pine (Pinus taeda L.) mature zygotic embryos usingAgrobacterium tumefaciens. The gene coding for the β-glucuronidase (GUS) gene was introduced into loblolly pine tissues and its transient expression
was detected with histochemical staining. The influences of different genotypes.Agrobacterum concentrations. and cocultivation time on GUS expression and kanamycin resistant callus and shoot regeneration were investigated.
The results showed that the highest GUS expression frequency (16.3%) and shoot regeneration frequency (78%) were obtained
from genotype 9–1003 withAgrobacterium concentration decreased 9 times and cocultivation time of 56 hours respectively. GUS expression was obtained in all genotypes
tested. The successful expression of the GUS gene in different genotypes suggested that it will be a useful transformation
system for loblolly pine.
(Responsible Editor: Chai Ruihai) 相似文献