用in planta方法转化甘蓝型油菜

用inplanta方法进行遗传转化极大地促进了模式植物拟南芥的功能基因组研究 ,但在农作物中仍少有inplanta的成功报道。我们采用inplanta方法 ,用含有质粒pSG5 2 9、pHTT6 1 3和ApSCO的农杆菌菌株转化中油 82 1、湘油 1 3、宁RS 1、S2 5 0 1 1、华双 3号等甘蓝型油菜品种。在适宜的条件下 ,将根癌农杆菌菌液浸染甘蓝型油菜花序 1 0d至 1 5d ,每天浸染 1至 2次 ,共收获约 1 0 0 0 0 0粒受处理的种子。将种子播撒在含有 2 70mg·L- 1 卡那霉素的选择培养基中选择转化植株。卡那霉素抗性植株经PCR分析检测 ,结果在 5个供试品种中均获得了转基因植株 ,平均转化效率 0 1 %左右 ,各甘蓝型油菜品种间的转化效率无明显差异。这种方法方便、快速且无须经过组织培养阶段即可获得上百株转基因植株。

农杆菌介导的油菜遗传转化研究进展

综述了农杆菌介导的油菜等芸薹属植物高效转化体系建立的影响因素,总结了基因工程在改良油 菜等芸薹属植物性状中的最新研究进展,评述了植物in Planta转化法在油菜中的应用。

Embryos - the evolutionary boundary of kingdoms of organisms

Bioevolution is still a main puzzle and attracts many scientists to research on it. Here I present that organisms have two impor- tant properties, definite structure and self-reproduction. Based on the number and connection of the structural units, organisms can be di- vided into three groups, unicellular, particellular and polycellular organisms. It can be called polycellular evolution that organisms evolve from unicellular, particellular to polycellular. Also it can be called diploid evolution that organisms evolve from haplobes to diplobes, two groups based on the reproductive differences. Some concepts like spore, zygote and embryos are redefined in the paper. Moreover, I present that embryos are the most important boundary of bioevolution and organisms can be divided into two evolutionary phases, the lower and the higher. The lower organisms, Kingdom Microbia (kingd. nov.), are inembryonate, which include Acytophyla (phyl. nov.), bacteria, protozoa, fungi and inembryonate algae. The higher organisms are embryonate and have two branches, Kingdom Plantae and Kingdom Animalia. Plantae are sessile and, autotrophic or sporogenic, which include higher plants and Nudembryophyta (phyl. nov.). Animalia are heterotrophic and, motile or gametogenic, which include all multicellular animals. The new system, which reflects the two important phases of bioevolu- tion and two branches of higher organisms, can really correct the problem of different kingdoms in different researches or by different re- searchers.

In planta Agrobacterium-Mediated Transformation of Rice

The floral-dip transformation, the simplest technique, is no requirement of tissue culture procedure, and can directly transfer the interest gene into plant reproductive cells. It has been successfully applied to various plant species. In this study, the optimal conditions of a floral-dip method for production of transgenic rice variety RD41 were explored. The simple and effective inoculation medium was composed of Murashige and Skoog(MS) medium, 5% sucrose, 44 nmol/L benzylaminopurine, and 0.075% surfactant Tween-20 with pH 5.7. The transformation efficiencies of Agrobacterium tumefaciens strains AGL1 and EHA105 were compared with the Agrobacterium density at OD_(600) = 0.8–1.0 and the co-cultivation at 25 ℃ for 48 h. A. tumefaciens strain EHA105 gave slightly higher transformation efficiency than AGL1, with statistically non-significant difference. The floral-drop transformation using the optimal floral-dip conditions showed higher transformation efficiency than the floral-dip method, but the dropped flowers turned brown and died within 2 d. Production of transgenic rice variety RD41 by the floral-dip method was achieved using A. tumefaciens strain EHA105 with the optimal conditions. Screening for the gus A gene by PCR using the gus A specific primers in the T_0 lines, there were 4 transgenic lines from 286 T_0 lines(1.4% transformation efficiency). However, histochemical glucuronidase(GUS) assay demonstrated that only three of four transgenic lines exhibited gus A expression. These results indicated that floral-dip transformation is a potential tool for production of the transgenic rice, which can be used for molecular breeding via genetic engineering in the future.

in planta法在亚麻转基因中的应用

为了建立简单快速、高效稳定的亚麻遗传转化体系,本试验用亚麻的种子胚作为外植体,采用原位转化(in planta)法对亚麻转基因的试验条件(预培养时间、共培养时间、As浓度、silwett浓度和抽真空时间)进行了系统优化。结果确定最佳组合为:预培养时间8 h,共培养时间72 h,As浓度100μmol/L,silwett浓度0.1%,抽真空时间5 min。摸索适宜的草丁膦最低致死剂量用于亚麻转化植株的抗药性筛选,并对存活的植株进行PCR鉴定,成功获得转基因阳性植株。本研究实现了in planta技术在亚麻种子胚的转基因体系优化,有较大的实际应用价值。

Transformation of Chinese Cabbage(Brassica rapa L. ssp. pekinensis)by Agrobacterium Micro-Injection into Flower Bud

We obtained two lines of Chinese head cabbage(Brassica rapa L. ssp. pekinensis)selfed progenies containing both an anti-sense gene of BcpLH and a gene for resistance to kanamycin by micro-injecting buds of their primary transformants(T0)with Agrobacterium tumefaciens strain LBA4404. 31 positive plants resistant to kanamycien were recovered. Southern blot analysis confirmed the presence of T-DNA in two transgenic plants. One(DHZ-13-1)exhibits the characteristics of out-toward rosette and cauline leaves, and nested flower model in which secondary complete flower developed from the base of the primary ovary and the third flower from the ovary in the secondary flower, and so on, while another(DHZ-6-1)has no phenotype change. ABA and IAA affected the root growth of progeny of DHZ-13-1, but 6-BA was insensitive to hypocotyl growth during its seedling development.

Rapid and convenient transformation of cotton (Gossypium hirsutum L.) using in planta shoot apex via glyphosate selection

Cotton plants are recalcitrant with regards to transformation and induced regeneration.In the present study,5-enolpyruvylshikimate-3-phosphate（EPSPS）,a glyphosate resistant gene from the bacterium Agrobacterium sp.strain CP4,was introduced into an elite Bt transgenic cotton cultivar with a modified technique involving in planta Agrobacteriummediated transformation of shoot apex.Primary transformants were initially screened using a 0.26%glyphosate spray and subsequently by PCR analysis.Five out of 4 000 transformants from T_1 seeds were obtained resulting in an in planta transformation rate of 0.125%.Four homozygous lines were produced by continuous self-fertilization and both PCR-based selection and glyphosate resistance.Transgene insertion was analyzed by Southern blot analysis.Gene transcription and protein expression levels in the transgenic cotton lines were further investigated by RT-PCR,Western blot,and ELISA methods.Transgenic T_3 plants were resistant to as much as 0.4% of glyphosate treatments in field trials.Our results indicate that the cotton shoot apex transformation technique which is both tissue-culture and genotype-independent would enable the exploitation of transgene technology in different cotton cultivars.Since this method does not require sterile conditions,the use of specialized growth media or the application of plant hormones,it can be conducted under the greenhouse condition.

Downregulation of the DST Transcription Factor Using Artificial microRNA to Increase Yield, Salt and Drought Tolerance in Rice

Abiotic stresses like salinity and drought directly affect plant growth and water availability, resulting in lower yield in rice. So, a combination of stress tolerance along with enhanced grain yield is a major focus of rice breeding. It was reported earlier that loss in function of the drought and salt tolerance (DST) gene results in increase in grain production through downregulating Gn1a/OsCKX2 expression. Moreover, dst mutants also showed enhanced drought and salt tolerance in rice by regulating genes involved in ROS homeostasis. In the present study, we proceeded to test these reports by downregulating DST using artificial microRNA technology in the commercial but salt sensitive, high-yielding, BRRIdhan 28 (BR28). This cultivar was transformed with DST_artificial microRNA (DST_amiRNA) driven by the constitutive CaMV35S promoter using tissue culture independent Agrobacterium mediated in planta transformation. DST_amiRNA transgenic plants were confirmed by artificial microRNA specific PCR. Transformed plants at T0 generation showed vigorous growth with significantly longer panicle length and higher primary branching resulting in higher yield, compared to the wild type (WT) BR28. Semi-quantitative RT PCR confirmed the decrease in DST expression in the BR28 transgenic plants compared to WT. T1 transgenic plants also showed improvement in a number of physiological parameters and greater growth compared to WT after 14 days of 120 mM salt (NaCl) stress at seedling stage. Therefore, DST downregulated transgenic plants showed both higher stress tolerance as well as better yields. Furthermore, stable inheritance of the improved phenotype of the DST_amiRNA transgenics will be tested in advanced generations.

近期Planta Medica高被引文章题录(英文)

Planta Medicais one of the leading international journals in the field of medicinal plants and natural products with original research papers,letters,rapid communications,reviews,minireviews and perspectives from researchers worldwide.Planta Medica publishes15issues per year.

Plant as Bioreactor for Producing Heterologous Proteins

Using plants to produce heterologous proteins makes it very attractive due to the potentially low costs. Using this procedure it is possible to produce medicinal protein for clinical applications with the plants bioreactors increasing gradually. The paper proposes the five major systems of the plant bioreactor as well as their advantage and disadvantage and the development of each system. Focuses on the five major systems of the plant bioreactor to produce vaccines, antibodies and medical protein and the research achievement at the present stage and the research on my laboratory. The key technology research of plant bioreactor such as new genes, new biological components, new technologies and new research methods related with plant bioreactor offer a work foundation for a long-term development in future.

Repairing small wounds around ankle by medial planta island flaps pedicled with anterior tibial artery perforator in front of inner malleolus

Objective To discuss the application of medial planta island flaps pedicled with anterior tibial artery perforator in front of inner malleolus for repairing small wounds around ankle Methods From Jan. 2005 to Jun. 2009,10 cases with small wounds around ankle

水稻白叶枯病菌在寄主体内外不同生长条件下致病基因差异表达的分析

To demonstrate the expression profiling of Xanthomonas oryzae pv.oryzae(Xoo) in vitro and in planta,DNA microarrays of 371 genes potentially associated with pathogenicity and virulence were used to compare the transcriptional level alteration of the wild-type strain PXO99A and gene deletion mutants ΔgacAxoo and ΔfleQxoo of Xoo grown in the rich medium NBY vs.hrp-inducing minimal medium XOM2 or leaf tissues of rice.Results indicated that 17 and 38 genes of PXO99A were differentially expressed in XOM2 and the leaf tissues of rice relative to NBY,respectively.Twenty-eight genes of ΔgacAxoo grown in XOM2 and 12 genes of ΔfleQxoo in NBY were differentially expressed relative to PXO99A.The identification of differentially-expressed genes,GacAxoo-and FleQxoo-regulons and novel candidate genes of Xoo strains would provide us the target genes for further functional analysis in pathogenesis of Xoo.

足底恶性黑色素瘤的整形外科治疗

目的探讨手术治疗足底恶性黑色素瘤的技术及效果。方法应用病灶切除,创面植皮或皮瓣修复的方法。结果5例患者,术后皮片、皮瓣全部成活,随访3个月～4年,无1例复发,患足功能正常。结论对足底恶性黑色素瘤的治疗,应用皮瓣修复可取得满意的临床效果。

Science Letters:Optimization of soybean (Glycine max (L.) Merrill) in planta ovary transformation using a linear minimal gus gene cassette

Soybean transformation by ovary-drip was improved by optimizing the length of the transformation pathway by cutting the styles. These modifications facilitated soybean transformation manipulation and improved transformation reproducibility and efficiency. Using a linear minimal gus gene cassette as the foreign DNA, a maximum transformation frequency of 11% was obtained in flowers of the soybean cultivar ‘Liaodou 14’ with their styles mostly removed, whereas removal of only the stigma, partial style cutting and partial ovary cutting gave transformation frequencies of 0%, 1%, and 2%, respectively. An average transformation frequency of 8.2% was obtained when 619 flowers from three soybean cultivars (‘Liaodou 14’, ‘Liaodou 13’, and ‘Tiefeng 29’) were transformed by this optimized method. Southern blotting analysis showed that the gus reporter gene (encoding β-glucuronidase) was stably inherited with a simple pattern. Reverse transcription-polymerase chain reaction (RT-PCR) and GUS staining confirmed the expression of the gus gene in transgenic plants.

La Rosa de Jericó:la planta de la resurrección

Es considerada sagrada y mágica por sus peculiares propiedades.LA Rosa de Jericó(Anastatica hierochuntica)es llamada oficialmente en China como juanbai,que en espanol quiere decir “hoja de ciprés arrollada”,pues sus hojas parecen las de ese árbol y también haber sido arrolladas.