香石竹植株再生及基因工程研究进展

本文对香石竹的再生体系、遗传转化体系及其分子育种现状作了较为系统的总结。香石竹的再生体系多以器官直接再生不定芽为主, 而通过愈伤组织和体细胞胚途径再生报道较少。用农杆菌介导法和基因枪法均可建立香石竹遗传转化体系, 但近年来的研究显示农杆菌介导法应用普遍且比较稳定。近年来以延长香石竹瓶插寿命为目标的分子育种研究已取得较大进展, 对其色、香和形等其他重要性状的分子育种也已经起步, 而有关香石竹抗性的分子育种有待进一步开拓。     

农杆菌介导的玉米遗传转化研究进展

农杆菌介导的转基因法是目前玉米遗传转化的主流方法之一。目前,模式玉米种质幼胚的转化体系已程式化,且开发了新筛选基因和获得不含筛选基因转基因玉米的方法,但是大多数育种骨干自交系转化频率低和转化受体基本上是幼胚。从农杆菌、受体及培养条件多方面各种因素对问题进行分析,多数研究认为针对特定基因型和受体材料建立好的受体再生系统,结合高效率农杆菌转化体系,获得多目的基因聚合(无其它外源片段)的转基因玉米将是农杆菌介导玉米转化体系研究的最终目标。本文主要从农杆菌介导(转基因)法应用于玉米遗传转化的历史、现状、问题等方面进行综述,为同领域的研究者提供一定的参考。     

农杆菌介导的葡萄转化研究进展

综述近年来以农杆菌介导法将外源基因转化葡萄的研究进展及基因型、再生途径、农杆菌菌株、菌液浓度、侵染时间、共培养及抗生素等对葡萄遗传转化的影响,并指出当前存在的问题及研究趋势。     

根癌农杆菌介导的黄瓜转基因研究进展

黄瓜Cucumis sativus是世界性的重要蔬菜作物。农杆菌介导的转基因技术是研究植物基因功能及品种改良的重要手段。为进一步加快黄瓜的转基因研究和育种进程,文中针对农杆菌介导的黄瓜遗传转化方法,从黄瓜再生能力的影响因素、遗传转化条件和过程中各类添加物质等方面,阐述了根癌农杆菌介导的黄瓜转基因研究进展及存在的问题,并对提高黄瓜遗传转化效率和安全筛选标记的应用等前景进行了展望,以期为黄瓜抗逆育种和果实品质改良等研究提供参考。     

能源植物麻疯树分子育种研究进展

麻疯树作为一种重要的能源植物,虽然在常规育种方面的研究比较多,但在分子育种上仍处于起步阶段。在详细综述了国内外能源植物麻疯树基因的克隆,组织培养再生体系的建立相关研究进展的基础上,重点介绍了农杆菌介导转化法、基因枪介导转化法和纳米载体转化法等几种新技术在麻疯树育种中的应用,提出了当前麻疯树分子育种中存在的问题,并对其前景进行了展望。     

苹果遗传转化的研究进展

苹果的遗传转化技术通过分子手段改良苹果,有助于缩短其育种周期。最近十年,在该领域的研究取得很大进展,涉及到一些重要的苹果基因型及有用的外源基因。迄今,苹果的遗传转化主要采用农杆菌介导法,侵染与转化材料的再生是影响其转化效率的关键过程,了解其影响因素,寻找有利因素以提高转化效率是目前苹果遗传转化研究的重点。     

甘蓝型油菜的遗传转化

甘蓝型油菜的各种外植体经遗传转化、组织培养后可以再生为转基因植株,但再生频率会因外植体的基因型、年龄、培养基添加成分和农杆菌共培养的不同而发生变化。转化方法包括农杆菌介导转化、基因枪法、花粉介导法、PEG介导法等,其应用前景非常广阔。甘蓝型油菜的遗传转化在其品质改良、抗逆性提高、雄性不育系的获得和一些特殊性状方面都取得了很大成就。简要介绍甘蓝型油菜的再生体系建立、转化方法及所取得的部分成就。     

根癌农杆菌介导的草莓遗传转化研究进展

随着植物离体再生技术的日益成熟和多种外源目的基因的分离与克隆,转基因技术以目的性强、周期短等优点成为草莓品种改良的重要途径。在草莓上采用的遗传转化方法主要是农杆菌介导法。概述了农杆菌介导法的转化机理以及基因型、农杆菌菌株、侵染时间和菌液浓度、共培养时间、酚类物质等对草莓遗传转化的影响,并从草莓果实贮藏保鲜、抗除草剂、抗病毒、抗虫、抗真菌、转基因疫苗等方面对近年来草莓的转基因研究进展进行了总结。     

红颜草莓叶盘再生及遗传转化体系的优化

草莓离体组织再生和遗传转化体系的研究对草莓分子育种至关重要。为了优化农杆菌介导草莓品种红颜(Fragaria×ananassa Duch.Benihoppe)的遗传转化体系,本研究以红颜草莓组培苗的离体叶盘作为试验材料,首先确定了其离体再生的最佳条件,对影响转化效率的因素进行优化,获得了稳定高效的再生及转化体系。结果表明,红颜草莓离体叶盘最佳暗培养时间为9 d,最佳愈伤诱导培养基为MS+TDZ(2. 0 mg/L)+IBA(0. 1 mg/L)+2,4-D(0. 1 mg/L),最佳愈伤分化培养基为MS+6-BA(0. 5 mg/L)+NAA(0. 1 mg/L),转化体系的最佳组合为预培养2 d、添加300μmol/L的AS、以OD600nm=0. 6的EHA105型的农杆菌浸染5 min、共培养5 d。经优化后的红颜草莓叶盘再生及遗传转化体系对该品种的进一步优化和选育有重要意义。     

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.     

Millets genetic engineering: the progress made and prospects for the future

Millets comprise a highly variable small-seeded group of Poaceae members that can grow in extreme environmental conditions of drought, high temperature and low soil fertility hence, recognized as climate-resilient. Among millets, the phylogenetic closeness of Setaria with other agronomically important grasses like maize, sugarcane, and sorghum helped in its adoption as a translational model plant. Established efficient gene transfer methodology is a prerequisite for embracing plant species as models. However, genetic engineering of some of the economically important millets has been started in the 1990s, but inadequate progress made this group lag behind other members of Poaceae as rice, maize and wheat. Genetic transformation in millets has generally been achieved by a physical method of microprojectile bombardment, recently Agrobacterium-mediated gene transfer technique has also established in some of the millets but with very few reports. The central hindrance in millet transformation is its recalcitrant nature to regeneration through tissue culture techniques. Optimization of highly efficient regeneration procedure for each millet species is thus, necessary to establish advanced transformation system for them. The possibility of alternative transformation approaches is also discussed. The establishment of robust gene transfer methods whether it’s conventional in-vitro tissue culture dependent or in-planta are important for functional validation studies and would enable development of crop improvement strategies. This review presents the progress made on millet genetic transformation, discussing the major challenges that need to be overcome and future opportunities of transgenic techniques in various millets.

     

Phaseolus vulgaris — Recalcitrant potential

Since the ability to genetically engineer plants was established, researchers have modified a great number of plant species to satisfy agricultural, horticultural, industrial, medicinal or veterinary requirements. Almost thirty years after the first approaches to the genetic modification of pulse crops, it is possible to transform many grain legumes. However, one of the most important species for human nutrition, Phaseolus vulgaris, still lacks some practical tools for genomic research, such as routine genetic transformation. Its recalcitrance towards in vitro regeneration and rooting significantly hampers the possibilities of improvement of the common bean that suffers from many biotic and abiotic constraints. Thus, an efficient and reproducible system for regeneration of a whole plant is desired. Although noticeable progress has been made, the rate of recovery of transgenic lines is still low. Here, the current status of tissue culture and recent progress in transformation methodology are presented. Some major challenges and obstacles are discussed and some examples of their solutions are presented.     

花生转基因研究进展

花生是世界上重要的油料和经济作物之一,是人们生活的植物脂肪和蛋白质来源。现代生物技术的不断发展为花生育种和种质创新提供了新的技术手段, 它可以直接将来自不同种属的异源目的基因插人到花生基因组, 使花生表达目标性状, 实现花生品种的遗传改良。近年来, 国内外花生转基因研究取得了重大进展。文章综述了花生转基因在抗虫、抗病、抗非生物逆境和品质改良等方面的最新进展,并总结了近年来人们对农杆菌介导法、基因枪法和不依赖组织培养的转化法等主要的花生遗传转化方法的改进和探索。     

Biotechnological applications in in vitro plant regeneration studies of broccoli (<Emphasis Type="Italic">Brassica oleracea</Emphasis> L. var. <Emphasis Type="Italic">italica</Emphasis>), an important vegetable crop

Biotechnology holds promise for genetic improvement of important vegetable crops. Broccoli (Brassica oleracea L. var. italica) is an important vegetable crop of the family Brassicaceae. However, various biotic and abiotic stresses cause enormous crop yield losses during commercial cultivation of broccoli. Establishment of a reliable, reproducible and efficient in vitro plant regeneration system with cell and tissue culture is a vital prerequisite for biotechnological application of crop improvement programme. An in vitro plant regeneration technique refers to culturing, cell division, cell multiplication, de-differentiation and differentiation of cells, protoplasts, tissues and organs on defined liquid/solid medium under aseptic and controlled environment. Recent progress in the field of plant tissue culture has made this area one of the most dynamic and promising in experimental biology. There are many published reports on in vitro plant regeneration studies in broccoli including direct organogenesis, indirect organogenesis and somatic embryogenesis. This review summarizes those plant regeneration studies in broccoli that could be helpful in drawing the attention of the researchers and scientists to work on it to produce healthy, biotic and abiotic stress resistant plant material and to carry out genetic transformation studies for the production of transgenic plants.     

Chilli peppers — A review on tissue culture and transgenesis

Biotechnology techniques involving plant tissue culture and recombinant DNA technologies are powerful tools that can complement conventional breeding and expedite Capsicum improvement. The rate of progress in Capsicum is relatively slower than other members of Solanaceae because of its high genotypic dependence and recalcitrant nature. Capsicum is a recalcitrant plant in terms of in vitro cell, tissue and organ differentiation, plant regeneration and genetic transformation which makes it difficult to apply recombinant DNA technologies aimed at genetic improvement against pests, diseases and abiotic stress. Despite this, application of tissue culture and genetic transformation have led to significant development in chilli pepper plants, and studies are underway to achieve the targets of pre-harvest improvement and post-harvest characterization for value addition to this crop. This review presents a consolidated account of in vitro propagation and focuses upon contemporary information on biotechnological advances made in Capsicum.     

Development of Efficient Plant Regeneration and Transformation System for Impatiens Using <Emphasis Type="Italic">Agrobacterium tumefaciens</Emphasis>and Multiple Bud Cultures as Explants

Background  

Impatiens (Impatiens walleriana) is a top selling floriculture crop. The potential for genetic transformation of Impatiens to introduce novel flower colors or virus resistance has been limited by its general recalcitrance to tissue culture and transformation manipulations. We have established a regeneration and transformation system for Impatiens that provides new alternatives to genetic improvement of this crop.     

促进植物高效遗传转化的发育调节因子及其在玉米中的应用进展*

高效遗传转化技术体系的建立对植物功能基因组学研究和作物新品种的培育均具有促进作用,目前,再生效率低下是限制许多植物高效遗传转化体系建立的主要技术屏障之一。随着对植物分生组织和体细胞胚形成过程研究的深入,鉴定到了一些关键调控基因,统称为发育调节因子。发育调节因子应用于植物遗传转化后,可以有效改善植物分生组织诱导和再生能力,为提高遗传转化效率提供了重要机遇。综述了7类发育调节因子在提高植物遗传转化效率中的研究进展,重点介绍了其中3类在促进玉米遗传转化中的应用,最后展望了建立植物高效遗传转化体系的发展方向。     

Efficient floral dip transformation method using Agrobacterium tumefaciens on Cosmos sulphureus Cav.

Yellow cosmos (Cosmos sulphureus Cav.) is a specific flowering plant and considered a suitable genetic engineering model. Agrobacterium-mediated plant transformation is commonly used for plant genetic engineering. Floral dip transformation is one of the plant genetic transformation methods, and it involves dipping flower buds into an Agrobacterium suspension. Studies on floral dip transformation of yellow cosmos have never been reported. Therefore, an efficient method in plant genetic engineering must be established. This study developed an effective and efficient floral dip transformation method for yellow cosmos.In this study, flower buds with sizes of 5–7 mm were used. Several parameters have been observed to optimize the floral dip method. These parameters included the optical density (OD₆₀₀) of Agrobacterium culture, concentration of surfactant, and duration of flower bud dipping into the Agrobacterium suspension.The results showed that the floral dip method was most efficient when the flower buds were dipped into Agrobacterium suspension with OD₆₀₀ = 0.8 and containing 5% sucrose and 0.1% Silwet L-77 for 30 s. This method enhanced the transformation efficiency at a rate of 12.78 ± 1.53%. The neomycin phosphotransferase II and green fluorescent protein genes with sizes of 550 and 736 bp, respectively, were confirmed by polymerase chain reaction. In addition, the transgenic plants were kanamycin resistant and fluorescent under ultraviolet light observation. This finding suggests that the proposed floral dip transformation provides new insights into efficient plant genetic engineering methods for yellow cosmos.     

Transgenic strawberry: state of the art for improved traits

Strawberry (Fragaria × ananassa Duch.), a member of the Rosaceae family, is one of the most important fruit crops cultivated worldwide. Strawberry is unique within the Rosaceae because it is a rapidly growing herbaceous perennial with a small genome, short reproductive cycle, and facile vegetative and generative propagation for genetic transformation. For these reasons, strawberry has been recognized as excellent germplasm for genetic and molecular studies for the Rosaceae family. Although traditional breeding methods have achieved steady improvement in agronomic traits, the lack of useful economic characters still remains a major challenge. Genetic transformation has opened a new era for greater creativity in strawberry breeding and germplasm by offering an effective method for creating new varieties that selectively targets a specific interested gene or a few heterologous traits. Enormous advances have been made in strawberry genetic transformation since the first transgenic strawberry plant was obtained in 1990. This paper reviews recent progress in genetic transformation of strawberry on increasing resistance to viruses, fungi, insects, herbicides, stress, and achieving better quality. Problems and prospects for future applications of genetic transformation in strawberry are also discussed.