香蕉基因工程研究进展

香蕉是世界上一种重要的作物.基因工程技术的发展为培育具有优良性状的香蕉品种提供了有效的手段.本文综述了近年来香蕉基因工程研究的最新进展,包括香蕉遗传转化受体系统的建立,香蕉作物中一些有益功能基因的克隆以及香蕉遗传转化的最新科研成果.并对各种香蕉遗传转化受体系统和目前主要的香蕉遗传转化进行了客观评价,提出了香蕉遗传转化研究中存在的问题,最后对通过基因工程手段改良香蕉品种进行了展望.     

人参的遗传改良*

遗传改良是人参育种的重要手段之一,而遗传转化和再生体系的建立是开展人参遗传改良工作的前提和基础。人参植株再生可以通过器官发生和体细胞胚发生,间接体细胞胚发生是人参植株再生的主要途径,从不同外植体,不同碳源,体细胞胚优化和无激素再生等方面进行了综述。在人参遗传转化方面,发根农杆菌和根癌农杆菌对人参的遗传转化均已成功,人参皂苷合成途径中的关键酶基因和抗除草剂基因也已陆续导入人参,得到了遗传改良的转化人参。发根培养系统可用于大量生产人参皂苷,讨论了rolC基因对人参发根诱导的作用,发根植株再生能力及生物反应器培养,最后指出了人参基因工程研究中存在的问题。     

葡萄遗传转化研究进展

本文从目的基因、遗传转化途径、再生系统以及筛选和鉴定四个方面评述了近10年来葡萄遗传转化的研究进展。与其它植物相比,葡萄遗传转化较困难,迄今为止仅获得了较少的转基因植株。葡萄再生频率低和葡萄组织对卡那霉素选择的敏感均是遗传转化困难的关键所在。     

葡萄遗传转化研究进展

本文从目的基因、遗传转化途径、再生系统以及筛选和鉴定四个方面评述了近１０年来葡萄遗传转化的研究进展,与其它植物相比,葡萄遗传转化较困难,迄今为止仅获得了较少的转基因植株,葡萄再生频率低和葡萄组织对卡那霉素选择的敏感均是遗传转化因素的关键所在。     

月季组织培养和遗传转化体系的研究进展

月季通过器官和体细胞胚发生途径都可以获得再生植株,在遗传转化中主要是利用体细胞胚作为转化受体。目前,利用农杆菌介导法和基因枪法已成功将外源基因如报告基因、抗病基因和改变花色的基因等导入月季基因组中。本文对近年来月季组织培养和转基因研究进展进行了综述,为建立月季高效遗传转化体系奠定了理论基础。     

84K杨愈伤组织再生体系和直接分化再生体系遗传转化的比较性研究

植物转基因必须以建立良好的植物转化受体系统为基础,而良好的受体系统必须以具有高频率的转化率及较强的再生能力为前提。实验以84K杨叶片为材料研究其在不同培养条件、不同种类及浓度的植物激素对其脱分化及再分化的影响,并建立了两种成熟的组织再生系统,得出组织培养体系与遗传转化体系的受体系统在激素配比方面存在一定差距,所以在此基础上进行了PCK1、PCK2两个基因的愈伤途径和不定芽途径途径的遗传转化,并对两种遗传转化体系进行了对比性分析并总结出两种体系的优缺点。     

大豆转基因体系的研究进展

从大豆的转基因方法和受体系统两个方面概述大豆转基因体系的最新研究进展,并讨论了大豆遗传转化的主要障碍及可能的解决途径。作者认为,以根癌农杆菌介导大豆子叶节和基因枪轰击大豆未成熟子叶是较有效的大豆遗传化系统。目前,在大豆遗传转化研究中存在着大豆组织培养体系有待于进一步完善、遗传转化率较低、重复性差、大豆受体基因型单一等问题,建立新的、高效和稳定的大豆组织培养体系,提高生产上栽培大豆品种的组织培养能力,改遗传转化现有的单一基因为多基因,可能是解决上述问题的有效途径。     

百合转基因的研究进展

综述了近年来国内外关于百合遗传转化系统的建立及其基因转化研究技术。着重讨论了百合基因转化受体系统建立的诱导培养基和抗生素浓度的筛选、遗传转化方法及基因调控表达等的研究动态。     

花生基因工程

建立花生遗传转化和高效植株再生系统是花生基因工再生技术和基因转化体系的完善,已获得抗病、抗虫和提高品质的转基因花生植株,取得了突破性进展.农杆菌介导法和微弹介导法是花生基因工程的主要方法.     

藻类基因工程的研究技术及方法

本文从藻类外源基因转移的载体系统、藻类基因的克隆、藻类的遗传转化三方面详细介绍了藻类基因工程的研究技术及方法,综述了藻类基因工程的研究进展,并对藻类基因工程的前景作了展望。     

Genetic transformation technology: Status and problems

Summary Transfer of genes from heterologous species provides the means of selectively introducing new traits into crop plants and expanding the gene pool beyond what has been available to traditional breeding systems. With the recent advances in genetic engineering of plants, it is now feasible to introduce into crop plants, genes that have previously been inaccessible to the conventional plant breeder, or which did not exist in the crop of interest. This holds a tremendous potential for the genetic enhancement of important food crops. However, the availability of efficient transformation methods to introduce foreign DNA can be a substantial barrier to the application of recombinant DNA methods in some crop plants. Despite significant advances over the past decades, development of efficient transformation methods can take many years of painstaking research. The major components for the development of transgenic plants include the development of reliable tissue culture regeneration systems, preparation of gene constructs and efficient transformation techniques for the introduction of genes into the crop plants, recovery and multiplication of transgenic plants, molecular and genetic characterization of transgenic plants for stable and efficient gene expression, transfer of genes to elite cultivars by conventional breeding methods if required, and the evaluation of transgenic plants for their effectiveness in alleviating the biotic and abiotic stresses without being an environmental biohazard. Amongst these, protocols for the introduction of genes, including the efficient regeneration of shoots in tissue cultures, and transformation methods can be major bottlenecks to the application of genetic transformation technology. Some of the key constraints in transformation procedures and possible solutions for safe development and deployment of transgenic plants for crop improvement are discussed.     

Efficient regeneration and <Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated stable transformation of perennial ryegrass

We utilized gene transfer technology for genetic perennial ryegrass improvement, efficient regeneration, and Agrobacterium-mediated transformation of phosphinothricin acetyltransferase gene (bar). Four growth regulator combinations were compared and intact seeds of six turf-type cultivars as mature embryo sources were tested to optimize the regeneration conditions. Callus formation and regeneration were observed in all seeds. The highest callus formation frequency was observed in the seeds cultured on MS medium supplemented with 9 mg/l 2,4-D, without benzyladenine. Cv. TopGun revealed the highest callus induction and regeneration frequencies of 96 and 48.9%, respectively. By using an optimized regeneration system, embryogenic calli were transformed by an Agrobacterium strain LBA4404 containing the plasmid pCAMBIA3301. After the selection of the potentially transgenic calli with phosphinothricin, a herbicide, 22 transgenic resistant plants were regenerated. With PCR, Southern-blot hybridizations, and GUS expression techniques, we confirmed that some regenerants were transgenic. Two of the tested transgenic plants showed herbicide resistance. Our results indicated that embryogenic calli from mature seeds can be directly used for perennial ryegrass efficient regeneration and transformation and this protocol is applicable for genetic engineering of herbicide-resistant plants. Published in Russian in Fiziologiya Rastenii, 2007, Vol. 54, No. 4, pp. 590–596. The text was submitted by the authors in English.     

Transgenic oil palm: production and projection

Oil palm is an important economic crop for Malaysia. Genetic engineering could be applied to produce transgenic oil palms with high value-added fatty acids and novel products to ensure the sustainability of the palm oil industry. Establishment of a reliable transformation and regeneration system is essential for genetic engineering. Biolistic was initially chosen as the method for oil palm transformation as it has been the most successful method for monocotyledons to date. Optimization of physical and biological parameters, including testing of promoters and selective agents, was carried out as a prerequisite for stable transformation. This has resulted in the successful transfer of reporter genes into oil palm and the regeneration of transgenic oil palm, thus making it possible to improve the oil palm through genetic engineering. Besides application of the Biolistics method, studies on transformation mediated by Agrobacterium and utilization of the green fluorescent protein gene as a selectable marker gene have been initiated. Upon the development of a reliable transformation system, a number of useful targets are being projected for oil palm improvement. Among these targets are high-oleate and high-stearate oils, and the production of industrial feedstock such as biodegradable plastics. The efforts in oil palm genetic engineering are thus not targeted as commodity palm oil. Due to the long life cycle of the palm and the time taken to regenerate plants in tissue culture, it is envisaged that commercial planting of transgenic palms will not occur any earlier than the year 2020.     

叶绿体基因工程：一种植物生物技术的新方法

以叶绿体转化为主的叶绿体基因工程,与传统的基因工程技术细胞核转化相比,在外源基因表达水平和转基因植物安全性等方面有明显的优势, 尤其是在控制转基因沉默和遗传稳定性方面,可以互补核转化带来的局限性。因此,叶绿体基因工程是一种很具有发展前景的植物转基因技术,并在未来工农业生物技术领域发挥重要作用。本文着重在叶绿体转化技术主要特点,应用领域及其未来的发展前景等方面进行了简单评述。     

转苏云金杆菌毒蛋白基因玉米植株的获得及其抗虫性分析

玉米( Zea mays L.)转化成功与否与基因型密切相关.在转化过程中,除少数模式品种能够形成再生频率较高且易转化的Ⅱ型愈伤组织外,大多数栽培品种往往只能够形成再生频率较低且不易转化的Ⅰ型愈伤组织.因此探索Ⅰ型愈伤组织的诱导及其转化条件,提高转化效率,对直接改良玉米优良自交系具有重要意义.应用基因枪转化技术将苏云金杆菌( Bacillus thuringiensis ) cry1Ac3基因导入玉米优良自交系E28及340的Ⅰ型胚性愈伤组织中,经过膦丝菌素(PPT)或潮霉素(HygB)筛选,获得了再生植株.经PCR检测、Southern blot分析及Bt毒蛋白ELISA检测证实,外源基因已整合到玉米基因组中,并已获得表达.抗虫性分析结果表明,部分转基因玉米植株对玉米螟虫有较强的抗性.还比较了PPT和HygB两种筛选剂的筛选效果,表明PPT筛选的抗性愈伤组织的再生频率要高于HygB筛选的再生频率.     

质体基因工程中选择标记基因研究进展

与细胞核基因工程相比,质体基因工程能更安全、精确和高效地对外源基因进行表达,作为下一代转基因技术已广泛用于基础研究和生物技术应用领域。与细胞核基因工程一样,质体基因工程中也需要合适的选择标记基因用于转化子的筛选和同质化,但基于质体基因组的多拷贝性和母系遗传特点,转化子的同质化需要一个长期的筛选过程,这就决定了质体基因工程中选择标记基因的选择标准将不同于细胞核基因工程中广泛使用的现行标准。目前,质体基因工程的遗传转化操作中使用较多的是抗生素选择标记基因,出于安全性考虑,需要找到可替换、安全的选择标记基因或有效的标记基因删除方法。本文在对质体基因工程研究的相关文献分析基础之上,对主要使用的选择标记基因及其删除体系进行了综述,并对比了其优缺点,同时探讨了质体基因工程中所使用的报告基因,以期为现有选择标记基因及其删除体系的改进和开发提供一定参考,进一步推动质体基因工程,尤其是单子叶植物质体基因工程的发展。     

大白菜生物技术研究进展

综述了国内外在大白菜(Brassica rapa ssp.pekinensis)单倍体培养、基因工程和分子标记方面的最新研究成果.重点讨论了大白菜小孢子培养的影响因素、取材要点、培养条件、植株再生和倍性鉴定以及大白菜的转基因载体选择、受体系统建立、遗传转化与转基因植株鉴定等.对小孢子培养及转基因研究的原理、方法和关键技术进行了总结.展望了大白菜生物技术研究的应用前景.     

大白菜生物技术研究进展

综述了国内外在大白菜(Brassica rapa ssp. pekinensis)单倍体培养、基因工程和分子标记方面的最新研究成果。重点讨论了大白菜小孢子培养的影响因素、取材要点、培养条件、植株再生和倍性鉴定以及大白菜的转基因载体选择、受体系统建立、遗传转化与转基因植株鉴定等。对小孢子培养及转基因研究的原理、方法和关键技术进行了总结。展望了大白菜生物技术研究的应用前景。     

水仙花生物技术研究进展(综述)

综述近年来水仙花生物技术的研究进展,包括水仙花离体植株再生、相关基因克隆与遗传转化及其在快繁、脱毒、遗传改良、种质资源保存以及次生代谢物质生产等方面的应用,并探讨该领域存在的问题。